期刊
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
卷 62, 期 5, 页码 606-614出版社
WILEY-BLACKWELL
DOI: 10.1002/bab.1303
关键词
cleavage; expression; molecular chaperone; plectasin; purification
资金
- Combinational Project of Guangdong Province and Education Ministry for Manufacturing, Education Research [2010B090400540]
Plectasin (PS) is the first defensin to be isolated from a fungus, the saprophytic ascomycete Pseudoplectania nigrella, and active against Streptococcus pneumoniae and S. aureus, including antibiotic-resistant pathogens. To establish a bacterium-based production system, we compared the efficiency of four molecular chaperones and corresponding cleavage to the expression and purification of plectasin. The results showed that the yield of plectasin combined with thioredoxin A (TrxA) and small ubiquitin-related modifier (SUMO) was at a higher level (0.0356 and 0.0358 g L-1, respectively) than that with intein (0.0238 g L-1) and glutathione-S-transferase (GST) (0.0243 g L-1). TrxA-plectasin, SUMO-plectasin, and 2-plectasin were cleaved at the correct site and purified, but their considerable amount was not cleaved and remained as a fusion peptide. The antimicrobial activity of plectasin cleaved from SUMOplectasin against methicillin-resistant Staphylococcus aureus (MRSA), penicillin-resistant S. pneumoniae (PRSP), and vancomycin-resistant enterococci (VRE)was stronger than ampicillin (Amp) for the same amount of substance (P 0.05). This is the first study to complete and compare the effect of different molecular chaperones and corresponding cleavage with the expression and purification of plectasin in the Escherichia coli expression system, which laid the foundation for future research and may develop the application and production of plectasin. (C) 2014 International Union of Biochemistry and Molecular Biology, Inc.
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