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  Subjects -> BIOLOGY (Total: 3149 journals)
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BIOTECHNOLOGY (237 journals)                  1 2 | Last

Showing 1 - 200 of 237 Journals sorted alphabetically
3 Biotech     Open Access   (Followers: 8)
Advanced Biomedical Research     Open Access  
Advances in Bioscience and Biotechnology     Open Access   (Followers: 14)
Advances in Genetic Engineering & Biotechnology     Hybrid Journal   (Followers: 8)
African Journal of Biotechnology     Open Access   (Followers: 6)
Algal Research     Partially Free   (Followers: 10)
American Journal of Biochemistry and Biotechnology     Open Access   (Followers: 64)
American Journal of Bioinformatics Research     Open Access   (Followers: 7)
American Journal of Polymer Science     Open Access   (Followers: 31)
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: 43)
Applied Bioenergy     Open Access  
Applied Biosafety     Hybrid Journal  
Applied Food Biotechnology     Open Access   (Followers: 3)
Applied Microbiology and Biotechnology     Hybrid Journal   (Followers: 63)
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: 8)
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)
Bio-Algorithms and Med-Systems     Hybrid Journal   (Followers: 2)
Bio-Research     Full-text available via subscription   (Followers: 3)
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  
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)
Biomarkers in Drug Development     Partially Free   (Followers: 1)
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)
Biosimilars     Open Access   (Followers: 1)
Biosurface and Biotribology     Open Access  
Biotechnic and Histochemistry     Hybrid Journal   (Followers: 2)
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: 5)
Biotechnology & Biotechnological Equipment     Open Access   (Followers: 4)
Biotechnology Advances     Hybrid Journal   (Followers: 33)
Biotechnology and Applied Biochemistry     Hybrid Journal   (Followers: 44)
Biotechnology and Bioengineering     Hybrid Journal   (Followers: 155)
Biotechnology and Bioprocess Engineering     Hybrid Journal   (Followers: 5)
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: 16)
Biotechnology Law Report     Hybrid Journal   (Followers: 4)
Biotechnology Letters     Hybrid Journal   (Followers: 34)
Biotechnology Progress     Hybrid Journal   (Followers: 39)
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  
Biotribology     Hybrid Journal   (Followers: 1)
BMC Biotechnology     Open Access   (Followers: 16)
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)
Contributions to Tobacco Research     Open Access   (Followers: 2)
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: 56)
Current Pharmaceutical Biotechnology     Hybrid Journal   (Followers: 9)
Current Research in Bioinformatics     Open Access   (Followers: 12)
Current Trends in Biotechnology and Chemical Research     Open Access   (Followers: 3)
Current trends in Biotechnology and Pharmacy     Open Access   (Followers: 8)
EBioMedicine     Open Access  
Electronic Journal of Biotechnology     Open Access  
Entomologia Generalis     Full-text available via subscription  
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  
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)
IIOAB Letters     Open Access  
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)
Industrial Biotechnology     Hybrid Journal   (Followers: 18)
International Biomechanics     Open Access  
International Journal of Bioinformatics Research and Applications     Hybrid Journal   (Followers: 13)
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: 3)
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)
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 Biosecurity, Biosafety and Biodefense Law     Hybrid Journal   (Followers: 3)
Journal of Biotechnology     Hybrid Journal   (Followers: 68)
Journal of Biotechnology and Strategic Health Research     Open Access  
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  
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 Essential Oil Research     Hybrid Journal   (Followers: 2)
Journal of Experimental Biology     Full-text available via subscription   (Followers: 24)
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: 16)
Journal of Integrative Bioinformatics     Open Access  
Journal of International Biotechnology Law     Hybrid Journal   (Followers: 3)
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: 11)
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: 11)
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)
Messenger     Full-text available via subscription  
Metabolic Engineering Communications     Open Access   (Followers: 4)
Metalloproteinases In Medicine     Open Access  
Microalgae Biotechnology     Open Access   (Followers: 2)
Microbial Biotechnology     Open Access   (Followers: 9)
MicroMedicine     Open Access   (Followers: 3)
Molecular and Cellular Biomedical Sciences     Open Access  
Molecular Biotechnology     Hybrid Journal   (Followers: 13)
Molecular Genetics and Metabolism Reports     Open Access   (Followers: 3)
Nanobiomedicine     Open Access  
Nanobiotechnology     Hybrid Journal   (Followers: 2)
Nanomaterials and Nanotechnology     Open Access  
Nanomaterials and Tissue Regeneration     Open Access  
Nanomedicine and Nanobiology     Full-text available via subscription  
Nanomedicine Research Journal     Open Access  
Nanotechnology Reviews     Hybrid Journal   (Followers: 5)
Nature Biotechnology     Full-text available via subscription   (Followers: 535)

        1 2 | Last

Journal Cover
New Biotechnology
Journal Prestige (SJR): 0.967
Citation Impact (citeScore): 4
Number of Followers: 3  
 
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 1871-6784
Published by Elsevier Homepage  [3163 journals]
  • Saccharification efficiencies of multi-enzyme complexes produced by
           aerobic fungi
    • Authors: Ajay Badhan; Jiangli Huang; Yuxi Wang; D. Wade Abbott; Marcos Di Falco; Adrian Tsang; Tim McAllister
      Pages: 1 - 6
      Abstract: Publication date: Available online 24 May 2018
      Source:New Biotechnology
      Author(s): Ajay Badhan, Jiangli Huang, Yuxi Wang, D. Wade Abbott, Marcos Di Falco, Adrian Tsang, Tim McAllister
      In the present study, we have characterized high molecular weight multi-enzyme complexes in two commercial enzymes produced by Trichoderma reesei (Spezyme CP) and Penicillium funiculosum (Accellerase XC). We successfully identified 146–1000 kDa complexes using Blue native polyacrylamide gel electrophoresis (BN-PAGE) to fractionate the protein profile in both preparations. Identified complexes dissociated into lower molecular weight constituents when loaded on SDS PAGE. Unfolding of the secondary structure of multi-enzyme complexes with trimethylamine (pH >10) suggested that they were not a result of unspecific protein aggregation. Cellulase (CMCase) profiles of extracts of BN-PAGE fractionated protein bands confirmed cellulase activity within the multi-enzyme complexes. A microassay was used to identify protein bands that promoted high levels of glucose release from barley straw. Those with high saccharification yield were subjected to LC-MS analysis to identify the principal enzymatic activities responsible. The results suggest that secretion of proteins by aerobic fungi leads to the formation of high molecular weight multi-enzyme complexes that display activity against carboxymethyl cellulose and barley straw.

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.05.003
      Issue No: Vol. 46 (2018)
       
  • Cleavage of poly(cis-1,4-isoprene) rubber as solid substrate by cultures
           of Gordonia polyisoprenivorans
    • Authors: R. Andler; S. Hiessl; O. Yücel; M. Tesch; A. Steinbüchel
      Pages: 6 - 12
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): R. Andler, S. Hiessl, O. Yücel, M. Tesch, A. Steinbüchel
      Potential biotechnological recycling processes for rubber products include the bacterial degradation of poly(cis-1,4-isoprene) (IR) in order to achieve its total biodegradation or its biotransformation into useful products. The actinomycete Gordonia polyisoprenivorans strain VH2 catalyzes the degradation of IR and enables its use as a sole carbon source via β-oxidation. The initial cleavage reaction is catalyzed by the extracellular latex clearing protein (Lcp). This dioxygenase is the key enzyme for the formation of oligo(cis-1,4-isoprene) molecules with different lengths, i.e., numbers of isoprene units. For the first time, IR was used as a solid substrate in 2-l fermenters. Two different particle size fractions (63–500 and 500–1000 μm) and three stirring rates (300, 400 and 500 rpm) were evaluated in the process. An increase of the cell concentration was achieved by using smaller particles and by using lower stirring rates, reaching a final biomass concentration of 0.52 g l−1 at 300 rpm after 12 days of cultivation. In order to enhance the formation of oligo(cis-1,4-isoprene) molecules, a transposon insertion mutant (TH5) of G. polyisoprenivorans strain VH2 that has lost the ability to transport the partial degradation products into the cells was used, thereby allowing the accumulation of the degradation products in the culture supernatants. Propionate, glucose and glycerol were evaluated as additional carbon sources besides IR, and the highest yields were observed on propionate. In 2-l bioreactors with pH control, different feeding regimes were performed during cultivation by the addition of propionate every 24 or 48 h for 16 days. After liquid-liquid extraction and a derivatization with Girard’s T reagent, the oligo(cis-1,4-isoprene) molecules were detected by ESI-MS. The mass distribution of the degradation products was affected by the selection of the extraction solvent, but no influence of longer cultivation periods was detected.

