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屎肠球菌JCM 5804T产生的肠球菌素的特性与鉴定

Characteristics and identification of enterocins produced by Enterococcus faecium JCM 5804T.

作者信息

Park S H, Itoh K, Fujisawa T

机构信息

Laboratory of Veterinary Public Health, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

出版信息

J Appl Microbiol. 2003;95(2):294-300. doi: 10.1046/j.1365-2672.2003.01975.x.

Abstract

AIMS

To screen bacteriocin-producing lactic acid bacteria (LAB) in 52 type and reference strains, which have not previously been studied, with respect to bacteriocins, and to characterize the presence of bacteriocins.

METHODS AND RESULTS

Only Enterococcus faecium JCM 5804T showed bacteriocin-like activity. It inhibited the growth of Lactobacillus spp., Enterococcus spp., Clostridium spp., Listeria monocytogenes, and vancomycin resistant Enterococcus (VRE). However, it was not effective against Gram-negative strains, Weisella spp., Leuconostoc spp., Lactococcus spp., or methicillin resistant Staphylococcus aureus (MRSA). The inhibitory activity of Ent. faecium JCM 5804T was inactivated by proteinase K, trypsin, alpha-chymotrypsin, and papain, but not by lysozyme, lipase, catalase, or beta-glucosidase. The inhibitory activity was stable at 100 degrees C for 30 min, and had a pH range from 2 to 10. The molecular weight of the partially purified bacteriocin(s) was approx. 4.5 kDa, according to tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Polymerase chain reaction and direct sequencing methods identified three different types of bacteriocins produced by Ent. faecium JCM 5804T, enterocin A, enterocin B, and enterocin P-like bacteriocin.

CONCLUSION

Enterococcus faecium JCM 5804T produced three different types of bacteriocins, and they inhibited LAB and pathogens.

SIGNIFICANCE AND IMPACT OF STUDY

This is the first report of enterocin A, enterocin B, and enterocin P-like bacteriocin, detected in Ent. faecium JCM 5804T among LAB type and reference strains.

摘要

目的

在52株此前未针对细菌素进行过研究的典型菌株和参考菌株中筛选产细菌素的乳酸菌(LAB),并对细菌素的存在情况进行表征。

方法与结果

仅粪肠球菌JCM 5804T表现出类细菌素活性。它抑制了乳酸杆菌属、肠球菌属、梭菌属、单核细胞增生李斯特菌和耐万古霉素肠球菌(VRE)的生长。然而,它对革兰氏阴性菌株、魏斯氏菌属、明串珠菌属、乳球菌属或耐甲氧西林金黄色葡萄球菌(MRSA)无效。粪肠球菌JCM 5804T的抑制活性被蛋白酶K、胰蛋白酶、α-胰凝乳蛋白酶和木瓜蛋白酶灭活,但未被溶菌酶、脂肪酶、过氧化氢酶或β-葡萄糖苷酶灭活。抑制活性在100℃下30分钟稳定,pH范围为2至10。根据十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,部分纯化的细菌素分子量约为4.5 kDa。聚合酶链反应和直接测序方法鉴定出粪肠球菌JCM 5804T产生的三种不同类型的细菌素,即肠球菌素A、肠球菌素B和类肠球菌素P。

结论

粪肠球菌JCM 5804T产生三种不同类型的细菌素,它们抑制乳酸菌和病原体。

研究的意义和影响

这是在LAB典型菌株和参考菌株中的粪肠球菌JCM 5804T中检测到肠球菌素A、肠球菌素B和类肠球菌素P的首次报道。

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