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BMC Biotechnology

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https://www.readbyqxmd.com/read/27908283/expression-of-a-fungal-manganese-peroxidase-in-escherichia-coli-a-comparison-between-the-soluble-and-refolded-enzymes
#1
Nan Wang, Kai Ren, Rong Jia, Wenting Chen, Ruirui Sun
BACKGROUND: Manganese peroxidase (MnP) from Irpex lacteus F17 has been shown to have a strong ability to degrade recalcitrant aromatic pollutants. In this study, a recombinant MnP from I. lacteus F17 was expressed in Escherichia coli Rosetta (DE3) in the form of inclusion bodies, which were refolded to achieve an active enzyme. Further, we optimized the in vitro refolding conditions to increase the recovery yield of the recombinant protein production. Additionally, we attempted to express recombinant MnP in soluble form in E...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27905914/development-of-a-gene-synthesis-platform-for-the-efficient-large-scale-production-of-small-genes-encoding-animal-toxins
#2
Ana Filipa Sequeira, Joana L A Brás, Catarina I P D Guerreiro, Renaud Vincentelli, Carlos M G A Fontes
BACKGROUND: Gene synthesis is becoming an important tool in many fields of recombinant DNA technology, including recombinant protein production. De novo gene synthesis is quickly replacing the classical cloning and mutagenesis procedures and allows generating nucleic acids for which no template is available. In addition, when coupled with efficient gene design algorithms that optimize codon usage, it leads to high levels of recombinant protein expression. RESULTS: Here, we describe the development of an optimized gene synthesis platform that was applied to the large scale production of small genes encoding venom peptides...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27903244/cryopreservation-of-dermal-fibroblasts-and-keratinocytes-in-hydroxyethyl-starch-based-cryoprotectants
#3
Yahaira Naaldijk, Adiv A Johnson, Annett Friedrich-Stöckigt, Alexandra Stolzing
BACKGROUND: Preservation of human skin fibroblasts and keratinocytes is essential for the creation of skin tissue banks. For successful cryopreservation of cells, selection of an appropriate cryoprotectant agent (CPA) is imperative. The aim of this study was to identify CPAs that minimize toxic effects and allow for the preservation of human fibroblasts and keratinocytes in suspension and in monolayers. RESULTS: We cryopreserved human fibroblasts and keratinocytes with different CPAs and compared them to fresh, unfrozen cells...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27881117/novel-prokaryotic-expression-of-thioredoxin-fused-insulinoma-associated-protein-tyrosine-phosphatase-2-ia-2-its-characterization-and-immunodiagnostic-application
#4
Luciano Lucas Guerra, Natalia Inés Faccinetti, Aldana Trabucchi, Bruno David Rovitto, Adriana Victoria Sabljic, Edgardo Poskus, Ruben Francisco Iacono, Silvina Noemí Valdez
BACKGROUND: The insulinoma associated protein tyrosine phosphatase 2 (IA-2) is one of the immunodominant autoantigens involved in the autoimmune attack to the beta-cell in Type 1 Diabetes Mellitus. In this work we have developed a complete and original process for the production and recovery of the properly folded intracellular domain of IA-2 fused to thioredoxin (TrxIA-2ic) in Escherichia coli GI698 and GI724 strains. We have also carried out the biochemical and immunochemical characterization of TrxIA-2icand design variants of non-radiometric immunoassays for the efficient detection of IA-2 autoantibodies (IA-2A)...
November 24, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27876044/generation-of-monoclonal-antibodies-against-native-viral-proteins-using-antigen-expressing-mammalian-cells-for-mouse-immunization
#5
Natalie Rose, Carlos Augusto Pinho-Nascimento, Alessia Ruggieri, Paola Favuzza, Marco Tamborrini, Hanna Roth, Marcia Terezinha Baroni de Moraes, Hugues Matile, Thomas Jänisch, Gerd Pluschke, Katharina Röltgen
BACKGROUND: Due to their rising incidence and progressive geographical spread, infections with mosquito-borne viruses, such as dengue (DENV), chikungunya and zika virus, have developed into major public health challenges. Since all of these viruses may cause similar symptoms and can occur in concurrent epidemics, tools for their differential diagnosis and epidemiological monitoring are of urgent need. RESULTS: Here we report the application of a novel strategy to rapidly generate monoclonal antibodies (mAbs) against native viral antigens, exemplified for the DENV nonstructural glycoprotein 1 (NS1)...
