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    Harnessing a Methane-Fueled, Sediment-Free Mixed Microbial Community for Utilization of Distributed Sources of Natural Gas.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Department of Organismic & Evolutionary Biology, Harvard University, Cambridge, MA 02138, USA.
    Harnessing the metabolic potential of uncultured microbial communities is a compelling opportunity for the biotechnology industry, an approach that would vastly expand the portfolio of usable feedstocks. Methane is particularly promising because it is abundant and energy-rich, yet the most efficient methane-activating metabolic pathways involve mixed communities of anaerobic methanotrophic archaea and sulfate reducing bacteria. These communities oxidize methane at high catabolic efficiency and produce chemically reduced by-products at a comparable rate and in near-stoichiometric proportion to methane consumption. Read More

    Peptide ligand-mediated endocytosis of nanoparticles to cancer cells: cell receptor-binding- vs. cell membrane-penetrating peptides.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
    The endocytosis-mediating performances of two types of peptide ligands, cell receptor binding peptide (CRBP) and cell membrane penetrating peptide (CMPP), were analyzed and compared using a common carrier of peptide ligands-human ferritin heavy chain (hFTH) nanoparticle. 24 copies of a CMPP(human immunodeficiency virus-derived TAT peptide) and/or a CRBP (peptide ligand with strong and specific affinity for either human integrin(αβ) or epidermal growth factor receptor I (EGFR) that is overexpressed on various cancer cells) were genetically presented on the surface of each hFTH nanopariticle. The quantitative level of endocytosis and intracellular localization of fluorescence dye-labeled CRBP- and CMPP-presenting nanoparticles were estimated in the in vitro cultures of integrin- and EGFR-overexpressing cancer and human dermal fibroblast cells(control). Read More

    Fluorescent Nanodiamond-bacteriophage Conjugates Maintain Host Specificity.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas, 77843, USA.
    Rapid identification of specific bacterial strains within clinical, environmental, and food samples can facilitate the prevention and treatment of disease. Fluorescent nanodiamonds (FNDs) are being developed as biomarkers in biology and medicine, due to their excellent imaging properties, ability to accept surface modifications, and lack of toxicity. Bacteriophages, the viruses of bacteria, can have exquisite specificity for certain hosts. Read More

    Engineered CRISPR/Cas9 System for Multiplex Genome Engineering of Polyploid Industrial Yeast Strains.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
    The CRISPR/Cas9 system has been widely used for multiplex genome engineering of Saccharomyces cerevisiae. However, its application in manipulating industrial yeast strains is less successful, probably due to the genome complexity and low copy numbers of gRNA expression plasmids. Here we developed an efficient CRISPR/Cas9 system for industrial yeast strain engineering by using our previously engineered plasmids with increased copy numbers. Read More

    Enhancing cytochrome P450-mediated non-natural cyclopropanation by mutation of a conserved second-shell residue.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Department of Chemistry, University of North Carolina at Chapel Hill, 125 South Road, CB. 3290, Chapel Hill, NC 27599.
    Engineered cytochrome P450s are emerging as powerful synthetic tools due to their ability catalyze non-native metallocarbenoid and -nitrenoid insertion reactions. P450-mediated cyclopropanation has garnered particular interest due to the high selectivity demonstrated by engineered scaffolds and their application towards the synthesis of therapeutic agents. We previously reported that mutation of a conserved, first-shell heme-ligating Cys to Ser led to significant improvements in cyclopropanation activity in a model enzyme, P450. Read More

    RNAi Assisted Genome Evolution Unveils Yeast Mutants with Improved Xylose Utilization.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Department of Chemical and Biomolecular Engineering and Carl R. Woese Institute for Genomic Biology, Zhejiang University, Hangzhou, 310027, China.
    Xylose is a major component of lignocellulosic biomass, one of the most abundant feedstocks for biofuel production. Therefore, efficient and rapid conversion of xylose to ethanol is crucial in the viability of lignocellulosic biofuel plants. In this study, RNAi Assisted Genome Evolution (RAGE) was used to improve the xylose utilization rate in SR8, one of the most efficient publicly available xylose utilizing Saccharomyces cerevisiae strains. Read More

