一株布氏乳杆菌所产类细菌素的初步纯化与部分特性

Purification and Biological Characteristics of Bacteriocin-like Substance Produced by Lactobacillus buchneri KLDS1.0364

从分离自内蒙古传统乳制品的67株乳酸菌中筛选得到一株产生类细菌素的布氏乳杆菌KLDS1.0364,对其所产类细菌素进行初步分离纯化,同时研究其所产类细菌素的生物学特性。KLDS1.0364无细胞发酵上清液经阳离子交换树脂纯化后,采用tricine-SDS-PAGE测定类细菌素分子量,并测定了类细菌素的部分特性。KLDS1.0364产生的类细菌素分子量约为21.6kD,对热和pH值稳定,可被多种蛋白酶失活,不能被过氧化氢酶和α-淀粉酶失活。KLDS1.0364产生的类细菌素的作用方式是杀菌,且抑菌谱广,可抑制多种革兰氏阳性菌、革兰氏阴性菌和真菌。

The purpose of this study is to obtain effective and stable bacteriocin-like substance from lactic acid bacteria. Lactobacillus buchneri KLDS 1.0364, which was isolated from fermented cream, a traditional dairy product in Inner Mongolia, could produce bacteriocin-like substance. The bacteriocin-like substance produced by KLDS 1.0364 was partially purified and the characteristics were studied. The bacteriocin-like substance was purified by SP-Sepharose fast flow cation exchange chromatography. The molecular weight of the bacteriocin-like substance was 21.6kD, as determined by tricine-SDS-PAGE. The bacteriocin-like substance remained stable after 30 min at 121 ℃ and after 2 h of incubation during pH 2-10. Complete inactivation of antimicrobial activity was observed after treatment of the bacteriocin-like substance with several proteinases. Treatment with catalase and a-amylase did not result in any changes of antimicrobial activity. The mode of action of the bacteriocin-like substance is bactericidal. It exhibited a broad spectrum of antago- nistic activity against various species of Gram-positive bacteria, Gram-negative bacteria and fungi.