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Biotechnology and Bioengineering

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https://www.readbyqxmd.com/read/29150961/modulation-of-igg1-immunoeffector-function-by-glycoengineering-of-the-gdp-fucose-biosynthesis-pathway
#1
Ronan M Kelly, Ronald L Kowle, Zhirui Lian, Beth A Strifler, Derrick R Witcher, Bhavin S Parekh, Tongtong Wang, Christopher C Frye
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...
November 18, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29150937/cgmp-production-and-analysis-of-bg505-sosip-664-an-extensively-glycosylated-trimeric-hiv-1-envelope-glycoprotein-vaccine-candidate
#2
Antu K Dey, Albert Cupo, Gabriel Ozorowski, Vaneet K Sharma, Anna-Janina Behrens, Eden P Go, Thomas J Ketas, Anila Yasmeen, Per J Klasse, Eddy Sayeed, Heather Desaire, Max Crispin, Ian A Wilson, Rogier W Sanders, Thomas Hassell, Andrew Ward, John P Moore
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. The successful translation of this prototype demonstrates the feasibility of producing similar immunogens on an appropriate scale and of an acceptable quality for Phase I experimental medicine clinical trials...
November 18, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29150933/design-of-a-novel-continuous-flow-reactor-for-low-ph-viral-inactivation
#3
Stephanie A Parker, Linus Amarikwa, Kevin Vehar, Raquel Orozco, Scott Godfrey, Jon Coffman, Parviz Shamlou, Cameron L Bardliving
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 187.7-375.5) tubular reactor for low pH viral inactivation with enhanced radial mixing via the incorporation of curvature and flow inversions. Towards this aim, the reactor described here, Jig in a Box (JIB), was designed with a flow path consisting of alternating 270° degree turns...
November 18, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29150930/photoautotrophic-production-of-macular-pigment-in-a-chlamydomonas-reinhardtii-strain-generated-by-using-dna-free-crispr-cas9-rnp-mediated-mutagenesis
#4
Kwangryul Baek, Jihyeon Yu, Jooyeon Jeong, Sang Jun Sim, Sangsu Bae, EonSeon Jin
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. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level...
November 18, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29143978/production-of-1-2-propanediol-in-photoautotrophic-synechocystis-is-linked-to-glycogen-turn-over
#5
Christian David, Andreas Schmid, Lorenz Adrian, Annegret Wilde, Katja Bühler
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. 1,2-propanediol could be successfully produced by Synechocystis, at an approximate rate of 55 µmol h(-1) gCDW(-1) ...
November 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29135022/a-fast-and-simple-method-to-estimate-relative-hyphal-tensile-strength-of-filamentous-fungi-used-to-assess-the-effect-of-autophagy
#6
Daniela Quintanilla, Cynthia Chelius, Sirasa Iambamrung, Sidney Nelson, Donnel Thomas, Krist V Gernaey, Mark R Marten
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. To overcome this issue, a method for fast and easy, statistically-verified quantification of relative hyphal tensile strength was developed. It involves off-line fragmentation in a high shear mixer followed by quantification of fragment size using laser diffraction...
November 14, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29131321/ty1-fused-protein-body-formation-for-spatial-organization-of-metabolic-pathways-in-saccharomyces-cerevisiae
#7
Jong Yun Han, Jae Myeong Song, Sung Hwa Seo, Chonglong Wang, Seung-Goo Lee, Hongweon Lee, Seon-Won Kim, Eui-Sung Choi
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...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29131314/electron-shuttling-to-ferrihydrite-selects-for-fermentative-rather-than-fe-3-reducing-biomass-in-xylose-fed-batch-reactors-derived-from-three-different-inoculum-sources
#8
Jovan Popovic, Kevin T Finneran
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...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29131308/glycosyltransferase-cascades-made-fit-for-chemical-production-integrated-biocatalytic-process-for-the-natural-polyphenol-c-glucoside-nothofagin
#9
Katharina Schmölzer, Martin Lemmerer, Bernd Nidetzky
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...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29131302/highly-active-spore-biocatalyst-by-self-assembly-of-co-expressed-anchoring-scaffoldin-and-multimeric-enzyme
#10
Long Chen, Megan Holmes, Elise Schaefer, Ashok Mulchandani, Xin Ge
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...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29131301/genome-wide-identification-of-tolerance-mechanisms-towards-p-coumaric-acid-in-pseudomonas-putida
#11
Patricia Calero, Sheila I Jensen, Klara Bojanovič, Rebecca Lennen, Anna Koza, Alex T Nielsen
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. P. putida was found to be significantly more tolerant towards three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found towards p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29105734/preferential-capture-of-epcam-expressing-extracellular-vesicles-on-solid-surfaces-coated-with-an-aptamer-conjugated-zwitterionic-polymer
#12
Mitsutaka Yoshida, Kazuhiro Hibino, Satoshi Yamamoto, Sachiko Matsumura, Yasutomo Yajima, Kiyotaka Shiba
Extracellular vesicles (EVs) collectively represent small vesicles that are secreted from cells and carry biomolecules (e.g., miRNA, lncRNA, mRNA, proteins, lipids, metabolites, etc.) that originate in those cells. Body fluids, such as blood and saliva, include large numbers of EVs, making them potentially a rich source of diagnostic information. However, these EVs are mixtures of vesicles released from diseased tissues as well as from normal cells. This heterogeneous nature therefore blurs the clinical information obtainable from EV-based diagnosis...
