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Journal of Biotechnology

Xin-Kai Zhang, Ming-Yue Nie, Jun Chen, Liu-Jing Wei, Qiang Hua
Lycopene has been broadly studied in recent decades due to its health benefits including cancer prevention, anti-atherogenic and anti-obesity effects, and modulation of the immune system. To obtain efficient synthesis of lycopene, extensive researches have been conducted in various microbial cells, including Yarrowia lipolytica, to heterologously produce lycopene using various genetic and metabolic engineering methods. In this study, the effects of copy numbers of lycopene synthesis genes, a variety of key central metabolic genes (especially AMP deaminase-encoding gene AMPD), and 5-L fermenter cultivation on lycopene production in Y...
November 15, 2018: Journal of Biotechnology
Cheng Zheng, Zhenjian Li, Haifeng Yang, Tianyi Zhang, Huanqing Niu, Dong Liu, Junzhi Wang, Hanjie Ying
Biocatalysis has become the main approach to produce cytidine diphosphate choline (CDP-choline), which has been applied for treatment of acute craniocerebral injury and consciousness after brain surgery. However, salt accumulates with the production and inhibits enzyme activity, and eventually reduces yield and product accumulation rate. Our work provided a possible solution to this problem by applying a computational designed halophilic choline kinase. The halotolerant CKI (choline kinase) was designed following a unique strategy considering the most variable residue positions on the protein surface among target enzymes from different sources...
November 13, 2018: Journal of Biotechnology
Diana Uhrich, Hyun-Young Jang, Jin-Byung Park, Jan von Langermann
This study presents the preparation and physical-chemical characterization of chemical resistant polyurethane-based compartments for biocatalytic application. The artificial compartments were prepared from an emulsion of polymer precursor and an aqueous phase that includes a biocatalytic reaction system. After curing, highly dispersed aqueous domains were obtained, which still contain the entire biocatalytic reaction system and remain fixed in the solid polymer preparation. The tensile and compression behavior of the prepared polymeric material is not significantly affected by the incorporation and facilitates excellent stability against various organic solvents and acid solutions...
November 12, 2018: Journal of Biotechnology
Dan Wu, Haifeng Zhu, Ju Chu, Jing Wu
Pichia pastoris is subjected to strong oxidative stress in the methanol induction phase. The oxidative stress inflicts severe injury to yeast cells, which causes cell death and reduces protein expression ability. N-acetyltransferase in Saccharomyces cerevisiae can protect yeast cells from damage caused by decreasing reactive oxygen species (ROS) in oxidative pressure environments such as ethanol treatment, freeze-thawing, or heat shock. In this study, N-acetyltransferase from P. pastoris (PpMpr1) was overexpressed for the first time to improve the anti-oxidative stress ability to protect cells from strong ROS damage during the methanol induction phase...
November 11, 2018: Journal of Biotechnology
Elena A Günter, Oxana V Popeyko, Elena I Istomina
The aims of this research were to obtain modified pectins of callus cultures using various culture conditions, to evaluate the relationship between the chemical characteristics of pectins, the swelling behavior and the release of prednisolone from calcium pectinate gel (CaPG) beads. An increase of the calcium concentration in the culture media correlated significantly with the rhamnogalacturonan I (RG-I) branching of the pectin. The beads from the pectin with a higher RG-I branching had the lower prednisolone release in a gastric fluid...
November 10, 2018: Journal of Biotechnology
Liang Zhang, Andreas Castan, Joanne Stevenson, Nathalie Chatzissavidou, Francisco Vilaplana, Veronique Chotteau
The glycosylation profile of therapeutic monoclonal antibodies (mAbs) is a crucial quality parameter for industrial Immunoglobulin G (IgG) production. Several alternative carbon sources, which function as glycosylation precursors, have been reported to impact the glycosylation pattern. Since the cells give priority to glucose uptake, the presence of this substrate can lower the effects of alternative sugars on the glycosylation. In order to get a better understanding of the influence of alternative sugars on the glycosylation and to investigate how they impact each other, combinations of mannose, fructose, galactose and fucose were fed to Chinese hamster ovary (CHO) cells in batch culture when the glucose became depleted and the lactate, accumulated in the culture, was used as carbon source...
