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https://www.readbyqxmd.com/read/28731580/rewiring-the-glucose-transportation-and-central-metabolic-pathways-for-overproduction-of-n-acetylglucosamine-in-bacillus-subtilis
#1
Yang Gu, Jieying Deng, Yanfeng Liu, Jianghua Li, Hyun-Dong Shin, Guocheng Du, Jian Chen, Long Liu
N-acetylglucosamine (GlcNAc) is an important amino sugar extensively used in the healthcare field. in a previous study, we constructed the recombinant Bacillus subtilis strain BSGN6-PxylA -glmS-pP43NMK-GNA1 (BN0-GNA1) for microbial production of GlcNAc by pathway design and modular optimization. Here, we further improved the production of GlcNAc by rewiring both the glucose transportation and central metabolic pathways. First, the phosphotransferase system (PTS) was blocked by deletion of three genes, yyzE (encoding the PTS system transporter subunit iiA YyzE), ypqE (encoding the PTS system transporter subunit iiA YpqE), and ptsG (encoding the PTS system glucose-specific EiiCBA component), resulting in 47...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28726163/phosphoenolpyruvate-glucose-phosphotransferase-system-modification-increases-the-conversion-rate-during-l-tryptophan-production-in-escherichia-coli
#2
Lina Liu, Sheng Chen, Jing Wu
Escherichia coli FB-04(pta1), a recombinant L-tryptophan production strain, was constructed in our laboratory. However, the conversion rate (L-tryptophan yield per glucose) of this strain is somewhat low. In this study, additional genes have been deleted in an effort to increase the conversion rate of E. coli FB-04(pta1). Initially, the pykF gene, which encodes pyruvate kinase I (PYKI), was inactivated to increase the accumulation of phosphoenolpyruvate, a key L-tryptophan precursor. The resulting strain, E...
July 19, 2017: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28720107/reversible-bacterial-immobilization-based-on-the-salt-dependent-adhesion-of-the-bacterionanofiber-protein-ataa
#3
Shogo Yoshimoto, Yuki Ohara, Hajime Nakatani, Katsutoshi Hori
BACKGROUND: Immobilization of microbial cells is an important strategy for the efficient use of whole-cell catalysts because it simplifies product separation, enables the cell concentration to be increased, stabilizes enzymatic activity, and permits repeated or continuous biocatalyst use. However, conventional immobilization methods have practical limitations, such as limited mass transfer in the inner part of a gel, gel fragility, cell leakage from the support matrix, and adverse effects on cell viability and catalytic activity...
July 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28716706/characterization-of-a-presymptomatic-stage-in-a-drosophila-parkinson-s-disease-model-unveiling-dopaminergic-compensatory-mechanisms
#4
Daniela Molina-Mateo, Nicolás Fuenzalida-Uribe, Sergio Hidalgo, Claudia Molina-Fernández, Jorge Abarca, Rafaella V Zárate, Marcela Escandón, Reinaldo Figueroa, María Florencia Tevy, Jorge M Campusano
Parkinson disease (PD) is a degenerative disorder characterized by several motor symptoms including shaking, rigidity, slow movement and difficult walking, which has been associated to the death of nigro-striatal dopaminergic neurons. >90% of PD patients also present olfactory dysfunction. Although the molecular mechanisms responsible for this disease are not clear, hereditary PD is linked to mutations in specific genes, including the PTEN-induced putative kinase 1 (PINK1). In this work we provide for the first time a thorough temporal description of the behavioral effects induced by a mutation in the PINK1 gene in adult Drosophila, a previously described animal model for PD...
July 14, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28699496/up-scaling-strategies-to-improve-the-industrial-production-of-bacitracin-at-largescale
#5
Chandra Kant Sharma, Monika Sharma
Bacitracin was discovered and named after a 7 year old American girl, Margaret Tracey in 1943 as Bacillus was isolated from her wounds. Bacillus licheniformis is usually present in soil and bird feathers. This bacterium is most commonly present around back plumage and chest of birds. There are different types of bacitracin but the one most potent is Bacitracin A. Bacitracin induced proteins are localized in bacterial membrane. Production of antibiotic initially stopped, resumed by induction of bacitracin induced protein but after few mitotic divisions microbes reverted to their vulnerable state...
