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    Cover Image, Volume 115, Number 1, January 2018.
    Biotechnol Bioeng 2018 Jan;115(1)
    Department of Chemical Engineering, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts.
    Cover Legend: The cover image, by Catherine B. Matthews et al., is based on the Article Development of a general defined medium for Pichia pastoris, DOI 10. Read More

    Modulation of IgG1 Immunoeffector Function by Glycoengineering of the GDP-Fucose Biosynthesis Pathway.
    Biotechnol Bioeng 2017 Nov 18. Epub 2017 Nov 18.
    Bioprocess Research and Development, Eli Lilly and Company, LTC-North, 1200 Kentucky Avenue, Indianapolis, IN 46225, United States.
    Cross-linking of the Fcγ receptors expressed on the surface of hematopoietic cells by IgG immune complexes triggers the activation of key immune effector mechanisms, including antibody-dependent cell mediated cytotoxicity (ADCC). A conserved N-glycan positioned at the N-terminal region of the IgG CH 2 domain is critical in maintaining the quaternary structure of the molecule for Fcγ receptor engagement. The removal of a single core fucose residue from the N-glycan results in a considerable increase in affinity for FcγRIIIa leading to an enhanced receptor-mediated immunoeffector function. Read More

    cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV-1 envelope glycoprotein vaccine candidate.
    Biotechnol Bioeng 2017 Nov 18. Epub 2017 Nov 18.
    Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10021, USA.
    We describe the properties of BG505 SOSIP.664 HIV-1 envelope glycoprotein trimers produced under current Good Manufacturing Practice (cGMP) conditions. These proteins are the first of a new generation of native-like trimers that are the basis for many structure-guided immunogen development programs aimed at devising how to induce broadly neutralizing antibodies (bNAbs) to HIV-1 by vaccination. Read More

    Design of a Novel Continuous Flow Reactor for Low pH Viral Inactivation.
    Biotechnol Bioeng 2017 Nov 18. Epub 2017 Nov 18.
    Keck Graduate Institute, Amgen Bioprocessing Center, Claremont, CA, 91711.
    Insufficient mixing in laminar flow reactors due to diffusion-dominated flow limits their use in applications where narrow residence time distribution (RTD) is required. The aim of this study was to design and characterize a laminar flow (Re 187.7-375. Read More

    Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis.
    Biotechnol Bioeng 2017 Nov 18. Epub 2017 Nov 18.
    Department of Life Science, Hanyang University, Seoul, South Korea.
    Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. Read More

    Production of 1,2-propanediol in photoautotrophic Synechocystis is linked to glycogen turn-over.
    Biotechnol Bioeng 2017 Nov 16. Epub 2017 Nov 16.
    Department Solar Materials, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
    We utilized a photoautotrophic organism to synthesize 1,2-propanediol from carbon dioxide and water fueled by light. A synthetic pathway comprising mgsA (methylglyoxal synthase), yqhD (aldehyde reductase), and adh (alcohol dehydrogenase) was inserted into Synechocystis sp. PCC6803 to convert dihydroxyacetone phosphate to methylglyoxal, which is subsequently reduced to acetol and then to 1,2-propanediol. Read More

    A fast and simple method to estimate relative, hyphal tensile-strength of filamentous fungi used to assess the effect of autophagy.
    Biotechnol Bioeng 2017 Nov 14. Epub 2017 Nov 14.
    Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County (UMBC), 1000 Hilltop Circle, Baltimore, MD 21250.
    Fungal hyphal strength is an important phenotype which can have a profound impact on bioprocess behavior. Until now, there is not an efficient method which allows its characterization. Currently available methods are very time consuming; thus, compromising their applicability in strain selection and process development. Read More