November 22, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27871321/biochemical-characterization-and-synergism-of-cellulolytic-enzyme-system-from-chaetomium-globosum-on-rice-straw-saccharification
#6
Wanwitoo Wanmolee, Warasirin Sornlake, Nakul Rattanaphan, Surisa Suwannarangsee, Navadol Laosiripojana, Verawat Champreda
BACKGROUND: Efficient hydrolysis of lignocellulosic materials to sugars for conversion to biofuels and chemicals is a key step in biorefinery. Designing an active saccharifying enzyme system with synergy among their components is considered a promising approach. RESULTS: In this study, a lignocellulose-degrading enzyme system of Chaetomium globosum BCC5776 (CG-Cel) was characterized for its activity and proteomic profiles, and synergism with accessory enzymes. The highest cellulase productivity of 0...
November 21, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27871279/biodegradation-of-lignin-and-nicotine-with-white-rot-fungi-for-the-delignification-and-detoxification-of-tobacco-stalk
#7
Yulong Su, He Xian, Sujuan Shi, Chengsheng Zhang, S M Nuruzzaman Manik, Jingjing Mao, Ge Zhang, Weihong Liao, Qian Wang, Haobao Liu
BACKGROUND: Tobacco stalk is one kind of abundant crop residues in China. The high lignification of tobacco stalk increases its reusing cost and the existing of nicotine will cause serious pollution. The biodegradation of lignocellulosic biomass has been demonstrated to be an environmental and economical approach for the utilization of plant stalk. Meanwhile, many nicotine-degrading microorganisms were found in nature. However, microorganisms which could degraded both nicotine and lignin haven't been reported...
November 21, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27855668/identification-and-characterization-of-two-new-5-keto-4-deoxy-d-glucarate-dehydratases-decarboxylases
#8
André Pick, Barbara Beer, Risa Hemmi, Rena Momma, Jochen Schmid, Kenji Miyamoto, Volker Sieber
BACKGROUND: Hexuronic acids such as D-galacturonic acid and D-glucuronic acid can be utilized via different pathways within the metabolism of microorganisms. One representative, the oxidative pathway, generates α-keto-glutarate as the direct link entering towards the citric acid cycle. The penultimate enzyme, keto-deoxy glucarate dehydratase/decarboxylase, catalyses the dehydration and decarboxylation of keto-deoxy glucarate to α-keto-glutarate semialdehyde. This enzymatic reaction can be tracked continuously by applying a pH-shift assay...
November 17, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27852252/a-novel-pyruvate-kinase-and-its-application-in-lactic-acid-production-under-oxygen-deprivation-in-corynebacterium-glutamicum
#9
Xin Chai, Xiuling Shang, Yu Zhang, Shuwen Liu, Yong Liang, Yun Zhang, Tingyi Wen
BACKGROUND: Pyruvate kinase (Pyk) catalyzes the generation of pyruvate and ATP in glycolysis and functions as a key switch in the regulation of carbon flux distribution. Both the substrates and products of Pyk are involved in the tricarboxylic acid cycle, anaplerosis and energy anabolism, which places Pyk at a primary metabolic intersection. Pyks are highly conserved in most bacteria and lower eukaryotes. Corynebacterium glutamicum is an industrial workhorse for the production of various amino acids and organic acids...
November 16, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27829414/repair-of-cartilage-defects-in-osteoarthritis-rats-with-induced-pluripotent-stem-cell-derived-chondrocytes
#10
Yanxia Zhu, Xiaomin Wu, Yuhong Liang, Hongsheng Gu, Kedong Song, Xuenong Zou, Guangqian Zhou
BACKGROUND: The incapacity of articular cartilage (AC) for self-repair after damage ultimately leads to the development of osteoarthritis. Stem cell-based therapy has been proposed for the treatment of osteoarthritis (OA) and induced pluripotent stem cells (iPSCs) are becoming a promising stem cell source. RESULTS: Three steps were developed to differentiate human iPSCs into chondrocytes which were transplanted into rat OA models induced by monosodium iodoacetate (MIA)...