July 11, 2017: Mini Reviews in Medicinal Chemistry
https://www.readbyqxmd.com/read/28695482/role-of-apolipoproteins-and-%C3%AE-synuclein-in-parkinson-s-disease
#6
REVIEW
Fatemeh Nouri Emamzadeh
Parkinson's disease (PD) is a progressive brain disorder that interferes with activities of normal life. The main pathological feature of this disease is the loss of more than 80% of dopamine-producing neurons in the substantia nigra (SN). Dopaminergic neuronal cell death occurs when intraneuronal, insoluble, aggregated proteins start to form Lewy bodies (LBs), the most important component of which is a protein called α-synuclein (α-syn). α-Syn structurally contains hexameric repeats of 11 amino acids, which are characteristic of apolipoproteins and thus α-syn can also be considered an apolipoprotein...
July 10, 2017: Journal of Molecular Neuroscience: MN
https://www.readbyqxmd.com/read/28693684/clostridium-tepidum-sp-nov-a-close-relative-of-clostridium-sporogenes-and-clostridium-botulinum-group-i
#7
Anatoly P Dobritsa, Kirthi K Kutumbaka, Kirsten Werner, Martin Wiedmann, Aaron Asmus, Mansour Samadpour
Obligately anaerobic, Gram-stain-positive, spore-forming bacteria indistinguishable by pulsed-field gel electrophoresis were isolated from non-dairy protein shakes in bloated bottles. One of the isolates, strain IEH 97212T, was selected for further study. The strain was closely related to Clostridium sporogenes and Clostridium botulinum Group 1 based on 16S rRNA gene sequence similarities. Phylogenetic analysis also showed that strain IEH 97212T and strain PE (=DSM 18688), a bacterium isolated from solfataric mud, had identical 16S rRNA gene sequences...
July 10, 2017: International Journal of Systematic and Evolutionary Microbiology
https://www.readbyqxmd.com/read/28688931/membrane-engineering-a-novel-strategy-to-enhance-the-production-and-accumulation-of-%C3%AE-carotene-in-escherichia-coli
#8
Tao Wu, Lijun Ye, Dongdong Zhao, Siwei Li, Qingyan Li, Bolin Zhang, Changhao Bi, Xueli Zhang
Carotenoids are a class of terpenes of commercial interest that exert important biological functions. While various strategies have been applied to engineer β-carotene production in microbial cell factories, no work has been done to study and improve the storage of hydrophobic terpene products inside the heterologous host cells. Although the membrane is thought to be the cell compartment that accumulates hydrophobic terpenes such as β-carotene, direct evidence is still lacking. In this work, we engineered the membrane of Escherichia coli in both its morphological and biosynthetic aspects, as a means to study and improve its storage capacity for β-carotene...
July 5, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28681607/n-glycosylation-engineering-to-improve-the-constitutive-expression-of-rhizopus-oryzae-lipase-in-komagataella-phaffii
#9
Xiaowei Yu, Min Yang, Chuanhuan Jiang, Xiaofeng Zhang, Yan Xu
Our previous studies demonstrated that the N-glycans in Rhizopus chinensis lipase (RCL) was important for its secretion. In order to improve the secretion of Rhizopus oryzae lipase (ROL) under the control of the GAP promoter in K. phaffii, two extra N-glycosylation sites were introduced in ROL according to the position of the N-glycosylation sites of RCL by sequence alignment. The results indicated that the secretion level of ROL was strongly improved by N-glycosylation engineering, and the highest value of extracellular enzyme activity was increased from 0...
July 6, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28653476/heterotrophic-production-of-chlorella-sp-tistr-8990-biomass-growth-and-composition-under-various-production-conditions
#10
Somruethai Bouyam, Wanna Choorit, Sarote Sirisansaneeyakul, Yusuf Chisti
The green microalga Chlorella sp. TISTR 8990 was grown heterotrophically in the dark using various concentrations of a basal glucose medium with a carbon-to-nitrogen mass ratio of 29:1. The final biomass concentration and the rate of growth were highest in the 5-fold concentrated basal glucose medium (25 g L(-1) glucose, 2.5 g L(-1) KNO3 ) in batch operations. Improving oxygen transfer in the culture by increasing the agitation rate and decreasing the culture volume in 500 mL shake flasks improved growth and glucose utilization...