      PubDate: 2018-03-20T03:48:14Z
      DOI: 10.1016/j.nbt.2018.03.002
      Issue No: Vol. 44 (2018)
       
  • Improved continuous fumaric acid production with immobilised Rhizopus
           oryzae by implementation of a revised nitrogen control strategy
    • Authors: Andre Naude; Willie Nicol
      Pages: 13 - 22
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): Andre Naude, Willie Nicol
      A novel fermentation system was employed whereby the mycelial mat of Rhizopus oryzae was attached to a polypropylene tube. Batch operation was used for growth, while continuous operation was employed during the fumaric acid production phase. A clear decrease in respiration, fumaric acid (FA) and ethanol production was observed when zero nitrogen was fed in the production phase, with FA productivity decreasing from an initial 0.7 g L−1 h−1 to 0.3 g L−1 h−1 after 150 h. With the addition of 0.625 mg L−1 h−1 of urea FA productivity dropped to only 0.4 g L−1 h−1 after 150 h and 0.3 g L−1 h−1 after 400 h. Under these conditions it was observed that the ethanol production rate decreased 20 times faster compared with the FA production rate, therefore resulting in high FA yields towards the end of the fermentation (instantaneous 0.96 g g−1 and average 0.81 g g−1 after 400 h). Increasing the urea feed rate to 1.875 mg L−1 h−1 resulted in a clear increase in FA production and respiration rates. This condition also resulted in a 25% increase in biomass after 150 h, while the decline in the ethanol production rate was seven times lower than in the 0.625 mg L−1 h−1 urea fermentation, resulting in lower FA yields.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.02.012
      Issue No: Vol. 44 (2018)
       
  • Quality and cost assessment of a recombinant antibody fragment produced
           from mammalian, yeast and prokaryotic host cells: A case study prior to
           pharmaceutical development
    • Authors: Kristell Lebozec; Martine Jandrot-Perrus; Gilles Avenard; Olivier Favre-Bulle; Philippe Billiald
      Pages: 31 - 40
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): Kristell Lebozec, Martine Jandrot-Perrus, Gilles Avenard, Olivier Favre-Bulle, Philippe Billiald
      Monoclonal antibody fragments (Fab) are a promising class of therapeutic agents. Fabs are aglycosylated proteins and so many expression platforms have been developed including prokaryotic, yeast and mammalian cells. However, these platforms are not equivalent in terms of cell line development and culture time, product quality and possibly cost of production that greatly influence the success of a drug candidate’s pharmaceutical development. This study is an assessment of the humanized Fab fragment ACT017 produced from two microorganisms (Escherichia coli and Pichia pastoris) and one mammalian cell host (CHO). Following low scale production and Protein L-affinity purification under generic conditions, physico-chemical and functional quality assessments were carried out prior to economic analysis of industrial scale production using a specialized software (Biosolve, Biopharm Services, UK). Results show higher titer production when using E. coli but associated with high heterogeneity of the protein content recovered in the supernatant. We also observed glycoforms of the Fab produced from P. pastoris, while Fab secreted from CHO was the most homogeneous despite a much longer culture time and slightly higher estimated cost of goods. This study may help inform future pharmaceutical development of this class of therapeutic proteins.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.04.006
      Issue No: Vol. 44 (2018)
       
  • Preparation of a robust immobilized biocatalyst of β-1,4-endoxylanase by
           surface coating with polymers for production of xylooligosaccharides from
           different xylan sources
    • Authors: Maria Romero-Fernández; Sonia Moreno-Perez; Sandro Martins de Oliveira; Ramón I. Santamaría; Jose M. Guisan; Javier Rocha-Martin
      Pages: 50 - 58
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): Maria Romero-Fernández, Sonia Moreno-Perez, Sandro Martins de Oliveira, Ramón I. Santamaría, Jose M. Guisan, Javier Rocha-Martin
      Xylooligosaccharides display interesting prebiotic effects on human health. The endoxylanase Xys1Δ, from Streptomyces halstedii JM8, was immobilized and stabilized on glyoxyl-agarose beads by multipoint covalent attachment using a novel strategy based on surface coating with a multilayer of polymers. The optimal modification consisted of surface coating with a bilayer formed by a layer of derived dextran polymers and a layer of polyethylenimine. The optimized biocatalyst was 550-fold more stable than one-point covalent immobilized Xys1Δ (at 70 °C, pH 7). This biocatalyst was tested for the production of xylooligosaccharides from soluble xylans from various sources. Hydrolysis of beechwood, wheat straw and corncob xylans was 93% in 4 h, 44% in 5 h and 100% in 1 h, respectively. Maximum values of xylooligosaccharides were found for beechwood at 20.6 mg/mL, wheat at 12.5 mg/mL and corncob at 30.4 mg/mL. The optimized biocatalyst was reused for 15 reaction cycles without affecting its catalytic activity.

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.04.007
      Issue No: Vol. 44 (2018)
       
  • Nanoreactors: Strategies to encapsulate enzyme biocatalysts in virus-like
           particles
    • Authors: Joshua W. Wilkerson; Seung-Ook Yang; Parker J. Funk; Steven K. Stanley; Bradley C. Bundy
      Pages: 59 - 63
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): Joshua W. Wilkerson, Seung-Ook Yang, Parker J. Funk, Steven K. Stanley, Bradley C. Bundy
      Enzyme-mediated biocatalysis is generally more selective and environmentally friendly and requires less energy than chemocatalysis. However, factors such as temperature, acidity and the presence of proteases can negate enzyme activity. Encapsulation in virus-like particles is one promising method to mitigate these difficulties. Encapsulation also can be used to create multi-reaction nanoreactors that increase process efficiency by isolating reaction intermediates. To successfully encapsulate enzymes, a variety of methods involving both non-covalent and covalent interactions have been developed. Here we review promising virus-like particle encapsulation strategies, their advantages and remaining challenges.

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.04.003
      Issue No: Vol. 44 (2018)
       
  • Overexpression of acetyl-CoA carboxylase in Aspergillus terreus to
           increase lovastatin production
    • Authors: Hanan Hasan; Muhammad Hafiz Abd Rahim; Leona Campbell; Dee Carter; Ali Abbas; Alejandro Montoya
      Pages: 64 - 71
      Abstract: Publication date: 25 September 2018
      Source:New Biotechnology, Volume 44
      Author(s): Hanan Hasan, Muhammad Hafiz Abd Rahim, Leona Campbell, Dee Carter, Ali Abbas, Alejandro Montoya
      The present work describes the application of homologous recombination techniques in a wild-type Aspergillus terreus (ATCC 20542) strain to increase the flow of precursors towards the lovastatin biosynthesis pathway. A new strain was generated to overexpress acetyl-CoA carboxylase (ACCase) by replacing the native ACCase promoter with a strong constitutive PadhA promoter from Aspergillus nidulans. Glycerol and a mixture of lactose and glycerol were used independently as the carbon feedstock to determine the degree of response by the A. terreus strains towards the production of acetyl-CoA, and malonyl-CoA. The new strain increased the levels of malonyl-CoA and acetyl-CoA by 240% and 14%, respectively, compared to the wild-type strain. As a result, lovastatin production was increased by 40% and (+)-geodin was decreased by 31% using the new strain. This study shows for the first time that the metabolism of Aspergillus terreus can be manipulated to attain higher levels of precursors and valuable secondary metabolites.
      Graphical abstract image

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.04.008
      Issue No: Vol. 44 (2018)
       
  • Microeukaryote community in a partial nitritation reactor prior to anammox
           and an insight into the potential of ciliates as performance bioindicators
           