    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    The Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, Washington 99164, USA.
    Biofilms alter their metabolism in response to environmental stress. This study explores the effect of a hyperosmotic agent-antibiotic treatment on the metabolism of Staphylococcus aureus biofilms through the use of nuclear magnetic resonance (NMR) techniques. To determine the metabolic activity of S. Read More

    A perfusion chamber for monitoring transepithelial NaCl transport in an in vitro model of the renal tubule.
    Biotechnol Bioeng 2018 Feb 20. Epub 2018 Feb 20.
    Instituto Aragonés de Ciencias de la Salud, IIS Aragón, Zaragoza, Spain.
    Transepithelial electrical measurements in the renal tubule have provided a better understanding of how kidney regulates electrolyte and water homeostasis through the reabsorption of molecules and ions (e.g., HO and NaCl). Read More

    Application of adaptive laboratory evolution to overcome a flux limitation in an Escherichia coli production strain.
    Biotechnol Bioeng 2018 Feb 19. Epub 2018 Feb 19.
    Department of Bioinfomatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
    Gene deletion strategies using flux balance analysis (FBA) have improved the growth-coupled production of various compounds. However, the productivities were often below the expectation because the cells failed to adapt to these genetic perturbations. Here, we demonstrate the productivity of the succinate of the designed gene deletion strain was improved by adaptive laboratory evolution (ALE). Read More

    Rapid eradication of bacterial phytopathogens by atmospheric pressure glow discharge generated in contact with a flowing liquid cathode.
    Biotechnol Bioeng 2018 Feb 19. Epub 2018 Feb 19.
    Wroclaw University of Science and Technology, Faculty of Chemistry, Department of Analytical Chemistry and Chemical Metallurgy, Wybrzeze St. Wyspianskiego 27, 50-370 Wroclaw, Poland.
    Diseases caused by phytopathogenic bacteria are responsible for significant economic losses, and these bacteria spread through diverse pathways including waterways and industrial wastes. It is therefore of interest to develop potent methods for their eradication. Here, antibacterial properties of direct current atmospheric pressure glow discharge (dc-APGD) generated in contact with flowing bacterial suspensions were examined against five species of phytopathogens. Read More

    Combination Drug Delivery via Multilamellar Vesicles Enables Targeting of Tumor Cells and Tumor Vasculature.
    Biotechnol Bioeng 2018 Feb 19. Epub 2018 Feb 19.
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90089.
    Blood vessel development is critical for the continued growth and progression of solid tumors and, therefore, makes an attractive target for improving cancer therapy. Indeed, vascular-targeted therapies have been extensively explored but they have shown minimal efficacy as monotherapies. Combretastatin A4 (CA-4) is a tubulin-binding vascular disrupting agent that selectively targets the established tumor endothelium, causing rapid vascular beak down. Read More

    Antibody Glycoengineering Strategies in Mammalian Cells.
    Biotechnol Bioeng 2018 Feb 19. Epub 2018 Feb 19.
    Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
    As a key parameter impacting functional and structural heterogeneity, protein glycosylation is a critical quality attribute for antibody biotherapeutic manufacturing. The glycan patterns on recombinant antibodies, particularly on the conserved fragment crystallizable (Fc) region, can have significant effects on an antibody's functional activities including clearance rate, antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC) and anti-inflammatory activity. In this review, we examined specific glycan attachments (fucosylation, sialylation, galactosylation, oligo-mannose and bisecting glycans) and their importance to antibody properties. Read More

    A coupled in vitro/in vivo approach for engineering a heterologous Type III PKS to enhance polyketide biosynthesis in Saccharomyces cerevisiae.
    Biotechnol Bioeng 2018 Feb 19. Epub 2018 Feb 19.
    Howard Hughes Medical Institute, Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
    Polyketides are attractive compounds for uses ranging from biorenewable chemical precursors to high-value therapeutics. In many cases, synthesis in a heterologous host is required to produce these compounds in industrially relevant quantities. The type III polyketide synthase 2-pyrone synthase (2-PS) from Gerbera hybrida was used for the production of triacetic acid lactone (TAL) in Saccharomyces cerevisiae. Read More