November 10, 2018: Journal of Biotechnology
Annesha Sengupta, Avinash Vellore Sunder, Sujata V Sohoni, Pramod P Wangikar
The light harvesting photosystem in cyanobacteria offers a potential pathway for the regeneration of the nicotinamide cofactor NADPH, thereby facilitating the application of cyanobacteria as excellent whole cell biocatalysts in oxidoreductase-mediated biotransformation. The use of cyanobacterial metabolism for cofactor recycling improves the atom economy of the process compared to the commonly employed enzyme-coupled cofactor recycling using enzymes such as glucose dehydrogenase. Here we report the asymmetric conversion of acetophenone to chiral 1-phenylethanol by recombinant Synechococcus elongatus PCC 7942 whole cell biocatalyst that expresses the NADPH dependent L...
November 6, 2018: Journal of Biotechnology
Jun Ren, Dokyun Na, Seung Min Yoo
Bacterial transformation is a fundamental technology to deliver engineered plasmids into bacterial cells, which is essential in industrial protein production, chemical production, etc. Previously, we developed a simple chemico-physical transformation method that can be applied to various bacterial species. Here, to accelerate the advance of bacteria biotechnology we optimize our method by combinatorially evaluating chemical compounds (rubidium chloride, lithium acetate, cesium chloride, dimethyl sulfoxide, and magnesium chloride) for increasing membrane permeability and nanomaterials (sepiolite, gold(III) chloride, multiwalled carbon nanotube, and chitosan) for piercing the membranes...
November 5, 2018: Journal of Biotechnology
Xiangwei Cui, Junxian Wan, Xing Zhang, Hui Wu, Zhimin Li, Qin Ye
Glutathione (GSH) is an important bioactive tripeptide and widely used in food, medicine and other industries. Recently, the bifunctional glutathione synthetase, GshF, has been applied for efficient GSH production under inducible promoter. In this study, the constitutive expression of GshF from Streptococcus sanguinis (GshFss) was investigated under four different constitutive promoters. Based on previous study, five genes in Escherichia coli JM109 were deleted to eliminate the degradation of precursors and GSH...
November 2, 2018: Journal of Biotechnology
Hao Chen, Yanqing Li, Chunhua Du, Yan Li, Junli Zhao, Xiaojing Zheng, Qinwen Mao, Haibin Xia
RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression...
November 1, 2018: Journal of Biotechnology
Gorawit Yusakul, Seiichi Sakamoto, Hiroyuki Tanaka, Satoshi Morimoto
Formation of disulfide bonds between heavy and light chains is challenge for production of recombinant immunoglobulin G (IgG) and fragment antigen-binding (Fab). Insufficient formation of the bond influences productive yield and quality of recombinant IgG and Fab. To investigate how amino acid sequences of the first heavy chain constant region (CH1) effect on assembly between heavy chain (VH-CH1 or Fd) and light chain (VL-CL), the CH1 gene sequence of an anti-paclitaxel fragment antigen-binding (anti-PT Fab IgG2a) was modified using the splicing by overlap extension-polymerase chain reaction (SOE-PCR) to be gene sequences encoding amino acid sequence of IgG1, IgG2b, and IgG3 subtypes...
October 31, 2018: Journal of Biotechnology
Alan Costello, Orla Coleman, Nga T Lao, Michael Henry, Paula Meleady, Niall Barron, Martin Clynes
miRNAs are potent molecular regulators of cellular behaviour. The manipulation of these small non-coding RNAs has been used to enhance industrially relevant phenotypes in Chinese Hamster Ovary (CHO) cells. We investigated the stable depletion of six miRNAs; miR-204-5p, 338-3p, 378-3p, 409-3p, 455-3p and 505-3p, robustly associated with cell growth rate from a previous profiling study. Inhibition of endogenous miR-378-3p function by miRNA-sponge-decoy improved peak cell density by 59%. Quantitative label free LC-MS/MS proteomic analysis of the fractionated cell cultures at day 4 and 8 of batch culture found 216 cytosolic and 114 membrane-associated proteins differentially expressed with stable miR-378-3p depletion...
October 31, 2018: Journal of Biotechnology
Dae Geun Kim, Changsu Lee, Yeoung-Sang Yun, Chang-Hee Hong, Yoon-E Choi
Plant factories have been developed to replace traditional agriculture, aiming to solve future problems of food availability. However, the nutrient solution in a plant factory is discharged after a single batch of plant cultivation, giving rise to large amounts of waste nutrient solution. Microalgae can be used to treat a wide variety of wastewater and effectively remove excessive nutrients from wastewater. Therefore, the incorporation of microalgal cultivation into a plant factory to treat waste nutrients would be a reasonable approach facilitating removal of waste nutrients with concomitant production of algal biomass...
