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  Subjects -> BIOLOGY (Total: 3190 journals)
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BIOTECHNOLOGY (244 journals)                  1 2 | Last

Showing 1 - 200 of 244 Journals sorted alphabetically
3 Biotech     Open Access   (Followers: 8)
Advanced Biomedical Research     Open Access  
Advances in Bioscience and Biotechnology     Open Access   (Followers: 17)
Advances in Genetic Engineering & Biotechnology     Hybrid Journal   (Followers: 9)
Advances in Regenerative Medicine     Open Access   (Followers: 3)
African Journal of Biotechnology     Open Access   (Followers: 6)
Algal Research     Partially Free   (Followers: 11)
American Journal of Biochemistry and Biotechnology     Open Access   (Followers: 69)
American Journal of Bioinformatics Research     Open Access   (Followers: 7)
American Journal of Polymer Science     Open Access   (Followers: 33)
Amylase     Open Access  
Anadolu University Journal of Science and Technology : C Life Sciences and Biotechnology     Open Access  
Animal Biotechnology     Hybrid Journal   (Followers: 8)
Annales des Sciences Agronomiques     Full-text available via subscription  
Applied Biochemistry and Biotechnology     Hybrid Journal   (Followers: 45)
Applied Biosafety     Hybrid Journal  
Applied Food Biotechnology     Open Access   (Followers: 3)
Applied Microbiology and Biotechnology     Hybrid Journal   (Followers: 67)
Applied Mycology and Biotechnology     Full-text available via subscription   (Followers: 4)
Arthroplasty Today     Open Access   (Followers: 1)
Artificial Cells, Nanomedicine and Biotechnology     Hybrid Journal   (Followers: 1)
Asia Pacific Biotech News     Hybrid Journal   (Followers: 2)
Asian Journal of Biotechnology     Open Access   (Followers: 9)
Asian Pacific Journal of Tropical Biomedicine     Open Access   (Followers: 2)
Australasian Biotechnology     Full-text available via subscription   (Followers: 1)
Banat's Journal of Biotechnology     Open Access  
BBR : Biochemistry and Biotechnology Reports     Open Access   (Followers: 5)
Beitr?ge zur Tabakforschung International/Contributions to Tobacco Research     Open Access   (Followers: 3)
Bio-Algorithms and Med-Systems     Hybrid Journal   (Followers: 2)
Bio-Research     Full-text available via subscription   (Followers: 4)
Bioactive Materials     Open Access   (Followers: 1)
Biocatalysis and Agricultural Biotechnology     Hybrid Journal   (Followers: 4)
Biocybernetics and Biological Engineering     Full-text available via subscription   (Followers: 5)
Bioethics UPdate     Hybrid Journal   (Followers: 1)
Biofuels     Hybrid Journal   (Followers: 11)
Biofuels Engineering     Open Access   (Followers: 1)
Biological & Pharmaceutical Bulletin     Full-text available via subscription   (Followers: 4)
Biological Cybernetics     Hybrid Journal   (Followers: 10)
Biomarkers and Genomic Medicine     Open Access   (Followers: 3)
Biomaterials Research     Open Access   (Followers: 4)
BioMed Research International     Open Access   (Followers: 4)
Biomédica     Open Access  
Biomedical and Biotechnology Research Journal     Open Access  
Biomedical Engineering Research     Open Access   (Followers: 6)
Biomedical Glasses     Open Access  
Biomedical Reports     Full-text available via subscription  
BioMedicine     Open Access  
Biomedika     Open Access  
Bioprinting     Hybrid Journal   (Followers: 1)
Bioresource Technology Reports     Hybrid Journal   (Followers: 1)
Bioscience, Biotechnology, and Biochemistry     Hybrid Journal   (Followers: 21)
Biosensors Journal     Open Access  
Biosimilars     Open Access   (Followers: 1)
Biosurface and Biotribology     Open Access  
Biotechnic and Histochemistry     Hybrid Journal   (Followers: 1)
BioTechniques : The International Journal of Life Science Methods     Full-text available via subscription   (Followers: 28)
Biotechnologia Acta     Open Access   (Followers: 1)
Biotechnologie, Agronomie, Société et Environnement     Open Access   (Followers: 2)
Biotechnology     Open Access   (Followers: 8)
Biotechnology & Biotechnological Equipment     Open Access   (Followers: 4)
Biotechnology Advances     Hybrid Journal   (Followers: 34)
Biotechnology and Applied Biochemistry     Hybrid Journal   (Followers: 44)
Biotechnology and Bioengineering     Hybrid Journal   (Followers: 159)
Biotechnology and Bioprocess Engineering     Hybrid Journal   (Followers: 6)
Biotechnology and Genetic Engineering Reviews     Hybrid Journal   (Followers: 13)
Biotechnology and Health Sciences     Open Access   (Followers: 1)
Biotechnology and Molecular Biology Reviews     Open Access   (Followers: 2)
Biotechnology Annual Review     Full-text available via subscription   (Followers: 5)
Biotechnology for Biofuels     Open Access   (Followers: 10)
Biotechnology Frontier     Open Access   (Followers: 2)
Biotechnology Journal     Hybrid Journal   (Followers: 17)
Biotechnology Law Report     Hybrid Journal   (Followers: 4)
Biotechnology Letters     Hybrid Journal   (Followers: 34)
Biotechnology Progress     Hybrid Journal   (Followers: 41)
Biotechnology Reports     Open Access  
Biotechnology Research International     Open Access   (Followers: 1)
Biotechnology Techniques     Hybrid Journal   (Followers: 10)
Biotecnología Aplicada     Open Access  
Bioteknologi (Biotechnological Studies)     Open Access  
BIOTIK : Jurnal Ilmiah Biologi Teknologi dan Kependidikan     Open Access  
Biotribology     Hybrid Journal   (Followers: 1)
BMC Biotechnology     Open Access   (Followers: 17)
Cell Biology and Development     Open Access  
Chinese Journal of Agricultural Biotechnology     Full-text available via subscription   (Followers: 4)
Communications in Mathematical Biology and Neuroscience     Open Access  
Computational and Structural Biotechnology Journal     Open Access   (Followers: 2)
Computer Methods and Programs in Biomedicine     Hybrid Journal   (Followers: 8)
Copernican Letters     Open Access   (Followers: 1)
Critical Reviews in Biotechnology     Hybrid Journal   (Followers: 20)
Crop Breeding and Applied Biotechnology     Open Access   (Followers: 3)
Current Bionanotechnology     Hybrid Journal  
Current Biotechnology     Hybrid Journal   (Followers: 4)
Current Opinion in Biomedical Engineering     Hybrid Journal   (Followers: 1)
Current Opinion in Biotechnology     Hybrid Journal   (Followers: 55)
Current Pharmaceutical Biotechnology     Hybrid Journal   (Followers: 9)
Current Research in Bioinformatics     Open Access   (Followers: 13)
Current Trends in Biotechnology and Chemical Research     Open Access   (Followers: 3)
Current trends in Biotechnology and Pharmacy     Open Access   (Followers: 8)
DNA and RNA Nanotechnology     Open Access  
EBioMedicine     Open Access  
Electronic Journal of Biotechnology     Open Access  
Entomologia Generalis     Full-text available via subscription   (Followers: 1)
Environmental Science : Processes & Impacts     Full-text available via subscription   (Followers: 4)
Experimental Biology and Medicine     Hybrid Journal   (Followers: 3)
Folia Medica Indonesiana     Open Access  
Food Bioscience     Hybrid Journal  
Food Biotechnology     Hybrid Journal   (Followers: 9)
Food Science and Biotechnology     Hybrid Journal   (Followers: 8)
Frontiers in Bioengineering and Biotechnology     Open Access   (Followers: 6)
Frontiers in Systems Biology     Open Access   (Followers: 2)
Fungal Biology and Biotechnology     Open Access   (Followers: 2)
GM Crops and Food: Biotechnology in Agriculture and the Food Chain     Full-text available via subscription   (Followers: 1)
GSTF Journal of BioSciences     Open Access  
HAYATI Journal of Biosciences     Open Access  
Horticultural Biotechnology Research     Open Access  
Horticulture, Environment, and Biotechnology     Hybrid Journal   (Followers: 11)
IEEE Transactions on Molecular, Biological and Multi-Scale Communications     Hybrid Journal   (Followers: 1)
IET Nanobiotechnology     Hybrid Journal   (Followers: 2)
IN VIVO     Full-text available via subscription   (Followers: 4)
Indian Journal of Biotechnology (IJBT)     Open Access   (Followers: 2)
Indonesia Journal of Biomedical Science     Open Access   (Followers: 2)
Indonesian Journal of Biotechnology     Open Access   (Followers: 1)
Indonesian Journal of Medicine     Open Access  
Industrial Biotechnology     Hybrid Journal   (Followers: 18)
International Biomechanics     Open Access  
International Journal of Bioinformatics Research and Applications     Hybrid Journal   (Followers: 14)
International Journal of Biomechatronics and Biomedical Robotics     Hybrid Journal   (Followers: 4)
International Journal of Biomedical Research     Open Access   (Followers: 2)
International Journal of Biotechnology     Hybrid Journal   (Followers: 5)
International Journal of Biotechnology and Molecular Biology Research     Open Access   (Followers: 4)
International Journal of Biotechnology for Wellness Industries     Partially Free   (Followers: 1)
International Journal of Environment, Agriculture and Biotechnology     Open Access   (Followers: 5)
International Journal of Functional Informatics and Personalised Medicine     Hybrid Journal   (Followers: 4)
International Journal of Medicine and Biomedical Research     Open Access   (Followers: 1)
International Journal of Nanotechnology and Molecular Computation     Full-text available via subscription   (Followers: 3)
International Journal of Radiation Biology     Hybrid Journal   (Followers: 4)
Iranian Journal of Biotechnology     Open Access  
ISABB Journal of Biotechnology and Bioinformatics     Open Access  
Italian Journal of Food Science     Open Access   (Followers: 1)
JMIR Biomedical Engineering     Open Access  
Journal of Biometrics & Biostatistics     Open Access   (Followers: 3)
Journal of Bioterrorism & Biodefense     Open Access   (Followers: 6)
Journal of Petroleum & Environmental Biotechnology     Open Access   (Followers: 1)
Journal of Advanced Therapies and Medical Innovation Sciences     Open Access  
Journal of Advances in Biotechnology     Open Access   (Followers: 5)
Journal Of Agrobiotechnology     Open Access  
Journal of Analytical & Bioanalytical Techniques     Open Access   (Followers: 7)
Journal of Animal Science and Biotechnology     Open Access   (Followers: 4)
Journal of Applied Biomedicine     Open Access   (Followers: 2)
Journal of Applied Biotechnology     Open Access   (Followers: 2)
Journal of Applied Biotechnology Reports     Open Access   (Followers: 2)
Journal of Applied Mathematics & Bioinformatics     Open Access   (Followers: 5)
Journal of Biologically Active Products from Nature     Hybrid Journal   (Followers: 1)
Journal of Biomaterials and Nanobiotechnology     Open Access   (Followers: 6)
Journal of Biomedical Photonics & Engineering     Open Access  
Journal of Biomedical Practitioners     Open Access  
Journal of Bioprocess Engineering and Biorefinery     Full-text available via subscription  
Journal of Bioprocessing & Biotechniques     Open Access  
Journal of BioScience and Biotechnology     Open Access  
Journal of Biosecurity Biosafety and Biodefense Law     Hybrid Journal   (Followers: 3)
Journal of Biotechnology     Hybrid Journal   (Followers: 63)
Journal of Biotechnology and Strategic Health Research     Open Access   (Followers: 1)
Journal of Chemical and Biological Interfaces     Full-text available via subscription   (Followers: 1)
Journal of Chemical Technology & Biotechnology     Hybrid Journal   (Followers: 9)
Journal of Chitin and Chitosan Science     Full-text available via subscription   (Followers: 1)
Journal of Colloid Science and Biotechnology     Full-text available via subscription  
Journal of Commercial Biotechnology     Full-text available via subscription   (Followers: 6)
Journal of Crop Science and Biotechnology     Hybrid Journal   (Followers: 3)
Journal of Ecobiotechnology     Open Access  
Journal of Essential Oil Research     Hybrid Journal   (Followers: 2)
Journal of Experimental Biology     Full-text available via subscription   (Followers: 25)
Journal of Genetic Engineering and Biotechnology     Open Access   (Followers: 5)
Journal of Ginseng Research     Open Access  
Journal of Industrial Microbiology and Biotechnology     Hybrid Journal   (Followers: 18)
Journal of Integrative Bioinformatics     Open Access  
Journal of Medical Imaging and Health Informatics     Full-text available via subscription  
Journal of Molecular Biology and Biotechnology     Open Access  
Journal of Molecular Microbiology and Biotechnology     Full-text available via subscription   (Followers: 13)
Journal of Nano Education     Full-text available via subscription  
Journal of Nanobiotechnology     Open Access   (Followers: 4)
Journal of Nanofluids     Full-text available via subscription   (Followers: 1)
Journal of Organic and Biomolecular Simulations     Open Access  
Journal of Plant Biochemistry and Biotechnology     Hybrid Journal   (Followers: 4)
Journal of Science and Applications : Biomedicine     Open Access  
Journal of the Mechanical Behavior of Biomedical Materials     Hybrid Journal   (Followers: 13)
Journal of Trace Elements in Medicine and Biology     Hybrid Journal   (Followers: 1)
Journal of Tropical Microbiology and Biotechnology     Full-text available via subscription  
Journal of Yeast and Fungal Research     Open Access   (Followers: 1)
Marine Biotechnology     Hybrid Journal   (Followers: 4)
Meat Technology     Open Access  
Messenger     Full-text available via subscription  
Metabolic Engineering Communications     Open Access   (Followers: 4)
Metalloproteinases In Medicine     Open Access  
Microbial Biotechnology     Open Access   (Followers: 10)
MicroMedicine     Open Access   (Followers: 3)
Molecular and Cellular Biomedical Sciences     Open Access   (Followers: 1)
Molecular Biotechnology     Hybrid Journal   (Followers: 13)
Molecular Genetics and Metabolism Reports     Open Access   (Followers: 3)
Nanobiomedicine     Open Access  
Nanobiotechnology     Hybrid Journal   (Followers: 2)

        1 2 | Last

Journal Cover
Journal of Genetic Engineering and Biotechnology
Journal Prestige (SJR): 0.375
Citation Impact (citeScore): 2
Number of Followers: 5  

  This is an Open Access Journal Open Access journal
ISSN (Print) 1687-157X
Published by Elsevier Homepage  [3160 journals]
  • Optimization of indole acetic acid production by isolated bacteria from
           Stevia rebaudiana rhizosphere and its effects on plant growth

    • Abstract: Publication date: Available online 8 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Sheela Chandra, Kazim Askari, Madhumita Kumari The ability to synthesize Indole-3-acetic acid (IAA) is widely associated with the plant growth promoting rhizobacteria (PGPR). The present work deals with isolation and characterization of such bacteria from the rhizosphere of medicinal plant Stevia rebaudiana and optimization of IAA production from its isolates. The optimization of IAA production was carried out at different pH and temperature with varied carbon and nitrogen sources of culture media. Out of different isolates obtained, three of them were screened as efficient PGPRs on the basis of different plant growth promoting attributes. Isolates CA1001 and CA2004 showed better production of IAA at pH 9 (91.7 µg ml−1) and at temperature 37 °C (81.7 µg ml−1). Dextrose (1%) was found to be the best carbon source for isolate CA1001 with 104 µg ml−1 IAA production. Isolate CA 2004 showed best production of IAA 36 µg ml−1 and 34 µg ml−1 at 1.5% and 1% Beef extract as nitrogen source respectively. Isolate CA 1001 showed 32 µg ml−1 IAA production at 0.5% nicotinic acid concentration. From the current study, CA1001 and CA2004 emerged as noble alternatives for IAA production further which also resulted in root and shoot biomass generation in crop plants, hence can be further used as bio-inoculants for plant growth promotion.
       
  • Cloning, transformation and expression of cell cycle-associated protein
           kinase OsWee1 in indica rice (Oryza sativa L.)

    • Abstract: Publication date: Available online 7 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Frengky H.H. Prasetyo, Bambang Sugiharto, Netty Ermawati The development process of seed in plants is a cycle of cells which occur gradually and regularly. One of the genes involved in controling this stage is the Wee1 gene. Wee1 encode protein kinase which plays an important role in phosphorylation, inactivation of cyclin-dependent kinase 1 (CDK1)-cyclin (CYC) and inhibiting cell division at mitotic phase. The Overexpression of Wee1 leads to delaying entry into mitotic phase, resulting in enlargement of cell size due to suppression of cell division. Accordingly, the cloning and overexpressing of Wee1 in rice plant is important aim of this research in achieving better quantity and quality of future rice. The main objective of this present study is to cloning and generate transgenic rice plants overexpressing of Wee1 gene. Wee1 was isolated from cDNA of indica rice (Oryza sativa), called OsWee1. The full length of OsWee1 was 1239 bp in size and successfully inserted into plant expression vector pRI101ON. Seven-day-old rice seedlings were prepared for transformation of OsWee1 gene using Agrobacterium-mediated transformation method. Four positive transgenic lines were identified through the presence of kanamycin resistance gene (nptII) using genomic PCR analysis. Southern blot analysis result provides evidence that four independent rice transformants contained one to three rearranged transgene copies. Further screening in transgenic rice generation is needed in order to obtain stable expression of OsWee1.
       