June 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28650068/rational-engineering-of-p-hydroxybenzoate-hydroxylase-to-enable-efficient-gallic-acid-synthesis-via-a-novel-artificial-biosynthetic-pathway
#11
Zhenya Chen, Xiaolin Shen, Jian Wang, Jia Wang, Qipeng Yuan, Yajun Yan
Gallic acid (GA) is a naturally occurring phytochemical that has strong antioxidant and antibacterial activities. It is also used as a potential platform chemical for the synthesis of diverse high-value compounds. Hydrolytic degradation of tannins by acids, bases or microorganisms serves as a major way for GA production, which however, might cause environmental pollution and low yield and efficiency. Here, we report a novel approach for efficient microbial production of GA. First, structure-based rational engineering of PobA, a p-hydroxybenzoate hydroxylase from Pseudomonas aeruginosa, generated a new mutant, Y385F/T294A PobA, which displayed much higher activity towards 3,4-dihydroxybenzoic acid (3,4-DHBA) than the wild type and any other reported mutants...
June 26, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28648546/process-intensification-for-high-yield-production-of-influenza-h1n1-gag-virus-like-particles-using-an-inducible-hek-293-stable-cell-line
#12
Alina Venereo-Sanchez, Melanie Simoneau, Stéphane Lanthier, Parminder Chahal, Lucie Bourget, Sven Ansorge, Rénald Gilbert, Olivier Henry, Amine Kamen
Influenza virus dominant antigens presentation using virus like particle (VLP) approach is attractive for the development of new generation of influenza vaccines. Mammalian cell platform offers many advantages for VLP production. However, limited attention has been paid to the processing of mammalian cell produced VLPs. Better understanding of the production system could contribute to increasing the yields and making large-scale VLP vaccine manufacturing feasible. In a previous study, we have generated a human embryonic kidney HEK-293 inducible cell line expressing Hemagglutinin (HA) and Neuraminidase (NA), which was used to produce VLPs upon transient transfection with a plasmid containing HIV-1 Gag...
June 22, 2017: Vaccine
https://www.readbyqxmd.com/read/28639328/immunogenicity-of-protein-aggregates-of-a-monoclonal-antibody-generated-by-forced-shaking-stress-with-siliconized-and-nonsiliconized-syringes-in-balb-c-mice
#13
Tomonobu Uchino, Yasunori Miyazaki, Takuto Yamazaki, Yoshiyuki Kagawa
OBJECTIVE: In this study, we aimed to investigate the immunogenicity of protein aggregates of monoclonal antibodies (mAbs), generated by forced shaking stress with siliconized and nonsiliconized syringes in a mouse model. METHODS: Samples were filled in siliconized and nonsiliconized syringes with shaking and headspace air. Characterization studies were performed using high-performance size-exclusion chromatography, nanoparticle tracking analysis, flow cytometry, micro-flow imaging and resonant mass measurement...
June 21, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/28633643/enhancement-of-thermoalkaliphilic-xylanase-production-by-pichia-pastoris-through-novel-fed-batch-strategy-in-high-cell-density-fermentation
#14
Tingting Shang, Dayong Si, Dongyan Zhang, Xuhui Liu, Longmei Zhao, Cong Hu, Yu Fu, Rijun Zhang
BACKGROUND: Xylanase degrades xylan into monomers of various sizes by catalyzing the endohydrolysis of the 1,4-β-D-xylosidic linkage randomly, possessing potential in wide industrial applications. Most of xylanases are susceptible to be inactive when suffering high temperature and high alkaline process. Therefore, it is necessary to develop a high amount of effective thermoalkaliphilic xylanases. This study aims to enhance thermoalkaliphilic xylanase production in Pichia pastoris through fermentation parameters optimization and novel efficient fed-batch strategy in high cell-density fermentation...