November 6, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29105733/engineering-escherichia-coli-for-malate-production-by-integrating-modular-pathway-characterization-with-crispri-guided-multiplexed-metabolic-tuning
#13
Cong Gao, Shihui Wang, Guipeng Hu, Liang Guo, Xiulai Chen, Peng Xu, Liming Liu
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...
November 4, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29105732/metabolic-profiles-analysis-of-1-3-propanediol-production-process-by-clostridium-butyricum-through-repeated-batch-fermentation-coupled-with-activated-carbon-adsorption
#14
Ai-Hui Zhang, Hao-Lin Liu, Shi-Yang Huang, You-Si Fu, Bai-Shan Fang
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...
November 4, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29105731/expression-of-tabersonine-16-hydroxylase-and-16-hydroxytabersonine-o-methyltransferase-in-catharanthus-roseus-hairy-roots
#15
Jiayi Sun, Le Zhao, Zengyi Shao, Jacqueline Shanks, Christie A M Peebles
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. roseus hairy roots do not produce vindoline due to the lack of expression of the seven-step pathway from tabersonine to vindoline (Murata and De Luca, 2005)...
November 4, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29080362/metabolic-phenotyping-of-cho-cells-varying-in-cellular-biomass-accumulation-and-maintenance-during-fed-batch-culture
#16
Alejandro Fernandez-Martell, Yusuf B Johari, David C James
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...
October 28, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29080352/dynamic-modelling-of-green-algae-cultivation-in-a-photobioreactor-for-sustainable-biodiesel-production
#17
Ehecatl Antonio Del Rio-Chanona, Jiao Liu, Jonathan L Wagner, Dongda Zhang, Yingying Meng, Song Xue, Nilay Shah
Biodiesel produced from microalgae has been extensively studied due to its potentially outstanding advantages over traditional transportation fuels. In order to facilitate its industrialisation and improve the process profitability, it is vital to construct highly accurate models capable of predicting the complex behaviour of the investigated biosystem for process optimisation and control, which forms the current research goal. Three original contributions are described in this paper. Firstly, a dynamic model is constructed to simulate the complicated effect of light intensity, nutrient supply and light attenuation on both biomass growth and biolipid production...
October 28, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29080350/identification-of-key-residues-modulating-the-stereoselectivity-of-nitrile-hydratase-towards-rac-mandelonitrile-by-semi-rational-engineering
#18
Zhongyi Cheng, Lukasz Peplowski, Wenjing Cui, Yuanyuan Xia, Zhongmei Liu, Jialei Zhang, Michihiko Kobayashi, Zhemin Zhou
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 towards rac-mandelonitrile (MAN)...
October 28, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29077207/a-plug-n-play-modular-metabolic-system-for-the-production-of-apocarotenoids
#19
Congqiang Zhang, Xixian Chen, Nic D Lindley, Heng-Phon Too
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. Here, an Escherichia coli-based modular system was developed to produce various apocarotenoids. Incorporation of enzyme engineering approaches (N-terminal truncation and protein fusion) into modular metabolic engineering strategy significantly improved α-ionone production from 0...
October 27, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29064570/effects-of-scaffold-microstructure-and-low-intensity-pulsed-ultrasound-on-chondrogenic-differentiation-of-human-mesenchymal-stem-cells
#20
Mitra Aliabouzar, Se-Jun Lee, Xuan Zhou, Grace Lijjie Zhang, Kausik Sarkar
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 1.5 MHz were found to be 100 mW/cm(2) and 20% duty cycle...
October 24, 2017: Biotechnology and Bioengineering
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