October 30, 2018: Journal of Biotechnology
Ju-Yong Park, Bit-Na Kim, Yang-Hoon Kim, Jiho Min
Rhodobacter sphaeroides consists of two chromosomes and many plasmids and incorporates many environmentally important functional gene. Rhodobacter sphaeroides MBTLJ-8 was derived from R. sphaeroides 2.4.1 using chemical mutagenesis and is characterized by enhanced production of physiological active compounds as well as improved carbon dioxide reduction capacity. We reported the complete genome sequence and characteristics based on genomic information of this bacteria. Therefore, this genome sequence provides elucidation for improved CO2 fixation and enhanced physiological active compounds production, and will be used as the efficient photosynthetic bacteria for the biological CO2 reduction system...
October 22, 2018: Journal of Biotechnology
Yimin Hu, Fang Nan, Sarah Wanjiku Maina, Jia Guo, Shenglu Wu, Zhihong Xin
Plipastatin, a cyclic lipopeptide, exhibits inhibitory activity against filamentous fungi and plays an important role in the prevention of plant diseases, and post-harvest preservation of fruits and vegetables. However, the application of plipastatin has been hampered by low yields in natural strains, while chemical synthesis is not feasible because of its complex chemical structure. In this study, a scarless genetic modification method was applied to construct a heterologous expression host (Bacillus subtilis 1A751 Δpps) by knocking out the natural plipastatin genes from B...
October 19, 2018: Journal of Biotechnology
Sebastian Andris, Matthias Rüdt, Jonas Rogalla, Michaela Wendeler, Jürgen Hubbuch
The conjugation reaction of monoclonal antibodies (mAbs) with small-molecule drugs is a central step during production of antibody-drug conjugates (ADCs). The ability to monitor this step in real time can be advantageous for process understanding and control. Here, we propose a method based on UV/Vis spectroscopy in conjunction with partial least squares (PLS) regression for non-invasive monitoring of conjugation reactions. In experiments, the method was applied to conjugation reactions with two surrogate drugs in microplate format as well as at 20 ml scale...
October 12, 2018: Journal of Biotechnology
Agnes Rodler, Christian Schuster, Eva Berger, Rupert Tscheließnig, Alois Jungbauer
Alternative separation methods operating in an aqueous environment are of increasing importance for further progress of molecular separation in life sciences and other industrial sectors working towards a biobased economy. By spincoating, membranes with thicknesses under 100 nm and 20 cm2 surface area were prepared from an epoxy based resin. For the first time such ultrathin epoxy films were used for the selective separation of small molecules and metabolites within an aqueous environment. Initially, selectivity is demonstrated by the separation of two dyes of similar size (0...
October 11, 2018: Journal of Biotechnology
R Román, M Farràs, M Camps, I Martínez-Monge, P Comas, M Martínez-Espelt, M Lecina, A Casablancas, J J Cairó
Although pH control at physiological levels is generally considered as the optimal culture condition, in some cases other strategies should be taken into account for their beneficial effects on process performance. pH and CO2 levels are chemical variables that have a major impact in cell growth and product titers in cell culture since their effect on key metabolic routes. HEK293 cells expressing recombinant hIFNγ showed different metabolic behavior when cultured in shake flask compared to pH-controlled bioreactors, in which a decrease in cell density and product titer were observed...
December 10, 2018: Journal of Biotechnology
Sabita Rangra, Mira Kabra, Vageesh Gupta, Preeti Srivastava
Biodesulfurization is an eco-friendly process for removing sulfur from petroleum fractions. The process could not be commercialized because of the inability of microorganisms to desulfurize a wide range of heterocyclic poly aromatic sulfur compounds like dibenzothiophene (DBT), 4, 6-Dimethyl DBT present in fuel and low desulfurization activity. In the present study, to improve the rates of conversion of dibenzothiophene to dibenzothiophene sulfone, the responsible enzyme dibenzothiophene monooxygenase DszC, is displayed on the surface of Escherichia coli...
December 10, 2018: Journal of Biotechnology
Wang Wen-Qiong, Zhang Xiao-Feng
The process of microbial transglutaminase (TG) covalent immobilization on an ultrafiltration membrane surface, the optimum immobilization conditions and the characteristics of the enzymatic membrane in a reactor were investigated. The process of TG immobilization on polyethersulfone (PES) membrane surfaces was analyzed by Fourier transform infrared and X-ray photoelectron spectroscopy. The optimal condition for TG immobilization was in pH 5.0 phosphate buffer containing TG (20 U/mL). The immobilized TG had a high affinity for the substrate according to the kinetic parameters and retained 50% activity until the twentieth day...
December 10, 2018: Journal of Biotechnology
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