    • Authors: Oriol Canals; Ramon Massana; Joan Lluís Riera; Vanessa Balagué; Humbert Salvadó
      Pages: 3 - 12
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Oriol Canals, Ramon Massana, Joan Lluís Riera, Vanessa Balagué, Humbert Salvadó
      An in-depth, long-term, multidisciplinary study was conducted in order to study the microeukaryote community in a partial nitritation (PN) reactor prior to anammox. The PN reactor operated with moving bed biofilm reactor (MBBR) technology, using plastic supports (carriers) for biofilm development. The microeukaryote community from the biofilm (BF) and the surrounding media (mixed liquor or ML) were analysed separately. Despite the physicochemical conditions under which the PN-MBBR operated (an average of 305.9±117mg TAN l−1 and 328.4±131.9mg N-NO2 − l−1), up to 24 microeukaryotic taxa were observed by microscope. Microeukaryote species showed an uneven distribution in the PN-MBBR, thus suggesting the existence of two habitats: the BF, preferred by species with specific structures for adhering to a substrate, such as the stalked Peritrichia, and the ML, preferred by free-swimming or non-substrate dependent species. The results indicated that most ciliate population dynamics mainly responded to the nitrous acid and free ammonia concentrations and, to a lesser extent, to sCOD values. In the BF, variations in the population of Epistylis camprubii and Opercularia coarctata suggest the existence of competition between these species due to niche overlap. A V4 18S rDNA molecular survey (Illumina) was carried out for some samples with the aim of obtaining maximum coverage of the main eukaryote species that were microscopically detected throughout the study. The diversity and abundance data provided by both detection methods were compared. The study helped identify broader tolerance ranges of the microeukaryote taxa to the physicochemical parameters analysed.
      Graphical abstract image

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.05.003
      Issue No: Vol. 43 (2018)
       
  • Optimization of washing conditions with biogenic mobilizing agents for
           marine fuel-contaminated beach sands
    • Authors: Alessia Arelli; Andrea Nuzzo; Claudia Sabia; Ibrahim M. Banat; Giulio Zanaroli; Fabio Fava
      Pages: 13 - 22
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Alessia Arelli, Andrea Nuzzo, Claudia Sabia, Ibrahim M. Banat, Giulio Zanaroli, Fabio Fava
      Washing is a rapid and effective treatment to remediate contaminated sands impacted by oil spills, although synthetic additives used to increase extraction efficiency may cause additional pollution issues due to their intrinsic toxicity and very often low biodegradability. In this study, different biogenic mobilizing agents (soybean lecithins, cyclodextrins, cholic acids, plant-derived cleaners, rhamnolipids and sophorolipids) were tested in the washing of beach sands artificially contaminated with the Intermediate Fuel Oil IFO-180. Among these, a de-oiled soybean lecithin (SL-1), hydroxypropyl-β-cyclodextrins (HPB-CD) and sophorolipids (SR) achieved hydrocarbon removals close to those attained with the synthetic surfactant Triton™ X-100 (TX) in preliminary washing tests carried out at constant mixing rate, water/sand ratio and IFO-180 contamination level using agents concentrations close to their critical micelle concentration (0.1% and 1% w/v for microbial and non-microbial agents, respectively). The effects of agent concentration, water/sand ratio, mixing rate and IFO-180 contamination on hydrocarbons removal were modelled using face-centred central composite design and ANOVA. Optimal washing parameters for sand contamination levels in the range 0.5–20 g/kg were identified with response surface methodology. While HPB-CD and SR performed equally to TX only at low sand contaminations, SL-1 attained hydrocarbon removal higher or equal to that of TX at any IFO-180 contamination and at lower application rates. SL-1 also outperformed TX when minimizing the water/sand ratio, i.e., the volume of water used. Considering its lower toxicity, higher biodegradability and higher hydrocarbon removal efficiencies, SL-1 is an effective and environmentally sustainable alternative to synthetic surfactants in washing treatments for marine fuel-contaminated sands.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.12.007
      Issue No: Vol. 43 (2018)
       
  • Biodegradation of mono-, di- and trifluoroacetate by microbial cultures
           with different origins
    • Authors: Diogo A.M. Alexandrino; Inês Ribeiro; Luís M. Pinto; Rafael Cambra; Rui S. Oliveira; Filipe Pereira; Maria F. Carvalho
      Pages: 23 - 29
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Diogo A.M. Alexandrino, Inês Ribeiro, Luís M. Pinto, Rafael Cambra, Rui S. Oliveira, Filipe Pereira, Maria F. Carvalho
      This work focused on the biodegradation of three structurally related fluoroacetates (FAs), mono- (MFA), di- (DFA) and trifluoroacetate (TFA), using as microbial inocula samples collected from a site with a long history of industrial contamination and activated sludge obtained from a municipal wastewater treatment plant. Biodegradation experiments were carried out under different modes of substrate supplementation, which included (i) FAs fed as sole carbon sources; (ii) FAs (only for DFA and TFA) fed in co-metabolism with sodium acetate; and (iii) mixtures of MFA with DFA or TFA. Biodegradation of the target compounds was assessed through fluoride ion release. Defluorination was obtained in the cultures fed with MFA, while DFA and TFA were recalcitrant in all tested conditions. When present in mixture, DFA was shown to inhibit biodegradation of MFA, while TFA had no effect. A total of 13 bacterial isolates obtained from MFA degrading cultures were found to degrade 20mgL−1 of this compound, as single strains, when supplemented as a sole carbon source. Sequencing of the 16S rRNA gene indicated that among these degrading bacteria only Delftia acidovorans had been previously reported to be able to degrade MFA. This work shows that, despite their similar chemical structures, biodegradation of the three tested FAs is very distinct and draws attention to the unknown impacts that the accumulation of DFA and TFA may have in the environment as a result of their high recalcitrance.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.08.005
      Issue No: Vol. 43 (2018)
       
  • Bacterial isolates degrading ritalinic acid—human metabolite of
           neuro enhancer methylphenidate
    • Authors: Marta Woźniak-Karczewska; Monika Čvančarová; Łukasz Chrzanowski; Philippe F.-X. Corvini; Danuta Cichocka
      Pages: 30 - 36
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Marta Woźniak-Karczewska, Monika Čvančarová, Łukasz Chrzanowski, Philippe F.-X. Corvini, Danuta Cichocka
      The consumption of nootropic drugs has increased tremendously in the last decade, though the studies on their environmental fate are still scarce. Nootropics are bioactive compounds designed to alter human's physiology therefore the adverse effects towards wildlife can be expected. In order to understand their environmental impact, the knowledge on their transformation pathways is necessary. Methylphenidate belongs to the most prescribed neuro-enhancers and is among the most favored smart drugs used in non-medical situations. It is metabolized in human liver and excreted as ritalinic acid. Here, we showed for the first time that ritalinic acid can be biodegraded and used as a sole carbon and nitrogen source by various microbial strains originating from different environmental samples. Five axenic strains were isolated and identified as: Arthrobacter sp. strain MW1, MW2 and MW3, Phycicoccus sp. strain MW4 and Nocardioides sp. strain MW5. Our research provides the first insight into the metabolism of ritalinic acid and suggests that it may differ depending on the strain and growth conditions, especially on availability of nitrogen. The isolates obtained in this study can serve as model organisms in further studies on the catabolism of ritalinic acid and methylphenidate but potentially also other compounds with similar structures. Our findings have important implication for the sewage epidemiology. We demonstrated that ritalinic acid is subject to quick and efficient biodegradation thus its use as a stable biomarker should be reconsidered.
      Graphical abstract image

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.08.009
      Issue No: Vol. 43 (2018)
       
  • Isolation of two Ochrobactrum sp. strains capable of degrading the
           nootropic drug—Piracetam
    • Authors: Marta Woźniak-Karczewska; Monika Čvančarová; Łukasz Chrzanowski; Boris Kolvenbach; Philippe F.-X. Corvini; Danuta Cichocka
      Pages: 37 - 43
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Marta Woźniak-Karczewska, Monika Čvančarová, Łukasz Chrzanowski, Boris Kolvenbach, Philippe F.-X. Corvini, Danuta Cichocka
      Piracetam (2-oxo-1-pyrrolidine acetamide) is a popular cognitive enhancer, which has recently been detected in waste and drinking water. Nootropic drugs are designed to affect human metabolism and act on the nervous system, but their environmental effects have yet to be the subject of detailed studies. In this report, we present the efficient biodegradation of the cognitive enhancer, piracetam. Two bacterial strains capable of using this compound as the sole carbon source were isolated and later identified as Ochrobactrum anthropi strain MW6 and Ochrobactrum intermedium strain MW7. The compound's mineralization and the cleavage of the heterocyclic ring were shown in the experiments with 14C-labeled piracetam. This is also the first report of a pharmaceutical's degradation by the Ochrobactrum genus. This study presents model microorganisms that can be used in further investigation of piracetam's degradation pathways as well as enzymes and genes involved in the process.
      Graphical abstract image