  • Transgenic approaches for genetic improvement in groundnut (Arachis
           hypogaea L.) against major biotic and abiotic stress factors

    • Abstract: Publication date: Available online 7 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Saikat Gantait, Suvendu Mondal Cultivated groundnut (Arachis hypogaea L.) is considered as one of the primary oilseed crops and a major fodder for cattle industry in most of the developing countries, owing to its rich source of protein. It is due to its geocarpic nature of growth that the overall yield performance of groundnut is hindered by several biotic and abiotic stress factors. Multidimensional attempts were undertaken to combat these factors by developing superior groundnut varieties, modified with integral mechanism of tolerance/resistance; however this approach proved to be futile, owing to inferior pod and kernel quality. As a superior alternative, biotechnological intervention like transformation of foreign genes, either directly (biolistic) or via Agrobacterium, significantly aided in the development of advanced groundnut genotypes equipped with integral resistance against stresses and enhanced yield attributing traits. Several genes triggered by biotic and abiotic stresses, were detected and some of them were cloned and transformed as major parts of transgenic programmes. Application of modern molecular biological techniques, in designing biotic and abiotic stress tolerant/resistant groundnut varieties that exhibited mechanisms of resistance, relied on the expression of specific genes associated to particular stress. The genetically transformed stress tolerant groundnut varieties possess the potential to be employed as donor parents in traditional breeding programmes for developing varieties that are resilient to fungal, bacterial, and viral diseases, as well as to draught and salinity. The present review emphasizes on the retrospect and prospect of genetic transformation tools, implemented for the enhancement of groundnut varieties against key biotic and abiotic stress factors.
       
  • In silico structural and functional modelling of Antifreeze protein (AFP)
           sequences of Ocean pout (Zoarces americanus, Bloch & Schneider 1801)

    • Abstract: Publication date: Available online 6 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Manojit Bhattacharya, Arpita Hota, Avijit Kar, Deep Sankar Chini, Ramesh Chandra Malick, Bidhan Chandra Patra, Basanta Kumar Das Antifreeze proteins (AFPs) are known to polypeptide components formed by certain plants, animals, fungi and bacteria which support to survive in sub-zero temperature. Current study highlighted the seven different antifreeze proteins of fish Ocean pout (Zoarces americanus), in which protein (amino acids sequence) were collected from National Centre for Biotechnology Information and finely characterized using several in silico tools. Such biocomputational techniques applied to figure out the physicochemical, functional and conformational characteristics of targeted AFPs. Multiple physicochemical properties such as Isoelectric Point, Extinction Coefficient and Instability Index, Aliphatic Index, Grand Average Hydropathy were calculated and analysed by ExPASy-ProtParam prediction web server. EMBOSS: pepwheel online tool was used to represent the protein sequences in a helical form. The primary structure analysis shows that most of the AFPs are hydrophobic in nature due to the high content of non-polar residues. The secondary structure of these proteins was calculated using SOPMA tool. SOSUI server and CYS_REC program also run for ideal prediction of transmembrane helices and disulfide bridges of experimental proteins respectively. The modelling of 3D structures of seven desired AFPs were executed by the homology modelling programmes; SWISS MODEL and ProSA web server. UCSF Chimera, Antheprot 3D, PyMOL and RAMPAGE were used to visualize and analysis of the structural variation of the predicted protein model. MEGA7.0.9 software used to know the phylogenetic relationship among these AFPs. These models offered excellent and reliable baseline information for functional characterization of the experimentally derived protein domain composition by using the advanced tools and techniques of Computational Biology.
       
  • In vitro differentiation of human multilineage differentiating
           stress-enduring (Muse) cells into insulin producing cells

    • Abstract: Publication date: Available online 5 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Ali M. Fouad, Mahmoud M. Gabr, Elsayed K. Abdelhady, Mahmoud M. Zakaria, Sherry M. Khater, Amani M. Ismail, Ayman F. Refaie Mesenchymal stem cells (MSCs) is a heterogeneous population. Muse cells is a rare pluripotent subpopulation within MSCs. This study aims to evaluate the pulirpotency and the ability of Muse cells to generate insulin producing cells (IPCs) after in vitro differentiation protocol compared to the non-Muse cells. Muse cells were isolated by FACSAria III cell sorter from adipose-derived MSCs and were evaluated for its pluripotency. Following in vitro differentiation, IPCs derived from Muse and non-Muse cells were evaluated for insulin production. Muse cells comprised 3.2 ± 0.7% of MSCs, approximately 82% of Muse cells were positive for anti stage-specific embryonic antigen-3 (SSEA-3). Pluripotent markers were highly expressed in Muse versus non-Muse cells. The percentage of generated IPCs by flow cytometric analysis was higher in Muse cells. Under confocal microscopy, Muse cells expressed insulin and c-peptide while it was undetected in non-Muse cells. Our results introduced Muse cells as a new adult pluripotent subpopulation, which is capable to produce higher number of functional IPCs.
       
  • Development and evaluation of latex agglutination test coating with
           recombinant antigen, LipL32 for serodiagnosis of human leptospirosis

    • Abstract: Publication date: Available online 3 December 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Kotchakorn Thongsukkaeng, Rerngwit Boonyom Leptospirosis is a widespread zoonotic disease caused by Leptospira interrogans. Symptoms of disease range from mild symptoms to serious complications including, jaundice, pulmonary hemorrhage, renal and hepatic failure, which may prove fatal. Clinical presentations of this disease are similar with other febrile illness. Therefore, rapid and appropriated laboratory diagnostic tests are needed to aid clinical case identification. As these reasons, objective of this study is to develop and evaluate a simple latex agglutination test coating with recombinant leptospiral antigens, LipL32 for serodiagnosis of human leptospirosis. Firstly, lipl32 gene was amplified from genomic DNA of Leptospira interogans serovar Pyrogenes. Then PCR product of lipl32 gene was ligated with pGEX-2T plasmid, generating pGRK32 recombinant plasmid. Recombinant GST-LipL32 protein was overexpressed and subsequently purified by using Glutathione-Agarose Resin. Recombinant GST-Lipl32 protein was coated on latex beads for development latex agglutination test (LAT). The relative sensitivity, specificity and accuracy of the developed LAT were compared with indirect immunofluorescences assay (IFA) for detection of anti-leptospiral antibodies in 30 human leptospirosis samples, 30 healthy blood donor samples, 10 dengue fever positive samples, 10 scrub typhus positive samples, and 10 melioidosis samples. Results showed that the developed LAT showed sensitivity, specificity and accuracy: 66.66%, 86.66%, and 80.00%, respectively, comparing with IFA method. Moreover, Kappa analysis showed agreement rate of the two methods were 0.421. It concluded that our developed gave compatible result with IFA. Additionally, Our LAT are simple, rapid and suitable for detection in the field. However, for better sensitivity, diagnostic specificity, positive predictive value, negative predictive value, accuracy and Cohen’s kappa comparison should be done in larger amounts of sera samples.
       
  • Callus mediated shoot organogenesis and regeneration of cytologically
           stable plants of Ledebouria revoluta: An ethnomedicinal plant with
           promising antimicrobial potency

    • Abstract: Publication date: Available online 12 November 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Sk Moquammel Haque, Avijit Chakraborty, Biswajit Ghosh Ledebouria revoluta are important ethnomedicinal plant found in India and South Africa. Micropropagation via indirect shoot organogenesis had been established from three types of explant (i.e. scale leaf, leaf lamina and root) of L. revoluta. Scale leaf was found superior as compared to leaf lamina and root explant with respect to their organogenic callus induction potentiality. Murashige and Skoog (1962) [MS] media supplemented with 3.0 mg L−1 2,4-dichlorophenoxyacetic acid, 0.75 mg L−1 β-naphthoxyacetic acid were best effective for inducing organogenic callus. Maximum 17.0 ± 0.52 bulblets were induced from about 500 mg of callus within 42–46 days sub-culturing on a medium containing 0.75 mg L−1 kinetin. The bulblets were matured (86.7% success) after one month culture on the same medium composition. The best result of in vitro root induction with 100% response and 8.4 ± 0.31 roots per bulb was achieved after 18 days of implantation on MS medium containing 2.0 mg L−1 indole-3-butyric acid. Plantlets were acclimatized with a 96.0% survival rate. Chromosomal studies revealed cytological stability of callus cells and all regenerants containing 2n = 30 chromosomes, same as parental plants. Antimicrobial activity of L. revoluta was tested against two Gram-positive bacteria, three Gram-negative bacteria and two fungi. The methanol and ethanol extract proved more effective against bacteria, whereas acetone and chloroform extract shows potential anti-fungal activities. Present protocol can be applied reliably to produce uniform planting materials in large scale. In addition, this efficient indirect regeneration pathway via callus culture opens a way for improvement through genetic transformation.
       
  • Population structure, morphological and genetic diversity within and among
           melon (Cucumis melo L.) landraces in Iran

    • Abstract: Publication date: Available online 28 August 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Masoud Maleki, Abdolali Shojaeiyan, Sajad Rashidi Monfared ISSR markers were applied to evaluate the genetic diversity and differentiation of 270 individuals of 27 Iranian C. melo landraces of various varietal groups include vars. inodorous, cantalupensis, reticulatus, ameri, dudaim. Genetic diversity among the studied genotypes obtained by GeneAlex analysis (H = 0.08, I = 0.12, Na = 0.77, PPL = 22.6%). Cluster analysis divided Iranian melon landraces into two main cluster. Non-sweet genotype (dudaim group) was well separated from sweet genotypes (inodorous, ameri, reticulatus, cantalupensis). The most similar genotypes were BANI and TONI (0.95) and the most dissimilar ones were GER and TS (0.58). AMOVA result showed that the percentage of genetic variation among and within Iranian melon is 69% and 31%, respectively. All landraces evaluated based on 10 morphological traits which revealed the diversity of melon varietal groups. Bayesian analysis assigned ten landraces to Pop 1, eight landraces to Pop 2 and nine melon landraces to Pop 3. Bayesian and UPGMA cluster analyses demonstrated the almost related results. Our results indicated that ISSR markers technique alongside polyacrylamide gel analysis could be helpful to discriminate varieties of melon.
       
  • Feline panleukopenia viral infection in cats: Application of some
           molecular methods used for its diagnosis

    • Abstract: Publication date: Available online 27 August 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Romane A. Awad, Wagdy K.B. Khalil, Ashraf G. Attallah Forty diseased cats and seven healthy control cats from different sex, ages and breeds had examined clinically to confirm presence or absence of clinical symptoms of Feline panleukopenia disease (FP). Several tools including ELISA, gene expression analysis (qRT-PCR), DNA fragmentation test and apoptosis assay were conducted to determine the Feline panleukopenia disease in cat tissues. Clinical symptoms in the form of depression, fever, anorexia, vomiting, diarrhea, dehydration, anaemia and leucopenia were recorded in the diseased cats while no clinical sings were observed in control healthy cats. ELISA results showed that all of diseased (n = 40) cats were positive while control cats (n = 7) were negative for FP viral antigen. After carrying out of ELISA assay, supportive treatment trials including fluid therapy, immunostimulant, antibiotics to overcome dehydration, restoring electrolytes imbalances, combating secondary bacterial infection were conducted but all diseased cats were died and control cats exposed to soft death. Gene expression analysis detected high levels of FP viral gene in several cat tissues in which ilium exhibited high viral expression levels compared with jejunum. Also, viral expression levels in jejunum were higher than in mesenteric lymph nodes. In addition, viral expression levels were not detected in tissues of control cats. The results of the DNA fragmentation assay observed that DNA extracted from different tissues of infected cats exhibited damaged DNA bands as compared with DNA of control cats. DNA fragmentation rates in infected tissues increased significantly (P 
       
  • Buffalo species identification and delineation using genetic barcoding
           markers

    • Abstract: Publication date: Available online 9 August 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Amal Ahmed Mohamed Hassan, Esraa Aly Balabel, Hanaa Abdel Sadek Oraby, Samy Anwar Darwish Enrichment of barcode databases with mitochondrial cytochrome c oxidase subunit I (COI) barcode sequences in different animal taxa has become important for identification of animal source in food samples to prevent commercial fraud. In this study, COI barcode sequence in seventy one river buffalo samples were determined, analyzed and deposited in Genbank barcode database and barcode of life database (BOLD) to contribute for construction of public reference library for COI barcode sequence in river buffalo. Moreover COI barcode sequence was used to identify the closely related buffalo groups: river buffalo, swamp buffalo, lowland anoa and African buffalo. Results indicated the success of the COI barcode in the identification of each of the tested groups. Whereas a suggested sequence of other mitochondrial segment representing two successive transfer RNA (tRNA) genes; tRNA-Threonine (MT-TT) and tRNA-Proline (MT-TP) was failed to be used as a barcode marker for differentiation between the tested buffalo groups.
       
  • Cytogenetic effects of silver and gold nanoparticles on Allium
           cepa
    roots

    • Abstract: Publication date: Available online 3 August 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Priyanka Debnath, Arghadip Mondal, Amita Hajra, Chittaranjan Das, Naba Kumar Mondal The present study evaluates the cytogenetic effects of both silver and gold nanoparticles on the root cells of Allium cepa. In this study, the root cells of Allium cepa were treated with both gold and silver nanoparticles of different concentrations (1 mg/L, 5 mg/L and 10 mg/L) along with control for 72 h. Experimental results revealed that after 72 h of exposure, a significant decrease in mitotic index (MI) from 68% (control) to 52.4% (1 mg/L), 47.3% (5 mg/L) and 41.4% (10 mg/L) for gold nanoparticles and 57.1% (1 mg/L), 53% (5 mg/l), 55.8% (10 mg/L) for silver nanoparticles. Through minute observation of the photograph, it was recorded that some specific chromosomal abnormalities such as stickiness of chromosome, chromosome breaks, nuclear notch, and clumped chromosome at different exposure conditions. Therefore, present results clearly suggest that Allium cepa root tip assay could be a viable path through which negative impact of both gold and silver nanoparticles can be demonstrated over a wide range of concentrations.
       
  • Valorisation of chicken feathers for xanthan gum production using
           Xanthomonas campestris MO-03

    • Abstract: Publication date: Available online 24 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Murat Ozdal, Esabi Basaran Kurbanoglu Xanthan gum is an important commercial polysaccharide produced by Xanthomonas species. In this study, xanthan production was investigated using a local isolate of Xanthomonas campestris MO-03 in medium containing various concentrations of chicken feather peptone (CFP) as an enhancer substrate. CFP was produced with a chemical process and its chemical composition was determined. The addition of CFP (1–8 g/l) increased the conversion of sugar to xanthan gum in comparison with the control medium, which did not contain additional supplements. The highest xanthan production (24.45 g/l) was found at the 6 g/l CFP containing control medium in 54 h. This value was 1.73 fold higher than that of control medium (14.12 g/l). Moreover, addition of CFP improved the composition of xanthan gum; the pyruvate content of xanthan was 3.86% (w/w), higher than that of the control (2.2%, w/w). The xanthan gum yield was also influenced by the type of organic nitrogen sources. As a conclusion, CFP was found to be a suitable substrate for xanthan gum production.
       
  • Biolytic extraction of poly(3-hydroxybutyrate) from Bacillus megaterium
           Ti3 using the lytic enzyme of Streptomyces albus Tia1

    • Abstract: Publication date: Available online 21 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Neetu Israni, Surabhi Thapa, Srividya Shivakumar The applicability of Streptomyces sp. cell lytic enzymes for devising a simple and competent biological polyhydroxyalkanoate (PHA) recovery approach from Bacillus megaterium cells was investigated. B. megaterium strain Ti3 produced 50% (w/w) PHA using glucose as carbon source. The intracellular PHA was recovered employing a non-PHA accumulating actinomycetes (Tia1) identified as Streptomyces albus, having potent lytic activity against living and heat inactivated B. megaterium. Interestingly, maximum biomass (2.53 ± 0.6 g/L by 24 h) of the lytic actinomycete was obtained in PHA production medium itself thus circumventing the prior actinomycete acclimatization just by co-inoculation with B. megaterium as an inducer. Maximum lytic activity was observed at pH 6.0, 40 °C, 220 mg of biomass and 33.3 mL of concentrated culture filtrate in a 100 mL reaction mixture. Preliminary biochemical investigations confirmed the proteolytic and caseinolytic nature of the lytic enzyme. PHA yield of 0.55 g/g by co-inoculation extraction approach was comparable with the conventional sodium hypochlorite based extraction method. Interestingly, S. albus also demonstrated a broad spectrum lytic potential against varied Gram-negative and Gram-positive PHA producers highlighting the extensive applicability of this biolytic PHA recovery approach. The lytic enzyme retained almost 100% relative activity on storage at −20 °C upto two months. 1H Nuclear magnetic resonance analysis of the extracted polymer confirmed it as a homopolymer composed of 3-hydroxybutyrate monomeric units. This is the first report on Streptomyces sp. based biological and eco-friendly, intracellular PHA recovery from Bacillus spp.
       
  • Purification and characterization of alkaline soda-bleach stable protease
           from Bacillus sp. APP-07 isolated from Laundromat soil

    • Abstract: Publication date: Available online 20 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): I.K. Shaikh, P.P. Dixit, T.M. Shaikh The detergent-compatible alkaline protease was produced from the bacterial strain Bacillus sp. APP-07 isolated from Laundromat soil of Solapur, Maharashtra, India. The culture was grown in 1000 ml capacity baffled flask with a working volume of 100 ml and incubated at 55 °C for 33 h on a rotary shaker. After incubation, alkaline protease was partially purified by the sequential method of acetone precipitation followed by nominal molecular weight limit (NMWL) cut-off ultrafiltration using 50 K and 10 K filters. Finally, Sephadex G-100 gel filtration chromatographic purification was performed to obtain 3.12 fold purified alkaline protease enzyme with a 66.67% final yield. The purified enzyme showed 31907.269 units (U) of enzyme activity containing 8741.718 U/mg of specific enzyme activity. The molecular weight of the enzyme was confirmed about 33.0 kDa (kDa) by the SDS-PAGE analysis. The purified enzyme was stable at higher pH and temperature range, with an optimum pH 10.5 and temperature 55 °C. The enzyme showed excellent stability and compatibility in various detergents, surfactants, bleach, and oxidizing agents. The enzyme activity enhanced in the presence of Ca2+, Cu2+, and surfactants, whereas; the phenylmethylsulphonyl fluoride (PMSF) and Diisopropyl fluorophosphate (DFP) completely inhibit the enzymatic activity, which pointed out that the enzyme affiliated to serine-centered metalloproteases family.In conclusion, the remarkable tolerance and stability of the enzyme explored the promising candidature for the several potential applications in the laundry detergents. The sustainability of the enzyme might serve several possible applications in the laundry detergents, leather industries, and other harsh industrial processes.
       