    Proteomics in biomanufacturing control: Protein dynamics of CHO-K1 cells and conditioned media during apoptosis and necrosis.
    Biotechnol Bioeng 2018 Feb 10. Epub 2018 Feb 10.
    NIBRT Characterisation and Comparability Laboratory, National Institute for Bioprocessing, Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, A94 × 099, Ireland.
    Cell viability has a critical impact on product quantity and quality during the biomanufacturing of therapeutic proteins. An advanced understanding of changes in the cellular and conditioned media proteomes upon cell stress and death is therefore needed for improved bioprocess control. Here, a high pH / low pH reversed phase data independent 2D-LC-MSdiscovery proteomics platform was applied to study the cellular and conditioned media proteomes of CHO-K1 apoptosis and necrosis models where cell death was induced by staurosporine exposure or aeration shear in a benchtop bioreactor, respectively. Read More

    Butyrated ManNAc analog improves protein expression in Chinese hamster ovary cells.
    Biotechnol Bioeng 2018 Feb 10. Epub 2018 Feb 10.
    Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
    The chemical additive sodium butyrate (NaBu) has been applied in cell culture media as a direct and convenient method to increase the protein expression in Chinese hamster ovary (CHO) and other mammalian cells. In this study, we examined an alternative chemical additive, 1,3,4-O-BuManNAc, for its effect on recombinant protein production in CHO. Supplementation with 1,3,4-O-BuManNAc for two stable CHO cell lines, expressing human erythropoietin or IgG, enhanced protein expression for both products with negligible impact on cell growth, viability, glucose utilization, and lactate accumulation. Read More

    Anodic Electro-Fermentation: Anaerobic production of L-Lysine by recombinant Corynebacterium glutamicum.
    Biotechnol Bioeng 2018 Feb 10. Epub 2018 Feb 10.
    Centre for Microbial Electrochemical Systems (CEMES), The University of Queensland, Brisbane, QLD, Australia.
    Microbial electrochemical technologies (MET) are promising to drive metabolic processes for the production of chemicals of interest. They provide microorganisms with an electrode as an electron sink or an electron source to stabilize their redox and/or energy state. Here, we applied an anode as additional electron sink to enhance the anoxic metabolism of the industrial bacterium Corynebacterium glutamicum through an anodic electro-fermentation. Read More

    Differentiation of 3D-shape-controlled mouse neural stem cell to neural tissues in closed agarose microchambers.
    Biotechnol Bioeng 2018 Feb 10. Epub 2018 Feb 10.
    Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, kanagawa 223-5822, Japan.
    This paper describes three-dimensional (3D) tissue shape control of mouse neural stem cell (mNSC) micro tissues by using closed agarose microchambers for effective differentiation induction of neurons in vitro. Our agarose microchambers, made by micromolding, can be sealed with an agarose sheet to form the mNSC tissues along the shape of microchambers. We constructed lane-shaped mNSC tissues with different width (∼60-210 µm) and thickness (∼25-95 µm) dimensions and induced differentiation to neurons with differentiation medium. Read More

    Highly sensitive detection of mutations in CHO cell recombinant DNA using multi-parallel single molecule real-time DNA sequencing.
    Biotechnol Bioeng 2018 Feb 10. Epub 2018 Feb 10.
    Department of Chemical and Biological Engineering, University of Sheffield, Sheffield S1 3JD, UK.
    High-fidelity replication of biologic-encoding recombinant DNA sequences by engineered mammalian cell cultures is an essential pre-requisite for the development of stable cell lines for the production of biotherapeutics. However, immortalized mammalian cells characteristically exhibit an increased point mutation frequency compared to mammalian cells in vivo, both across their genomes and at specific loci (hotspots). Thus unforeseen mutations in recombinant DNA sequences can arise and be maintained within producer cell populations. Read More