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.07.006
      Issue No: Vol. 43 (2018)
       
  • Biotransformation of ritalinic acid by laccase in the presence of mediator
           TEMPO
    • Authors: Aza Kobakhidze; Vladimir Elisashvili; Philippe F.-X. Corvini; Monika Čvančarová
      Pages: 44 - 52
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Aza Kobakhidze, Vladimir Elisashvili, Philippe F.-X. Corvini, Monika Čvančarová
      Methylphenidate is widely used as a medication for the treatment of attention deficit hyperactivity disorder (ADHD) in children. Less than 1% of methylphenidate is excreted unchanged in urine, while 80% of an oral dose is excreted as ritalinic acid (which is reportedly poorly degradable). This study aims to investigate the biotransformation of ritalinic acid by free and immobilized enzymes. The influence of various laccase mediators on biotransformation efficiency has been tested. Formation of the main transformation products has been monitored and their potential structures suggested. The effective transformation of ritalinic acid was observed only in the presence of 2,2,6,6-tetramethylpiperidine 1-oxyl mediator (TEMPO). The most effective enzyme was the laccase of T. versicolor 159. The main transformation product was an N-methyl derivative of ritalinic acid. Ritalinic acid was also reduced to aldehyde and alcohol, and a broad spectrum of intermediate complexes with oxoammonium ion of TEMPO were detected. This is the first time the biotransformation of ritalinic acid has been investigated in detail.
      Graphical abstract image

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.08.008
      Issue No: Vol. 43 (2018)
       
  • Biodegradation of endocrine disruptors in urban wastewater using Pleurotus
           ostreatus bioreactor
    • Authors: Zdena Křesinová; Lucie Linhartová; Alena Filipová; Martin Ezechiáš; Pavel Mašín; Tomáš Cajthaml
      Pages: 53 - 61
      Abstract: Publication date: 25 July 2018
      Source:New Biotechnology, Volume 43
      Author(s): Zdena Křesinová, Lucie Linhartová, Alena Filipová, Martin Ezechiáš, Pavel Mašín, Tomáš Cajthaml
      The white rot fungus Pleurotus ostreatus HK 35, which is also an edible industrial mushroom commonly cultivated in farms, was tested in the degradation of typical representatives of endocrine disrupters (EDCs; bisphenol A, estrone, 17β-estradiol, estriol, 17α-ethinylestradiol, triclosan and 4-n-nonylphenol); its degradation efficiency under model laboratory conditions was greater than 90% within 12 days and better than that of another published strain P. ostreatus 3004. A spent mushroom substrate from a local farm was tested for its applicability in various batch and trickle-bed reactors in degrading EDCs in model fortified and real communal wastewater. The reactors were tested under various regimes including a pilot-scale trickle-bed reactor, which was finally tested at a wastewater treatment plant. The result revealed that the spent substrate is an efficient biodegradation agent, where the fungus was usually able to remove about 95% of EDCs together with suppression of the estrogenic activity of the sample. The results showed the fungus was able to operate in the presence of bacterial microflora in wastewater without any substantial negative effects on the degradation abilities. Finally, a pilot-scale trickle-bed reactor was installed in a wastewater treatment plant and successfully operated for 10days, where the bioreactor was able to remove more than 76% of EDCs present in the wastewater.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2017.05.004
      Issue No: Vol. 43 (2018)
       
  • Economic feasibility and environmental impact of synthetic spider silk
           production from escherichia coli
    • Authors: Alan M. Edlund; Justin Jones; Randolph Lewis; Jason C. Quinn
      Pages: 12 - 18
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Alan M. Edlund, Justin Jones, Randolph Lewis, Jason C. Quinn
      Major ampullate spider silk represents a promising protein-based biomaterial with diverse commercial potential ranging from textiles to medical devices due to its excellent physical and thermal properties. Recent advancements in synthetic biology have facilitated the development of recombinant spider silk proteins from Escherichia coli (E. coli). This study specifically investigates the economic feasibility and environmental impact of synthetic spider silk manufacturing. Pilot scale data was used to validate an engineering process model that includes all of the required sub-processing steps for synthetic fiber manufacture: production, harvesting, purification, drying, and spinning. Modeling was constructed modularly to support assessment of alternative downstream processing technologies. The techno-economic analysis indicates a minimum sale price from pioneer and optimized E. coli plants of $761 kg−1 and $23 kg−1 with greenhouse gas emissions of 572 kg CO2-eq. kg−1 and 55 kg CO2-eq. kg−1, respectively. Elevated costs and emissions from the pioneer plant can be directly tied to the high material consumption and low protein yield. Decreased production costs associated with the optimized plant includes improved protein yield, process optimization, and an Nth plant assumption. Discussion focuses on the commercial potential of spider silk, the production performance requirements for commercialization, and the impact of alternative technologies on the system.

      PubDate: 2018-02-16T00:16:22Z
      DOI: 10.1016/j.nbt.2017.12.006
      Issue No: Vol. 42 (2018)
       
  • Scientific underpinnings of biotechnology regulatory frameworks
    • Authors: Savannah Gleim; Stuart J. Smyth
      Pages: 26 - 32
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Savannah Gleim, Stuart J. Smyth
      Part of what is presently missing at domestic regulatory levels (and that is important at the international level as well) is a detailed understanding of what the rules of, and for, regulation should be, who the actors, stakeholders and major decision makers are and finally, how to get agreement about the rules. Greater insights into the system of rules that underpin regulatory frameworks for agri-food and biotechnology products in genetically modified (GM) crop- adopting nations will provide value by clarifying the evidence used to commercialize these technologies. This article examines the public documents available from Canada, the United States, the European Union and the Organisation for Economic Cooperation and Development regarding the development of regulatory risk assessment frameworks for products of biotechnology to determine what science grounds these frameworks. The documentation used to provide the initial structure to the existing regulatory frameworks identifies the linkages, connections and relationships that exist between science, risk assessment and regulatory policy. The relationship between risk and regulation has never been more critical to the commercialization of innovative agricultural products. Documenting the role of science-based risk assessment in regulations and how this has changed over the 20 years of experience in regulating GM crops will identify changes in the risk/regulation relationship.

      PubDate: 2018-02-16T00:16:22Z
      DOI: 10.1016/j.nbt.2018.01.004
      Issue No: Vol. 42 (2018)
       
  • Biotechnologically produced chitosan for nanoscale products. A legal
           analysis
    • Authors: Greet Smets; Patrick Rüdelsheim
      Pages: 42 - 47
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Greet Smets, Patrick Rüdelsheim
      Conventionally, chitosans are derived from shrimp and other crustacean shells. Biotechnology offers an alternative route to produce chitosans and more importantly, specific chitosan structures tailored to the needs of a diversity of industries. However, for biotech chitosans and products thereof to be commercialised, legislation should not create a burden. Here, the requirements of the EU regulatory framework have been analysed for the entire chain from research to development and production of several potential applications including nanomaterials. The animal or biotechnological origin leads to specific requirements in production of the raw material. No EU legislation dedicated to nanomaterials has been adopted. Instead, products are governed under the respective existing product legislation subject to extra requirements for safety assessment. While a knowledge gap exists on hazards related to nanomaterials in general, there is a need to establish realistic regulatory study designs to assess the safety of specific products. Furthermore, as many of the existing chitosan applications are not considered nanomaterials, it would be discriminatory to treat biotechnology derived products differently.