    Ty1-fused Protein-body Formation for Spatial Organization of Metabolic Pathways in Saccharomyces cerevisiae.
    Biotechnol Bioeng 2017 Nov 13. Epub 2017 Nov 13.
    Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Cheongju, 28116, Republic of Korea.
    Metabolite production through a multistep metabolic pathway can often be increased by efficient substrate channeling created by spatial sequestration of the metabolic reactions. Here, Tya, a structural component in the Ty1 retrotransposon element that forms virus-like particles (VLPs) in Saccharomyces cerevisiae, was used to spatially organize enzymes involved in a metabolic pathway into a multi-enzyme protein body in yeast. As a proof of principle, Tya fusion to three key enzymes involved in biosynthesis of the isoprenoids farnesene and farnesol was tested to assess its potential to improve productivity. Read More

    Electron shuttling to ferrihydrite selects for fermentative rather than Fe(3+) -reducing biomass in xylose-fed batch reactors derived from three different inoculum sources.
    Biotechnol Bioeng 2017 Nov 13. Epub 2017 Nov 13.
    Department of Environmental Engineering and Earth Sciences, Clemson University, 105 Collings St. Clemson, SC, USA.
    Reports suggest that ferric iron and electron shuttling molecules will select for Fe(3+) -reducer dominated microbial biomass. We investigated the influence of the redox mediators anthraquinone-2,6-disulfonate (AQDS) and riboflavin using xylose as the sole fermentation substrate, with or without ferric iron. Electron shuttling to insoluble ferrihydrite enhanced solventogenesis, acidogenesis, hydrogen production, and xylose consumption, relative to the cells plus xylose controls in fermentations inoculated with woodland marsh sediment, wetwood disease, or raw septic liquid, over multiple transfers in 15-day batch fermentations. Read More

    Glycosyltransferase Cascades Made Fit for Chemical Production: Integrated Biocatalytic Process for the Natural Polyphenol C-Glucoside Nothofagin.
    Biotechnol Bioeng 2017 Nov 13. Epub 2017 Nov 13.
    Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria.
    Glycosyltransferase cascades are promising tools of biocatalysis for natural product glycosylation, but their suitability for actual production remains to be shown. Here, we demonstrate at a scale of 100 g isolated product the integrated biocatalytic production of nothofagin, the natural 3'-C-β-D-glucoside of the polyphenol phloretin. A parallel reaction cascade involving coupled C-glucosyltransferase and sucrose synthase was optimized for the one-pot glucosylation of phloretin from sucrose via an UDP/UDP-glucose shuttle. Read More

    Highly active spore biocatalyst by self-assembly of co-expressed anchoring scaffoldin and multimeric enzyme.
    Biotechnol Bioeng 2017 Nov 13. Epub 2017 Nov 13.
    Department of Chemical and Environmental Engineering, University of California, Riverside, California.
    We report a spore-based biocatalysis platform capable of producing and self-assembling active multimeric enzymes on a spore surface with a high loading density. This was achieved by co-expressing both a spore surface-anchoring scaffoldin protein containing multiple cohesin domains and a dockerin-tagged enzyme of interest in the mother cell compartment during Bacillus subtilis sporulation. Using this method, tetrameric β-galactosidase was successfully displayed on the spore surface with a loading density of 1. Read More

    Genome-wide identification of tolerance mechanisms towards p-coumaric acid in Pseudomonas putida.
    Biotechnol Bioeng 2017 Nov 13. Epub 2017 Nov 13.
    Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800 Kgs. Lyngby, Denmark.
    The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 towards eleven toxic chemical compounds was investigated. Read More

    Preferential capture of EpCAM-expressing extracellular vesicles on solid surfaces coated with an aptamer-conjugated zwitterionic polymer.
    Biotechnol Bioeng 2017 Nov 6. Epub 2017 Nov 6.
    Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Koto, Tokyo, Japan.
    Extracellular vesicles (EVs) collectively represent small vesicles that are secreted from cells and carry biomolecules (e.g., miRNA, lncRNA, mRNA, proteins, lipids, metabolites, etc. Read More

