National Engineering Laboratory for High efficiency Enzyme Expression

Fuzhou, China

National Engineering Laboratory for High efficiency Enzyme Expression

Fuzhou, China

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Li R.-K.,Fuzhou University | Li R.-K.,National Engineering Laboratory for High efficiency Enzyme Expression | Fu C.-L.,Fuzhou University | Chen P.,National Engineering Laboratory for High efficiency Enzyme Expression | And 3 more authors.
Environmental Toxicology and Pharmacology | Year: 2013

Production of a sika deer Cu/Zn-SOD was achieved in Pichia pastoris after the reconstituted expression vector pPIC9K was transformed into the strain GS115. By employing Saccharomyces cerevisiae secretion signal peptide (α-factor) under the regulation of the methanol-inducible promoter of the gene of alcohol oxidase 1 (AOX1), sika deer Cu/Zn-SOD with a molecular mass of 16. kDa was expressed while recombinant sika deer Cu/Zn-SOD with an activity of 3500. U/mL was obtained from a 5. L bioreactor. After two successive steps of chromatography on DEAE-650. C and Superdex75, recombinant sika deer Cu/Zn-SOD was obtained with 13.8% yield, 14.5-fold purification, and a specific activity of 3447. U/mg. Its optimum temperature and optimum pH were 40°C and 7.0, respectively. © 2012.


Wang G.,Fuzhou University | Wang G.,National Engineering Laboratory for High efficiency Enzyme Expression | Huang X.,Fuzhou University | Ng T.B.,Chinese University of Hong Kong | And 4 more authors.
PLoS ONE | Year: 2014

Soda lakes are one of the most stable naturally occurring alkaline and saline environments, which harbor abundant microorganisms with diverse functions. In this study, culture-independent molecular methods were used to explore the genetic diversity of glycoside hydrolase (GH) family 10 and GH11 xylanases in Lake Dabusu, a soda lake with a pH value of 10.2 and salinity of 10.1%. A total of 671 xylanase gene fragments were obtained, representing 78 distinct GH10 and 28 GH11 gene fragments respectively, with most of them having low homology with known sequences. Phylogenetic analysis revealed that the GH10 xylanase sequences mainly belonged to Bacteroidetes, Proteobacteria, Actinobacteria, Firmicutes and Verrucomicrobia, while the GH11 sequences mainly consisted of Actinobacteria, Firmicutes and Fungi. A full-length GH10 xylanase gene (xynAS10-66) was directly cloned and expressed in Escherichia coli, and the recombinant enzymes showed high activity at alkaline pH. These results suggest that xylanase gene diversity within Lake Dabusu is high and that most of the identified genes might be novel, indicating great potential for applications in industry and agriculture. Copyright: © 2014 Wang et al.


Chen P.,National Engineering Laboratory for High efficiency Enzyme Expression | Fu X.,National Engineering Laboratory for High efficiency Enzyme Expression | Ng T.B.,Chinese University of Hong Kong | Ye X.-Y.,National Engineering Laboratory for High efficiency Enzyme Expression | Ye X.-Y.,Fuzhou University
Biotechnology Letters | Year: 2011

A β-glucosidase gene (bglI) from Trichoderma reesei was cloned into the pPIC9 vector and integrated into the genome of Pichia pastoris GS115. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter and using Saccharomycescerevisiae secretory signal peptide (α-factor), the recombinant β-glucosidase was expressed and secreted into the culture medium. The maximum recombinant β-glucosidase activity achieved was 60 U/ml, and β-glucosidase expression reached 0.3 mg/ml. The recombinant 76 kDa β-glucosidase was purified 1.8-fold with 26% yield and a specific activity of 197 U/mg. It was optimally active at 70°C and pH 5.0. © 2011 Springer Science+Business Media B.V.


PubMed | National Engineering Laboratory for High efficiency Enzyme Expression
Type: Journal Article | Journal: Biotechnology letters | Year: 2011

A -glucosidase gene (bglI) from Trichoderma reesei was cloned into the pPIC9 vector and integrated into the genome of Pichia pastoris GS115. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter and using Saccharomyces cerevisiae secretory signal peptide (-factor), the recombinant -glucosidase was expressed and secreted into the culture medium. The maximum recombinant -glucosidase activity achieved was 60 U/ml, and -glucosidase expression reached 0.3 mg/ml. The recombinant 76 kDa -glucosidase was purified 1.8-fold with 26% yield and a specific activity of 197 U/mg. It was optimally active at 70 C and pH 5.0.

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