      PubDate: 2018-02-16T00:16:22Z
      DOI: 10.1016/j.nbt.2018.02.005
      Issue No: Vol. 42 (2018)
       
  • One-step integrated clarification and purification of a monoclonal
           antibody using Protein A Mag Sepharose beads and a cGMP-compliant
           high-gradient magnetic separator
    • Authors: Moritz Ebeler; Ola Lind; Nils Norrman; Ronnie Palmgren; Matthias Franzreb
      Pages: 48 - 55
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Moritz Ebeler, Ola Lind, Nils Norrman, Ronnie Palmgren, Matthias Franzreb
      Monoclonal antibodies are a dominant component of today's biopharmaceutical market and are typically purified by classical platform processes. However, high costs and rising demands are drivers for the development of new, efficient and flexible integrated purification processes. Currently, high-gradient magnetic separation as a direct capturing tool for protein purification suffers from the lack of suitable GMP-compliant separation equipment for industrial scale. As a solution for this bottleneck, we present a purification process for a monoclonal antibody directly from CHO cell culture by use of protein A-functionalized magnetic particles together with the first pilot-scale GMP-compliant ‘rotor-stator’ high-gradient magnetic separator. Five consecutive purification cycles were performed, achieving consistent yields of over 85% and purities of over 95%. Stable cell viabilities during the magnetic separation process enable integration of the device as an in situ product removal tool. A comparison with state-of-the-art protein A column-based purification processes reveals a 3-times higher process productivity per mL of applied resin and demonstrates the great potential of magnetic separation in downstream processing.
      Graphical abstract image

      PubDate: 2018-02-26T01:13:15Z
      DOI: 10.1016/j.nbt.2018.02.007
      Issue No: Vol. 42 (2018)
       
  • Enhanced bioproduction of 2-phenylethanol in a biphasic system with
           rapeseed oil
    • Authors: Karolina Chreptowicz; Jolanta Mierzejewska
      Pages: 56 - 61
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Karolina Chreptowicz, Jolanta Mierzejewska
      Increasing demand for natural fragrance ingredients and products has led to their global market growth. 2-Phenylethanol (2-PE) is a volatile substance widely used in food and cosmetics manufacturing. It is generally known that yeast can metabolize l-phenylalanine (l-Phe) to produce 2-PE. However, because the product exhibits an inhibitory effect on yeast cells, simple batch cultivation is uneconomic. The aim of this study was to enhance 2-PE productivity using in situ product removal. Here we present a new method of 2-PE production by yeast in a biphasic system with rapeseed oil as the second phase. The chosen solvent is safe, inexpensive and suitable for the extraction of 2-PE. In addition, rapeseed oil appeared to be a valuable source of intermediates for 2-PE synthesis as its presence in the yeast culture significantly enhanced productivity. The process is an environmentally friendly route and gives two final products that can be considered natural: rapeseed oil with a rose odor and pure 2-PE. Both may be subsequently used as food or cosmetics additives. The results obtained are competitive with previously reported values, as it was possible to enhance the overall concentration of 2-PE by 2.7-fold. The total 2-PE concentration in the biphasic system in the 4.5-L biofermentor used was increased to 9.79 g/L, while the 2-PE concentration in the organic phase attained a value of 18.50 g/L.
      Graphical abstract image

      PubDate: 2018-02-26T01:13:15Z
      DOI: 10.1016/j.nbt.2018.02.009
      Issue No: Vol. 42 (2018)
       
  • Rosa hybrida orcinol O-methyl transferase-mediated production of
           pterostilbene in metabolically engineered grapevine cell cultures
    • Authors: Ascensión Martínez-Márquez; Jaime A. Morante-Carriel; Javier Palazon; Roque Bru-Martínez
      Pages: 62 - 70
      Abstract: Publication date: 25 May 2018
      Source:New Biotechnology, Volume 42
      Author(s): Ascensión Martínez-Márquez, Jaime A. Morante-Carriel, Javier Palazon, Roque Bru-Martínez
      Stilbenes are naturally scarce high-added-value plant compounds with chemopreventive, pharmacological and cosmetic properties. Bioproduction strategies include engineering the metabolisms of bacterial, fungal and plant cell systems. Strikingly, one of the most effective strategies consists in the elicitation of wild grapevine cell cultures, which leads to vast stilbene resveratrol accumulation in the extracellular medium. The combination of both cell culture elicitation and metabolic engineering strategies to produce resveratrol analogs proved more efficient for the hydroxylated derivative piceatannol than for the dimethylated derivative pterostilbene, for which human hydroxylase HsCYP1B1- and grapevine O-methyltransferase VvROMT-transformed cell cultures were respectively used. Rose orcinol O-methyltransferase (OOMT) displays enzymatic properties, which makes it an appealing candidate to substitute VvROMT in the combined strategy to enhance the pterostilbene production level by engineered grapevine cells upon elicitation. Here we cloned a Rosa hybrida OOMT gene, and created a genetic construction suitable for Agrobacterium-mediated plant transformation. OOMT’s ability to catalyze the conversion of resveratrol into pterostilbene was first assessed in vitro using protein extracts of agroinfiltrated N. benthamiana leaves and transformed grapevine callus. The grapevine cell cultures transformed with RhOOMT produced about 16 mg/L culture of pterostilbene and reached an extracellular distribution of up to 34% of total production at the best, which is by far the highest production reported to date in a plant system. A bonus large resveratrol production of ca. 1500–3000 mg/L was simultaneously obtained. Our results demonstrate a viable successful metabolic engineering strategy to produce pterostilbene, a resveratrol analog with enhanced pharmacological properties.

      PubDate: 2018-03-20T03:48:14Z
      DOI: 10.1016/j.nbt.2018.02.011
      Issue No: Vol. 42 (2018)
       
  • Plant Biotechnology: Green for Good IV
    • Authors: Petr Tarkowski; Ivo Frébort
      Abstract: Publication date: Available online 26 May 2018
      Source:New Biotechnology
      Author(s): Petr Tarkowski, Ivo Frébort


      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.05.198
       
  • Development of microreactors with surface-immobilized biocatalysts for
           continuous transamination
    • Authors: Nataša Miložič; Gorazd Stojkovič; Andreas Vogel; Dominique Bouwes; Polona Žnidaršič-Plazl
      Abstract: Publication date: Available online 18 May 2018
      Source:New Biotechnology
      Author(s): Nataša Miložič, Gorazd Stojkovič, Andreas Vogel, Dominique Bouwes, Polona Žnidaršič-Plazl
      The industrial importance of optically pure compounds has thrown a spotlight on ω-transaminases that have shown a high potential for the synthesis of bioactive compounds with a chiral amine moiety. The implementation of biocatalysts in industrial processes relies strongly on fast and cost effective process development, including selection of a biocatalyst form and the strategy for its immobilization. Here, microscale reactors with selected surface-immobilized amine-transaminase (ATA) in various forms are described as platforms for high-throughput process development. Wild type ATA (ATA-wt) from a crude cell extract, as well as Escherichia coli cells intracellularly overexpressing the enzyme, were immobilized on the surfaces of meander microchannels of disposable plastics by means of reactor surface silanization and glutaraldehyde bonding. In addition, a silicon/glass microchannel reactor was used for immobilization of an ATA-wt, genetically engineered to contain a silica-binding module (SBM) at the N-terminus (N-SBM-ATA-wt), leading to immobilization on the non-modified inner microchannel surface. Microreactors with surface-immobilized biocatalysts were coupled with a quenching system and at-line HPLC analytics and evaluated based on continuous biotransformation, yielding acetophenone and l-alanine. E. coli cells and N-SBM-ATA-wt were efficiently immobilized and yielded a volumetric productivity of up to 14.42 g L−1 h−1, while ATA-wt small load resulted in two orders of magnitude lower productivity. The miniaturized reactors further enabled in-operando characterization of biocatalyst stability, crucial for successful transfer to a production scale.
      Graphical abstract image

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.05.004
       
  • Effects of copper and arsenic stress on the development of Norway spruce
           somatic embryos and their visualization with the environmental scanning
           electron microscope
    • Authors: Biljana Đorđević; Vilém Neděla; Eva Tihlaříková; Václav Trojan; Ladislav Havel
      Abstract: Publication date: Available online 18 May 2018
      Source:New Biotechnology
      Author(s): Biljana Đorđević, Vilém Neděla, Eva Tihlaříková, Václav Trojan, Ladislav Havel
      Somatic embryogenesis is an important biotechnological technique which can be used in studies associated with environmental stress. Four embryogenic cell lines of Norway spruce were grown on media enriched with copper and arsenic in concentration ranges 50–500 μM and 10–50 μM, respectively. The effects were observed during subsequent stages of somatic embryogenesis, the characteristics evaluated being proliferation potential, average number of somatic embryos obtained per g/fresh weight, morphology of developed somatic embryos, metal uptake, and microanalysis of macro- and micronutrients uptake. Copper and arsenic at higher concentrations significantly reduced the growth of early somatic embryos. In almost all treatments, the cell line V-1-3 showed the best performance compared with the other lines tested. Environmental scanning electron microscopy was used to visualize and identify morphological abnormalities in the development of somatic embryos. Abnormalities observed were classified into several categories: meristemless somatic embryos, somatic embryos with disrupted meristem, reduced number of cotyledons, single cotyledon and fused cotyledons. With the application of a low temperature method for the environmental scanning electron microscope, samples were stabilized and whole meristems could be investigated in their native state. As far as we are aware, this is the first report of the effect of copper and arsenic during the process of somatic embryogenesis and the first to evaluate the content of macro and micronutrients uptake in Norway spruce.

