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Molecular chaperones (TrxA, SUMO, Intein, and GST) mediating expression, purification, and antimicrobial activity assays of plectasin in Escherichia coli
Author(s) -
Chen Xin,
Shi Jiawei,
Chen Rui,
Wen Yaoan,
Shi Yu,
Zhu Zhe,
Guo Songwen,
Li Ling
Publication year - 2015
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1303
Subject(s) - intein , escherichia coli , thioredoxin , microbiology and biotechnology , staphylococcus aureus , antimicrobial , biology , fusion protein , defensin , recombinant dna , bacteria , biochemistry , chemistry , enzyme , gene , rna , genetics , rna splicing
Abstract 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 SUMO—plectasin 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.