    Engineering Escherichia coli for malate production by integrating modular pathway characterization with CRISPRi-guided multiplexed metabolic tuning.
    Biotechnol Bioeng 2017 Nov 4. Epub 2017 Nov 4.
    State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China.
    The application of rational design in reallocating metabolic flux to overproduce desired chemicals is always restricted by the native regulatory network. Here, we demonstrated that in vitro modular pathway optimization combined with in vivo multiplexed combinatorial engineering enables effective characterization of the bottleneck of a complex biosynthetic cascade and improves the output of the engineered pathway. As a proof of concept, we systematically identified the rate-limiting step of a five-gene malate biosynthetic pathway by combinatorially tuning the enzyme loads of a reconstituted biocatalytic reaction in a cell-free system. Read More

    Metabolic profiles analysis of 1,3-propanediol production process by Clostridium butyricum through repeated batch fermentation coupled with activated carbon adsorption.
    Biotechnol Bioeng 2017 Nov 4. Epub 2017 Nov 4.
    Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
    1,3-propanediol production by Clostridium butyricum is a low productivity process due to the long time seed cultivation and thus hinders its industrial scale production. In the present study, repeated batch fermentation coupled with activated carbon adsorption strategy was first established which conduced not only to saving the time of seed cultivation and enhancing the productivity, but also to reducing the costs for the seed cultivation to achieve the purpose of 1,3-propanediol continuous production. The concentration of 1,3-propanediol from first to fourth cycle was 42. Read More

    Expression of tabersonine 16-hydroxylase and 16-hydroxytabersonine-O-methyltransferase in Catharanthus roseus hairy roots.
    Biotechnol Bioeng 2017 Nov 4. Epub 2017 Nov 4.
    Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado.
    The monoterpene indole alkaloids vindoline and catharanthine, which are exclusively synthesized in the medicinal plant Catharanthus roseus, are the two important precursors for the production of pharmaceutically important anti-cancer medicines vinblastine and vincristine. Hairy root culture is an ideal platform for alkaloids production due to its industrial scalability, genetic and chemical stability, and availability of genetic engineering tools. However, C. Read More

    Metabolic Phenotyping of CHO Cells Varying in Cellular Biomass Accumulation and Maintenance during Fed-Batch Culture.
    Biotechnol Bioeng 2017 Oct 28. Epub 2017 Oct 28.
    Department of Chemical and Biological Engineering, University of Sheffield, Mappin St., Sheffield, S1 3JD, UK.
    CHO cell lines capable of high-level recombinant protein product biosynthesis during fed-batch culture are still generally obtained by intensive empirical screening of transfected cells rather than knowledge-guided cellular engineering. In this study, we investigate how CHO cell lines create and maintain cellular biosynthetic capacity during fed-batch culture to achieve the optimal combination of rapid exponential proliferation and extended maintenance of high cell biomass concentration. We perform a comparative meta-analysis of mitochondrial and glycolytic functions of 22 discrete parental CHO cell lineages varying in fed-batch culture performance to test the hypotheses that (i) "biomass-intensive" CHO cells exhibit conserved differences in metabolic programming and (ii) it is possible to isolate parental CHO cell lines with a biomass-intensive phenotype to support fed-batch bioproduction processes. Read More

    Dynamic modeling of green algae cultivation in a photobioreactor for sustainable biodiesel production.
    Biotechnol Bioeng 2017 Oct 28. Epub 2017 Oct 28.
    Marine Bioengineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
    Biodiesel produced from microalgae has been extensively studied due to its potentially outstanding advantages over traditional transportation fuels. In order to facilitate its industrialization and improve the process profitability, it is vital to construct highly accurate models capable of predicting the complex behavior of the investigated biosystem for process optimization and control, which forms the current research goal. Three original contributions are described in this paper. Read More

    Identification of key residues modulating the stereoselectivity of nitrile hydratase toward rac-mandelonitrile by semi-rational engineering.
    Biotechnol Bioeng 2017 Oct 28. Epub 2017 Oct 28.
    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
    Optically pure compounds are important in the synthesis of fine chemicals. Using directed evolution of enzymes to obtain biocatalysts that can selectively produce high-value chiral chemicals is often time-, money-, and resource-intensive; traditional semi-rational designs based on structural data and docking experiments are still limited due to the lack of accurate selection of hot-spot residues. In this study, through ligand-protein collision counts based on steered molecular dynamics simulation, we accurately identified four residues related to improving nitrile hydratase stereoselectivity toward rac-mandelonitrile (MAN). Read More