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.05.005
       
  • Generating a recombinant phosphothreonine-binding domain for a
           phosphopeptide of the human transcription factor, c-Myc
    • Authors: Leon A. Venegas; Stefanie L. Kall; Oluwadamilola Bankole; Arnon Lavie; Brian K. Kay
      Abstract: Publication date: Available online 12 May 2018
      Source:New Biotechnology
      Author(s): Leon A. Venegas, Stefanie L. Kall, Oluwadamilola Bankole, Arnon Lavie, Brian K. Kay
      Transcription factor c-Myc is an oncoprotein that is regulated at the post-translational level through phosphorylation of two conserved residues, Serine 62 (Ser62) and Threonine 58 (Thr58). A highly specific tool capable of recognizing Myc via pThr58 is needed to monitor activation and localization. Through phage display, we have isolated 10 engineered Forkhead-associated (FHA) domains that selectively bind to a phosphothreonine (pThr)-containing peptide (53-FELLPpTPPLSPS-64) segment of human c-Myc. One domain variant was observed to bind to the Myc-pThr58 peptide with a KD value of 800 nM and had >1000-fold discrimination between the phosphorylated and non-phosphorylated peptide. The crystal structure of the engineered FHA Myc-pThr-binding domain (Myc-pTBD) was solved in complex with its cognate ligand. The Myc-pTBD was observed to be structurally similar to the yeast Rad9 FHA1 domain, except that its β4-β5 and β10-β11 loops form a hydrophobic pocket to facilitate the interaction between the domain and the peptide ligand. The Myc-pTBD’s specificity for its cognate ligand was demonstrated to be on a par with 3 commercial polyclonal antibodies, suggesting that this recombinant reagent is a viable alternative to antibodies for monitoring Myc regulation.

      PubDate: 2018-05-31T15:42:19Z
      DOI: 10.1016/j.nbt.2018.05.001
       
  • Core element characterization of Rhodococcus promoters and development of
           a promoter-RBS mini-pool with different activity levels for efficient gene
           expression
    • Authors: Song Jiao; Huimin Yu; Zhongyao Shen
      Abstract: Publication date: Available online 22 April 2018
      Source:New Biotechnology
      Author(s): Song Jiao, Huimin Yu, Zhongyao Shen
      To satisfy the urgent demand for promoter engineering that can accurately regulate the metabolic circuits and expression of specific genes in the Rhodococcus microbial platform, a promoter-ribosome binding site (RBS) coupled mini-pool with fine-tuning of different activity levels was successfully established. Transcriptome analyses of R. ruber TH revealed several representative promoters with different activity levels, e.g., Pami, Pcs, Pnh, P50sl36, PcbiM, PgroE and Pniami. β-Galactosidase (LacZ) reporter measurement demonstrated that different gene expression levels could be obtained with these natural promoters combined with an optimal RBS of ami. Further use of these promoters to overexpress the nitrile hydratase (NHase) gene with RBSami in R. ruber THdAdN produced different expression levels consistent with the transcription analyses. The −35 and −10 core elements of different promoters were further analyzed, and the conserved sequences were revealed to be TTGNNN and (T/C)GNNA(A/C)AAT. By mutating the core elements of the strong promoters, Pnh and Pami, into the above consensus sequence, two even stronger promoters, PnhM and PamiM, were obtained with 2.2-fold and 7.7-fold improvements in transcription, respectively. Integrating several strategies, including transcriptome promoter screening, −35 and −10 core element identification, core element point-mutation, RBS optimization and diverse reporter verification, a fine-tuning promoter-RBS combination mini-pool with different activity levels in Rhodococcus strains was successfully established. This development is significant for broad applications of the Rhodococcus genus as a microbial platform.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.04.005
       
  • Engineering low-cadmium rice through stress-inducible expression of
           OXS3-family member genes
    • Authors: Changhu Wang; Weili Guo; Xingzhe Cai; Ruyu Li; David W. Ow
      Abstract: Publication date: Available online 21 April 2018
      Source:New Biotechnology
      Author(s): Changhu Wang, Weili Guo, Xingzhe Cai, Ruyu Li, David W. Ow
      Cadmium (Cd) as a carcinogen poses a great threat to food security and public health through plant-derived foods such as rice, the staple for nearly half of the world’s population. We have previously reported that overexpression of truncated gene fragments derived from the rice genes OsO3L2 and OsO3L3 could reduce Cd accumulation in transgenic rice. However, we did not test the full length genes due to prior work in Arabidopsis where overexpression of these genes caused seedling lethality. Here, we report on limiting the overexpression of OsO3L2 and OsO3L3 through the use of the stress- inducible promoter RD29B. However, despite generating 625 putative transformants, only 7 lines survived as T1 seedlings and only 1 line of each overexpressed OsO3L2 or OsO3L3-produced T2 progeny. The T2 homozygotes from these 2 lines showed the same effect of reducing accumulation of Cd in root and shoot as well as in T3 grain. As importantly, the concentrations of essential metals copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn) were unaffected. Analysis of the expression profile suggested that low Cd accumulation may be due to high expression of OsO3L2 and OsO3L3 in the root tip region. Cellular localization of OsO3L2 and OsO3L3 indicate that they are histone H2A interacting nuclear proteins in vascular cells and especially in the root tip region. It is possible that interaction with histone H2A modifies chromatin to regulate downstream gene expression.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.04.004
       
  • Caught in-between: System for in-flow inactivation of enzymes as an
           intermediary step in “plug-and-play” microfluidic platforms
    • Authors: Ana C. Fernandes; Benjamin Petersen; Lars Møller; Krist V. Gernaey; Ulrich Krühne
      Abstract: Publication date: Available online 20 April 2018
      Source:New Biotechnology
      Author(s): Ana C. Fernandes, Benjamin Petersen, Lars Møller, Krist V. Gernaey, Ulrich Krühne
      The need for fast and comprehensive characterization of biocatalysts has pushed the development of new screening platforms based on microfluidics, capable of monitoring several parameters simultaneously, with new configurations of liquid handling, sample treatment and sensing. Modular microfluidics allows the integration of these newly developed approaches in a more flexible way towards increasing applicability of the microfluidic chips to different types of biocatalysts and reactions. A highly relevant operation in such a system is biocatalyst inactivation, which can enable the precise control of reaction time by avoiding the continuation of the reaction in another module or connecting tubes. Such control is important when different modules of reactors and/or sensing units are used and changed frequently. Here we describe the development, characterization and application of a module for rapid enzyme inactivation. The thermal inactivation platform developed is compared with a standard benchtop ThermoMixer in terms of inactivation efficiency for glucose oxidase and catalase. A higher activity loss was observed for enzyme inactivation under flow conditions (inactivation achieved at 120 s residence time at 338 K and 20 s residence time at 353 K) which indicated a high heat transfer to the fluid under dynamic conditions. Moreover, partial deactivation of the enzymes was observed for the continuous thermal inactivation module, when activity measurements were performed after 1 and 2 days following inactivation. The thermal inactivation unit presented can be easily integrated into modular microfluidic platforms and can be a useful addition for enzyme characterization and screening.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.04.002
       
