Partial characterization and cloning of leuconocin J, a bacteriocin produced by Leuconostoc sp. J2 isolated from the Korean fermented vegetable Kimchi
Partial characterization and cloning of leuconocin J, a bacteriocin produced by Leuconostoc sp. J2 isolated from the Korean fermented vegetable Kimchi
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DOI：
10.1046/j.1365-2672.1999.00471.x
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发表时间：
1999-02-01
影响因子：
4
通讯作者：
Yoon, SS
中科院分区：
文献类型：
--
作者：
Choi, HJ;Lee, HS;Yoon, SS
明串珠菌属J2是从自然发酵泡菜中分离出来的细菌素，命名为明串珠菌素J。该细菌素对多种乳酸菌和一些食源性病原体具有抑制活性。抗菌物质在对数生长后期分泌到培养基中。它似乎是蛋白质，因为它的活性被一系列蛋白水解酶完全灭活，并且它也是相对热稳定的。透析后，通过硫酸铵沉淀部分纯化细菌素。经Tricine-SDS-PAGE活性测定，部分纯化的细菌素的表观分子量为2.5- 3.5kDa。将经EcoRI酶切的明串珠菌J2质粒DNA克隆到pUC 118中，转化大肠杆菌DH 5 α。在携带pULBJ 5.5的转化体中检测到细菌素生产的表型表达。最后，用2.3 lib插入片段作为针对明串珠菌J2的质粒引物的探针进行Southern印迹，表明克隆的外源DNA来自明串珠菌J2。
Leuconostoc sp. J2, isolated from naturally fermented Kimchi, produced a bacteriocin which was named leuconocin J. This bacteriocin exhibited an inhibitory activity against several lactic acid bacteria and some foodborne pathogens. The antimicrobial substance was secreted into the medium during the late log phase. It appears to be proteinaceous since its activity was completely inactivated by a range of proteolytic enzymes, and it was also relatively heat-stable. The bacteriocin was partially purified by ammonium sulphate precipitation, following dialysis. The apparent molecular mass of partially purified bacteriocin, as indicated by activity detection after Tricine-SDS-PAGE, was 2.5-3.5 kDa. Leuconostoc sp. J2 plasmid DNA digested by EcoRI was cloned into pUC118 and transformed into Escherichia coli DH5 alpha. Phenotypic expression of the bacteriocin production was detected in transformants harbouring pULBJ5.5. Finally, Southern blotting with the 2.3 lib insert as a probe against plasmid digests of Leuconostoc sp. J2 revealed that the cloned foreign DNA originated from Leuconostoc sp. J2.