  • Influence of cold pretreatment on shoot regeneration from callus in date
           palm (Phoenix dactylifera L.) cv. ‘Barhee’

    • Abstract: Publication date: Available online 10 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Ahmed Madi Waheed Al-Mayahi, Abdulminam Hussien Ali, Hussein J. Shareef Mass propagation of date palm through indirect somatic embryogenesis or organogenesis has attracted the interest of commercial producers. But, this technique still faces some problems that hindered the production of date palm plantlets in vitro. Tissue browning is one of the serious problems that reduce callus growth and shoot regeneration. So the objective of the present study is to investigate the effect of cold pretreatment on callus growth, shoot regeneration, and polyphenol oxidase (PPO) activity during the callus culture. Results showed that a high survival rate of callus cultures (100%) were obtained when cultures were incubated in low temperature (cold treatment) for 45 and 75 days. On the other hand, total amount on phenolic compounds was also reduced to 0.47 and 0.53 mg GAE/g after same period of incubation (45 and 75 days respectively) at low temperature. In additional, our results showed that the highest frequency of shoot formation (66.67 and 73.34, %) and the highest shoot numbers (7.8 and 8.6 shoots/100 mg) were obtained from callus treated with low temperature for 45 and 75 days, respectively.
       
  • Quorum sensing intervened bacterial signaling: Pursuit of its cognizance
           and repression

    • Abstract: Publication date: Available online 7 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Kayeen Vadakkan, Abbas Alam Choudhury, Ramya Gunasekaran, Janarthanam Hemapriya, Selvaraj Vijayanand Bacteria communicate within a system by means of a density dependent mechanism known as quorum sensing which regulate the metabolic and behavioral activities of a bacterial community. This sort of interaction occurs through a dialect of chemical signals called as autoinducers synthesized by bacteria. Bacterial quorum sensing occurs through various complex pathways depending upon specious diversity. Therefore the cognizance of quorum sensing mechanism will enable the regulation and thereby constrain bacterial communication. Inhibition strategies of quorum sensing are collectively called as quorum quenching; through which bacteria are incapacitated of its interaction with each other. Many virulence mechanism such as sporulation, biofilm formation, toxin production can be blocked by quorum quenching. Usually quorum quenching mechanisms can be broadly classified into enzymatic methods and non-enzymatic methods. Substantial understanding of bacterial communication and its inhibition enhances the development of novel antibacterial therapeutic drugs. In this review we have discussed the types and mechanisms of quorum sensing and various methods to inhibit and regulate density dependent bacterial communication.
       
  • In silico structural homology modeling of nif A protein of rhizobial
           strains in selective legume plants

    • Abstract: Publication date: Available online 7 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Sadam D.V. Satyanarayana, M.S.R. Krishna, Pindi Pavan Kumar, Sirisha Jeereddy Symbiosis is a complex genetic regulatory biological evolution which is highly specific pertaining to plant species and microbial strains. Biological nitrogen fixation in legumes is a functional combination of nodulation by nod genes and regulation by nif, fix genes. Three rhizobial strains (Rhizobium leguminosarum, Bradyrhizobium japonicum, and Mesorhizobium ciceri) that we considered for in silico analysis of nif A are proved to be the best isolates with respect to N2 fixing for ground nut, chick pea and soya bean (in vitro) out of 47 forest soil samples. An attempt has been made to understand the structural characteristics and variations of nif genes that may reveal the factors influencing the nitrogen fixation. The primary, secondary and tertiary structure of nif A protein was analyzed by using multiple bioinformatics tools such as chou-Fasman, GOR, ExPasy ProtParam tools, Prosa -web. Literature shows that the homology modeling of nif A protein have not been explored yet which insisted the immediate development for better understanding of nif A structure and its influence on biological nitrogen fixation. In the present predicted 3D structure, the nif A protein was analyzed by three different software tools (Phyre2, Swiss model, Modeller) and validated accordingly which can be considered as an acceptable model. However further in silico studies are suggested to determine the specific factors responsible for nitrogen fixing in the present three rhizobial strains.
       
  • Production of biomass and flavonoid of Gynura procumbens (Lour.) Merr
           shoots culture in temporary immersion system

    • Abstract: Publication date: Available online 5 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Ayu Dewi Pramita, Alfinda Novi Kristanti, Sugiharto, Edy Setiti Wida Utami, Yosephine Sri Wulan Manuhara Gynura procumbens (Lour.) Merris one of medicinal plant which was carried out used as antioxidant, anticancer, anti-inflammatory, hepatoprotective, and antimicrobial. Many strategies were used to increase the production of biomass and valuable compounds. This study was to investigate the variation effect of growth regulators and immersion frequency on production of biomass and flavonoid contained of G. procumbens shoots culture in temporary immersion bioreactor. Stem nodes were used as an explants and induction of shoots were done in solid MS medium supplemented with many kinds of growth regulator. The best treatments were used to produce biomass and flavonoid compounds in temporary immersion bioreactor; there are combination of IAA 2 mg/L and BA 4, 6, 8 mg/L and immersion frequency (5 min each 3 h; 15 min each 12 h). Results showed that the growths of G. procumbens shoots in solid MS medium were influenced by supplementation of growth regulators. MS medium supplemented with single cytokinine (6 mg/L kinetin) and combination of auxin (IAA) and cytokinine (BA) caused increasing of shoots growth. Production of biomass of G. procumbens in temporary immersion bioreactor was achieved in long immersion interval (12 h) and highest flavonoid production was obtained in combination treatment of immersion frequency 15 min each 12 h and MS medium supplemented with IAA 2 mg/L, BA 8 mg/L.
       
  • Phytochemical analysis, antioxidant and antimicrobial activity of wild and
           in vitro derived plants of Ceropegia thwaitesii Hook – An endemic
           species from Western Ghats, India

    • Abstract: Publication date: Available online 3 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): S. Muthukrishnan, T. Senthil Kumar, A. Gangaprasad, F. Maggi, M.V. Rao Ceropegia thwaitesii Hook (Asclepiadaceae), an endemic plant species, due to habitat destruction and over exploitation has a very restricted distribution in the Western Ghats of Tamil Nadu, India. The present wrok aimed to determine the chemical composition, the total phenolic (TPC), flavonoid (TFC) and tannin content (TEC), and to assess the antioxidant properties of various extracts of in vivo plants (IVP) and in vitro regenerated plants (IRP) of C. thwaitesii. Some phenolic compounds like gallic acid, cathechol, vanillin and salicylic acid were identified and quantified by HPLC. All the extracts possessed relevant radical scavenging activity on DPPH, Superoxide radical scavenging activity, and Nitric oxide radicals as well as total antioxidant ability. DPPH assay of in vitro methanol stems extracts and ethanol leaves extracts revealed the best antioxidant properties with important IC50 values of 0.248 ± 0.45 µg/mL and 0.397 ± 0.67 µg/mL, respectively, whereas in vivo chloroform stems extracts showed a lower antioxidant activity (IC50 of 10.99 ± 0.24 µg/mL). The IRP methanol extracts of stem and leaves had good inhibitory activity against all tested microorganisms in a dose-dependent manner. These results suggested that in vitro raised plants of C. thwaitesii are an excellent source of antioxidant compounds to be exploited on an industrial level as food additive.
       
  • Expression, purification and biological characterisation of recombinant
           human irisin (12.5 kDa)

    • Abstract: Publication date: Available online 2 July 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Kalpana Panati, Venkata Ramireddy Narala, Vydyanath R. Narasimha, Madhavi Derangula, Venkat R.R. Arva Tatireddigari, Suneetha Yeguvapalli Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of energy expenditure markers PGC-1α, UCP-1, besides abundance of beige adipocytes in WAT. Though varied sizes of irisin were reported in humans and rodents it is not yet clear about the actual size of the irisin produced physiologically. Hence, we cloned and expressed human irisin (32–143 aa of FNDC5) in Escherichia coli based on the proposed cleavage site that yields 12.5 kDa peptide to study its antigenicity and other biological functions in vitro. We purified recombinant human irisin (rh-irisin) to 95% homogeneity with simple purification method with a yield of 25 mg/g wet cell pellet. rh-irisin has been detected by commercially available antibodies from different sources with similar antigenicity. Biological activity of the rh-irisin was confirmed by using 3T3-L1 pre-adipocyte differentiation by Oil red O staining. Further, rh-irisin treatment on pre-adipocytes showed increased expression of markers associated with energy expenditure. As it is involved in energy expenditure process, it could be considered as potential therapeutic option for various metabolic diseases.
       
  • Phylogenetic diversity and biotechnological potentials of marine bacteria
           from continental slope of eastern Arabian Sea

    • Abstract: Publication date: Available online 29 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Arakkaveettil Kabeer Farha, Thasneem TR, Aswathy Purushothaman, Jaseetha Abdul Salam, Abdulla Mohamed Hatha Marine environments are substantially untapped source for the isolation of bacteria with the capacity to produce various extracellular hydrolytic enzymes, which have important ecological roles and promising biotechnological applications. Hydrolases constitute a class of enzymes widely distributed in nature from bacteria to higher eukaryotes. Marine microbial communities are highly diverse and have evolved during extended evolutionary processes of physiological adaptations under the influence of a variety of ecological conditions and selection pressures. A number of marine hydrolases have been described, including amylases, lipases and proteases, which are being used extensively for biotechnological applications. The present study was carried out to isolate marine bacteria from continental slope sediments of the eastern Arabian Sea and explore their biotechnological potential. Among the 119 isolates screened, producers of amylases (15%), caseinases (40%), cellulases (40%), gelatinases (60%), lipases (26%), ligninases (33%), phytase (11%) and Malachite Green dye degraders (16%) were detected. Phylogenetic analysis based on 16S rRNA gene sequencing showed that predominant marine sediment bacteria possessing more than four enzymatic activities belonged to the phyla Firmicutes and Proteobacteria, was assigned to the genera Bacillus, Planococcus, Staphylococcus, Chryseomicrobium, Exiguobacterium and Halomonas. Biodegradation of the dye Malachite Green using the liquid decolorization assay showed that both the individual cultures (Bacillus vietnamensis, Planococcus maritimus and Bacillus pumilus) and their consortium were able to decolorize more than 70% of dye within 24 h of incubation. This is the first report on diversity and extracellular hydrolytic enzymatic activities and bioremediation properties of bacteria from continental slope sediment of eastern Arabian Sea.
       
  • Improvement of cellulose degradation by cloning of endo-β-1, 3-1, 4
           glucanase (bgls) gene from Bacillus subtilis BTN7A strain

    • Abstract: Publication date: Available online 28 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Wafaa K. Hegazy, Mohamed S. Abdel-Salam, Azhar A. Hussain, Hoda H. Abo-Ghalia, Safa S. Hafez The aim of this study is to construct a new recombinant strain able to degrade cellulose efficiently. The endo-β-1, 3-1, 4 glucanase (bgls) gene was cloned from Bacillus subtilis BTN7A strain by using PCR technique. The specific primers of bgls gene were deduced. Optimization of PCR mixture and program were identified. The nucleotide sequence of bgls was placed in the public domain (GenBank accession number KM009051.1). The obtained bgls DNA was cloned with pGEM®-T Easy Vector. The recombinant plasmid designated as Bgls-NRC-1 was transformed into E. coli DH5α. The successful cloning of the bgls gene was tested either by PCR or by evaluating its expression in its new bacterial host. The bgls gene was expressed efficiently in E. coli and the enzyme activity of the transformant was compared to the enzyme activity of the donor bacterial strain. The new constructs produce much higher enzyme yields than the donor bacterial strain, they produce about 29% and about 57% higher cellulase specific activity at 37 °C and 55 °C respectively. Optimization of cellulolytic activity of the new recombinant strain were described. The effect of minimal medium supplemented with CMC or cellulose, or complete medium (LB) on bgls expression were tested, the order of cellulase activity production was CMC27.2 > cellulose 21.9 > LB 19.8 U/mg protein, respectively at 24 h. CMC was proved to be the best medium for cellulase production. Results also showed that double the initial inoculum resulted in more cellulase activities in all media.
       
  • Screening of plant growth promoting traits in heavy metals resistant
           bacteria: Prospects in phytoremediation

    • Abstract: Publication date: Available online 28 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): N. Tirry, N. Tahri Joutey, H. Sayel, A. Kouchou, W. Bahafid, M. Asri, N. El Ghachtouli Phytoremediation is considered as a novel environmental friendly technology, which uses plants to remove or immobilize heavy metals. The use of metal-resistant plant growth-promoting bacteria (PGPB) constitutes an important technology for enhancing biomass production as well as tolerance of the plants to heavy metals. In this study, we isolated twenty seven (NF1-NF27) chromium resistant bacteria. The bacteria were tested for heavy metals (Cr, Zn, Cu, Ni, Pb and Co) resistance, Cr(VI) reduction and PGPB characters (phosphate solubilization, production of IAA and siderophores). The results showed that the bacterial isolates resist to heavy metals and reduce Cr(VI), with varying capabilities. 37.14% of the isolates have the capacity of solubilizing phosphate, 28.57% are able to produce siderophores and all isolates have the ability to produce IAA. Isolate NF2 that showed high heavy metal resistance and plant growth promotion characteristics was identified by 16S rDNA sequence analysis as a strain of Cellulosimicrobium sp.. Pot culture experiments conducted under greenhouse conditions showed that this strain was able to promote plant growth of alfalfa in control and in heavy metals (Cr, Zn and Cu) spiked soils and increased metal uptake by the plants. Thus, the potential of Cellulosimicrobium sp. for both bioremediation and plant growth promotion has significance in the management of environmental pollution.
       
  • Synthesis of silver nanoparticles by Bacillus clausii and computational
           profiling of nitrate reductase enzyme involved in production

    • Abstract: Publication date: Available online 28 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Koel Mukherjee, Rashmi Gupta, Gourav Kumar, Sarita Kumari, Saptaswa Biswas, Padmini Padmanabhan Biogenic synthesis of silver nanoparticles using microorganisms has found interest recently since last decade because of their prospect to synthesize nanoparticles of various size, shape and morphology which are eco-friendly. Here, an eco-friendly method for production of silver nanoparticles from Bacillus clausii cultured from Enterogermina is explored. Along with the biosynthesis and conformity test, in silico studies was done on NADPH dependent nitrate reductase enzymes from the view point of designing a rational enzymatic strategy for the synthesis. The detailed characterization of the nanoparticles was carried out using UV-Vis spectroscopy, Dynamic Light Scattering (DLS) particle size analysis, Transmission Electron Microscopy (TEM), X-Ray Diffraction (XRD) analysis. Computational profiling and in silico characterization of NADH dependent enzymes was carried out based on literature and work done so far. Nitrate reductase sequence was retrieved from NCBI for characterization. Secondary structure was evaluated and verified by JPred as well as SOPMA Tool. Tertiary structure was also modeled by MODELLER and ITASSER parallel and the best structure was selected based on energy values. Structure validation was done by GROMACS and RMSD, RMSF, temperature variation plot were also plotted. Interactions graphs between nitrate reductase and ligand silver nitrate was done through molecular docking using Hex.
       
  • Evaluation of the alleviative role of Chlorella vulgaris and Spirulina
           platensis extract against ovarian dysfunctions induced by monosodium
           glutamate in mice

    • Abstract: Publication date: Available online 28 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Sekena H Abdel-Aziem, Heba A.M. Abd El-Kader, Faten M. Ibrahim, Hafiza A Sharaf, Aida I. El makawy Microalgae provide a wealthy natural resource of bioactive compounds, which have many biological activities. Monosodium glutamate is a food additive that acts either as food preservatives or as tastiness enhancer. It was confirmed that monosodium glutamate poses a serious responsibility in the pathogenesis of anovulatory infertility. Therefore, the idea of this research was directed to reveal efficiency of Chlorella vulgaris and Spirulina platensis extracts against the ovarian dysfunction resulted due to monosodium glutamate consumption. Adult female albino mice were gavages orally monosodium glutamate alone or with either Chlorella vulgaris or Spirulina platensis aqueous extracts for 28 days. Female mice were subjected to superovulation to study the oocytes nuclear maturation stages. Histological and quantitative investigation was carried on ovaries. Biochemical assessment to measure the sex hormones level and ovarian enzymatic antioxidants was done. In addition, ovarian antioxidant mRNA genes were determined using quantitative PCR and Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. The result revealed that monosodium glutamate reduced the oocytes quality and maturation rate, while, both algae improve the oocyte quality and maturation rate than in monosodium glutamate group. Chlorella vulgaris and Spirulina platensis improved the monosodium glutamate ovarian tissue histological alteration, sex hormones content and raised the ovarian enzymatic antioxidants level. In addition, monosodium glutamate markedly diminished the Glutathione peroxidase, superoxide dismutase and catalase mRNA expressions, However, Chlorella vulgaris or Spirulina platensis upregulated the expression of genes close to control. In conclusion, Chlorella vulgaris and Spirulina platensis showed potential alleviative role against the monosodium glutamate ovarian dysfunction.
       
  • Antibacterial activity of soil bacteria isolated from Kochi, India and
           their molecular identification

    • Abstract: Publication date: Available online 14 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Davis Gislin, Dorairaj Sudarsanam, Gnanaprakasam Antony Raj, Kathirvelu Baskar The present study, deal about the antibiosis activity of soil bacteria, isolated from 10 different locations of rhizosphere and diverse cultivation at Kochi, Kerala, India. The bacteria were isolated by standard serial dilution plate techniques. Morphological characterization of the isolate was done by Gram’s staining and found that all of them gram positive. Isolated bacteria were tested against 6 human pathogens viz., Escherichia coli, Enterococcus sp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus and Acinetobacter sp. Primary screening was carried out by perpendicular streaking and seed overlay method. Based on the result of primary screening most potential isolates of S1A1 and S7A3 were selected for secondary screening. Both the isolates showed positive results against Enterococcus sp. and S.aureus. The maximum antagonistic activity of 20.98 and 27.08 mm zone of inhibition was recorded at S1A1 against Enterococcus sp. and S. aureus respectively, at 180 µl concentration. Molecular identification was carried out by 16S rRNA sequence. The 16S rRNA was amplified from the DNA samples by using PCR. The amplified 16S rRNA PCR products were purified and sequenced. The sequences were subjected to NCBI BLAST. The isolates S1A1 and S7A3 BLAST results showed 99% and 95% respectively, similarity with the available database sequence of Bacillus amyloliquefaciens. The sequences were deposited in GenBank and the accession numbers KY864390 (S1A1) and KY880975 (S7A3) were obtained.
       