  • Agrobacterium rhizogenes-mediated transformation of a dioecious plant
           model Silene latifolia
    • Authors: Vojtech Hudzieczek; Radim Cegan; Tomas Cermak; Nela Bacovska; Zuzana Machalkova; Karel Dolezal; Lucie Plihalova; Daniel Voytas; Roman Hobza; Boris Vyskot
      Abstract: Publication date: Available online 12 April 2018
      Source:New Biotechnology
      Author(s): Vojtech Hudzieczek, Radim Cegan, Tomas Cermak, Nela Bacovska, Zuzana Machalkova, Karel Dolezal, Lucie Plihalova, Daniel Voytas, Roman Hobza, Boris Vyskot
      Silene latifolia serves as a model species to study dioecy, the evolution of sex chromosomes, dosage compensation and sex-determination systems in plants. Currently, no protocol for genetic transformation is available for this species, mainly because S. latifolia is considered recalcitrant to in vitro regeneration and infection with Agrobacterium tumefaciens. Using cytokinins and their synthetic derivatives, we markedly improved the efficiency of regeneration. Several agrobacterial strains were tested for their ability to deliver DNA into S. latifolia tissues leading to transient and stable expression of the GUS reporter. The use of Agrobacterium rhizogenes strains resulted in the highest transformation efficiency (up to 4.7% of stable transformants) in hairy root cultures. Phenotypic and genotypic analyses of the T1 generation suggested that the majority of transformation events contain a small number of independent T-DNA insertions and the transgenes are transmitted to the progeny in a Mendelian pattern of inheritance. In short, we report an efficient and reproducible protocol for leaf disc transformation and subsequent plant regeneration in S. latifolia, based on the unique combination of infection with A. rhizogenes and plant regeneration from hairy root cultures using synthetic cytokinins. A protocol for the transient transformation of S.latifolia protoplasts was also developed and applied to demonstrate the possibility of targeted mutagenesis of the sex linked gene SlAP3 by TALENs and CRISPR/Cas9.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.04.001
       
  • Generation and validation of murine monoclonal and camelid recombinant
           single domain antibodies specific for human pancreatic glycoprotein 2
    • Authors: Anja Schlör; Pamela Holzlöhner; Martin Listek; Cindy Grieß; Monique Butze; Burkhard Micheel; Christian Hentschel; Mandy Sowa; Dirk Roggenbuck; Peter Schierack; Jonas Füner; Erik Schliebs; Alexander Goihl; Dirk Reinhold; Katja Hanack
      Abstract: Publication date: Available online 7 April 2018
      Source:New Biotechnology
      Author(s): Anja Schlör, Pamela Holzlöhner, Martin Listek, Cindy Grieß, Monique Butze, Burkhard Micheel, Christian Hentschel, Mandy Sowa, Dirk Roggenbuck, Peter Schierack, Jonas Füner, Erik Schliebs, Alexander Goihl, Dirk Reinhold, Katja Hanack
      Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified as a major autoantigenic target in Crohn’s disease patients. It was reported recently that a long (GP2a) and a short (GP2b) isoform of GP2 exist and that in the outcome of inflammatory bowel diseases (IBD) GP2-specific autoantibodies probably appear as new serological markers for diagnosis and therapeutic monitoring. To investigate this further and in order to establish diagnostic tools for the discrimination of both GP2 isoforms, a set of different murine monoclonal and camelid recombinant single domain antibodies (camelid VHH) was generated and validated in various enzyme-linked immunosorbent assay (ELISA) formats, immunofluorescence on transgenic cell lines and immunohistochemistry on monkey pancreas tissue sections. Out of six binders identified, one was validated as highly specific for GP2a. This murine monoclonal antibody (mAb) was used as capture antibody in construction of a sandwich ELISA for the detection of GP2a. Camelid VHHs or a second murine mAb served as detection antibodies in this system. All antibodies were also able to stain GP2a or GP2b on transgenic cell lines as well as on pancreatic tissue in immunohistochemistry. The KD values measured for the camelid VHHs were between 7 nM and 23pM. This set of specific binders will enable the development of suitable diagnostic tools for GP2-related studies in IBD.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.03.006
       
  • Optimization of aqueous two-phase systems for the production of
           6-aminopenicillanic acid in integrated microfluidic reactors-separators
    • Authors: Lucie Vobecká; Alexandr Romanov; Zdeněk Slouka; Pavel Hasal; Michal Přibyl
      Abstract: Publication date: Available online 31 March 2018
      Source:New Biotechnology
      Author(s): Lucie Vobecká, Alexandr Romanov, Zdeněk Slouka, Pavel Hasal, Michal Přibyl
      Aqueous two-phase systems (ATPSs) were screened for the production of 6-aminopenicillanic acid (6-APA) catalyzed by penicillin acylase, followed by the extractive separation of 6-APA from the reaction mixture. The key point of this study was to find an ATPS exhibiting a large difference in the partition coefficients of the biocatalyst and reaction products. Several ATPSs based on polyethylene glycol (PEG)/phosphate, PEG/citrate, and PEG/dextran were tested. We found that an ATPS consisting of 15 wt% of PEG 4000, 10 wt% of phosphates, 75 wt% of water (pH value 8.0 after dissolution) provided optimal separation of 6-APA from the enzyme. While the 6-APA was mainly found in the top PEG phase, the free enzyme favored the bottom salt-rich phase. This ATPS also fulfils other important requirements: (i) high buffering capacity, reducing an undesirable pH decrease due to the dissociation of phenylacetic acid (the side product of the reaction), (ii) a relatively low cost of the ATPS components, (iii) the possibility of electrophoretic transport of fine droplets as well as the reaction products for both the acceleration of phase separation and the enhancement of 6-APA concentration in the product stream. Extraction experiments in microcapillary and batch systems showed that the transport of 6-APA formed in the salt-rich phase to the corresponding PEG phase could occur within 30 s. The experimental results described form a base of knowledge for the development of continuously operating integrated microfluidic reactors-separators driven by an electric field for the efficient production of 6-APA.

      PubDate: 2018-04-25T01:54:23Z
      DOI: 10.1016/j.nbt.2018.03.005
       
  • Relationship between microbial community, operational factors and ammonia
           inhibition resilience in anaerobic digesters at low and moderate ammonia
           background concentrations
    • Authors: Y. Lu; R. Liaquat; S. Astals; P.D. Jensen; D.J. Batstone; S. Tait
      Abstract: Publication date: Available online 3 March 2018
      Source:New Biotechnology
      Author(s): Y. Lu, R. Liaquat, S. Astals, P.D. Jensen, D.J. Batstone, S. Tait
      The relationship between anaerobic digestion operational conditions and (i) microbial community, (ii) acetoclastic methanogenic activity and (iii) free ammonia (NH3) inhibition resilience was investigated. Thirteen inocula were obtained from full and pilot scale digesters fed with different substrates, digester configurations, operating temperatures and NH3 concentrations (0.1–241 mgN·L−1). Substrate type and temperature were the primary factors influencing microbial community composition. Methanogenic activity ranged from 0.04 to 0.14 gCOD-CH4·g−1VS·day−1, and was significantly correlated with archaeal relative abundance and archaeal community PC2. The variability of NH3 resilience among inocula was moderate, with inhibition threshold values (KI50) ranging between 32 and 175 mgNH3-N·L−1. No microbial or operational factors correlated with NH3 resilience. However, the slopes of inhibition threshold curves were influenced by some environmental factors, namely substrate type, digester temperature and NH3 concentration. Overall, these results indicate that low and moderate background NH3 concentrations is not a key determinant of microbial community nor NH3 resilience.

      PubDate: 2018-03-08T02:44:51Z
      DOI: 10.1016/j.nbt.2018.02.013
       
  • Potential use of methane fermentation digested slurry as a low-cost,
           environmentally-friendly nutrient for bioethanol production from crude
           glycerol by Klebsiella variicola TB-83D
    • Authors: Kohei Seta; Toshihiro Suzuki; Keiji Kiyoshi; Toshiya Shigeno; Toshiaki Nakajima-Kambe
      Abstract: Publication date: Available online 26 February 2018
      Source:New Biotechnology
      Author(s): Kohei Seta, Toshihiro Suzuki, Keiji Kiyoshi, Toshiya Shigeno, Toshiaki Nakajima-Kambe
      A methane fermentation digested slurry (MFDS) was evaluated as a substitute for the commercial nutrient, yeast extract (YE), in ethanol production from glycerol by Klebsiella variicola strain TB-83D. In pH-controlled fed-batch cultures, partial replacement of YE by MFDS did not reduce ethanol productivity significantly. However, non-sterilized MFDS had negative effects on glycerol fermentation by this strain. Although ethanol production decreased when YE was completely replaced by sterilized MFDS, the use of crude glycerol and sterilized MFDS achieved a yield of 14.6 g/L ethanol. This is the first study to report the use of MFDS as the sole nutrient for ethanol production from glycerol, which contributes to the development of a low-cost glycerol biorefinery derived from the biodiesel fuel industry.

