Expression of plectasin in Bacillus subtilis using SUMO technology by a maltose-inducible vector
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Plectasin, the first fungus defensin, is especially efficient against Gram-positive bacteria. To explore an effective approach for expressing plectasin in Bacillus subtilis, the sequence encoding plectasin fused with the small ubiquitin-like modifier (SUMO) gene, the 6 × His gene and the signal peptide of SacB were cloned into an E. coli–B. subtilis shuttle vector pGJ148 in which the maltose utilization operon promoter Pglv directed the expression. The fusion protein successfully secreted in culture and approximately, 41 mg of the recombinant fusion protein SUMO-plectasin was purified per liter of culture supernatant. After purification by Ni-NTA resin column and digestion by SUMO protease, 5.5 mg of plectasin with a purity of 94 % was obtained from 1 L fermentation culture. Recombinant plectasin was found inhibition activity against S. pneumoniae, S. aureus and S. epidermidis. These results indicate that the maltose-induced expression system may be a safe and efficient way for the large-scale production of soluble peptides in B. subtilis.
KeywordsAntimicrobial peptide Plectasin Bacillus subtilis Maltose-inducible promoter SUMO
We gratefully acknowledge financial support from the National Natural Science Foundation of China (31472104), the National Basic Research Program (2012CB124703), the National Science and Technology Support Program (2013BAD10B03), the China Agriculture Research System (CARS-36), and the Program for Universities in Heilongjiang Province (1254CGZH22).
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