Purification and Functional Analysis of Recombinant Nisin Resistance Protein (NSR) Expressed in Escherichia coli
- VernacularTitle:大肠杆菌重组乳链菌肽抗性蛋白(NSR)的表达纯化及其功能分析
- Author:
Jiale LIU
;
Zhizeng SUN
;
Yiwei LIU
;
Xueling GAO
;
Jin ZHONG
- Publication Type:Journal Article
- Keywords:
Nisin, Nisin resistance protein, Tail-specific protease, GST fusion protein
- From:
Microbiology
2008;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Nisin is a cationic antimicrobial peptide produced by some lactic acid bacteria. However, expression of nisin resistance protein (NSR) could confer nisin resistance on some non-nisin-producing Lactococcus lactis. To deeply elucidate molecular mechanism underlying NSR-mediated nisin resistance, an NSR mutant with N-terminal 38 amino acid residues deleted (NSR?38) was overexpressed in Escherichia coli by fusion with GST. Purified NSR?38 was obtained through glutathione (GSH) affinity chromatography followed by cleavage of GST tag. Putative proteolytic activity of NSR?38 was determined in vitro against nisin. Antimicrobial activity analysis revealed that nisin lost its bactericidal activity after incubation with NSR?38. Further reversed-phase high performance liquid chromatography (RP-HPLC) analysis indicated that NSR?38 displayed proteolytic activity against nisin, thus inactivating the antimicrobial peptide. The current study paves the way for in-depth functional studies on NSR.
