食品科学2025，Vol.46Issue(3)：83-89,7.DOI:10.7506/spkx1002-6630-20240513-101
副溶血性弧菌噬菌体vB_VpP_1裂解酶的生物信息学分析、原核表达及生物活性鉴定
Bioinformatics Analysis,Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1
摘要
Abstract
To study the in vitro cleavage effect of a recombinant lysin from bacteriophage vB_VpP_1 on Vibrio parahaemolyticus,the gp32 gene fragment of vB_VpP_1 was identified as a lysin gene according to the results of whole-genome sequencing(WGS)and functional analysis.The amino acid sequence and structure of gp32 were analyzed by various tools such as Expasy.Primers were designed using Primer 5.0 software and cloned into the pET-28a(+)vector for prokaryotic expression.The lytic activity of the purified lysin against the host bacterium before and after being treated with ethylene diamine tetraacetic acid(EDTA)was measured.Tertiary structure analysis showed that the lysin was a spherical hydrophilic protein,which was predicted to contain two catalytic domains.This was consistent with the basic characteristics of Gram-negative phage lysins.The lysin had no transmembrane region or signal peptide.The purified bacteriophage vB_VpP_1 had a lysin activity of approximately(1 487±182)U/mg,which showed a strong capacity to lyse V.parahaemolyticus with damaged cell walls but not V.parahaemolyticus with intact cell walls.The prokaryotic expression vector for bacteriophage vB_VpP_1 lysin was successfully constructed,and the expressed and purified lysin could lyse V.parahaemolyticus with damaged cell walls.关键词
副溶血性弧菌/噬菌体/裂解酶/生物抗菌剂/食品安全Key words
Vibrio parahaemolyticus/bacteriophage/lysin/biocontrol agents/food safety分类
轻工业引用本文复制引用
张德福,李学鹏,励建荣,杨雯静,刘可,刘青青,白梧桐,李凡,吕欣然,柏雪,檀茜倩..副溶血性弧菌噬菌体vB_VpP_1裂解酶的生物信息学分析、原核表达及生物活性鉴定[J].食品科学,2025,46(3):83-89,7.基金项目
渤海大学海洋研究院项目(BDHYYJY2024003) （BDHYYJY2024003）
渤海大学食品科学与工程学院预研项目 （）
