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Large-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli.

Byline: Minjing Zhang (1,2,4), Xuebin Jiang (3), Zhijian Su (1,2,4), Jiancong Lin (3), Qi Xiang (2), Zhan Yang (2), Yadong Huang (1,2), Xiaokun Li (4) Keywords: Human fibroblast growth factor 21; Expression; Purification; Glucose uptake activity Abstract: As a novel important regulator of glucose an... Full description

Journal Title: Applied microbiology and biotechnology January 2012, Vol.93(2), pp.613-621
Main Author: Zhang, Minjing
Other Authors: Jiang, Xuebin , Su, Zhijian , Lin, Jiancong , Xiang, Qi , Yang, Zhan , Huang, Yadong , Li, Xiaokun
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-011-3427-8
Link: http://search.proquest.com/docview/916151250/?pq-origsite=primo
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title: Large-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli.
format: Article
creator:
  • Zhang, Minjing
  • Jiang, Xuebin
  • Su, Zhijian
  • Lin, Jiancong
  • Xiang, Qi
  • Yang, Zhan
  • Huang, Yadong
  • Li, Xiaokun
subjects:
  • Bioreactors–Methods
  • Biotechnology–Methods
  • Chromatography, Liquid–Genetics
  • Cloning, Molecular–Metabolism
  • Escherichia Coli–Genetics
  • Fibroblast Growth Factors–Isolation & Purification
  • Gene Expression–Metabolism
  • Genes, Synthetic–Metabolism
  • Genetic Vectors–Genetics
  • Glucose–Isolation & Purification
  • Humans–Metabolism
  • Plasmids–Metabolism
  • Recombinant Proteins–Metabolism
  • Recombinant Proteins
  • Fibroblast Growth Factor 21
  • Fibroblast Growth Factors
  • Glucose
ispartof: Applied microbiology and biotechnology, January 2012, Vol.93(2), pp.613-621
description: Byline: Minjing Zhang (1,2,4), Xuebin Jiang (3), Zhijian Su (1,2,4), Jiancong Lin (3), Qi Xiang (2), Zhan Yang (2), Yadong Huang (1,2), Xiaokun Li (4) Keywords: Human fibroblast growth factor 21; Expression; Purification; Glucose uptake activity Abstract: As a novel important regulator of glucose and lipid metabolism homeostasis, human fibroblast growth factor 21 (hFGF21) has become a potential drug candidate for the treatment of metabolic diseases including obesity, and type 2 diabetes, as well as non-alcoholic fatty liver disease. To improve the production of recombinant hFGF21 to meet the increasing demand in clinical applications, an artificial gene encoding its mature peptide sequence was constructed, cloned into vector pET-3c and then expressed in Escherichia coli Origami B (DE3). Under optimal conditions in a 50-L fermentor, the average bacterial yield and the soluble expression level of recombinant hFGF21 of six batches attained 1750+-185 g and 32+-1.5%, respectively. The target protein was purified by the combination of nickel-nitrilotriacetic acid affinity chromatography and Sephadex S-100 resin. 5% (w/v) trehalose solution was able to prevent rhFGF21 from degradation effectively. The purity of rhFGF21 was higher than 97%, and the yield was 213+-17 mg/L. The preliminary biochemical characterization of rhFGF21 was confirmed using Western blot and peptide map finger analysis. Based on the glucose oxidase--peroxidase assay, the EC50 of glucose uptake activity of the purified rhFGF21 was 22.1 nM. Author Affiliation: (1) Institute of Life and Health Engineering, and National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, 510632, People's Republic of China (2) Guangdong Provincial Key Laboratory of Bioengineering Medicine, Jinan University, Guangzhou, 510632, People's Republic of China (3) School of Life Sciences, Guangzhou University, Guangzhou, 510006, People's Republic of China (4) Wenzhou Medical College, Wenzhou, 325035, People's Republic of China Article History: Registration Date: 13/06/2011 Received Date: 12/04/2011 Accepted Date: 03/06/2011 Online Date: 29/06/2011 Article note: Authors Minjing Zhang and Xuebin Jiang contributed equally to this study.
language: eng
source:
identifier: E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-011-3427-8
fulltext: fulltext
issn:
  • 14320614
  • 1432-0614
url: Link


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titleLarge-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli.
creatorZhang, Minjing ; Jiang, Xuebin ; Su, Zhijian ; Lin, Jiancong ; Xiang, Qi ; Yang, Zhan ; Huang, Yadong ; Li, Xiaokun
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identifierE-ISSN: 1432-0614 ; DOI: 10.1007/s00253-011-3427-8
subjectBioreactors–Methods ; Biotechnology–Methods ; Chromatography, Liquid–Genetics ; Cloning, Molecular–Metabolism ; Escherichia Coli–Genetics ; Fibroblast Growth Factors–Isolation & Purification ; Gene Expression–Metabolism ; Genes, Synthetic–Metabolism ; Genetic Vectors–Genetics ; Glucose–Isolation & Purification ; Humans–Metabolism ; Plasmids–Metabolism ; Recombinant Proteins–Metabolism ; Recombinant Proteins ; Fibroblast Growth Factor 21 ; Fibroblast Growth Factors ; Glucose
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titleLarge-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli.
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