      PubDate: 2018-03-08T02:44:51Z
      DOI: 10.1016/j.nbt.2018.02.014
       
  • Potential use of ricotta cheese whey for the production of lactobionic
           acid by Pseudomonas taetrolens strains
    • Authors: Stefania De Giorgi; Noura Raddadi; Angelo Fabbri; Tullia Gallina Toschi; Fabio Fava
      Abstract: Publication date: Available online 21 February 2018
      Source:New Biotechnology
      Author(s): Stefania De Giorgi, Noura Raddadi, Angelo Fabbri, Tullia Gallina Toschi, Fabio Fava
      Lactobionic acid (LBA) is a fine chemical largely applied in the food, chemical, cosmetics and pharmaceutical industries. Here, its production from ricotta cheese whey (RCW), or scotta, the main by-product obtained from ricotta cheese production process and currently employed mainly for cattle feed, was evaluated. Among seven bacterial species tested, only two Pseudomonas taetrolens strains were selected after preliminary screening in shake-flasks. When autoclaved RCW was used, a lactobionic acid titer of 34.25 ± 2.86 g/l, with a conversion yield (defined as mol LBA/mol of consumed lactose %) of up to 85 ± 7.0%, was obtained after 48 h of batch fermentation in 3 L stirred tank bioreactor. This study is a preliminary investigation on the potential industrial use of scotta as a substrate for bacterial growth and lactobionic acid production that details the possible biotechnological valorization pathways and feasibility of the process.

      PubDate: 2018-02-26T01:13:15Z
      DOI: 10.1016/j.nbt.2018.02.010
       
  • Deciphering the growth pattern and phytohormonal content in Saskatoon
           berry (Amelanchier alnifolia) in response to in vitro cytokinin
           application
    • Authors: Mack Moyo; Adeyemi O. Aremu; Lenka Plačková; Lucie Plíhalová; Aleš Pěnčík; Ondřej Novák; Jan Holub; Karel Doležal; Johannes Van Staden
      Abstract: Publication date: Available online 15 February 2018
      Source:New Biotechnology
      Author(s): Mack Moyo, Adeyemi O. Aremu, Lenka Plačková, Lucie Plíhalová, Aleš Pěnčík, Ondřej Novák, Jan Holub, Karel Doležal, Johannes Van Staden
      Clonal propagation plays a critical integral role in the growth and success of a global multi-billion dollar horticulture industry through a constant supply of healthy stock plants. The supply chain depends on continuously improving the micropropagation process, thus, understanding the physiology of in vitro plants remains a core component. We evaluated the influence of exogenously applied cytokinins (CKs, N 6-benzyladenine = BA, isopentenyladenine = iP, meta-topolin = mT, 6-(3-hydroxybenzylamino)-9-(tetrahydropyran-2-yl)purine = mTTHP) in Murashige and Skoog (MS)-supplemented media on organogenic response and accumulation of endogenous CK and indole-3-acetic acid (IAA) metabolites. The highest shoot proliferation (30 shoots/explant) was obtained with 20 μM mT treatment. However, the best quality regenerants were produced in 10 μM mT treatment. Rooting of Amelanchier alnifolia in vitro plantlets was observed at the lowest CK concentrations, with the highest root proliferation (3 roots/explant) in 1 μM mTTHP regenerants. Similar to the organogenic response, high levels of endogenous bioactive CK metabolites (free bases, ribosides, and nucleotides) were detected in mT and mTTHP-derived regenerants. The level of O-glucosides was also comparatively high in these cultures. All CK-treated plants had high levels of endogenous free IAA compared to the control. This may suggest an influence of CKs on biosynthesis of IAA.

      PubDate: 2018-02-16T00:16:22Z
      DOI: 10.1016/j.nbt.2018.02.001
       
  • A SEP tag enhances the expression, solubility and yield of recombinant TEV
           protease without altering its activity
    • Authors: Kalpana Nautiyal; Yutaka Kuroda
      Abstract: Publication date: Available online 12 February 2018
      Source:New Biotechnology
      Author(s): Kalpana Nautiyal, Yutaka Kuroda
      Tobacco Etch Virus (TEV) protease is used in the purification of recombinant proteins, but its use is often hampered by solubility issues. Here, we report a short, 12-residue solubility enhancing peptide (SEP) tag attached at the C-terminus of TEV (TEV-C9R). We assessed the effects of the C9R tag on the biophysical and biochemical characteristics of TEV. The yield of HPLC purified TEV-C9R expressed in E. coli grown in 200 mL LB or TB media was between 10 and 13 mg, which was up to 6.5 times higher than the yield of the untagged TEV. TEV-C9R was active over a pH range of 5–8, which was wider than that of the commonly used thrombin, and it remained active upon incubation at 60 °C much longer than the untagged TEV, which aggregated at this temperature. Static and dynamic light scattering demonstrated the higher solubility of purified TEV-C9R. Furthermore, the thermal unfolding of TEV-C9R, as assessed by circular dichroism at pH 4.7, was almost perfectly reversible, in contrast to that of untagged TEV, which aggregated at high temperature. These results demonstrate the improved biophysical and biochemical characteristics of TEV-C9R originating from higher solubility and provide another example of how SEP tags can enhance enzyme solubility without altering its activity.

      PubDate: 2018-02-16T00:16:22Z
      DOI: 10.1016/j.nbt.2018.02.006
       
  • Laccase catalyzed elimination of morphine from aqueous systems
    • Authors: Daniela Huber; Klaus Bleymaier; Alessandro Pellis; Robert Vielnascher; Andreas Daxbacher; Katrin J. Greimel; Georg M. Guebitz
      Abstract: Publication date: Available online 6 January 2018
      Source:New Biotechnology
      Author(s): Daniela Huber, Klaus Bleymaier, Alessandro Pellis, Robert Vielnascher, Andreas Daxbacher, Katrin J. Greimel, Georg M. Guebitz
      Pharmaceuticals contaminate the environment for several reasons, including metabolic excretion after intake, industrial waste and improper disposal. The narcotic drug morphine is commonly utilized for chronic pain management, and the distribution of morphine in aqueous systems and in waste waters is of high concern. Here. the removal of morphine by a laccase from Myceliophthora thermophila both in its free form as well as immobilized on Accurel MP1000 beads was investigated. Complete morphine elimination was achieved within 30 min for the free and the immobilized enzyme (70% bound protein) for concentrations between 1 and 1,000 mg L−1 according to LC-TOF mass spectrometry analysis. Higher morphine concentrations up to 60 g L−1 were also tested and total elimination was achieved within 6 h. Therefore, laccases are ideal candidates for removing morphine from aqueous systems.

      PubDate: 2018-01-09T16:46:25Z
      DOI: 10.1016/j.nbt.2018.01.003
       
  • Production of polyhydroxybutyrates and carbohydrates in a mixed
           cyanobacterial culture: effect of nutrients limitation and photoperiods
    • Authors: Dulce María Arias; Enrica Uggetti; María Jesús García-Galán; Joan García
      Abstract: Publication date: Available online 3 January 2018
      Source:New Biotechnology
      Author(s): Dulce María Arias, Enrica Uggetti, María Jesús García-Galán, Joan García
      In the present study, different photoperiods and nutritional conditions were applied to a mixed wastewater-borne cyanobacterial culture in order to enhance the intracellular accumulation of polyhydroxybutyrates (PHBs) and carbohydrates. Two different experimental set-ups were used. In the first, the culture was permanently exposed to illumination, while in the second it was submitted to light/dark alternation (12 h cycles). In both cases, two different nutritional regimes were also evaluated, N-limitation and P-limitation. Results showed that the highest PHB concentration (104 mg L−1) was achieved under P limited conditions and permanent illumination, whereas the highest carbohydrate concentration (838 mg L−1) was obtained under N limited condition and light/dark alternation. With regard to bioplastics and biofuel generation, this study demonstrates that the accumulation of PHBs (bioplastics) and carbohydrates (potential biofuel substrate) is favored in wastewater-borne cyanobacteria under conditions where nutrients are limited.
      Graphical abstract image

      PubDate: 2018-01-05T13:38:25Z
      DOI: 10.1016/j.nbt.2018.01.001
       
 
 
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