    A "plug-n-play" modular metabolic system for the production of apocarotenoids.
    Biotechnol Bioeng 2018 Jan;115(1):174-183
    Biotransformation Innovation Platform (BioTrans), Agency for Science, Technology, and Research (A*STAR), Singapore, Singapore.
    Apocarotenoids, such as α-, β-ionone, and retinol, have high commercial values in the food and cosmetic industries. The demand for natural ingredients has been increasing dramatically in recent years. However, attempts to overproduce β-ionone in microorganisms have been limited by the complexity of the biosynthetic pathway. Read More

    Cover Image, Volume 114, Number 12, December 2017.
    Biotechnol Bioeng 2017 Dec;114(12)
    Department of Chemical Engineering, University of Massachusetts Lowell, One University Avenue, Lowell, Massachusetts.
    The cover image, by Zhuangrong Huang et al., is based on the Review Quantitative intracellular flux modeling and applications in biotherapeutic development and production using CHO cell cultures, DOI: 10.1002/bit. Read More

    Effects of scaffold microstructure and low intensity pulsed ultrasound on chondrogenic differentiation of human mesenchymal stem cells.
    Biotechnol Bioeng 2017 Oct 24. Epub 2017 Oct 24.
    Department of Mechanical and Aerospace Engineering, The George Washington University, Washington DC.
    The effects of low intensity pulsed ultrasound (LIPUS) on proliferation and chondrogenic differentiation of human mesenchymal stem cells (hMSCs) seeded on 3D printed poly-(ethylene glycol)-diacrylate (PEG-DA) scaffolds with varying pore geometries (square and hexagonal channels) were investigated. The scaffold with square pores resulted in higher hMSC growth and chondrogenic differentiation than a solid or a hexagonally porous scaffold. The optimal LIPUS parameters at 1. Read More

    Effect of culture density on biomass production and light utilization efficiency of Synechocystis sp. PCC 6803.
    Biotechnol Bioeng 2017 Oct 24. Epub 2017 Oct 24.
    Schoolof Sustainable Engineering and the Built Environment, Arizona State University, Tempe, Arizona.
    The viability of large-scale microalgae cultivation depends on providing optimal growth conditions, for which a key operational parameter is culture density. Using Synechocystis sp. PCC 6803, we conducted a series of fixed-density, steady-state experiments and one batch-growth experiment to investigate the role of culture density on biomass production and light utilization efficiency. Read More

    Directed evolution of polypropylene and polystyrene binding peptides.
    Biotechnol Bioeng 2017 Oct 24. Epub 2017 Oct 24.
    RWTH Aachen University, Worringerweg 3, Aachen, Germany.
    Surface functionalization of biological inert polymers (e.g., polypropylene PP; polystyrene PS) with material binding peptides facilitates an efficient immobilization of enzymes, bioactive peptides or antigens at ambient temperature in water. Read More

    Comparison of purification strategies for antibodies used in a broad spectrum host cell protein immunoassay.
    Biotechnol Bioeng 2017 Oct 24. Epub 2017 Oct 24.
    Analytical Research and Development, Biotherapeutics Pharmaceutical Sciences, Pfizer Inc., Chesterfield, Missouri.
    Host cell proteins (HCPs) are a heterogeneous mixture of impurities that should be minimized in bulk preparations of biotechnologically produced medicines. Immunoassays are commonly used to detect and measure HCPs in therapeutic products, and a successful assay is directly dependent on the quality of the polyclonal antibodies (pAbs) used. These pAbs are enriched from antisera of animals immunized with a broad mixture of HCPs, but there is limited information regarding the best strategy for purification of these critical reagents. Read More