    Purification, Characterization, and N-glycosylation of Recombinant Butyrylcholinesterase from Transgenic Rice Cell Suspension Cultures.
    Biotechnol Bioeng 2018 Feb 7. Epub 2018 Feb 7.
    Department of Chemical Engineering, University of California, Davis, CA, USA.
    Recombinant butyrylcholinesterase produced in a metabolically-regulated transgenic rice cell culture (rrBChE) was purified to produce a highly pure (95%), active form of enzyme. The developed downstream process uses common manufacturing-friendly operations including tangential flow filtration, anion-exchange chromatography, and affinity chromatography to obtain a process recovery of 42% active rrBChE. The purified rrBChE was then characterized to confirm its comparability to the native human form of the molecule (hBChE). Read More

    Split trehalase as a versatile reporter for a wide range of biological analytes.
    Biotechnol Bioeng 2018 Feb 7. Epub 2018 Feb 7.
    Faculty of Veterinary Medicine, Department of Production Animal Health, University of Calgary, Calgary, Alberta, Canada.
    In health care, biosensors are envisioned as universal diagnostic devices with AAAA characteristics (i.e., available for anything, anywhere, anytime, to anyone). Read More

    Photo-switchable Microbial Fuel-Cells.
    Biotechnol Bioeng 2018 Feb 7. Epub 2018 Feb 7.
    Department of Life Sciences and Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
    Regulation of Biosystems in a clean, simple and efficient way is important for the design of smart bio-interfaces and bioelecronic devices. Light as a non-invasive mean to control the activity of a protein enables spatial and temporal control far superior to other chemical and physical methods. The ability to regulate the activity of a catalytic enzyme in a biofuel-cell reduces the waste of resources and energy and turns the fuel-cell into a smart and more efficient device for power generation. Read More

    Improving methyl ketone production in E. coli by heterologous expression of NADH-dependent FabG.
    Biotechnol Bioeng 2018 Feb 7. Epub 2018 Feb 7.
    Joint BioEnergy Institute (JBEI), 5885 Hollis Street, Emeryville, CA 94608.
    We previously engineered E. coli to overproduce medium- to long-chain saturated and monounsaturated methyl ketones, which could potentially be applied as diesel fuel blending agents or in the flavor and fragrance industry. Recent efforts at strain optimization have focused on cofactor balance, as fatty acid-derived pathways face the systematic metabolic challenge of net NADPH consumption [in large part, resulting from the key fatty acid biosynthetic enzyme FabG (β-ketoacyl-ACP reductase)] and net NADH production. Read More

    Improving the recombinant human erythropoietin glycosylation using microsome supplementation in CHO cell-free system.
    Biotechnol Bioeng 2018 Jan 31. Epub 2018 Jan 31.
    Center for Advanced Sensor Technology (CAST) and Department of Chemical Biochemical and Environmental Engineering (CBEE), University of Maryland Baltimore County, Baltimore, Maryland.
    Cell-Free Protein Synthesis (CFPS) offers many advantages for the production of recombinant therapeutic proteins using the CHO cell-free system. However, many complex proteins are still difficult to express using this method. To investigate the current bottlenecks in cell-free glycoprotein production, we chose erythropoietin (40% glycosylated), an essential endogenous hormone which stimulates the development of red blood cells. Read More

    Multi-facet implications of PEGylated lysozyme stabilized-silver nanoclusters loaded recombinant PTEN cargo in cancer theranostics.
    Biotechnol Bioeng 2018 Jan 31. Epub 2018 Jan 31.
    Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, India.
    Amalgamation of delivery and tracking of therapeutically relevant moieties on a single platform is made possible by the application of metal nanoclusters, an innovative class of luminescent nanomaterials. Metal nanoclusters, possessing molecule-like attributes, display extraordinary size and shape tunable properties befitting theranostic applications. Herein, we report successful assembly of therapeutically significant phosphatase protein PTEN and fluorescent lysozyme-stabilized silver nanoclusters to accomplish delivery and tracking of the protein. Read More

    Application of hydrocarbon and perfluorocarbon oxygen vectors to enhance heterologous production of hyaluronic acid in engineered Bacillus subtilis.
    Biotechnol Bioeng 2018 Jan 31. Epub 2018 Jan 31.
    Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario, N2L 5B6, Canada.
    In microbial cultivations for hyaluronic acid (HA) production, oxygen can be a limiting substrate due to its poor solubility in aqueous medium and the substantial increase in culture viscosity at relatively low HA titers. Shear stress due to the high agitation and aeration rates required to overcome oxygen limitation may reduce the quality (i.e. Read More