  • In silico analysis of squalene synthase in Fabaceae family using
           bioinformatics tools

    • Abstract: Publication date: Available online 6 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Zahra Aminfar, Masoud Tohidfar Triterpenoid saponins are a diverse group of bioactive compounds, which are used for possessing of many biomedical and pharmaceutical products. Generally, squalene synthase (SQS) is defined as an emerging and essential branch point enzyme far from the major pathway of isoprenoids biosynthetic and a latent adjusting point, which manages carbon flux into triterpenes biosynthesis and sterols. The present study deals with the detailed characterization of SQS by bioinformatics approaches to evaluate physicochemical properties, structural characteristics including secondary and 3D structure prediction and functional analysis from eight plants related to Fabaceae family and Arabidopsis thaliana. Bioinformatics analysis revealed that SQS proteins have two transmembrane regions in the C-terminal. The predicted motifs were used to design universal degenerate primers for PCR analysis and other molecular applications. Phylogenetic analysis showed conserved regions at different stretches with maximum homology in amino acid residues within all SQSs. The secondary structure prediction results showed that the amino acid sequence of all squalene synthases had α helix and random coil as the main components. The reliability of the received model was confirmed using the ProSA and RAMPAGE programs. Determining of active site by CASTp proposes the possibility of using this protein as probable medication target. The findings of the present study may be useful for further assessments on characterization and cloning of squalene synthase.
       
  • Healthcare-associated (HA) and community-associated (CA) methicillin
           resistant Staphylococcus aureus (MRSA) in Bangladesh – Source, diagnosis
           and treatment

    • Abstract: Publication date: Available online 5 June 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Md. Anowar Khasru Parvez, Rabeya Nahar Ferdous, Md. Shahedur Rahman, Sohidul Islam Methicillin-resistant Staphylococcus aureus (MRSA) has long been a common pathogen in healthcare facilities, but now, it has emerged as a problematic pathogen in the community setting as well. This study reported source, diagnosis and treatment of HA-MRSA and CA-MRSA.A total of sixty-five clinical samples (urine, pus, wound swab) were collected from clinical origin of Dhaka city, Bangladesh. All the isolates were tested phenotypically by conventional methods and genotypically by PCR targeting nuc, pvl and mecA genes. Finally sequencing was carried out for pvl gene to know the mutagenic variation or any amino acid changes in pvl gene. Chi square test was employed for statistical analysis. Patients of age group 51–60 years are more susceptible (46.15%) to MRSA, CA-MRSA or HA-MRSA infection. Female are (32.30%) more susceptible to MRSA infection. Among 65 isolates 53 isolates identified phenotypically as S. aureus. These were positive for amplification of nuc (270 bp) gene of S. aureus. Moreover, among 53 isolates 33 phenotypically considered as MRSA and 38 (72%) showed positive amplification for mecA (162 bp) gene. Among 38 MRSA isolates 22 (57.89%) confirmed as CA-MRSA and 16 (42.10%) as HA-MRSA. Finally, sequence analysis for lukS/F-PV genes from 4 representative isolates detected a new single nucleotide polymorphism in comparison with the control sequence. However, no amino acid changes were found. Statistical analysis showed HA-MRSA isolates were more commonly found in urine sample and CA-MRSA in pus and wound swab. CA-MRSA isolates were more resistant to tested antibiotics than HA-MRSA.
       
  • Production and characterisation of exopolysaccharide from Streptomyces
           carpaticus isolated from marine sediments in Egypt and its effect on
           breast and colon cell lines

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Manal S. Selim, Shaimaa K. Amer, Sahar S. Mohamed, Marwa M. Mounier, Hala M. Rifaat Twenty streptomycete strains were isolated from marine sediment samples collected from Nabq area, Sharm El-Sheikh, Red Sea Coast, Egypt. Four of them produce exopolysaccharides (EPS) showing marked in vitro antitumor activities. Morphological and cultural characteristics of the most significant strain (No. 3) were shown. Moreover, the sequence of this strain showed similarity with Streptomyces carpaticus. The results reveal that EPS produced by Streptomyces carpaticus No. 3 had high cytotoxicity reaching 51.7% and 59.1% against human tumor cells of breast and colon lines respectively. A chemical analysis of EPS indicated that the composing monosaccharides were galactouronic acid, glucose, xylose, galactose, mannose, and fructose with relative ratio of 3:1:1:2:2:1 respectively, with an average molecular weight (Mw) 1.180 × 105 g/mol and of a number average molecular weight (Mn) 1.052 × 105 g/mol. Also the EPS contained uronic acid (0.5072%) and monosaccharide sulphates (21.753%).
       
  • Identification of Mx gene nucleotide dimorphism (G/A) as genetic marker
           for antiviral activity in Egyptian chickens

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Mohamed S. Hassanane, Amal A.M. Hassan, Fatma M. Ahmed, Esteftah M. El-Komy, Khaled M. Roushdy, Nagwa A. Hassan Egyptian chickens, representing 2 breeds and 7 strains, were genotyped using the PCR-RFLP and sequencing techniques for detection of a non-synonymous dimorphism (G/A) in exon 14 of chicken Myxovirus resistance (Mx) gene. This dimorphic position is responsible for altering Mx protein’s antiviral activity. Polymerase Chain reactions were performed using Egyptian chickens DNA and specific primer set to amplify Mx DNA fragments of 299 or 301 bp, containing the dimorphic position. Amplicons were cut with restriction enzyme Hpy81. Genotype and allele frequencies for the resistant allele A and sensitive allele G were calculated in all the tested chickens. Results of PCR-RFLP were confirmed by sequencing. The three genotypes AA, AG, GG at the target nucleotide position in Mx gene were represented in all the studied Egyptian chicken breeds and strains except Baladi strain which showed only one genotype AA. The average allele frequency of the resistant A allele in the tested birds (0.67) was higher than the sensitive G allele average frequency in the same birds (0.33). Appling PCR-RFLP technique in the breeding program can be used to select chickens carrying the A allele with high frequencies. This will help in improving poultry breeding in Egypt by producing infectious disease-resistant chickens.
       
  • Molecular detection and PCR-RFLP analysis using Pst1 and Alu1 of multidrug
           resistant Klebsiella pneumoniae causing urinary tract infection in women
           in the eastern part of Bangladesh

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Golam Mahmudunnabi, Al Nahian Khan Majlish, Farhana Momtaz, Md Javed Foysal, Md Mahbubur Rahman, Kamrul Islam Klebsiella pneumoniae is the second leading causative agent of UTI. In this study, a rapid combined polymerase chain reaction and restriction fragment length polymorphism analysis was developed to identify K. pneumoniae in women, infected with urinary tract infection in the Sylhet city of Bangladesh. Analysis of 11 isolates from women at the age range of 20–55 from three different hospitals were done firstly by amplification with K. pneumoniae specific ITS primers. All of the 11 collected isolates were amplified in PCR and showed the expected 136 bp products. Then, restriction fragment length polymorphism analysis of 11 isolates were conducted after PCR amplification by 16s rRNA universal primers, followed by subsequent digestion and incubation with two restriction enzymes, Pst1 and Alu1. Seven out of 11 isolates were digested by Pst1 restriction enzymes, six isolates digested by Alu1, and while others were negative for both enzymes. Data results reveal that, women at age between 25 and 50 were digested by both enzymes. A woman aged over than 50 was negative while bellow 20 was digested by only Pst1. The results could pave the tactic for further research in the detection of K. pneumoniae from UTI infected women.
       
  • The sensitivity and efficacy method of PIK3CA exon 9 E545A as a high
           diagnostic accuracy in breast cancer

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Desriani, Fuad Al-Ahwani The phosphatidylinositol 3-kinases (PIK3s) are lipid kinases. Mutation in the exon 9 and exon 20 determined as a predictive factor in anti-HER-2 therapy. In some countries, such as Singapore, China, and Peru, PIK3CA exon 9 E545A was reported to produce the highest rate of mutation. In this research, we developed and optimized PIK3CA exon 9 E545A detection methods with intercalating dye SYBR Green I based on the Tm Shift approach by using prepared recombinant plasmid pGEMT-easy PIK3CA exon 9 and PIK3CA exon 9 E545A. Recombinant plasmid was used due to the limited number of samples.MethodsRecombinant plasmid was prepared based on manufactured procedures, and this process was then followed by Tm prediction with Poland software, Tm Shift SYBR Green I development, and its characterization (reproducibility, repeatability, sensitivity, qPCR efficiency, and qPCR amplification), respectively.ResultA method for PIK3CA E545A detection based on TM shift SYBR Green I has been successfully developed. The melting temperature for PIK3CA exon 9 was 78.1 ± 0.1 °C, while that for PIK3CA exon E545A was 80.20 °C. The Tm of mutant was the same as that predicted using Polland Software. The reproducibility of the methods was high, with the coefficient values for inter and intra assays were below 10% with a high sensitivity at 1%, while R2 0.99 and PCR efficiency was 97.75%.ConclusionThe results presented here demonstrate that the PIK3CA exon 9 E545A detection method has a good sensitivity and efficacy assay, which proves that the method has a high diagnostic accuracy in breast cancer.
       
  • Differentially expressed genes: OCT-4, SOX2, STAT3, CDH1 and CDH2, in
           cultured mesenchymal stem cells challenged with serum of women with
           endometriosis

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Ehab Salama, Ghada Nour Eldeen, Mazen Abdel Rasheed, Sahar Abdel Atti, Amr Elnoury, Tamer Taha, Osama Azmy Endometriosis is a common chronic gynecological disorder defined as the presence of ectopic functional endometrial tissues, outside uterine cavity, primarily on the pelvic peritoneum and the ovaries. Several studies revealed a correlation between aberrant stem-cell activity in the endometrium and endometriosis. Yet the molecular and cellular behaviors of mesnchymal stem cells in development of endometriosis are hampered by lack of invitro experiments. Our aim was to explore morphological and molecular changes associated with mesenchymal stem cells (MSCs) exposition to serum derived from women with severe endometriosis. Two cell cultures of MSCs isolated from endometrial tissues of two endometriosis-free women. Each cell culture was treated individually with the serum of women with endometriosis (experimental group/n = 7), and serum of women without endometriosis (control group/ n = 4) for 14 days. Quantitative Real-Time PCR was performed later to reveal expression of OCT-4, CDH1 and CDH2, STAT3 and SOX2 genes. Morphologically, cells showed no significant changes. However from molecular point of view, we found increased expression in OCT-4, CDH1 and CDH2. For STAT3 and SOX2 we did not find a significant difference. This study shows that endometriosis serum induced molecular changes in human endometrial MSCs (EnMSCs) that might be related to altered cell behavior which may be a step in differentiation that may be completed invivo by other factors to complete the process of transition. Further researches are needed for optimization to reach differentiation.
       
  • Effect of vitamin A deficiency on thymosin-β4 and CD4 concentrations

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Salwa Refat El-Zayat, Hiba Sibaii, Nermine N. Mahfouz, Sara F. Sallam, Reham F. Fahmy, Azza Abd El-Shaheed Vitamins are evaluated for their role in immunity. Recently, vitamin A received a particular attention as a critical micronutrient for regulating immune system. Therefore, the present study aimed to search for new about vitamin A. Forty-eight Egyptian adults aged from 18 to 42 years old from both sexes were subjected to clinical examination and nutrition questionnaire and were screened for vitamin A by using ELISA method. Forty subjects were selected and subdivided into two groups. Group 1 with vitamin A at level>200 µg/dl consists of 10 healthy subjects. Group 2 with vitamin A deficiency at level
       
  • Down-regulation of circulating microRNA let-7a in Egyptian smokers

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Sanaa A. Rizk, Fateheya M. Metwally, Asmaa M. Elfiky, Asmaa A. Mahmoud, Nadia A. Badawi, Nevin E. Sharaf, Mahmoud M. Elhefnawi Altered miRNAs were associated with cigarette smoking. The study aimed to examine the gene expression level of plasma let-7a among healthy smokers and compared it with the non-smokers. Forty subjects were recruited for the present study and classified into 21 smokers and 19 non-smokers, age, and sex were matched. The software that used to design functional primers was MIRprimer. Quantitative real-time PCR was employed to compare the relative expression of plasma let-7a. Results showed that the level of let-7a was down-regulated in smokers to 0.34fold (p = 0.006) that of the non-smokers. Plasma let-7a showed an area under curve (AUC) of 0.749 with sensitivity 43% and specificity 100%. In conclusion, plasma let-7a was significantly down-regulated in the smokers, and it might be considered a candidate biomarker to discriminate between smokers and non-smokers.
       
  • Impact of Apo E gene polymorphism on HCV therapy related outcome in a
           cohort of HCV Egyptian patients

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Howayda E. Gomaa, Mohamed Mahmoud, Nevine E. Saad, Amal Saad-Hussein, Somaia Ismail, Eman H. Thabet, Hebatallah Farouk, Dina Kandil, Ahmed Heiba, Wael Hafez The functional apolipoprotein E (Apo E) gene polymorphism could be used as a determinant of outcome of HCV infection. This study aimed to demonstrate the impact of Apo E genotype on the response to HCV combined therapy. Material and methods: The study has been implemented on 125 individuals with persistent HCV infection and 120 cases with sustained virologic response (SVR). All participants were genotyped for ApoE gene polymorphism by a real-time quantitative PCR (qPCR). Results: Statistically significant differences were demonstrated regarding the Apo E genotypes between the two groups (P-value 
       
  • Isolation, partial purification, biochemical characterization and
           detergent compatibility of alkaline protease produced by Bacillus
           subtilis, Alcaligenes faecalis and Pseudomonas aeruginosa obtained from
           sea water samples

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Sarika Kedar Marathe, Manisha Arun Vashistht, Aishwarya Prashanth, Nikhat Parveen, Shailayee Chakraborty, Sindhu S. Nair In the current study, bacteria isolated from sea water samples of Murdeshwar, Karnataka, were screened for the production of alkaline protease by culturing them onto skim milk agar media. Of the isolated bacteria, Bacillus subtilis, Pseudomonas aeruginosa and Alcaligenes faecalis showed distinct zones of hydrolysis due to enzyme production. They were each inoculated into enzyme production media under submerged fermentation conditions at 37 °C for 48 h with a constant agitation of 120 rpm. Partial purification of alkaline protease was carried out by isoelectric precipitation. Enzyme activity was determined under varying conditions of pH, incubation temperature, different substrates, carbon and nitrogen sources and salt concentrations using sigma’s universal protease activity assay. Enzyme immobilization was carried out using 2% Sodium alginate and 0.1 M ice cold CaCl2 and its activity under varying pH, temperature conditions and detergent compatibility was assayed. Efficacy of enzyme in stain removal was tested and haemolysis was observed within of 60 s which resulted in removal of the stain. Among the three organisms, enzyme from Bacillus subtilis showed highest activity in all cases indicating that it was the most ideal organism for enzyme production.
       
  • Partial purification and characterization of serine protease produced
           through fermentation of organic municipal solid wastes by Serratia
           marcescens A3 and Pseudomonas putida A2

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Asif Iqbal, Al Hakim, Md. Saddam Hossain, Mohammad Rejaur Rahman, Kamrul Islam, Md. Faisal Azim, Jahed Ahmed, Md. Assaduzzaman, Md. Mozammel Hoq, Abul Kalam Azad Proteolytic bacteria isolated from municipal solid wastes (MSW) were identified as Serratia marcescens A3 and Pseudomonas putida A2 based on 16S rDNA sequencing. Protease produced through fermentation of organic MSW by these bacteria under some optimized physicochemical parameters was partially purified and characterized. The estimated molecular mass of the partially purified protease from S. marcescens and P. putida was approximately 25 and 38 kDa, respectively. Protease from both sources showed low Km 0.3 and 0.5 mg ml−1 and high Vmax 333 and 500 µmole min−1 at 40 °C, and thermodynamics analysis suggested formation of ordered enzyme-substrate (E-S) complexes. The activation energy (Ea) and temperature quotient (Q10) of protease from S. marcescens and P. putida were 16.2 and 19.9 kJ/mol, and 1.4 and 1.3 at temperature range from 20 to 40 °C, respectively. Protease of the both bacterial isolates was serine and cysteine type. The protease retained approximately 97% of activity in the presence of sodium dodecyl sulphate. It was observed that the purified protease of S. marcescens could remove blood stains from white cotton cloth and degrade chicken flesh remarkably. Our study revealed that organic MSW can be used as raw materials for bacterial protease production and the protease produced by S. marcescens A3 might be potential for applications.
       
  • Influence of PEG induced drought stress on molecular and biochemical
           constituents and seedling growth of Egyptian barley cultivars

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): F.A. Hellal, H.M. El-Shabrawi, M. Abd El-Hady, I.A. Khatab, S.A.A. El-Sayed, Chedly Abdelly In order to investigate the effects of drought stress on germination components of barley cultivars, a laboratory experiment was conducted in a factorial randomized complete design with four replications. The controlled experiment included ten of Egyptian barley cultivars namely; (Giza 123, 124, 125, 126, 127, 129, 130, 134, 135 and 2000) as first factor. The second factor included 4 levels of drought stress inducer by applying 0, 5, 10 and 20% of polyethylene glycol-6000 (PEG) which is equivalent to four osmotic potential levels including −0.001, −0.27, −0.54 and −1.09 MPa, respectively. The results showed that, the highest reduction was related to the drought level of 20% PEG among the barley cultivars. The best cultivars in terms of germination traits were Giza 134, Giza 127, and Giza 126 this indicate their tolerance to drought stress and Giza 130, 135, 2000 cultivars was moderately tolerance and remaining is less tolerance. The protein band 27 kDa and 78 kDa showed high intensity after stress in almost all cultivars. Those two protein bands their exciting was very clear in treated barley leaf tissue. It could be related to dehydrine and oxygen evolving enhancer protein 2 (OEE2) which involved in drought stress tolerance response. Cultivars Giza 127, 130 and 134 showed highest tolerance response under drought stress. The antioxidant enzymes PAGE pattern of Peroxidase (POX), Sodium dismutase (SOD) and Ascorbate peroxidase (APX) for Barley cultivars under drought stress revealed a high activities for Giza 126, 127, 134, 136 and 2000 under −0.5 MPa osmotic stress by PEG in most of their isoforms. Based on similarity coefficient values the highest values were 1.0 with 100% similarly between tolerant cultivars Giza 130 and Giza 127. Similarly between the susceptible cultivars 125 and Giza 129 was 60%.These data confirmed by the growth parameters which we ranked as tolerant to drought stress.
       
