Document Detail

Expression of soluble recombinant transglutaminase from Zea mays in Pichia pastoris.
MedLine Citation:
PMID:  23296916     Owner:  NLM     Status:  Publisher    
Transglutaminases (TGases) catalyze post-translational protein modifications by ε-(γ-glutamyl) links and covalent amide bonds. In plant, this enzyme is poorly studied and only the Zea mays TGase gene (tgz) has been cloned. The tgz had been expressed in Escherichia coli, but the recombinant protein was mainly present in inclusion bodies. Therefore, to obtain active, soluble protein, we optimized its coding sequence according to the codon bias of Pichia pastoris and synthesized the sequence with SOEing-PCR. The optimized fragment was successfully transformed into P. pastoris GS115 by electroporation. The optimal conditions for expression were under a final concentration of 0.5 % methanol and a time-course of 96 h. The synthesized recombinant Zea mays transglutaminase (TGZs) was purified by affinity method, its production was 4.4 mg/L, and the specific activity was 0.889 U/mg under optimal expression condition. Optimal activity for TGZs was observed at 37 °C and a pH of 8.0, respectively. The cross-linking reaction of TGZs to the casein was studied, and the result was same as the reaction of casein by microbial transglutaminase. These results indicated that an effective procedure for expressing and purifying TGZs in P. pastoris GS115 was established.
Hongbo Li; Lanwei Zhang; Yanhua Cui; Xue Luo; Chaohui Xue; Shumei Wang
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-1-8
Journal Detail:
Title:  World journal of microbiology & biotechnology     Volume:  -     ISSN:  1573-0972     ISO Abbreviation:  World J. Microbiol. Biotechnol.     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-1-8     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9012472     Medline TA:  World J Microbiol Biotechnol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
School of Food Science and Engineering, Harbin Institute of Technology, Harbin, 150090, China.
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