    Glutamine synthetase gene knockout-human embryonic kidney 293E cells for stable production of monoclonal antibodies.
    Biotechnol Bioeng 2018 Jan 23. Epub 2018 Jan 23.
    Department of Biological Sciences, KAIST, Yuseong-gu, Daejeon, Republic of Korea.
    Previously, it was inferred that a high glutamine synthetase (GS) activity in human embryonic kidney (HEK) 293E cells results in elevated resistance to methionine sulfoximine (MSX) and consequently hampers GS-mediated gene amplification and selection by MSX. To overcome this MSX resistance in HEK293E cells, a GS-knockout HEK293E cell line was generated using the CRISPR/Cas9 system to target the endogenous human GS gene. The GS-knockout in the HEK293E cell line (RK8) was confirmed by Western blot analysis of GS and by observation of glutamine-dependent growth. Read More

    Correlation of simulation/finite element analysis to the separation of intrinsically magnetic spores and red blood cells using a microfluidic magnetic deposition system.
    Biotechnol Bioeng 2018 Jan 16. Epub 2018 Jan 16.
    William G. Lowrie Department of Chemical and Biomolecular Engineering Director, Analytical Cytometry Shared Resource, The OSU Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio.
    Magnetic separation of cells has been, and continues to be, widely used in a variety of applications, ranging from healthcare diagnostics to detection of food contamination. Typically, these technologies require cells labeled with antibody magnetic particle conjugate and a high magnetic energy gradient created in the flow containing the labeled cells (i.e. Read More

    Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture.
    Biotechnol Bioeng 2018 Jan 16. Epub 2018 Jan 16.
    Division of Bioengineering, Incheon National University, Incheon, Yeonsu-gu, Republic of Korea.
    Animal cell culture technology for therapeutic protein production has shown significant improvement over the last few decades. Chinese hamster ovary (CHO) cells have been widely adapted for the production of biopharmaceutical drugs. In the biopharmaceutical industry, it is crucial to develop cell culture media and culturing conditions to achieve the highest productivity and quality. Read More

    Effective role of medium supplementation in microalgal lipid accumulation.
    Biotechnol Bioeng 2018 Jan 16. Epub 2018 Jan 16.
    Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
    The present study investigated the interaction between starch and lipid accumulation in a green microalgae enrichment culture. The objective was to optimize the lipid content by manipulation of the medium in regular batch culture. Two medium designs were evaluated: First a high ortho-P concentration with vitamin supplementary (Pi-vitamins supplemented medium), second normal growth medium (control). Read More

    Role of respiratory terminal oxidases in the extracellular electron transfer ability of cyanobacteria.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Nano Electrochemistry Laboratory, College of Engineering, University of Georgia, Athens, Georgia.
    Cyanobacteria are used as anode catalysts in photo-bioelectrochemical cells to generate electricity in a sustainable, economic, and environmental friendly manner using only water and sunlight. Though cyanobacteria (CB) possess unique advantage for solar energy conversion by virtue of its robust photosynthesis, they cannot efficiently perform extracellular electron transfer (EET). The reasons being, unlike dissimilatory metal reducing bacteria (that are usually exploited in microbial fuel cells to generate electricity), (1) CB do not possess any special features on their outer membrane to carry out EET and, (2) the electrons generated in photosynthetic electron transport chain are channeled into competing respiratory pathways rather than to the anode. Read More

    Isotype dependent on-column non-reversible aggregation of monoclonal antibodies.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Biopharm Process Development, GlaxoSmithKline, GSK Medicine Research Centre, Stevenage, United Kingdom.
    Monoclonal antibodies of the IgG2 and IgG4 isotype were found to exhibit an increased propensity for displaying two-peak elution profiles during cation exchange chromatography. In some cases, this two-peak elution profile also resulted in the formation of non-reversible mAb aggregates. Comparison of IgG1, IgG2, and IgG4 molecules with the same variable region reveals that the two-peak behaviour is predominantly mediated by the constant region and most likely the lower CH1, hinge and upper CH2 regions of the mAb. Read More