  • Bioprospecting of indigenous resources for the exploration of
           exopolysaccharide producing lactic acid bacteria

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Syeda Bushra Zafar, Nadir Naveed Siddiqui, Faiza Shahid, Shah Ali Ul Qader, Afsheen Aman Exploration of biodiversity lead towards the discovery of novel exopolysaccharide (EPS) producing microbes that have multiple applications. The safety compatibility status of lactic acid bacteria (LAB) makes it an attractive candidate for the production of EPS in industries. Therefore, new bacterial isolates are continuously being identified from different habitats. Current research was conducted to explore indigenous biodiversity for the production of dextransucrase, which is involved in the synthesis of dextran. Dextran is an EPS which is used in different industries. In this study, thirty-nine LAB were isolated from different food samples. The isolates were identified as genus Leuconostoc, Weissella and Streptococcus based on genotypic and phenotypic characteristics. Screening revealed that only eight isolates can produce dextransucrase in high titres. Fermentation conditions of dextran producing LAB was optimized. The results indicated that Weissella confusa exhibited maximum specific activity (1.50 DSU mg−1) in 8 h at 25 °C with pH 7.5. Dextran produced from Weissella proved to be a useful alternative to commercially used dextran produced by Leuconostoc mesenteroides in industries for various applications.
       
  • Augmented cellulase production by Bacillus subtilis strain MU S1 using
           different statistical experimental designs

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): C.P. Sreena, Denoj Sebastian The production of cellulase by Bacillus subtilis MU S1, a strain isolated from Eravikulam National Park, was optimized using one-factor-at-a-time (OFAT) and statistical methods. Physical parameters like incubation temperature and agitation speed were optimized using OFAT and found to be 40 °C and 150 rpm, respectively, whereas, medium was optimized by statistical tools. Plackett-Burman design (PBD) was employed to screen the significant variables that highly influence cellulase production. The design showed carboxymethyl cellulose (CMC), yeast extract, NaCl, pH, MgSO4 and NaNO3 as the most significant components that affect cellulase production. Among these CMC, yeast extract, NaCl and pH showed positive effect whereas MgSO4 and NaNO3 were found to be significant at their lower levels. The optimum levels of the components that positively affect enzyme production were determined using response surface methodology (RSM) based on central composite design (CCD). Three factors namely CMC, yeast extract and NaCl were studied at five levels whilst pH of the medium was kept constant at 7. The optimal levels of the components were CMC (13.46 g/l), yeast extract (8.38 g/l) and NaCl (6.31 g/l) at pH 7. The maximum cellulase activity in optimized medium was 566.66 U/ml which was close to the predicted activity of 541.05 U/ml. Optimization of physical parameters and medium components showed an overall 3.2-fold increase in activity compared to unoptimized condition (179.06 U/ml).
       
  • Omics based approach for biodiscovery of microbial natural products in
           antibiotic resistance era

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): N. Chandra Mohana, H.C. Yashavantha Rao, D. Rakshith, P.R. Mithun, B.R. Nuthan, S. Satish The need for a new antibiotic pipeline to confront threat imposed by resistant pathogens has become a major global concern for human health. To confront the challenge there is a need for discovery and development of new class of antibiotics. Nature which is considered treasure trove, there is re-emerged interest in exploring untapped microbial to yield novel molecules, due to their wide array of negative effects associated with synthetic drugs. Natural product researchers have developed many new techniques over the past few years for developing diverse compounds of biopotential. Taking edge in the advancement of genomics, genetic engineering, in silico drug design, surface modification, scaffolds, pharmacophores and target-based approach is necessary. These techniques have been economically sustainable and also proven efficient in natural product discovery. This review will focus on recent advances in diverse discipline approach from integrated Bioinformatics predictions, genetic engineering and medicinal chemistry for the synthesis of natural products vital for the discovery of novel antibiotics having potential application.
       
  • A common neighbor based technique to detect protein complexes in PPI
           networks

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Mokhtarul Haque, Rosy Sarmah, Dhruba K. Bhattacharyya Detection of protein complexes by analyzing and understanding PPI networks is an important task and critical to all aspects of cell biology. We present a technique called PROtein COmplex DEtection based on common neighborhood (PROCODE) that considers the inherent organization of protein complexes as well as the regions with heavy interactions in PPI networks to detect protein complexes. Initially, the core of the protein complexes is detected based on the neighborhood of PPI network. Then a merging strategy based on density is used to attach proteins and protein complexes to the core-protein complexes to form biologically meaningful structures. The predicted protein complexes of PROCODE was evaluated and analyzed using four PPI network datasets out of which three were from budding yeast and one from human. Our proposed technique is compared with some of the existing techniques using standard benchmark complexes and PROCODE was found to match very well with actual protein complexes in the benchmark data. The detected complexes were at par with existing biological evidence and knowledge.
       
  • Optimization of germination, callus induction, and cell suspension culture
           of African locust beans Parkia biglobosa (Jacq.) Benth

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Mohamed S. Abbas, Hattem M. El-Shabrawi, Amira Sh. Soliman, Mai A. Selim The present study was carried out to determine the best pre-sowing treatments that can enhance the germination and seedling growth of Parkia biglobosa (Jacq.) Also, to establish and long-term maintenance of calli and cell suspension cultures . The result of various pre-sowing treatments showed that seeds soaked in concentrated H2SO4 treatment appeared the highest germination percentage, higher value of plant height, number of leaves, number of branches and stem girth. The MS medium containing 1mg/l 2, 4-D was the best for callus induction of stem explants. The addition of 50 mg /l citric acid to the MS medium was effective for reducing browning of callus than other treatments. However, the viability percent recorded the maximum (87.76%) on the 9th day while the concentration of viable cells per ml reached the higher record (137.5 viable cell/ml) at the 12th and cell viability remains (≈ 68.39%) throughout 18 days of culture
       
  • Utilization of horticultural waste (Apple Pomace) for multiple
           carbohydrase production from Rhizopus delemar F2 under solid state
           fermentation

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Shruti Pathania, Nivedita Sharma, Shweta Handa The brown rot fungus Rhizopus delemar F2 was shown to produce extracellular thermostable and multiple carbohydrase enzymes. The potential of Rhizopus delemar F2 in utilizing apple pomace under solid state fermentation (SSF) is the purpose of the study. Solid state fermentation (SSF) is a very effective technique opposed to submerged fermentation in various aspects. Enhanced production of multiple carbohydrases 18.20 U g−1 of cellulose, 158.30 U g−1 of xylanase, 61.50 U g−1 of pectinase and amylase 21.03 U g−1 was released by microwave pretreatment of apple pomace at 450 W for 1 min and then by incubation the culture thus obtained at 30 °C for 6 days with moisture content of 1:4.5. Apple pomace can serve as a potential source of raw material for the production of multiple carbohydrases. Besides, it can find great commercial significance in production of bioethanol and various industries like textile, fruit juice, paper and pulp industry.
       
  • Effect of plant growth regulators on indirect shoot organogenesis of Ficus
           religiosa through seedling derived petiole segments

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Mohsen Hesami, Mohammad Hosein Daneshvar, Mohsen Yoosefzadeh-Najafabadi, Milad Alizadeh Ficus religiosa is known as a long-lived multipurpose forest tree. The tree plays an important role for religious, medicinal, and ornamental purposes. However, the propagation rate of Ficus religiosa is low in natural habitat so the plant tissue culture techniques are an applicable method for multiplication of this valuable medicinal plants. Thus, the aim of this study is to understand the effect of different auxin/cytokinin ratios on indirect shoot organogenesis of this plant. According to our results, the maximum callus induction frequency (100%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) plus 0.05 mg/l 6-benzylaminopurine (BAP) from petiole segments. For shoot induction purpose, the yellow-brownish, friable, organogenic calli were inoculated on shoot induction medium. On MS medium supplemented with 1.5 mg/l BAP and 0.15 mg/l Indole-3-butyric acid (IBA), 96.66% of the petiole-derived calli responded with an average number of 3.56 shoots per culture. The highest root formation frequency (96.66%), root number (5.5), and root length (4.83 cm) were achieved on MS medium containing 2.0 mg/l IBA plus 0.1 mg/l Naphthaleneacetic acid (NAA). The rooted shoots were successfully transferred to field condition and the substrate with the mixture of cocopeat and perlite (1:1) had the highest survival rate (96.66%). This is the first report of an effective in vitro organogenesis protocol for F. religiosa by indirect shoot organogenesis through axenic seedling derived petiole explants, which can be efficiently employed for conservation of this important medicinal plant species as well as the utilization of active biomolecules.
       
  • Molecular insights into the genetic and haplotype diversity among four
           populations of Catla catla from Madhya Pradesh revealed through mtDNA cyto
           b gene sequences

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): R.K. Garg, Vaishali Mishra In the present investigation, the genetic structure of four populations of Catla catla, sequences of mitochondrial gene, cytochrome b (cyto b) from four populations were sequenced and analyzed. The sequences of mitochondrial regions revealed high haplotype diversity and low nucleotide diversity. The lowest 249 polymorphic sites and 0.00 parsimony informative sites were detected in populations of Fish Federation Pond (CCFFB) whereas highest 330 polymorphic sites and 56 parsimony informative sites were detected in populations of Narmada River (CCNRH) in the cyto b gene sequences in Catla catla populations. The twelve different haplotypes were detected among the four populations studied, lowest population specific haplotype as 2.00 was observed in Fish Federation Pond (CCFFB) and highest was in Population of Narmada River and Tighra reservoir. Sequencing of cyto b gene revealed 12 number of haplotypes (h) with haplotype (gene) diversity (Hd) 0.8736 and nucleotide diversity (π) 0.6474. These data clearly indicated that, feral/wild population showing highest values of polymorphisms, parsimony, haplotype diversity showing good, healthy habitat is lotic water (Narmada River) and lentic water body (Tighra reservoir). The results also concluded that the partial cyto b is polymorphic and can be a potential marker to determine ecological habitat based genetic differentiation among the populations.
       
  • Genetic variation of salt-stressed durum wheat (Triticum turgidum subsp.
           durum Desf.) genotypes under field conditions and gynogenetic capacity

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Olfa Ayed-Slama, Imen Bouhaouel, Zoubeir Chamekh, Youssef Trifa, Ali Sahli, Nadhira Ben Aissa, Hajer Slim-Amara Agriculture has new challenges against the climate change: the preservation of genetic resources and the rapid creation of new varieties better adapted to abiotic stress, specially salinity. In this context, the agronomic performance of 25 durum wheat (Triticum turgidum subsp. durum Desf.) genotypes (nineteen landraces and six improved varieties), cultivated in two semi-arid regions in the center area of Tunisia, were assessed. These sites (Echbika, 2.2 g l−1; Barrouta, 4.2 g l−1) differ by their degree of salinity of the water irrigation. The results showed that most of the agronomic traits (e.g. spike per meter square, thousand kernels weight and grain yield) were reduced by salinity. Durum wheat landraces, Mahmoudi and Hmira, and improved varieties, Maali and Om Rabia showed the widest adaptability to different quality of irrigation water. Genotypes including Jneh Kotifa and Arbi were estimated as stable genotypes under adverse conditions. Thereafter, salt-tolerant (Hmira and Jneh Khotifa) and the most cultivated high-yielding (Karim, Razzak and Khiar) genotypes were tested for their gynogenetic ability to obtain haploids and doubled haploid lines. Genotypes with good induction capacity had not necessarily a good capacity of regeneration of haploid plantlets. In our conditions, Hmira and Khiar exhibited the best gynogenetic ability (3.1% and 2.9% of haploid plantlets, respectively).
       
  • Assessment of genetic diversity in Colletotrichum falcatum Went accessions
           based on RAPD and ISSR markers

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Prittesh Patel, B.K. Rajkumar, Preeti Parmar, Rusabh Shah, R. Krishnamurthy Sugarcane is susceptible to red rot disease caused by phytopathogenic fungus Colletotrichum falcatum Went which ultimately affect the economy of farmers as well as sugar based industry. One of the various ways to control this devastating disease is to develop disease resistance sugarcane cultivar and this requires the complete understanding of genetic makeup of pathogen. Although South Gujarat is well known sugarcane cultivating area, less published data can be found about PCR-based genetic diversity in prevalent C. falcatum accessions. So, present investigation aims at finding molecular variation among the ten accessions of C. falcatum using RAPD and ISSR molecular markers. A total of 35 RAPD and 39 ISSR primers were screened across 10 C. falcatum accessions, of which 15 RAPD and 21 ISSR primers have showed consistent amplification. Statistics related to genetic variation were estimated using NTSYS-PC by means of Dice’s coefficient. The results revealed 80.6% and 68.07% polymorphism and similarity coefficient ranged from 0.43 to 0.91 and 0.73 to 0.93 in RPAD and ISSR analysis respectively. The dendrogram generated using RAPD, ISSR and combined RAPD-ISSR grouped accessions into different clusters which reveal considerable level molecular variation among the C. falcatum accessions. It is also evident from PCA plots that accessions are rather dispersed with tested marker systems indicating good genetic base. So, in nut shell, we found considerable genetic variation and relatedness within C. falcatum accessions collected from different areas of south Gujarat, India using RAPD and ISSR markers.
       
  • Evaluation of genetic diversity in wild populations of Peganum harmala L.,
           a medicinal plant

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Ranya EL-Bakatoushi, Dalia Gamil Aseel Ahmed Peganum harmala L. is a perennial herbaceous plant and can be a future drug due to its wide medicinal purposes. Despite its economic importance, the molecular genetics of P. harmal have not yet been studied in detail. Genetic diversity of 12 P. harmala genotypes were investigated by using Inter-Simple Sequence Repeats (ISSR), PCR-RFLP of rDNA-ITS, PCR-SSCP of rDNA-ITS and Simple Sequence Repeat (SSR) markers. The level of polymorphism revealed by ITS-SSCP is the lowest, followed by ITS-RFLP then ISSR and the highest polymorphism level was reported for SSR marker. The AMOVA analysis implied that most of the variation occurred within the Populations. A value of inbreeding coefficient Fis estimated by the three co-dominant markers was nearly equal and offer an indication of the partial out-crossing reproductive system of P. harmala. Principal Coordinate Analysis (PCOA) plot revealed a clear pattern of clustering based on the locations of collected plants which coincide with the isolation by distance. The study revealed that ITS-SSCP and ISSR markers respectively were more informative than the other used markers in the assessment of genetic diversity of P. harmala. The results reflect the great diversity of P. harmala and data obtained from this study can be used for future collecting missions.
       
  • Inter and intraspecific genetic diversity (RAPD) among three most frequent
           species of macrofungi (Ganoderma lucidum, Leucoagricus sp. and Lentinus
           sp.) of Tropical forest of Central India

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Sandhya Dwivedi, Surendra Singh, U.K. Chauhan, Mahendra Kumar Tiwari In present study seven RAPD primers were used to access the diversity within and among twelve populations of three mushroom species Ganoderma lucidum, leucoagaricus sp. and Lentinus sp. Total of 111 bands were scored by 7 RAPD primers in 30 accessions of three mushroom species collected from different sampling sites of central India. Total 111 bands were generated using seven primers which were F-1, OPG-06, OPC-07, OPD-08, OPA-02, OPD-02, OPB-10. All 111 bands were polymorphic in nature (100%). Therefore, it revealed that the used primers had sufficient potency for population studies and 30 accessions had higher genetic differences among each other. In best of the knowledge, this is the first report, which accesses the genetic diversity between three mushroom species (Gd Ganoderma lucidum, Lg Leucoagaricus sp., Ls Lentinus). The polymorphic percentage ranged from 3.60 to 23% within twelve populations, while polymorphic percentage among group was 40.56, among population within groups was 41.12 and within population was 18.32. This indicated that the genetic diversity within the population was very low, but slightly higher in the populations of three species. Among three groups representing Gd., Lg and Ls, Among populations within groups shown highest percentage of variation (Pv = 41.12) while within populations, the lowest percentage of variation (18.32) was observed. This result also support that the highest genetic variation was present among groups in comparison to among the population within a species and lowest genetic variation was observed within the population.
       
  • Overexpression of rice thaumatin-like protein (Ostlp) gene in transgenic
           cassava results in enhanced tolerance to Colletotrichum gloeosporioides f.
           sp. manihotis

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Patroba Odeny Ojola, Evans N. Nyaboga, Paul N. Njiru, George Orinda Cassava (Manihot esculenta Crantz) is the most important staple food for more than 300 million people in Africa, and anthracnose disease caused by Colletotrichum gloeosporioides f. sp. manihotis is the most destructive fungal disease affecting cassava production in sub-Saharan Africa. The main objective of this study was to improve anthracnose resistance in cassava through genetic engineering. Transgenic cassava plants harbouring rice thaumatin-like protein (Ostlp) gene, driven by the constitutive CaMV35S promoter, were generated using Agrobacterium-mediated transformation of friable embryogenic calli (FEC) of cultivar TMS 60444. Molecular analysis confirmed the presence, integration, copy number of the transgene all the independent transgenic events. Semi-quantitative RT-PCR confirmed high expression levels of Ostlp in six transgenic lines tested. The antifungal activity of the transgene against Colletotrichum gloeosporioides pathogen was evaluated using the leaves and stem cuttings bioassay. The results demonstrated significantly delayed disease development and reduced size of necrotic lesions in leaves and stem cuttings of all transgenic lines compared to the leaves and stem cuttingss of non-transgenic control plants. Therefore, constitutive overexpression of rice thaumatin-like protein in transgenic cassava confers enhanced tolerance to the fungal pathogen C. gloeosporioides f. sp. manihotis. These results can therefore serve as an initial step towards genetic engineering of farmer-preffered cassava cultivars for resistance to anthracnose disease.
       