November 9, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27825339/improvement-of-alkalophilicity-of-an-alkaline-xylanase-xyn11a-lc-from-bacillus-sp-sn5-by-random-mutation-and-glu135-saturation-mutagenesis
#11
Wenqin Bai, Yufan Cao, Jun Liu, Qinhong Wang, Zhenhu Jia
BACKGROUND: Family 11 alkaline xylanases have great potential economic applications in the pulp and paper industry. In this study, we would improve the alkalophilicity of family 11 alkaline xylanase Xyn11A-LC from Bacillus sp. SN5, for the better application in this field. RESULTS: A random mutation library of Xyn11A-LC with about 10,000 clones was constructed by error-prone PCR. One mutant, M52-C10 (V116A and E135V), with improved alkalophilicity was obtained from the library...
November 8, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27825338/sequential-and-counter-selectable-cassettes-for-fission-yeast
#12
Hanna Amelina, Vera Moiseeva, Laura Catharine Collopy, Siân Rosanna Pearson, Christine Anne Armstrong, Kazunori Tomita
BACKGROUND: Fission yeast is one of the most commonly used model organisms for studying genetics. For selection of desirable genotypes, antibiotic resistance cassettes are widely integrated into the genome near genes of interest. In yeasts, this is achieved by PCR amplification of the cassette flanked by short homology sequences, which can be incorporated by homology directed repair. However, the currently available cassettes all share the same tef promoter and terminator sequences. It can therefore be challenging to perform multiple genetic modifications by PCR-based targeting, as existing resistance cassettes in strains can be favored for recombination due to shared homology between the cassettes...
November 8, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27793152/improving-the-active-expression-of-transglutaminase-in-streptomyces-lividans-by-promoter-engineering-and-codon-optimization
#13
Song Liu, Miao Wang, Guocheng Du, Jian Chen
BACKGROUND: Transglutaminases (TGase), which are synthesized as a zymogen (pro-TGase) in Streptomyces sp., are important enzymes in the food industry. Because this pro-peptide is essential for the correct folding of Streptomyces TGase, TGase is usually expressed in an inactive pro-TGase form, which is then converted to active TGase by the addition of activating proteases in vitro. In this study, Streptomyces hygroscopicus TGase was actively produced by Streptomyces lividans through promoter engineering and codon optimization...
October 28, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27784303/event-specific-detection-of-transgenic-potato-av43-6-g7-using-real-time-and-digital-pcr-methods
#14
Hongwei Gao, Xiaofan Yu, Tingting Deng, Min Sun, Xizhi Xiao, Xin Huang, Ying Chen, Ronggui Li
BACKGROUND: The isolation of unknown DNA sequences flanked by known sequences is an important task in the event-specific detection of GMOs. None of event-specific detection method was developed based on the junction sequence of an exogenous integrant in the transgenic potato AV43-6-G7. RESULTS: The flanking sequence between the exogenous fragment and recombinant chromosome of this potato was successfully acquired through exogenous gene 5'-RACE. The event-specific primers and Taqman probe were designed to amplify fragments spanning the exogenous DNA and potato genomic DNA...
October 27, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27770795/replacement-of-carbohydrate-binding-modules-improves-acetyl-xylan-esterase-activity-and-its-synergistic-hydrolysis-of-different-substrates-with-xylanase
#15
Shiping Liu, Shaojun Ding
BACKGROUND: Acetylation of the xylan backbone was a major obstacle to enzymatic decomposition. Removal of acetyl groups by acetyl xylan esterases (AXEs) is essential for completely enzymatic hydrolysis of xylan. Appended carbohydrate binding modules (CBMs) can promote the enzymatic deconstruction of plant cell walls by targeting and proximity effects. Fungal acetyl xylan esterases are strictly appended to cellulose-specific CBM1. It is still unclear whether xylan-specific CBMs have a greater advantage than CBM1 in potentiating the activity of fungal deacetylating enzymes and its synergistic hydrolysis of different substrates with xylanase...