    Using extensional flow to reveal diverse aggregation landscapes for three IgG1 molecules.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, West Yorkshire, UK.
    Monoclonal antibodies (mAbs) currently dominate the biopharmaceutical sector due to their potency and efficacy against a range of disease targets. These proteinaceous therapeutics are, however, susceptible to unfolding, mis-folding, and aggregation by environmental perturbations. Aggregation thus poses an enormous challenge to biopharmaceutical development, production, formulation, and storage. Read More

    Ultra scale-down approaches to study the centrifugal harvest for viral vaccine production.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Dept. of Biochemical Engineering, Bernard Katz building, University College London, Gordon Street, London, WC1H 0AH.
    Large scale continuous cell-line cultures promise greater reproducibility and efficacy for the production of influenza vaccines, and adenovirus for gene therapy. This paper seeks to use an existing validated ultra scale-down tool, which is designed to mimic the commercial scale process environment using only millilitres of material, to provide some initial insight into the performance of the harvest step for these processes. The performance of industrial scale centrifugation and subsequent downstream process units is significantly affected by shear. Read More

    Development of a temperature-responsive yeast cell factory using engineered Gal4 as a protein switch.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, P.R. China.
    Conflict between cell growth and product accumulation is frequently encountered in biosynthesis of secondary metabolites. Herein, a temperature-dependent dynamic control strategy was developed by modifying the GAL regulation system to facilitate two-stage fermentation in yeast. A temperature-sensitive Gal4 mutant Gal4M9 was created by directed evolution, and used as a protein switch in ΔGAL80 yeast. Read More

    Enrichment of high affinity subclasses and glycoforms from serum-derived IgG using FcγRs as affinity ligands.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Thayer School of Engineering, Dartmouth College, Hanover, New Hampshire.
    As antibodies continue to gain predominance in drug discovery and development pipelines, efforts to control and optimize their activity in vivo have matured to incorporate sophisticated abilities to manipulate engagement of specific Fc binding partners. Such efforts to promote diverse functional outcomes include modulating IgG-Fc affinity for FcγRs to alternatively potentiate or reduce effector functions, such as antibody-dependent cellular cytotoxicity and phagocytosis. While a number of natural and engineered Fc features capable of eliciting variable effector functions have been demonstrated in vitro and in vivo, elucidation of these important functional relationships has taken significant effort through use of diverse genetic, cellular and enzymatic techniques. Read More

    Consolidated bioprocessing of lignocellulosic biomass to lactic acid by a synthetic fungal-bacterial consortium.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Laboratory of Biofuels and Biochemicals, School of Agricultural, Forest and Food Sciences, Bern University of Applied Sciences (BFH), Zollikofen, Switzerland.
    Consolidated bioprocessing (CBP) of lignocellulosic feedstocks to platform chemicals requires complex metabolic processes, which are commonly executed by single genetically engineered microorganisms. Alternatively, synthetic consortia can be employed to compartmentalize the required metabolic functions among different specialized microorganisms as demonstrated in this work for the direct production of lactic acid from lignocellulosic biomass. We composed an artificial cross-kingdom consortium and co-cultivated the aerobic fungus Trichoderma reesei for the secretion of cellulolytic enzymes with facultative anaerobic lactic acid bacteria. Read More

    Selective production of decanoic acid from iterative reversal of β-oxidation pathway.
    Biotechnol Bioeng 2018 Jan 8. Epub 2018 Jan 8.
    Department of Chemical and Biomolecular Engineering, Rice University, Houston, Texas.
    Decanoic acid is a valuable compound used as precursor for industrial chemicals, pharmaceuticals, and biofuels. Despite efforts to produce it from renewables, only limited achievements have been reported. Here, we report an engineered cell factory able to produce decanoic acid as a major product from glycerol, and abundant and renewable feedstock. Read More

    Modulation of saturation and chain length of fatty acids in Saccharomyces cerevisiae for production of cocoa butter-like lipids.
    Biotechnol Bioeng 2018 Jan 4. Epub 2018 Jan 4.
    Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
    Chain length and degree of saturation plays an important role for the characteristics of various products derived from fatty acids, such as fuels, cosmetics, and food additives. The seeds of Theobroma cacao are the source of cocoa butter, a natural lipid of high interest for the food and cosmetics industry. Cocoa butter is rich in saturated fatty acids that are stored in the form of triacylglycerides (TAGs). Read More