  • Green synthesis of zero valent colloidal nanosilver targeting A549 lung
           cancer cell: In vitro cytotoxicity

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Minakshi Jha, Navinchandra G. Shimpi An eco-friendly green approach was proposed to synthesise stable, cytotoxic colloidal silver nanoparticles (AgNPs) using Momordica charantia (M. charantia) fruit extract. Bioinspired green method adopted for fabrication of AgNPs because of easy, fast, low-cost and benign bioprocess. Phytocomponents played the crucial role in capping, stabilisation and inherent cytotoxic potential of colloidal nanosilver. The physiochemical, crystalline, optical and morphological properties of AgNPs were characterized using UV-vis, FT-IR, XRD, SEM, TEM, EDX and AFM. FT-IR reveals the presence of carbonyl, methyl, polyphenol (flavonoid), primary and secondary amine (protein), carboxyl group, ester as major functional groups over the surface of nanomaterials. Mechanistic pathway for formation and stabilisation of colloidal nanosilver has been discussed. Average crystalline size of AgNPs was found to be 12.55 nm from XRD. TEM shows AgNPs nanosphere with size range 1–13.85 nm. Consistency in spherical morphology was also confirmed through Atomic Force Microscopy (AFM). AFM measurement provided image Rq value 3.62, image Ra 2.47, roughness Rmax 36.4 nm, skewness 1.99 and kurtosis 9.87. The SRB assay revealed substantial in vitro noticeable anti-cancer activity of colloidal nanosilver on A549 and HOP-62 human lung cancer cells in a dose dependent manner with IC50 value of 51.93 µg/ml and 76.92 µg/ml. In addition, M. charantia capped AgNPs were found to be more biocompatible in comparison to M. charantia FE. Our study demonstrated the integration of green chemistry principle in nanomaterials fabrication and focused on the potential use of M. charantia fruit extract as an efficient precursor for biocompatible AgNPs anodrug formulation with improved cytotoxic applications.Graphical abstractGraphical abstract for this article
       
  • Study the influence of culture conditions on rennin production by
           Rhizomucor miehei using solid-state fermentations

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Houthail Alahmad Aljammas, Hassan Al Fathi, Walid Alkhalaf Investigations were conducted on the production of Rennin enzyme from the fungi Rhizomucor miehei 3420 NRRL using Solid-State fermentation. Wheat bran was used as a substrate. The influence of moisture content, incubation temperature, and the initial pH of fermentation medium were studied. The protein content, milk clotting activity (MCA), specific activity, proteolytic activity (PA), and (MCA/PA) ratio of the extracted enzyme were calculated after 4 days of incubation to evaluate the quality of the enzyme. The results showed that the optimal conditions for production were as follows: incubation temperature of 40 °C, moisture content of 60%, and pH of (3). Under these conditions, a production process of Rennin enzyme was established, and the values of protein content, milk clotting activity, specific activity, proteolytic activity, and (MCA/PA) ratio reached to 4 mg/mL, 600 SU/mL, 150 SU/mg, 45 PU/mL, 13.3 respectively.
       
  • Biosynthesis of silver nanoparticles formation from Caesalpinia
           pulcherrima stem metabolites and their broad spectrum biological
           activities

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Pooja Moteriya, Sumitra Chanda The present work illustrates eco-friendly, rapid and cost effective method of AgNPs synthesis using C. pulcherrima stem extract. Initially, various physico chemical factors were optimized. Characterization was done by different spectroscopic and microscopic analysis. AgNPs were spherical in shape with an average size of 8 nm. AgNPs showed good synergistic antimicrobial, antibiofilm and antioxidant activity. The cytotoxicity effect against HeLa cancer cell line was dose dependent while genotoxic study revealed the non toxic nature of AgNPs at lower concentration. The results suggest that AgNPs from C. pulcherrima stem extract have great potential in biomedical applications.
       
  • Production and purification of IgY antibodies from chicken egg yolk

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Wala Ahmad Amro, Wael Al-Qaisi, Fawzi Al-Razem The isolation of polyclonal antibodies from the serum of immunized mammals has significantly contributed to scientific research and diagnosis. The fact that recent technologies allow the production of antibodies in the yolk of eggs laid by hens, has led to the development of an alternative method for antibody generation that is less stressful to animals. As hens are kept under almost all their natural conditions, antibodies are isolated from the collected eggs; this technology is expected to become an interesting alternative to the conventionally serum-based techniques that eventually require to sacrifice the animal.Here we present a modified protocol for the isolation of IgY antibodies from immunized chickens and provide comparison between two chicken breeds in relative to IgY yield per egg. Our results show the possibility of generating large quantities of highly pure IgY from chicken eggs and also show large differences in the yield of IgY production between the two studied breeds. The results of this work indicate that IgY technology can be used for the production of primary antibodies for immunological work and disease diagnosis.
       
  • Genetic variability of myostatin and prolactin genes in popular goat
           breeds in Egypt

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Sekena H. Abdel-Aziem, K.F. Mahrous, M.A.M. Abd El-Hafez, M. Abdel Mordy The genetic polymorphisms of two functional genes named: myostatin (MSTN) and prolactin (PRL) were investigated in three goat breeds (Barki, Damascus and Zaraibi) using Sanger nucleotide sequence and restriction fragment length polymorphism (RFLP) methods, in order to differentiate between these breeds. Nucleotide sequencing of 337 bp MSTN gene detected five SNPs in Barki breed, two SNPs in Damascus breed, while the Zaraibi breed did not show any SNPs. Moreover, MSTN-HaeIII/PCR-RFLP gave a single Genotype BB was found in all the studied breeds. Meanwhile, Nucleotide sequencing of 196 bp PRL gene showed two SNPs in Damascus breed, one SNPs in Zaraibi breed, while the Barki breed did not show any SNPs. Moreover, PRL-Eco24I/PCR-RFLP showed three genotypes (AA, AB and BB). The genotype AB showed the maximum frequency in all the studied breeds (0.75, 0.85, and 0.90 for Damascus, Barki and Zaraibi breeds, respectively). Observed heterozygosity (Ho) value was higher than expected heterozygosity (He) value all studied breeds. In addition, the values of both Ho and He were the highest in Zaraibi breed (0.90 and 0.51 respectively). Chi-square (χ2) value revealed a significant variation Hardy-Weinberg equilibrium (P < .05) in the three studied breeds. It is the highest in Zaraibi goats and lowest in Damascus breed. The results demonstrated that the PRL-Eco24I/PCR-RFLP polymorphism may be utilized as effective marker for genetic differentiation between goat breeds, but MSTN-HaeIII/PCR-RFLP revealed no polymorphism or variation, thus it is not recommended in the selection program. Moreover, these results open up interesting prospects for future selection programs, especially marker assisted selection. In addition, the results established that PCR-RFLP method is a suitable tool for calculating genetic variability.
       
  • Detecting protein complexes based on a combination of topological and
           biological properties in protein-protein interaction network

    • Abstract: Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Pooja Sharma, D.K. Bhattacharyya, J.K. Kalita Protein complexes are known to play a major role in controlling cellular activity in a living being. Identifying complexes from raw protein protein interactions (PPIs) is an important area of research. Earlier work has been limited mostly to yeast. Such protein complex identification methods, when applied to large human PPIs often give poor performance. We introduce a novel method called CSC to detect protein complexes. The method is evaluated in terms of positive predictive value, sensitivity and accuracy using the datasets of the model organism, yeast and humans. CSC outperforms several other competing algorithms for both organisms. Further, we present a framework to establish the usefulness of CSC in analyzing the influence of a given disease gene in a complex topologically as well as biologically considering eight major association factors.
       
  • In silico studies on bacterial xylanase enzyme: Structural and
           functional insight

    • Abstract: Publication date: Available online 31 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Bhramar Dutta, Aparna Banerjee, Priyanka Chakraborty, Rajib Bandopadhyay Xylans are the second most abundant form of hemicelluloses and are the second most abundant polysaccharide in nature after cellulose. To degrade xylan, microbes produce mainly xylanase enzyme. Wide range of microorganisms like fungi, bacteria, yeast, marine algae etc. are capable of producing xylanase. Main source of xylanase is fungi but industrial production of bacterial xylanase is low cost, easy downstream process and high production rate. To understand primary, secondary and tertiary structure of xylanase, in silico composition of amino acids, basic physiological characteristics; viz., pI, molecular weight, instability index, GRAVY, molar extinction coefficient, secondary structure, presence of functional domain and motifs, phylogenetic tree, salt bridge compositions are determined. In silico study of xylanase focused on 36 different bacterial sources are performed by retrieving FASTA and PDB sequences using RCSB PDB. FASTA and PDB files are proceed further in ExPASy-ProtParam, RAMPAGE, QMEAN, MEME, PSIPRED, InterProScan, MOTIF scan, ERRAT, Peptide cutter, ESBRI and MEGA 7. The instability index range (16.90–38.78) clearly indicates that the protein is highly stable. α-helix mean value (27.11%) infers the protein is dominated by α-helix region. The aliphatic index (39.80–90.68) gives information that the protein is highly thermostable, prevalence by alanine amino acid in aliphatic side chain. No transmembrane domain was found in the protein which confirms the enzyme is extracellular in nature. Ancestor chart analysis confirmed that it is a part of carbohydrate metabolic process and more specifically a member of glycoside hydrolase super family.
       
  • Increased level of B cell differentiation factor in systemic lupus
           erythematosus patients

    • Abstract: Publication date: Available online 30 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Hala Zaki Raslan, Hiba Sibaii, Salwa Refat El- Zayat, Hagar Hassan, Mahitab El- Kassaby Most autoimmune disease are driven by a dysfunction in T and B cells, but B cells are still an interesting area of research, perturbations in their development are implicated in autoimmune diseases. B cell differentiating factor (BCDF) plays a part in the differentiation of B cells. The aim was To assess the levels of BCDF, IgM and IgG in SLE patients and whether they have any peculiarity in the clinical context of SLE. Thirty six patients with SLE and 24 healthy volunteers as control were enrolled in the study. BCDF was measured using Sandwich ELISA, total human IgM and IgG were measured by calorimetric methods. The mean concentrations of BCDF and IgM were significantly higher in patients with SLE as compared with controls (P 
       
  • Biodegradation of feather waste by keratinase produced from newly isolated
           Bacillus licheniformis ALW1

    • Abstract: Publication date: Available online 28 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Azza M. Abdel-Fattah, Mamdouh S. El-Gamal, Siham A. Ismail, Mohamed A. Emran, Amal M. Hashem Keratinase are proteolytic enzymes which have gained much attention to convert keratinous wastes that cause huge environmental pollution problems. Ten microbial isolates were screened for their keratinase production. The most potent isolate produce 25.2 U/ml under static condition and was primarily identified by partial 16s rRNA gene sequence as Bacillus licheniformis ALW1. Optimization studies for the fermentation conditions increased the keratinase biosynthesis to 72.2 U/ml (2.9-fold). The crude extracellular keratinase was optimally active at pH 8.0 and temperature 65 °C with 0.7% soluble keratin as substrate. The produced B. licheniformis ALW1 keratinase exhibited a good stability over pH range from 7 to 9 and over a temperature range 50–60 °C for almost 90 min. The crude enzyme solution was able to degrade native feather up to 63% in redox free system.
       
  • Purification and characterization of alkaline protease with novel
           properties from Bacillus cereus strain S8

    • Abstract: Publication date: Available online 26 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): B.K.M Lakshmi, D. Muni Kumar, K.P.J Hemalatha Proteases are the hydrolytic enzymes which hydrolyzes peptide bond between proteins with paramount applications in pharmaceutical and industrial sector. Therefore production of proteases with efficient characteristics of biotechnological interest from novel strain is significant. Hence, in this study, an alkaline serine protease produced by Bacillus cereus strain S8 (MTCC NO 11901) was purified and characterized. The alkaline protease was purified by ammonium sulfate precipitation (50%), ion exchange (DEAE-Cellulose) and gel filtration (Sephadex G-100) chromatographic techniques. As a result of this purification, a protein with specific activity of 300U/mg protein was obtained with purification fold 17.04 and recovery percentage of 34.6%. The molecular weight of the purified protease was determined using SDS-PAGE under non-reducing (71 kDa) and reducing conditions (35 kDa and 22 kDa). Zymogram analysis revealed that proteolytic activity was only associated with 22 kDa. These results indicate that existence of the enzyme as dimer in its native state. The molecular weight of the protease (22 kDa) was also determined by gel filtration (Sephadex G-200) chromatography and it was calculated as 21.8 kDa. The optimum activity of the protease was observed at pH 10.0 and temperature 70 °C with great stability towards pH and temperature with casein as a specific substrate. The enzyme was completely inhibited by PMSF and TLCK indicating that it is a serine protease of trypsin type. The enzyme exhibits a great stability towards organic solvents, oxidizing and bleaching agents and it is negatively influenced by Li2+ and Co2+ metal ions. The purified protein was further characterized by Matrix Assisted Laser Desorption Ionization/Mass Spectroscopy (MALDI/MS) analysis which reveals that total number of amino acids is 208 with isoelectric point 9.52.
       
  • Enhancement of nematicidal potential through cloning and expression of
           chitinase gene from Bacillus subtilis subsp. Subtilis BTN7A strain

    • Abstract: Publication date: Available online 23 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Mohamed S. Abdel-Salam, Hoda H. Ameen, Abdallah S.M. Kassab, Ahmed E.A. Mahgoob, Usama S. Elkelany A gene encoding chitinase from B. subtilis has been isolated after optimization of PCR conditions. It was cloned with two different prometers, T7 promoter of the pJET1.2/blunt cloning vector and the SP6 promoter of pGEM®-T Easy vector. After transforming E. coli DH5α, two transformants were selected, CHI-NRC-4 from the first vector and T-CHI-NRC-6 from the second vector, and used for further studies. The complete CDS sequence of chitinase gene was determined and submitted to GenBank with the accession number KX268692.1. Culture supernatants of E. coli (CHI-NRC-4) and E. coli (T-CHI-NRC-6) were investigated for their inhibitory effect on M. javanica egg hatch under laboratory conditions. Result showed up to 96% inhibition in egg hatching due to both E. coli transformants as compared to control which reflect the same expression efficiency of both used prometers. A greenhouse experiment was carried out to evaluate the nematicidal effect of culture supernatants of the two transformts E. coli (CHI-NRC-4) and E. coli (T-CHI-NRC-6) against M. javanica infected eggplant. Obtained results showed a significant reduction in nematode population in soil and roots and enhancement in eggplant growth parameters as compared to control.
       
  • Molecular diversity of internal transcribed spacer among the monoconidial
           isolates of Magnaporthe oryzae isolated from rice in Southern Karnataka,
           India

    • Abstract: Publication date: Available online 19 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): D. Jagadeesh, M.K. Prasanna Kumar, R. Chandrakanth, N.S. Devaki Blast disease of rice plant is caused by Magnaporthe oryzae (anamorph Pyricularia oryzae). This disease is recognized to be one of the most serious diseases of rice crop around the world. A total of 72 monoconidial isolates of M. oryzae obtained from blast disease samples collected around Southern Karnataka were characterized using internal transcribed spacers of the ribosomal DNA sequences. These were analyzed by comparing with already deposited sequences in GenBank database. It helped in diagnosing the invasive pathogen in all locations. Variability of rDNA sequences was found to be highly polymorphic with 0.068962 nucleotide diversity showing 6 distinct clades. 33 haplotype groups were identified with haplotype diversity of 0.8881 and Tajima's neutrality test with a D value of −1.96827 with P 
       
  • Study on the potential of cold-active lipases from psychrotrophic fungi
           for detergent formulation

    • Abstract: Publication date: Available online 7 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Sanjay Sahay, Deepak Chouhan Lipases from psychrotrophic fungal isolates BPF4 and BPF6 identified as Penicilium canesense and Pseudogymnoascus roseus respectively were characterized for their compatibility towards laundry detergent. BPF4 and BPF6 lipases showed maximum activity at pH 11 and 9 respectively and at 40 °C. The residual activities at 20 °C and 4 °C of BPF4 lipase were 35% and 20% and of BPF6 lipase were 70% and 20 °C respectively. Both the enzymes were stable at 4 °C, 20 °C and 40 °C for 2 h losing at the most 20% of activities. Both the enzymes were metalloenzymes with activity enhancement by nearly threefold by Ca2+. Contrary to BPF6 lipase, BPF4 enzyme was not stimulated by EDTA nor inhibited, rather stimulated by SDS and Triton X-100 by 125% and 330% respectively. Both the lipases showed minor to moderate inhibition by NaClO3 and H2O2, and exhibited nearly 90% residual activity after 1 h of incubation in selected detergent brands thus indicating potential for their inclusion in detergent formulation thereby facilitating cold-washing as a step towards mitigation of climate change.
       
  • Isolation and characterization of Bacillus sp. strain BC01 from soil
           displaying potent antagonistic activity against plant and fish pathogenic
           fungi and bacteria

    • Abstract: Publication date: Available online 7 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Md Javed Foysal, Asura Khanam Lisa Fungal and bacterial pathogens infect a diverse range of hosts including various plant and animal species. Fungal and bacterial diseases, especially of plants and aquatic animals, such as fish, lead to significant damage to crops and aquaculture, respectively, worldwide. The present study was conducted to isolate and characterize potent Bacillus strains with significant antagonistic activity against the major plant and fish pathogenic fungi and bacteria. We randomly collected 22 isolates of Bacillus from the soil, rhizosphere, and sediment from different parts of Bangladesh. Initial characterization, based on in vitro antagonistic activity on the culture plate, resulted in the selection of four gram-positive Bacillus sp. isolates. Among these, the isolate BC01, obtained from soil demonstrated the highest broad-spectrum anti-bacterial and anti-fungal activities. We confirmed the genus of BC01 to be Bacillus by morphological and biochemical tests as well as using molecular data analysis tools, including the study of 16s rDNA, phylogenetic relationship, and evolutionary divergence scores. The isolate significantly inhibited the mycelial growth of the plant pathogen, Penicillium digitatum and fish pathogen, Aphanomyces invadans in vitro. The anti-bacterial effect of the isolate was also evaluated against Pseudomonas spp. and Xanthomonas spp., the two deadliest plant pathogens, and Aeromonas veronii, Pseudomonas fluorescens, and Streptococcus iniae, three major fish pathogens that are primarily responsible for global aquaculture loss. The results of the present study could pave the way for developing potent drugs to combat microbial infection of plants and fish.
       