June 21, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28627778/engineering-cho-cells-with-an-oncogenic-kit-improves-cells-growth-resilience-to-stress-and-productivity
#15
Mohamed Mahameed, Boaz Tirosh
An optimized biomanufacturing process in mammalian cells is contingent on the ability of the producing cells to reach high viable cell densities. In addition, at the peak of growth, cells need to continue producing the biological entity at a consistent quality. Thus, engineering cells with robust growth performance and resilience to variable stress conditions is highly desirable. The tyrosine kinase receptor, KIT, plays a key role in cell differentiation and the survival of several immune cell types. Its oncogenic mutant, D816V, endows cells with high proliferation capacity and resistance to kinase inhibitors...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28619018/efficient-soluble-expression-of-disulfide-bonded-proteins-in-the-cytoplasm-of-escherichia-coli-in-fed-batch-fermentations-on-chemically-defined-minimal-media
#16
Anna Gąciarz, Narendar Kumar Khatri, M Lourdes Velez-Suberbie, Mirva J Saaranen, Yuko Uchida, Eli Keshavarz-Moore, Lloyd W Ruddock
BACKGROUND: The production of recombinant proteins containing disulfide bonds in Escherichia coli is challenging. In most cases the protein of interest needs to be either targeted to the oxidizing periplasm or expressed in the cytoplasm in the form of inclusion bodies, then solubilized and re-folded in vitro. Both of these approaches have limitations. Previously we showed that soluble expression of disulfide bonded proteins in the cytoplasm of E. coli is possible at shake flask scale with a system, known as CyDisCo, which is based on co-expression of a protein of interest along with a sulfhydryl oxidase and a disulfide bond isomerase...
June 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28606738/engineering-high-level-production-of-fatty-alcohols-by-saccharomyces-cerevisiae-from-lignocellulosic-feedstocks
#17
Leo d'Espaux, Amit Ghosh, Weerawat Runguphan, Maren Wehrs, Feng Xu, Oliver Konzock, Ishaan Dev, Melissa Nhan, Jennifer Gin, Amanda Reider Apel, Christopher J Petzold, Seema Singh, Blake A Simmons, Aindrila Mukhopadhyay, Héctor García Martín, Jay D Keasling
Fatty alcohols in the C12-C18 range are used in personal care products, lubricants, and potentially biofuels. These compounds can be produced from the fatty acid pathway by a fatty acid reductase (FAR), yet yields from the preferred industrial host Saccharomyces cerevisiae remain under 2% of the theoretical maximum from glucose. Here we improved titer and yield of fatty alcohols using an approach involving quantitative analysis of protein levels and metabolic flux, engineering enzyme level and localization, pull-push-block engineering of carbon flux, and cofactor balancing...
June 10, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28606596/mini-review-recombinant-production-of-tailored-bio-pharmaceuticals-in-different-bacillus-strains-and-future-perspectives
#18
REVIEW
Antonia Lakowitz, Thibault Godard, Rebekka Biedendieck, Rainer Krull
Bio-pharmaceuticals like antibodies, hormones and growth factors represent about one-fifth of commercial pharmaceuticals. Host candidates of growing interest for recombinant production of these proteins are strains of the genus Bacillus, long being established for biotechnological production of homologous and heterologous proteins. Bacillus strains benefit from development of efficient expression systems in the last decades and emerge as major industrial workhorses for recombinant proteins due to easy cultivation, non-pathogenicity and their ability to secrete recombinant proteins directly into extracellular medium allowing cost-effective downstream processing...
June 9, 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/28603019/friability-testing-as-a-new-stress-stability-assay-for-biopharmaceuticals
#19
Tetsuo Torisu, Takahiro Maruno, Saki Yoneda, Yoshinori Hamaji, Shinya Honda, Tadayasu Ohkubo, Susumu Uchiyama
A cycle of dropping and shaking a vial containing antibody solution was reported to induce aggregation. In this study, antibody solutions in glass prefillable syringes with or without silicone oil lubrication were subjected to the combined stresses of dropping and shaking, using a friability testing apparatus. Larger numbers of subvisible particles were generated, regardless of silicone oil lubrication, upon combination stress exposure than that with shaking stress alone. Nucleation of antibody molecules upon perturbation by an impact of the dropping, and/or adsorption of antibody molecules to the syringe surface followed by film formation and antibody film desorption were considered key steps in the particle formation promoted by combination stress...
June 8, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28579488/effective-cell-free-drug-screening-protocol-for-protein-protein-interaction
#20
Shaked Ashkenazi, Alexander Plotnikov, Anat Bahat, Rivka Dikstein
Specific protein-protein interaction (PPI) is an essential feature of many cellular processes however, targeting these interactions by small molecules is highly challenging due to the nature of the interaction interface. Thus, screening for PPI inhibitors requires enormous number of compounds. Here we describe a simple and improved protocol designed for a search of direct PPI inhibitors. We engineered a bacterial expression system for the split-Renilla luciferase (RL) complementation assay that monitors PPI...
June 1, 2017: Analytical Biochemistry
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