    From lab to market: An integrated bioprocess design approach for new-to-nature biosurfactants produced by Starmerella bombicola.
    Biotechnol Bioeng 2017 Dec 30. Epub 2017 Dec 30.
    Centre for Industrial Biotechnology and Biocatalysis (, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
    Glycolipid microbial biosurfactants, such as sophorolipids (SLs), generate high industrial interest as 100% biobased alternatives for traditional surfactants. A well-known success story is the efficient SL producer Starmerella bombicola, which reaches titers well above 200 g/L. Recent engineering attempts have enabled the production of completely new types of molecules by S. Read More

    A robust protocol for directed aryl sulfotransferase evolution toward the carbohydrate building block GlcNAc.
    Biotechnol Bioeng 2017 Dec 30. Epub 2017 Dec 30.
    DWI-Leibniz-Institut für Interaktive Materialien e.V., Aachen, Germany.
    Bacterial aryl sulfotransferases (AST) utilize p-nitrophenylsulfate (pNPS) as a phenolic donor to sulfurylate typically a phenolic acceptor. Interest in aryl sulfotransferases is growing because of their broad variety of acceptors and cost-effective sulfuryl-donors. For instance, aryl sulfotransferase A (ASTA) from Desulfitobacterium hafniense was recently reported to sulfurylate d-glucose. Read More

    Gold nanorod-photosensitizer conjugates with glutathione-sensitive linkages for synergistic cancer photodynamic/photothermal therapy.
    Biotechnol Bioeng 2017 Dec 30. Epub 2017 Dec 30.
    Graduate School of Energy Science and Technology, Chungnam National University, Daejeon, Korea.
    Recently, photodynamic therapy (PDT) has been intensively investigated as a useful modality for the treatment of various cancers. In addition, near infrared (NIR) photothermal therapy (PTT) using gold nanocarriers has attracted particular interest as a hyperthermia strategy. In this study, gold nanorod (AuNR)-photosensitizer conjugates with glutathione-sensitive linkages were designed for PDT and PTT. Read More

    High titer oncolytic measles virus production process by integration of dielectric spectroscopy as online monitoring system.
    Biotechnol Bioeng 2017 Dec 30. Epub 2017 Dec 30.
    Institute of Bioprocess Engineering Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Giessen, Germany.
    Oncolytic viruses offer new hope to millions of patients with incurable cancer. One promising class of oncolytic viruses is Measles virus, but its broad administration to cancer patients is currently hampered by the inability to produce the large amounts of virus needed for treatment (10-10virus particles per dose). Measles virus is unstable, leading to very low virus titers during production. Read More

    Modeling and optimization of lipid accumulation by Yarrowia lipolytica from glucose under nitrogen depletion conditions.
    Biotechnol Bioeng 2017 Dec 30. Epub 2017 Dec 30.
    LISBP, Université de Toulouse, CNRS, INRA, INSA, Toulouse, France.
    Oleaginous yeasts have been seen as a feasible alternative to produce the precursors of biodiesel due to their capacity to accumulate lipids as triacylglycerol having profiles with high content of unsaturated fatty acids. The yeast Yarrowia lipolytica is a promising microorganism that can produce lipids under nitrogen depletion conditions and excess of the carbon source. However, under these conditions, this yeast also produces citric acid (overflow metabolism) decreasing lipid productivity. Read More

    Precision-porous templated scaffolds of varying pore size drive dendritic cell activation.
    Biotechnol Bioeng 2017 Dec 27. Epub 2017 Dec 27.
    Department of Bioengineering, University of Washington, Seattle, WA.
    Scaffold based systems have shown significant potential in modulating immune responses in vivo. While there has been much attention on macrophage interactions with tissue engineered scaffolds for tissue regeneration, fewer studies have looked at the effects of scaffold design on the response of immune cells-that is, dendritic cells (DCs). Here, we present the effects of varying pore size of poly (2-hydroxyethyl methacrylate) (pHEMA) and poly(dimethylsiloxane) (PDMS, silicone) scaffolds on the maturation and in vivo enrichment of DCs. Read More