    Comparative genomics and transcriptomics analysis-guided metabolic engineering of Propionibacterium acidipropionici for improved propionic acid production.
    Biotechnol Bioeng 2017 Oct 24. Epub 2017 Oct 24.
    Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.
    Acid stress induced by the accumulation of organic acids during the fermentation of propionibacteria is a severe limitation in the microbial production of propionic acid (PA). To enhance the acid resistance of strains, the tolerance mechanisms of cells must first be understood. In this study, comparative genomic and transcriptomic analyses were conducted on wild-type and acid-tolerant Propionibacterium acidipropionici to reveal the microbial response of cells to acid stress during fermentation. Read More

    Enhanced pyruvate production in Candida glabrata by carrier engineering.
    Biotechnol Bioeng 2017 Oct 16. Epub 2017 Oct 16.
    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
    Pyruvate is an important organic acid that plays a key role in the central metabolic pathway. Manipulating transporters is an efficient strategy to enhance production of target organic acids and a means to understand the effects of altered intracellular pyruvate content on global metabolic networks. Efforts have been made to manipulate mitochondrial pyruvate carrier (MPC) to transport pyruvate into different subcellular compartments in Candida glabrata to demonstrate the effects of the subcellular distribution of pyruvate on central carbon metabolism. Read More

    Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy.
    Biotechnol Bioeng 2017 Oct 14. Epub 2017 Oct 14.
    Genome Science & Technology Program, Vancouver, Canada.
    Cell death is the ultimate cause of productivity loss in bioreactors that are used to produce therapeutic proteins. We investigated the ability of Raman spectroscopy to detect the onset and types of cell death for Chinese Hamster Ovary (CHO) cells-the most widely used cell type for therapeutic protein production. Raman spectroscopy was used to compare apoptotic, necrotic, autophagic, and control CHO cells. Read More

    Metabolic reduction of resazurin; location within the cell for cytotoxicity assays.
    Biotechnol Bioeng 2017 Oct 14. Epub 2017 Oct 14.
    Nanyang Environment & Water Research Institute, Advanced Environmental Biotechnology Centre, Nanyang Technological University, Singapore.
    Resazurin is widely used as a metabolic indicator for living cells, however, there has been considerable debate in the literature with regards to the specific location in the cell where the non-fluorescent resazurin is reduced to the strongly fluorescent resorufin. This lack of clarity about the reduction site makes the use of resazurin reduction data in cytotoxicity studies difficult to interpret. In this study, E. Read More

    Overproduction of MCL-PHA with high 3-hydroxydecanoate Content.
    Biotechnol Bioeng 2017 Oct 14. Epub 2017 Oct 14.
    Chemical Engineering, Queen's University, Kingston, Ontario, Canada.
    Methods of producing medium-chain-length poly-3-hydroxyalkanoate (mcl-PHA) with high content of the dominant subunit, 3-hydroxydecanoate (HD), were examined with an emphasis on a high yield of polymer from decanoic acid. High HD content was achieved by using a β-oxidation knockout mutant of Pseudomonas putida KT2440 (designated as P. putida DBA-F1) or by inhibiting β-oxidation with addition of acrylic acid (Aa) to wild type P. Read More

    Impact of carbon monoxide partial pressures on methanogenesis and medium chain fatty acids production during ethanol fermentation.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    Swette Center for Environmental Biotechnology, The Biodesign Institute, Arizona State University, Tempe, Arizona.
    Medium-chain fatty acids (MCFA) are important biofuel precursors. Carbon monoxide (CO) is a sustainable electron and carbon donor for fatty acid elongation, since it is metabolized to MCFA precursors, it is toxic to most methanogens, and it is a waste product generated in the gasification of waste biomass. The main objective of this work was to determine if the inhibition of methanogenesis through the continuous addition of CO would lead to increased acetate or MCFA production during fermentation of ethanol. Read More

    One-step affinity capture and precipitation for improved purification of an industrial monoclonal antibody using Z-ELP functionalized nanocages.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware.
    Protein A chromatography has been identified as a potential bottleneck in the monoclonal antibody production platform, leading to increased interest in non-chromatographic capture technologies. Affinity precipitation using environmentally responsive, Z-domain-elastin-like polypeptide (Z-ELP) fusion proteins has been shown to be a promising alternative. However, elevated temperature and salt concentrations necessary for precipitation resulted in decreased antibody monomer content and reduced purification capacity. Read More