  • Assessment of genetic diversity in Salvadora persica L. based on inter
           simple sequence repeat (ISSR) genetic marker

    • Abstract: Publication date: Available online 3 May 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Mohammad Asadi Monfared, Davood Samsampour, Gholam Reza Sharifi-Sirchi, Fatemeh Sadeghi Studies on the genetic variation in marginal populations and differentiation between them are essential for assessment of best gene conservation strategies and sampling schemes. In this study, ISSR markers were used to establish the level of genetic relationships and polymorphism 50 genotypes of Salvadora persica collected from 6 different regions of Hormozgan province. The ISSR analysis with 9 anchored primers also generated 105 scorable loci, of which 85 were polymorphic (80.95%). Parameters of genetic diversity and its partitioning were calculated. The genetic analysis demonstrated that S. persica maintain relatively high genetic diversity (PIC was 0.63, Na was 1.27 and Ho and He were 0.15 and 0.17 respectively). The coefficient of genetic differentiation among populations based on FST equaled 0.20. Genetic identities between population's pairs were high (mean I = 0.88). These values are high as compared with other widespread congener species. Cluster analysis based on the Unweighted Pair Group Method with Arithmetic Averages (UPGMA) revealed 3 main clusters for the ISSR data. The levels of genetic diversity maintained within populations of S. persica indicate that an appropriate sampling design for ex situ safeguarding should capture the majority of genetic diversity found within these taxa to help ensure the long term viability of this species. Furthermore, it could be inferred that ISSR markers are suitable tools for the evaluation of genetic diversity and relationships within the Salvadora persica.
       
  • Optimization of novel halophilic lipase production by Fusarium solani
           strain NFCCL 4084 using palm oil mill effluent

    • Abstract: Publication date: Available online 27 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Kiptoo Geoffry, Rajeshwara N. Achur Among different sources of lipases, fungal lipases have continued to attract a wide range of applications. Further, halophilic lipases are highly desirable for biodiesel production due to the need to mitigate environmental pollution caused as result of extensive use of fossil fuels. However, currently, the high production cost limits the industrial application of lipases. In order to address this issue, we have attempted to optimize lipase production by Fusarium solani NFCCL 4084 and using palm oil mill effluent (POME) based medium. The production was optimized using a combinatory approach of Plackett-Burman (PB) design, one factor at a time (OFAT) design and face centred central composite design (FCCCD). The variables (malt extract, (NH4)2SO4, CaCl2, MgSO4, olive oil, peptone, K2HPO4, NaNO3, Tween-80, POME and pH) were analyzed using PB design and the variables with positive contrast coefficient were found to be K2HPO4, NaNO3, Tween-80, POME and pH. The significant variables selected were further analyzed for possible optimum range by using OFAT approach and the findings revealed that K2HPO4, NaNO3, and Tween-80 as the most significant medium components, and thus were further optimized by using FCCCD. The optimum medium yielded a lipase with an activity of 7.8 U/ml, a significant 3.2-fold increase compared to un-optimized medium. The present findings revealed that POME is an alternative and suitable substrate for halophilic lipase production at low cost. Also, it is clearly evident that the combinatory approach employed here proved to be very effective in producing high activity halophilic lipases, in general.
       
  • Detection of myostatin gene MSTN in some goat breeds (Capra
           hircus
    )

    • Abstract: Publication date: Available online 26 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Y.A. Dowidar, M.A. El-Sayed, Aly M. Elrefy, Hytham E. Shoura Till now not information about myostatin MSTN gene in Egyptian goat breeds. Here we show more information about MSTN in some Egyptian goat breeds to enrich the database with new sequences for Egyptian goat breeds. Our conducted study focused on detection and identifying the MSTN gene as a candidate gene of the muscles growth trait in three goat breeds (Zaraibi, Baladi and Damascus). We found the similarity between the registered sequences with the accession numbers KY463684 for Zaraibi and KY463685 for Baladi and Chinese goat breeds of the MSTN gene deposited with international gene banks by up to 99% and some other species including sheep, cows and bull breeds with percentages of 95 to 97% and between 95 to 99%, respectively. There is also a correlation between the sequences of the registered pieces of Baladi with KY463686 and Damascus and Chinese breeds with KY441464 of MSTN deposited with international gene banks by up to 99% and some other species including sheep and bull breeds at a ratio of 99% for two pieces. Results demonstrated the deposited sequences of object are part of intron 1, exon 2 is fully sequenced with Zaraibi and Baladi breeds; the intron 1, exon 1 with Baladi breed; and the intron 2, part of exon 3 with Damascus breed. Therefore, the Egyptian goat breeds consider national wealth can be used to develop breeding and improvement programs which helps in more applicable scopes like biotechnology, genetic engineering and molecular biology with the help of bioinformatics tools.
       
  • Cloning and expression of MPT83 gene from Mycobacterium tuberculosis in E.
           coli BL21 as vaccine candidate of tuberculosis: A preliminary study

    • Abstract: Publication date: Available online 13 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Ahyar Ahmad, Rosana Agus, Muh. Nasrum Massi, Rosdiana Natzir, Radha Madhyastha, Harish Kumar Madhyastha, Masugi Maruyama The appearance of Mycobacterium tuberculosis strains leading to drug resistance has caused new problems in TB treatment in various parts of the world and forces WHO to declare TB as a global emergency. With the increase of TB drug resistance, it is convinced that a more effective vaccine development will stop the epidemic of TB. Some M. tuberculosis antigens, one of which is MPT83, have been examined as TB vaccine candidate. MPT83 antigen, which is very immunogenic in lipoprotein micro bacteria, is identified as surface cell interrelated to antigen with cytometry circulation. Having TB resistance from BCG vaccine, MPT83 is considered TB vaccine candidate that can protect people against TB at adult age. The purpose of this research is to conduct amplification of MPT83 antigen cloning, and expression of its antigen on E. coli bacteria. From the result of the research, it is expected that raw material to produce TB vaccine as well as a high-quality antigen can be obtained. The band of DNA in PCR product is 660 bp, while the one in pGEMT-Easy-Mpt83 recombinant plasmid is 3678 bp. This is expressed in E. coli BL21 strain and produces 48 kDa protein as well as GST-MPT83 fusion protein.
       
  • High level expression and purification of recombinant flounder growth
           hormone in E. coli

    • Abstract: Publication date: Available online 9 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Tae-Jin Choi, Temesgen Tola Geletu Recombinant flounder growth hormone was overproduced in E. coli by using codon optimized synthetic gene and optimized expression conditions for high level production. The gene was cloned into PET-28a expression vector and transformed into E. coli BL21 (DE3). Induction at lower temperature, lower IPTG concentrations and richer growth media during expression resulted in increased expression level. The protein expression profile was analyzed by SDS-PAGE, the authenticity was confirmed by western blotting and the concentration was determined by Bradford assay. In addition, several attempts were made to produce soluble product and all resulted in insoluble product. The overexpressed protein was efficiently purified from inclusion bodies by moderate speed centrifugation after cell lysis. Among the solubilization buffers examined, buffer with 1% N-lauroylsarcosine in the presence of reducing agent DTT at alkaline pH resulted in efficient solubilization and recovery. The denaturant was removed by filtration and dialysis. The amount of the growth hormone recovered was significantly higher than previous reports that expressed native growth hormone genes in E. coli. The methodology adapted in this study, can be used to produce flounder growth hormone at large scale level so that it can be used in aquaculture. This approach may also apply to other proteins if high level expression and efficient purification is sought in E. coli.
       
  • Micropropagation protocol for Antigonon leptopus an important ornamental
           and medicinal plant

    • Abstract: Publication date: Available online 5 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Zenna Fawzia Ghareeb, Lobna S. Taha The effect of some factors on in vitro consecutive micropropagation behavior of Antigonon leptopus was examined including those of culture establishment, shootlets multiplication, rooting and acclimatization stages. The highest percent of aseptic cultures and survival of explants (100%) were obtained as a result of using Clorox 10% for 3 min followed by MC 0.1% for 2 min while, using each of them individually (Clorox 20% or MC 0.1%) for 5 min caused the highest percent of shoot formation. During the multiplication stage, the highest percent of shoot formation was reached to 100% with repeating culture of explants (two times) on MS medium supplemented with 2ip at 1.0 and IBA at 0.2 mg/l. The highest numbers of shootlets/explant were obtained when 2.0 mg/l of BAP or 0.5 mg/l BA + 0.2 mg/l of IBA were added to MS culture medium. Culturing the explants on MS medium supplemented with 2ip at 0.5 or 1.0 mg/l each combined with 0.2 mg/l of IBA showed the longest shootlets. Reducing the strength of culture media to ½ or ¾ had promotion effect on rooting formation of shootlets. The best results of plant acclimatization (survival percent, plant height and root length) were obtained by using sand or peat moss soil. The amplified DNA fragments using B7, B9 and C19 primers for mother and micropropagated plants showed that the produced pattern by primer B7 had a maximum number of 10 bands of DNA fragments with molecular size ranging between 1025.57 and 176.36 bp, micropropagated plants showed 95.2% similarity in relation to mother plant.
       
  • Immobilization of thermostable exo-inulinase from mutant thermophilic
           Aspergillus tamarii-U4 using kaolin clay and its application in inulin
           hydrolysis

    • Abstract: Publication date: Available online 4 April 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Emmanuel O. Garuba, Abiodun, A. Onilude In this study, attempts were made to immobilize purified exo-inulinase from mutant thermophic Aspergillus tamarii-U4 onto Kaolinite clay by covalent bonding cross-linked with glutaraldehyde with an immobilization yield of 66% achieved. The free and immobilized inulinases were then characterized and characterization of the enzymes revealed that temperature and pH optima for the activity of the free and immobilized enzymes were both 65 °C and pH 4.5 respectively. The free inulinase completely lost its activity after incubation at 65 °C for 6 h while the immobilized inulinase retained 16.4% of its activity under the same condition of temperature and incubation time. The estimated kinetic parameters Km and Vmax for the free inulinase as estimated from Lineweaver-Burk plots were 0.39 mM and 4.21 µmol/min for the free inulinase and 0.37 mM and 4.01 µmol/min for the immobilized inulinase respectively. Inulin at 2.5% (w/v) and a flow rate of 0.1 mL was completely hydrolysed for 10 days at 60 °C in a continuous packed bed column and the operational stability of the system revealed that the half-life of the immobilized inulinase was 51 days. These properties make the immobilized exo-inulinase from Aspergillus tamarii-U4 a potential candidate for the production of fructose from inulin hydrolysis.
       
  • Screening of potential probiotic lactic acid bacteria and production of
           amylase and its partial purification

    • Abstract: Publication date: Available online 27 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Tallapragada Padmavathi, Rayavarapu Bhargavi, Purushothama Rao Priyanka, Naige Ranganath Niranjan, Pogakul Veerabhadrappa Pavitra Probiotics are the healthy living bacteria when administered in adequate amounts confers health benefits in the host. The main objective of present study was to screen the bacteria for potential probiotic characters and enzyme production. The probiotic characters like tolerance to low pH, bile salts, antibiotic sensitivity, hydrophobicity and auto-aggregation properties were evaluated. Among all isolates Lactobacillus fermentum and Lactobacillus sp G3_4_1TO2 showed maximum potential probiotic characters and produced amylase enzyme by observing the halo zone around the colonies with the diameter 0.9 mm and 1.23 mm. Lactobacillus sp G3_4_1TO2 produced maximum amylase when compared with Lb. fermentum. The protein yield was 55.4% with the specific activity of 88.9 U/mg and obtained 40.8% purification fold. The molecular weight of amylase enzyme determined by SDS PAGE was 95,000 Da. From the present study it was considered that Lactobacillus sp G3_4_1TO2 was a potential probiotic bacteria producing maximum amylase enzyme.
       
  • Screening of anti-inflammatory phytocompounds from Crateva adansonii leaf
           extracts and its validation by in silico modeling

    • Abstract: Publication date: Available online 23 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Rathinavel Thirumalaisamy, Subramanian Ammashi, Govarthanan Muthusamy Anti-inflammatory phytocompounds from Crateva adansonii DC leaf extracts were identified by GCMS analysis and its anti-inflammatory potential was evaluated by in silico molecular docking study against inflammatory molecular targets. Three different (Aqueous, Methanol and Petroleum ether) dried leaf extracts of Crateva adansonii were obtained from soxhlet extraction method. Preliminary phytoconstituents analysis of three different leaf extracts of C. adansonii confirmed the presence of various major classes of bioactive phytoconstituents such as polyphenols (tannins and flavonoids), steroids, alkaloid, coumarin, carbohydrate and terpenoids. Among three leaf extracts, methanolic leaf extract possess highest total phenolic content of 77 ± 1.65 µg gallic acid equivalent (GAE)/g of dry weight of leaf extract, subsequently methanolic leaf extract also shows maximal in vitro antioxidant activity (DPPH scavenging activity) of 71.22 ± 1.32% among three different leaf extracts. GC–MS analysis of petroleum ether leaf extract revealed the presence of nine phytocompounds representing 95.43% peak area percentage, among nine identified phytocompounds three phytocompounds of C. adansonii possess anti-inflammatory property namely phytol, 1-Hexyl-2-Nitrohexane and 2-Isopropyl-5-Methylcyclohexyl 3-(1-(4-Chlorophenyl)-3-Oxobutyl)-Coumarin-4-Yl Carbonate were chosen for in silico molecular docking study against four inflammatory receptor targets (COX-2, TNFα, IL-1β and IL-6) and they shows less binding energy with highest docking score ranging from −15.9500 to 5.0869. The present study substantially indicated and proven that anti-inflammatory potential of phytocompounds from C. adansonii leaf extracts which can be exploited for commercial designing of novel anti-inflammatory drug to treat various inflammatory disorders.
       
  • Total phenolic and flavonoid contents and antioxidant activity of ginger
           (Zingiber officinale Rosc.) rhizome, callus and callus treated with some
           elicitors

    • Abstract: Publication date: Available online 21 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Ammar Mohammed Ahmed Ali, Mawahib ElAmin Mohamed El-Nour, Sakina Mohamed Yagi The present study was aimed at determining total phenolic and flavonoid contents and studying the antioxidant activity of ginger (Zingiber officinale Rosc.) rhizome and callus, 6-gingerol and 6-shogaol and callus treated with elicitors. Petroleum ether (PE) and chloroform: methanol (1:1, v/v) (CM) extracts were prepared by maceration. Highest total phenolic content was obtained from the CM extract (60.34 ± 0.43 mg gallic acid/g) of rhizome while callus showed lower content detected in the CM extract (33.6 ± 0.07 mg gallic acid/g). Flavonoids were only detected in rhizome (CM extract 40.25 ± 0.21 mg quercetin/g). Both rhizome extracts exhibited good antioxidant activity with higher activity recorded in PE extract (IC50 value 8.29 ± 1.73 μg/mL). Callus extracts revealed lower antioxidant activity (IC50 value 1265.49 ± 59.9 μg/mL obtained from CM extract). 6-gingerol and 6-shogaol displayed high antioxidant activity in both assays with IC50 4.85 + 0.58DPPH and 5.35 ± 0.33ABTS μg/mL for the former and IC50 7.61 ± 0.81DPPH and IC50 7.05 ± 0.23ABTS μg/mL for the latter. Treatment of callus with elicitors showed significant (p 
       
  • In vitro biotechnological advancements in Malabar nut (Adhatoda vasica
           Nees): Achievements, status and prospects

    • Abstract: Publication date: Available online 19 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Saikat Gantait, Jitendriya Panigrahi Adhatoda vasica Nees, belonging to family Acanthaceae, is a well-known medicinal plant. It is endorsed for its pyrroloquinazoline alkaloids and its derivatives, such as vasicine and vasicinone. Germinating A. vasica seeds is a tedious task; on that account, vegetative propagation is the preferred method for its multiplication. For rapid and large-scale multiplication, germplasm conservation as well as secondary metabolites production, in vitro culture of A. vasica was preferred over conventional propagation by several researchers; however, some major applications of this tissue culture technique are still awaiting to undergo extensive research. The present review, for the first time, illustrates all the major achievements associated with in vitro regeneration of A. vasica, reported till date and highlights the future prospects.
       
  • Assessment of Ki-67 as a potential biomarker in patients with breast
           cancer

    • Abstract: Publication date: Available online 10 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Halla Mohamed Ragab, Nervana Samy, Mie Afify, Nabila Abd El Maksoud, HebatAllah Mohamed Shaaban Breast cancer is the most common cancer in females, it accounts for one third of all malignancies affecting women. Appropriate biomarkers play significant role in predicting the prognosis and decide the specific therapy to each patient. In this study we aimed at evaluating the value of Ki-67 as a prognostic marker in breast cancer patients and to analyze the associations between Ki-67 and their clinicopathological parameters. This study included 92 patients with developed non metastatic breast cancer and 10 women had benign breast tumor served as controls. We measured the serum level by ELISA technique and tissue expression of Ki-67 by immunohistochemical technique. Our results showed that there were no statistically significant differences in serum Ki-67 levels between the two studied groups. As for Ki-67expression in breast cancer cells, the score increases with increase of tumor size, grade, premenopausal, Ki-67 expression in estrogen and progesterone receptor positive tumors showed lower values than estrogen and progesterone negative tumors, while higher Ki-67 expression was more frequently associated with HER2-positive. In conclusion; our study supports the finding that tissue Ki-67 expression may add prognostic information to that obtained from classical prognostic factors and can provide data of significant value to other important prognostic indicators such as pathological grading, and axillary lymph node involvement.
       