    Methanol independent induction in Pichia pastoris by simple derepressed overexpression of single transcription factors.
    Biotechnol Bioeng 2017 Dec 27. Epub 2017 Dec 27.
    Institute of Molecular Biotechnology, NAWI Graz, Graz University of Technology, Graz, Austria.
    Carbon source regulated promoters are well-studied standard tools for controlling gene expression. Acquiring control over the natural regulation of promoters is important for metabolic engineering and synthetic biology applications. In the commonly used protein production host Komagataella phaffii (Pichia pastoris), methanol-inducible promoters are used because of their tight regulation and exceptional strength. Read More

    Identification of process conditions influencing protein aggregation in Chinese hamster ovary cell culture.
    Biotechnol Bioeng 2017 Dec 27. Epub 2017 Dec 27.
    Institute of Applied Biotechnology, Biberach University of Applied Sciences, Biberach, Germany.
    Protein aggregation of monoclonal antibodies (mAbs) is a common phenomenon associated with the production of these biopharmaceuticals. These aggregates can lead to adverse side effects in patients upon administration, thus expensive downstream processing steps to remove the higher molecular weight species are inevitable. A preferable approach is to reduce the level of aggregation during bioprocessing by a careful adjustment of critical process parameters. Read More

    Protein-engineering of chitosanase from Bacillus sp. MN to alter its substrate specificity.
    Biotechnol Bioeng 2017 Dec 27. Epub 2017 Dec 27.
    Institute for Biology and Biotechnology of Plants, University of Münster, Münster, Germany.
    Partially acetylated chitosan oligosaccharides (paCOS) have various potential applications in agriculture, biomedicine, and pharmaceutics due to their suitable bioactivities. One method to produce paCOS is partial chemical hydrolysis of chitosan polymers, but that leads to poorly defined mixtures of oligosaccharides. However, the effective production of defined paCOS is crucial for fundamental research and for developing applications. Read More

    Toward industrial production of isoprenoids in Escherichia coli: Lessons learned from CRISPR-Cas9 based optimization of a chromosomally integrated mevalonate pathway.
    Biotechnol Bioeng 2017 Dec 26. Epub 2017 Dec 26.
    Joint BioEnergy Institute (JBEI), Emeryville, California.
    Escherichia coli has been the organism of choice for the production of different chemicals by engineering native and heterologous pathways. In the present study, we simultaneously address some of the main issues associated with E. coli as an industrial platform for isoprenoids, including an inability to grow on sucrose, a lack of endogenous control over toxic mevalonate (MVA) pathway intermediates, and the limited pathway engineering into the chromosome. Read More

    Central pathway engineering for enhanced succinate biosynthesis from acetate in Escherichia coli.
    Biotechnol Bioeng 2017 Dec 26. Epub 2017 Dec 26.
    State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
    Acetate, a non-food based substrate obtained from multiple biological and chemical ways, is now being paid great attention in bio-manufacturing and have a strong potential to compete with sugar-based carbon source. In this study, acetate can be efficiently converted to succinate by engineered Escherichia coli strains via the combination of several metabolic engineering strategies, including reducing OAA decarboxylation, engineering TCA cycle, enhancement of acetate assimilation pathway and increasing aerobic ATP supply through cofactor engineering. The engineered strain HB03(pTrc99a-gltA, pBAD33-Trc-fdh) accumulated 30. Read More

    Manipulation of the sodium-potassium ratio as a lever for controlling cell growth and improving cell specific productivity in perfusion CHO cell cultures.
    Biotechnol Bioeng 2017 Dec 26. Epub 2017 Dec 26.
    Process Science, Boehringer Ingelheim, Fremont, California.
    Perfusion processes typically require removal of a continuous or semi-continuous volume of cell culture in order to maintain a desired target cell density. For fast growing cell lines, the product loss from this stream can be upwards of 35%, significantly reducing the overall process yield. As volume removed is directly proportional to cell growth, the ability to modulate growth during perfusion cell culture production thus becomes crucial. Read More

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