    A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β-carotene.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    BIMLip, Biologie Intégrative du Métabolisme Lipidique Team, Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France.
    The increasing market demands of β-carotene as colorant, antioxidant and vitamin precursor, requires novel biotechnological production platforms. Yarrowia lipolytica, is an industrial organism unable to naturally synthesize carotenoids but with the ability to produce high amounts of the precursor Acetyl-CoA. We first found that a lipid overproducer strain was capable of producing more β-carotene than a wild type after expressing the heterologous pathway. Read More

    Biocatalytic conversion of cycloalkanes to lactones using an in-vivo cascade in Pseudomonas taiwanensis VLB120.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    Department of Solar Materials, Helmholtz-Centre for Environmental Research-UFZ GmbH, Leipzig, Germany.
    Chemical synthesis of lactones from cycloalkanes is a multi-step process challenged by limitations in reaction efficiency (conversion and yield), atom economy (by-products) and environmental performance. A heterologous pathway comprising novel enzymes with compatible kinetics was designed in Pseudomonas taiwanensis VLB120 enabling in-vivo cascade for synthesizing lactones from cycloalkanes. The respective pathway included cytochrome P450 monooxygenase (CHX), cyclohexanol dehydrogenase (CDH), and cyclohexanone monooxygenase (CHXON) from Acidovorax sp. Read More

    Enzymatic synthesis of chiral amino-alcohols by coupling transketolase and transaminase-catalyzed reactions in a cascading continuous-flow microreactor system.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    Department of Biochemical Engineering, University College London, London, United Kingdom.
    Rapid biocatalytic process development and intensification continues to be challenging with currently available methods. Chiral amino-alcohols are of particular interest as they represent key industrial synthons for the production of complex molecules and optically pure pharmaceuticals. (2S,3R)-2-amino-1,3,4-butanetriol (ABT), a building block for the synthesis of protease inhibitors and detoxifying agents, can be synthesized from simple, non-chiral starting materials, by coupling a transketolase- and a transaminase-catalyzed reaction. Read More

    Metabolic engineering of Bacillus subtilis for production of D-lactic acid.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida.
    Poly lactic acid (PLA) based plastics is renewable, bio-based, and biodegradable. Although present day PLA is composed of mainly L-LA, an L- and D- LA copolymer is expected to improve the quality of PLA and expand its use. To increase the number of thermotolerant microbial biocatalysts that produce D-LA, a derivative of Bacillus subtilis strain 168 that grows at 50°C was metabolically engineered. Read More

    Comparison of platform host cell protein ELISA to process-specific host cell protein ELISA.
    Biotechnol Bioeng 2017 Oct 7. Epub 2017 Oct 7.
    CMC Paradigm LLC, San Anselmo, California.
    During expression of biotherapeutic proteins, complex mixtures of additional proteins are also produced by normal expression machinery of the host cell (termed "host cell proteins," or HCP). HCPs pose a potential impact to patient safety and product efficacy, and therefore must be well-characterized and the ability of the process to clear these proteins must be demonstrated. Due to the complexity of HCP, the method(s) used for monitoring must be demonstrated to provide sufficient information about relevant proteins. Read More

    Effect of NADPH availability on free fatty acid production in Escherichia coli.
    Biotechnol Bioeng 2017 Oct 4. Epub 2017 Oct 4.
    Department of Bioengineering, Rice University, Houston, Texas.
    Microbial conversion of renewable carbon sources to free fatty acids has attracted significant attention in recent years. Accumulation of free fatty acids in Escherichia coli by overexpression of an acyl-ACP thioesterase which can break the fatty acid elongation has been well established. Various efforts have been made to increase fatty acid production in E. Read More