  • Partial purification and characterization of exoinulinase produced from
           Bacillus sp.

    • Abstract: Publication date: Available online 7 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): R. Ramapriya, A. Thirumurugan, T. Sathishkumar, D.R. Manimaran Inulinase are industrial food enzymes which have gained much attention in recent scenario. In this study, Inulinase producing eight bacterial colonies were isolated and screened from three different plant root tubers soil sample. Among 8 inulinase producing colonies, the higher yielding colony was selected with 25.10 U/mL for further studies. The best inulinase producing colony was identified by partial 16S rRNA gene sequence as Bacillus sp. The crude inulinase was purified by using ammonium sulphate precipitation, dialysis and ion exchange chromatography on DEAE – sephacel and obtained 1.9 purification fold with total activity 293 U. The purified enzyme was subjected to characterization studies and it was found to be stable at 30–60 °C and optimum temperature was at 55 °C. The enzyme was stable at pH 3.0–7.0 and optimum pH was at 6.5. The Km and Vmax value for inulinase was found to be 0.117 mg/mL and 4.45 μmol min mg−1 respectively, demonstrate its greater affinity. Hence, this enzyme can be widely used for the production of fructose, and fructooligosaccharides, which are important ingredients in food and pharmaceutical industry.
       
  • Elevated carotenoids in staple crops: The biosynthesis, challenges and
           measures for target delivery

    • Abstract: Publication date: Available online 6 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Adebanjo Ayobamidele Badejo Poverty eradication and global food security are among the targets of world leaders, most especially combating the scourge of hidden hunger. Provitamin A carotenoids cannot be synthesized de novo by human and so it must be taken as part of the diet. The deficiency of which is causing almost 6000 sights to be lost daily in most developing countries because of the monotonous starchy diets lacking substantial amount of carotenoid. Conventional breeding as well as genetic engineering have been used to increase the level of carotenoid in many staples including rice, potato, maize and cassava. While products from genetic engineering are still been subjected to strict regulatory measures preventing the delivery of the products to target consumers, some of the products from conventional breeding are already on the table of consumers. Interestingly, both technologies are crucial to tackling micronutrient deficiencies. This review discusses the role of carotenoid in human, the biosynthesis in plant and some of the staple crops that have been modified for increased carotenoid. Some measures expected of the leaders of the countries in need of these products for safe delivery to the target population after two decades is also highlighted.
       
  • Effect of vitamins and cell constructions on the activity of microbial
           fuel cell battery

    • Abstract: Publication date: Available online 3 March 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Dena Z. Khater, K.M. El-Khatib, Rabeay Y.A. Hassan Construction of efficient performance of microbial fuel cells (MFCs) requires certain practical considerations. In the single chamber microbial fuel cell, there is no border between the anode and the cathode, thus the diffusion of the dissolved oxygen has a contrary effect on the anodic respiration and this leads to the inhibition of the direct electron transfer from the biofilm to the anodic surface. Here, a fed-batch single chambered microbial fuel cells are constructed with different distances 3 and 6 cm (anode- cathode spacing), while keeping the working volume is constant. The performance of each MFC is individually evaluated under the effects of vitamins & minerals with acetate as a fed load. The maximum open circuit potential during testing the 3 and 6 cm microbial fuel cells is about 946 and 791 mV respectively. By decreasing the distance between the anode and the cathode from 6 to 3 cm, the power density is decreased from 108.3 mW m−2 to 24.5 mW m−2. Thus, the short distance in membrane-less MFC weakened the cathode and inhibited the anodic respiration which affects the overall performance of the MFC efficiency. The system is displayed a maximum potential of 564 and 791 mV in absence & presence of vitamins respectively. Eventually, the overall functions of the acetate single chamber microbial fuel cell can be improved by the addition of vitamins & minerals and increasing the distance between the cathode and the anode.
       
  • In vitro culture, transformation and genetic fidelity of Milk
           Thistle

    • Abstract: Publication date: Available online 23 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): M.R. Rady, M.M. Saker, M.A. Matter This review article presents a consolidated explanation and provides a comprehensive description of various studies, carried out on in vitro culture and hairy root cultures of S. marianum which can be consider an alternative source of flavonolignans. To overcome the constrains of conventional propagation of silybum plant, tissue culture and advanced biotechnology proved to be an influential tool that can complement conventional breeding and accelerate silybum development. The present review is focused on biotechnological tools like in vitro culture, hairy root cultures and genetic fidelity of S. marianum which can be a potent tool for production of secondary metabolites from these cultures.
       
  • Rice straw fermentation by Schizophyllum commune ARC-11 to produce high
           level of xylanase for its application in pre-bleaching

    • Abstract: Publication date: Available online 23 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Archana Gautam, Amit Kumar, Amit Kumar Bharti, Dharm Dutt Rice straw is valuable resource that has been used as substrate for cost effective production of xylanase under solid-state fermentation by a newly isolated white rot fungi, S. commune ARC-11. Out of eleven carbon sources tested, rice straw was found most effective for the induction of xylanase that produced 4288.3 IU/gds of xylanase by S. commune ARC-11. Maximum xylanase production (6721.9 IU/gds) was observed on 8th day of incubation at temperature (30 °C), initial pH (7.0) and initial moisture content (70.0%). The supplementation of ammonium sulphate (0.08% N, as available nitrogen) enhanced the xylanase production up to 8591.4 IU/gds. The xylanase production by S. commune ARC-11 was further improved by the addition of 0.10%, (w/v) of Tween-20 as surfactant. The maximum xylanase activities were found at pH 5.0 and temperature 55 °C with a longer stability (180 min) at temperature 45, 50 and 55 °C. This xylanase preparation was also evaluated for the pre-bleaching of ethanol-soda pulp from Eulaliopsis binata. An enzyme dosage of 10 IU/g of xylanase resulted maximum decrease in kappa number (14.51%) with a maximum improvement 2.9% in ISO brightness compared to control.
       
  • Agrobacterium tumefaciens-mediated transformation of Dendrobium
           lasianthera J.J.Sm: An important medicinal orchid

    • Abstract: Publication date: Available online 22 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Edy Setiti Wida Utami, Sucipto Hariyanto, Yosephine Sri Wulan Manuhara A protocol for genetic transformation mediated by Agrobacterium tumefaciens and production of transgenic Dendrobium lasianthera has been developed for the first time. The 8-week-old protocorm explants were used as target of transformation with Agrobacterium tumefaciens strain LBA4404 carrying plasmid pG35SKNAT1. Several parameters such as infection period, Agrobacterium density, concentration of acetosyringone, and co-cultivation period were evaluated for the transformation efficiency. The data were analyzed using one-way analysis of variance (ANOVA) and Duncan's Multiple Range Test (DMRT) with p 
       
  • Decolorization of Textile Reactive Dyes by Bacterial Monoculture and
           Consortium Screened from Textile Dyeing Effluent

    • Abstract: Publication date: Available online 22 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Md. Ekramul Karim, Kartik Dhar, Md. Towhid Hossain Dyeing effluents have become a vital source of water pollution. Due to the xenobiotic properties and toxicity to all life forms including humans, removal of undesirable color and associated toxicity is crucial. In this study, five dye decolorizing bacteria were isolated from dyeing effluent using selective enrichment culture in Bushnell-Haas (BH) medium amended with co-substrate (i.e. glucose, yeast extract) and 100 mg L−1 of each commercially available reactive dyes viz. Novacron Orange FN-R, Novacron Brilliant Blue FN-R, Novacron Super Black G, Bezema Yellow S8-G and Bezema Red S2-B. The isolated bacteria were identified and assigned as Neisseria sp., Vibrio sp., Bacillus sp., Bacillus sp. and Aeromonas sp. based on their phenotypic (cultural, morphological, physiological and biochemical characteristic) observation. The dye decolorization efficiency was estimated spectrophotometrically up to 6 days of static incubation at 37 °C and observed that all of the isolates were unable to induce decolorization in absence of co-substrate. In case of monoculture, decolorization percentage varies from no visible decolorization (Bezema Red S2-B by Ek-5) to highest 90% decolorization (Novacron Brilliant Blue FN-R by Ek-13) whereas the decolorization percentage of bacterial consortium varies from 65% (Bezema Yellow S8-G) to 90% (Novacron Brilliant Blue FN-R and Novacron Super Black G). The study outlines the co-substrates mediated decolorization process where bacterial consortium proved as efficient dye decolorizer than that of the monocultures. This finding confers possibility of using novel microbial consortium for biological treatment of disreputable dyeing effluents.
       
  • Kolaviron and selenium reduce hydrogen peroxide-induced alterations of the
           inflammatory response

    • Abstract: Publication date: Available online 17 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Tebekeme Okoko The abilities of kolaviron and selenium (either separately or in combination) to prevent hydrogen peroxide-induced alterations in cell viability and activation were investigated. The cell line U937 was incubated with the antioxidants (i.e. kolaviron or selenium) for 24 h before exposure to hydrogen peroxide and cell viability was assessed via trypan blue dye exclusion assay. The U937 cells were also transformed to the macrophage form, incubated with the antioxidants before exposure to hydrogen peroxide. Subsequently, production of nitric oxide and pro-inflammatory cytokines were assessed as indices of macrophage activation. The myoblast cell line H9c2 was also incubated with Se and kolaviron for 24 h before exposure to hydrogen peroxide. Cell viability and generation of reactive oxygen species (ROS) were assessed via MTT and DCHF assays. The results revealed that hydrogen peroxide significantly reduced (p 
       
  • Optimization of quorum quenching mediated bacterial attenuation of Solanum
           torvum root extract by response surface modelling through Box-Behnken
           approach

    • Abstract: Publication date: Available online 7 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Kayeen Vadakkan, Selvaraj Vijayanand, Abbas Alam Choudhury, Ramya Gunasekaran, Janarthanam Hemapriya The present study was intended to optimize the quorum sensing inhibitory action of Solanum torvum root extract against Chromobacterium violaceum. Factors such as bacterial density, frequency of administration and concentration of extract were analysed. Plant samples were collected from Thrissur District, Kerala, India. Response surface modelling of factors by Box-Behnken approach was employed for optimizing quorum quenching activity of extract. The adequacy of mathematical model was verified by ANOVA and Cook’s distance table. Results revealed that quorum quenching property of Solanum torvum root extract is highly influenced by variables studied whereas maximum activity was found during administration of 300 µg/ml extract thrice in a day. It was also understood that extract does not possess any bactericidal activity wherein it only silence its quorum sensing mediated functions. This observations can be further used in quorum quenching studies.
       
  • Expression of Leptospira membrane proteins Signal Peptidase (SP) and
           Leptospira Endostatin like A (Len A) in BL-21(DE3) is toxic to the host
           cells

    • Abstract: Publication date: Available online 2 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Padikara K. Satheeshkumar, Prasannan V. Anu, Mohmed I. Junaida, Madathiparambil G. Madanan, Tennison Jebasingh, Ananthakrishnan J. Nair, Gangaprasad A. Nair, Govinda Pillai M. Nair, Perumana R. Sudhakaran Heterologous expression of Integral Membrane Proteins (IMPs) is reported to be toxic to the host system in many studies. Even though there are reports on various concerns like transformation efficiency, growth properties, protein toxicity, inefficient expression and protein degradation in IMP overexpression, no studies so far addressed these issues in a comprehensive way. In the present study, two transmembrane proteins of the pathogen Leptospira interrogans, namely Signal peptidase (SP), and Leptospira Endostatin like A (Len-A) were taken along with a cytosolic protein Hydrolase (HYD) to assess the differences in transformation efficiency, protein toxicity, and protein stability when over expressed in Escherichia coli (E. coli). Bioinformatics analysis to predict the transmembrane localization indicated that both SP and Len are targeted to the membrane. The three proteins were expressed in full length in the E. coli expression strain, BL 21 (DE3). Significant changes were observed for the strains transformed with IMP genes under the parameters analysed such as, the transformation efficiency, survival of colonies on IPTG-plate, culture growth kinetics and protein expression compared to the strain harbouring the cytosolic protein gene.
       
  • Scenedesmus obliquus: Antioxidant and antiviral activity of proteins
           hydrolyzed by three enzymes

    • Abstract: Publication date: Available online 1 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Abd El-Moneim M.R. Afify, Gamal S. El Baroty, Farouk K. El Baz, Hanaa H. Abd El Baky, Soha A. Murad PurposeTo obtain protein hydrolysates from fresh water green algae Scenedesmus obliquus by three different enzymes and evaluate its antioxidant and antiviral activity.MethodsEnzymatic hydrolysates of green algae Scenedesmus obliquus protein were prepared by treatment with: 1.2% solution of pepsin, trypsin or papain. Protein was extracted from S. obliquus by three different extraction methods. Protein extracts and hydrolysates were assessed from stained gels following SDS–PAGE of samples. Antioxidant activity of protein hydrolysates was investigated.ResultsS. obliquus cells and protein extracts were rich in Arg, Lys, Asp, Ala, and His. Protein hydrolyzed by papain (Sd1pa) and protein hydrolyzed by trypsin (Sd2Try) induced highest antioxidant activity based on 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging (41.41% and 40.62%) respectively, and on 2,2′-azinobis 3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical (87.03% and 45.12%) respectively, at 150 µg/ml. The inhibitory effect and mode of action of protein hydrolysates were evaluated against Coxsackie B3 virus (CVB3). Protein hydrolyzed by papain (Sd2pa) and protein hydrolyzed by pepsin (Sd1pep) at 100 µg/ml exhibited antiviral activity (66.2% and 57.6%, respectively), against (CVB3) from all protein hydrolysates.ConclusionS. obliquus protein hydrolysates have a potential as antioxidative neutraceutical ingredients and a potential therapeutic agent against CVB3.
       
  • Interaction of rs316019 variants of SLC22A2 with metformin and other
           drugs- an in silico analysis

    • Abstract: Publication date: Available online 1 February 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Abu Ashfaqur Sajib, Tasmia Islam, Nilanjana Paul, Sabina Yeasmin Metformin is one of the first-line and most widely prescribed drugs to treat type 2 diabetes (T2D). Its clearance from circulation is mostly facilitated by SLC22A2 (OCT2) in the renal cells. SLC22A2 is a polyspecific organic cation transporter and mediate transport of structurally unrelated endogenous and exogenous compounds including many drugs. rs316019 (p.270A > S) is the most common variant of SLC22A2 with a frequency as high as 15% or more in many populations. The 270S form of SLC22A2 clears metformin from circulation at much reduced level compared to the 270A form. If accumulated, metformin increases plasma lactate level in a concentration-dependent manner which can lead to a condition known as metformin-associated lactic acidosis (MALA). MALA is a potentially life-threatening complication with a mortality rate of 30–50%. Pre-existing clinical conditions, such as renal impairment, sepsis, anoxia, etc may make individuals more prone to MALA. In this study, we used computational approaches to investigate the effect of 270A > S change in SLC22A2 on interaction with metformin and other drugs. Based on the structural models, all substrates bind to the same pocket of SLC22A2. The substrates fit better to the binding site of 270A form of SLC22A2. The binding site has a few core interacting residues, among which SER358 appears to be the most important. It is an in silico prediction that the T2D patients, who are under metformin regimen, should be cautious in taking ranitidine (an over-the-counter sold drug) on a regular basis as it may lead to metformin associated lactate accumulation in blood.
       
  • Statistical optimization of crude oil bio-degradation by a local marine
           bacterium isolate Pseudomonas sp. sp48

    • Abstract: Publication date: Available online 6 January 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): Soha Farag, Nadia A. Soliman, Yasser R. Abdel-Fattah Pseudomonas sp. sp48, a marine bacterium isolated from Bahary area (Alexandria, Egypt), showed a high potency for oil degradation up to 1.5%. Additionally, it showed an ability to consume aromatic hydrocarbons (phenol & naphthalene) and aliphatic (pentadecane) reaching to 79; 73; 62%, respectively. In the current study, Plackett-Burman factorial design was applied to evaluate culture conditions affecting the degradation potency. Analysis of Plackett-Burman design results revealed that, the most significant variables affecting oil removal were magnesium sulfate, inoculum size, glucose and Triton X-100. To optimize the levels of these significant variables Response Surface Methodology (RSM) was followed. In this respect, the three-level Box–Behnken design was employed and a polynomial model was created to correlate the relationship between the three variables and oil removal. The optimal combinations of the major constituents of media that was evaluated from the non-linear optimization algorithm of EXCEL-Solver was as follows: (w/v%) 1 crude oil, 0.5 peptone, 0.5 yeast-extract, 1 ammonium chloride, 0.7418 D-glucose, 0.5 MgSO4·7H2O, 0.1 Triton X-100 and inoculums size 4.18 ml% in natural sea water at pH 7; 30 °C incubation temperature, 200 rpm for 6 days. The predicted optimum oil removal was 89%, which is 2.4 times more than the basal medium.
       
  • Influence of bioprocess variables on the production of extracellular
           chitinase under submerged fermentation by Streptomyces pratensis strain
           KLSL55

    • Abstract: Publication date: Available online 3 January 2018Source: Journal of Genetic Engineering and BiotechnologyAuthor(s): A. Shivalee, K. Lingappa, Divatar Mahesh Chitinases are the enzymes which are capable of hydrolyzing chitin to its monomer N-acetyl glucosamine (GlcNac). Present study emphasizes on the impact of critical process variables on the production of chitinase from Streptomyces pratensis strain KLSL55. Initially the isolate was noticed to produce 84.67 IU chitinase in basal production medium. At optimization of bioprocess variables, the physical parameters pH of 8.00, 40 °C of incubation temperature, agitation speed of 160 rpm and 1.25 mL of spore suspension were found optimum for improved production of chitinase. Further, formulated production medium with 1.5% colloidal chitin, 1.25% fructose greatly influenced the chitinase production. At all described optimum conditions with formulated production media, a total of 14.30-fold increment was achieved in the chitinase production with final activity of 1210.67 IU when compared to the initial fermentation conditions in basal production medium.
       
 
 
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