October 22, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27756365/plant-expression-of-cocaine-hydrolase-fc-fusion-protein-for-treatment-of-cocaine-abuse
#16
Guojun Wang, Ting Zhang, Haifeng Huang, Shurong Hou, Xiabin Chen, Fang Zheng, Chang-Guo Zhan
BACKGROUND: A recently reported cocaine hydrolase (CocH3) fused with fragment crystallizable (Fc) region of human immunoglobulin G1, denoted as CocH3-Fc, is known as a promising therapeutic candidate for the treatment of cocaine overdose and addiction. A challenge for practical therapeutic use of this enzyme exists in the large-scale protein production and, therefore, it is interesting to identify a low-cost and feasible, sustainable source of CocH3-Fc production. RESULTS: CocH3-Fc was transiently expressed in plant Nicotiana benthamiana leaves...
October 19, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27756363/efficient-biosynthesis-of-ethyl-r-4-chloro-3-hydroxybutyrate-using-a-stereoselective-carbonyl-reductase-from-burkholderia-gladioli
#17
Xiang Chen, Zhi-Qiang Liu, Chao-Ping Lin, Yu-Guo Zheng
BACKGROUND: Ethyl (R)-4-chloro-3-hydroxybutyrate ((R)-CHBE) is a versatile chiral precursor for many pharmaceuticals. Although several biosynthesis strategies have been documented to convert ethyl 4-chloro-3-oxobutanoate (COBE) to (R)-CHBE, the catalytic efficiency and stereoselectivity are still too low to be scaled up for industrial applications. Due to the increasing demand of (R)-CHBE, it is essential to explore more robust biocatalyst capable of preparing (R)-CHBE efficiently. RESULTS: A stereoselective carbonyl reductase toolbox was constructed and employed into the asymmetric reduction of COBE to (R)-CHBE...
October 18, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27756290/evaluating-the-use-of-a-cpg-free-promoter-for-long-term-recombinant-protein-expression-stability-in-chinese-hamster-ovary-cells
#18
Steven C L Ho, Esther Y C Koh, Benjamin P C Soo, Mariati, Sheng-Hao Chao, Yuansheng Yang
BACKGROUND: Methylated CpG dinucleotides in promoters are associated with the loss of gene expression in recombinant Chinese hamster ovary (CHO) cells during large-scale commercial manufacturing. We evaluated a promoter devoid of CpG dinucleotides, CpGfree, in parallel with a similar CpG containing promoter, CpGrich, for their ability to maintain the expression of recombinant enhanced green fluorescent protein (EGFP) after 8 weeks of culturing. RESULTS: While the promoters gave similar transient expression levels, CpGfree clones had significantly higher average stable expression possibly due to increased resistance to early silencing during integration into the chromosome...
October 18, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27717369/overexpression-of-atoxr-gene-improves-abiotic-stresses-tolerance-and-vitamin-c-content-in-arabidopsis-thaliana
#19
Yuanyuan Bu, Bo Sun, Aimin Zhou, Xinxin Zhang, Testuo Takano, Shenkui Liu
BACKGROUND: Abiotic stresses are serious threats to plant growth, productivity and result in crop loss worldwide, reducing average yields of most major crops. Although abiotic stresses might elicit different plant responses, most induce the accumulation of reactive oxygen species (ROS) in plant cells leads to oxidative damage. L-ascorbic acid (AsA, vitamin C) is known as an antioxidant and H2O2-scavenger that defends plants against abiotic stresses. In addition, vitamin C is also an important component of human nutrition that has to be obtained from different foods...
October 7, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27515683/high-efficiency-crispr-cas9-multiplex-gene-editing-using-the-glycine-trna-processing-system-based-strategy-in-maize
#20
Weiwei Qi, Tong Zhu, Zhongrui Tian, Chaobin Li, Wei Zhang, Rentao Song
BACKGROUND: CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice. RESULTS: We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter...
August 11, 2016: BMC Biotechnology
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