    A novel, smaller scaffold for Affitins: Showcase with binders specific for EpCAM.
    Biotechnol Bioeng 2017 Oct 4. Epub 2017 Oct 4.
    CRCINA, Inserm, CNRS, Université d'Angers, Université de Nantes, Nantes, France.
    Affitins are highly stable engineered affinity proteins, originally derived from Sac7d and Sso7d, two 7 kDa DNA-binding polypeptides from Sulfolobus genera. Their efficiency as reagents for intracellular targeting, enzyme inhibition, affinity purification, immunolocalization, and various other applications has been demonstrated. Recently, we have characterized the 7 kDa DNA-binding family, and Aho7c originating from Acidianus hospitalis was shown to be its smallest member with thermostability comparable to those of Sac7d and Sso7d. Read More

    Enhancement of biomass and lipid productivity by overexpression of a bZIP transcription factor in Nannochloropsis salina.
    Biotechnol Bioeng 2017 Oct 4. Epub 2017 Oct 4.
    Department of Chemical and Biomolecular Engineering, Daejeon, Republic of Korea.
    Microalgae are considered as excellent platforms for biomaterial production that can replace conventional fossil fuel-based fuels and chemicals. Genetic engineering of microalgae is prerequisite to maximize production of materials and to reduce costs for the production. Transcription factors (TFs) are emerging as key regulators of metabolic pathways to enhance production of molecules for biofuels and other materials. Read More

    Cover Image, Volume 114, Number 11, November 2017.
    Biotechnol Bioeng 2017 Nov;114(11)
    Department of Chemical Engineering, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts.
    Cover Legend: The cover image, by Catherine B. Matthews et al., is based on the Review Reexamining opportunities for therapeutic protein production in eukaryotic microorganisms, DOI 10. Read More

    Intracellular response to process optimization and impact on productivity and product aggregates for a high-titer CHO cell process.
    Biotechnol Bioeng 2018 Jan;115(1):126-138
    Cell Culture and Fermentation Sciences, MedImmune LLC, Gaithersburg, Maryland.
    A key goal in process development for antibodies is to increase productivity while maintaining or improving product quality. During process development of an antibody, titers were increased from 4 to 10 g/L while simultaneously decreasing aggregates. Process development involved optimization of media and feed formulations, feed strategy, and process parameters including pH and temperature. Read More

    Engineering of cell membrane to enhance heterologous production of hyaluronic acid in Bacillus subtilis.
    Biotechnol Bioeng 2018 Jan 23;115(1):216-231. Epub 2017 Oct 23.
    Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario, Canada.
    Hyaluronic acid (HA) is a high-value biopolymer used in the biomedical, pharmaceutical, cosmetic, and food industries. Current methods of HA production, including extraction from animal sources and streptococcal cultivations, are associated with high costs and health risks. Accordingly, the development of bioprocesses for HA production centered on robust "Generally Recognized as Safe (GRAS)" organisms such as Bacillus subtilis is highly attractive. Read More

    Reconstruction of lignin and hemicelluloses by aqueous ethanol anti-solvents to improve the ionic liquid-acid pretreatment performance of Arundo donax Linn.
    Biotechnol Bioeng 2018 Jan 23;115(1):82-91. Epub 2017 Oct 23.
    Beijing Key Laboratory of Lignocellulosic Chemistry, Beijing Forestry University, Beijing, PR China.
    Ionic liquid (IL)-acid pretreatment is known to not only enhance the enzymatic hydrolysis efficiency of lignocellulose but also to generate deposits on the surface of fiber by conventional water regeneration, which retard the increment. In this study, ethanol aqueous solution regeneration was developed as a new method to change the substrates characteristics for IL-acid pretreatment and their effects on the enzymatic hydrolysis were evaluated. Following the IL-acid reaction, the biomass slurry was subjected to ethanol aqueous solution at various concentration. Read More

