title: Expression of plectasin in Bacillus subtilis using SUMO technology by a maltose-inducible vector.
abstract: 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.
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Licong
authors: L
authors: Zhang
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Xiaodan
authors: X
authors: Li
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Dandan
authors: D
authors: Wei
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Jue
authors: J
authors: Wang
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China. asshan@neau.edu.cn.
authors: Anshan
authors: A
authors: Shan
authors: Institute of Animal Nutrition, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Zhongyu
authors: Z
authors: Li
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D008320:Maltose / Q000378:metabolism / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D010447:Peptide Hydrolases / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics* / Q000378:metabolism; D012995:Solubility; D013210:Staphylococcus / Q000187:drug effects; D013296:Streptococcus pneumoniae / Q000187:drug effects
chemical_list: D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; C504857:plectasin; D008320:Maltose; D010447:Peptide Hydrolases
keywords: Antimicrobial peptide; Bacillus subtilis; Maltose-inducible promoter; Plectasin; SUMO