一株产细菌素乳杆菌的鉴定及其细菌素编码基因的获得

应用双层平板打孔法，从自制樱桃酒里分离的乳杆菌中筛选到一株对革兰氏阳性细菌和革兰氏阴性细菌均有明显抑制作用的菌株，命名为LD 1.0008。通过API 50 CHL糖发酵产酸实验、16S rDNA基因序列分析及其特异性recA基因多重聚合酶链式反应（polymerase chain reaction，PCR），鉴定LD 1.0008为植物乳杆菌。排除有机酸的干扰，用多种蛋白酶处理后，其抑菌活性均有明显降低，而用过氧化氢酶处理后其抑菌活性基本不变，从而确定LD 1.0008所产生的抑菌物质为细菌素。使用已报道的10 对植物乳杆菌细菌素基因片段设计的引物对菌株LD 1.0008进行PCR扩增，发现其至少含有4 个植物乳杆菌素相关编码基因，即plnD、plnO、plnV和plnW。

Identification of a Bacteriocin-Producing Lactobacillus and Acquisition of Bacteriocin-Encoding Genes

One Lactobacillus strain isolated from homemade cherry wine revealed significant inhibitory activity
against Gram-positive and Gram-negative bacteria using double plate punching method, which was named LD 1.0008.
LD 1.0008 was identified as Lactobacillus plantarum by its carbohydrate fermentation pattern using API 50 CHL test kits,
16S rDNA sequence analysis and recA-based polymerase chain reaction (PCR) test. The antibacterial activity was
significantly decreased after protease treatment, whereas it remained stable after treatment with catalase. The antibacterial
substances were confirmed as bacteriocin after excluding the interference of organic acid. PCR amplification was carried out
using previously reported 10-pair plantaricin primers. It was indicated that at least 4 bacteriocin-encoding genes could be
produced by LD 1.0008, including plnD, plnO, plnV and plnW. It is of great significance for the separation and application of
plantaricins of LD 1.0008.