    Advancements in the design and scalable production of viral gene transfer vectors.
    Biotechnol Bioeng 2018 Jan 30;115(1):25-40. Epub 2017 Oct 30.
    Department of Bioengineering, McGill University, Montreal, Quebec, Canada.
    The last 10 years have seen a rapid expansion in the use of viral gene transfer vectors, with approved therapies and late stage clinical trials underway for the treatment of genetic disorders, and multiple forms of cancer, as well as prevention of infectious diseases through vaccination. With this increased interest and widespread adoption of viral vectors by clinicians and biopharmaceutical industries, there is an imperative to engineer safer and more efficacious vectors, and develop robust, scalable and cost-effective production platforms for industrialization. This review will focus on major innovations in viral vector design and production systems for three of the most widely used viral vectors: Adenovirus, Adeno-Associated Virus, and Lentivirus. Read More

    Burkholderia cepacia lipase: A versatile catalyst in synthesis reactions.
    Biotechnol Bioeng 2018 Jan 30;115(1):6-24. Epub 2017 Oct 30.
    Planta Piloto de Ingeniería Química (PLAPIQUI), Universidad Nacional del Sur, CONICET, Bahía Blanca, Argentina.
    The lipase from Burkholderia cepacia, formerly known as Pseudomonas cepacia lipase, is a commercial enzyme in both soluble and immobilized forms widely recognized for its thermal resistance and tolerance to a large number of solvents and short-chain alcohols. The main applications of this lipase are in transesterification reactions and in the synthesis of drugs (because of the properties mentioned above). This review intends to show the features of this enzyme and some of the most relevant aspects of its use in different synthesis reactions. Read More

    Formic acid as a secondary substrate for succinic acid production by metabolically engineered Mannheimia succiniciproducens.
    Biotechnol Bioeng 2017 Dec 19;114(12):2837-2847. Epub 2017 Sep 19.
    Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus program), BioProcess Engineering Research Center, Institute for the BioCentury, Yuseong-gu, Daejeon, Republic of Korea.
    There has been much effort exerted to reduce one carbon (C1) gas emission to address climate change. As one promising way to more conveniently utilize C1 gas, several technologies have been developed to convert C1 gas into useful chemicals such as formic acid (FA). In this study, systems metabolic engineering was utilized to engineer Mannheimia succiniciproducens to efficiently utilize FA. Read More

    Combinatorial genome and protein engineering yields monoclonal antibodies with hypergalactosylation from CHO cells.
    Biotechnol Bioeng 2017 Dec 19;114(12):2848-2856. Epub 2017 Sep 19.
    Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland.
    One of the key quality attributes of monoclonal antibodies is the glycan pattern and distribution. Two terminal galactose residues typically represent a small fraction of the total glycans from antibodies. However, antibodies with defined glycosylation properties including enhanced galactosylation have been shown to exhibit altered properties for these important biomedical modalities. Read More

    One pot synthesis of GDP-mannose by a multi-enzyme cascade for enzymatic assembly of lipid-linked oligosaccharides.
    Biotechnol Bioeng 2018 Jan 23;115(1):192-205. Epub 2017 Oct 23.
    Max Planck Institute for Dynamics of Complex Technical Systems, Bioprocess Engineering, Magdeburg, Germany.
    Glycosylation of proteins is a key function of the biosynthetic-secretory pathway in the endoplasmic reticulum (ER) and Golgi apparatus. Glycosylated proteins play a crucial role in cell trafficking and signaling, cell-cell adhesion, blood-group antigenicity, and immune response. In addition, the glycosylation of proteins is an important parameter in the optimization of many glycoprotein-based drugs such as monoclonal antibodies. Read More

    Improved performance of Pseudomonas putida in a bioelectrochemical system through overexpression of periplasmic glucose dehydrogenase.
    Biotechnol Bioeng 2018 Jan 6;115(1):145-155. Epub 2017 Oct 6.
    Centre for Microbial Electrochemical Systems (CEMES), The University of Queensland, St Lucia, Brisbane, Australia.
    It was recently demonstrated that a bioelectrochemical system (BES) with a redox mediator allowed Pseudomonas putida to perform anoxic metabolism, converting sugar to sugar acids with high yield. However, the low productivity currently limits the application of this technology. To improve productivity, the strain was optimized through improved expression of glucose dehydrogenase (GCD) and gluconate dehydrogenase (GAD). Read More

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