食品与发酵工业 >

2025 , Vol. 51 >Issue 11: 150 - 162

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.039758

研究报告

三株副干酪乳酪杆菌蛋白酶基因的差异性分析

  • 郝燕萍 ,
  • 刘振民 ,
  • 张欢畅 ,
  • 韩瑨 ,
  • 贾向飞 ,
  • 吴涵清 ,
  • 徐杏敏 ,
  • 郑远荣 ,
  • 吴正钧
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  • 1(上海海洋大学 食品学院, 上海, 201306)
    2(乳业生物技术国家重点实验室, 上海乳业生物工程技术研究中心, 光明乳业股份有限公司乳业研究院, 上海, 200436)
    3(江南大学 食品学院, 江苏 无锡, 214122)
第一作者:硕士研究生(刘振民教授级高级工程师为通信作者,E-mail:liuzhenmin@brightdairy.com)

收稿日期: 2024-05-01

  修回日期: 2024-07-10

  网络出版日期: 2025-06-19

基金资助

十四五国家重点研发计划项目(2022YFD2100704);上海市国资委企业创新发展与能级提升项目(2022013)

收起

Differential analysis of protease genes of three strains of Lacticaseibacillus paracasei

  • HAO Yanping ,
  • LIU Zhenmin ,
  • ZHANG Huanchang ,
  • HAN Jin ,
  • JIA Xiangfei ,
  • WU Hanqing ,
  • XU Xingmin ,
  • ZHENG Yuanrong ,
  • WU Zhengjun
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  • 1(College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China)
    2(State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Center of Dairy Biotechnology, Dairy Research Institute, Bright Dairy&Food Co. Ltd, Shanghai 200436, China)
    3(School of Food Science, Jiangnan University, Wuxi 214122, China)

Received date: 2024-05-01

  Revised date: 2024-07-10

  Online published: 2025-06-19

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摘要

为探索3株副干酪乳酪杆菌BDⅡ、J4、J17蛋白酶基因特征与表型的联系和差异,采用透明圈法和福林法筛选出蛋白酶活力较高的3株副干酪乳酪杆菌,然后对其进行全基因组测序,并结合15株已公开的副干酪乳酪杆菌和约氏乳杆菌进行比较基因组分析,重点研究影响副干酪乳酪杆菌产蛋白酶能力的关键基因。结果显示,BDⅡ、J4、J17的蛋白酶活力相对较高,其中BDⅡ的蛋白酶活力最高,为22.017 U/mL;全基因组测序分析表明3株菌GC含量为46.14%~46.64%,基因组大小为2.3~3.2 Mb。通过CAZys注释,发现BDⅡ含有与细菌细胞壁的合成密切相关的特有蛋白家族GT51(间接参与蛋白水解调控)和特有的肽酶pepX(分解肽),因此推测肽酶pepX和GT51的存在是BDⅡ蛋白酶活力最高的原因。该研究通过表型与基因组结合的方法筛选出1株高产蛋白酶的副干酪乳酪杆菌BDⅡ,阐明了蛋白水解机制和蛋白水解相关基因的调控机制,发现BDⅡ在功能性乳制品中具有巨大的应用潜力。

关键词： 蛋白酶活力; 基因组; 副干酪乳酪杆菌BDⅡ

本文引用格式

郝燕萍 , 刘振民 , 张欢畅 , 韩瑨 , 贾向飞 , 吴涵清 , 徐杏敏 , 郑远荣 , 吴正钧 . 三株副干酪乳酪杆菌蛋白酶基因的差异性分析[J]. 食品与发酵工业, 2025 , 51(11) : 150 -162 . DOI: 10.13995/j.cnki.11-1802/ts.039758

Abstract

To explore the linkage and differences between the protease gene characteristics and phenotypes of three strains of Lacticaseibacillus paracasei BDⅡ, J4, and J17, three strains of Lacticaseibacillus paracasei with high protease viability were screened by using the clear circle method and the Folin method, and then they were subjected to whole-genome sequencing, and combined with the comparative genome analyses of 15 publicly available strains of Lacticaseibacillus paracasei and Lactobacillus johannes, with emphasis on key genes affecting the protease-producing ability of Lacticaseibacillus paracasei.Results showed that BDⅡ, J4, and J17 had relatively high protease activity, with BDⅡ having the highest protease activity of 22.017 U/mL.Whole-genome sequencing analysis indicated that the GC content of the three strains ranged from 46.14% to 46.64%, and the genome sizes were located between 2.3 Mb and 3.2 Mb.By CAZys annotation, BDⅡ was found to contain the unique protein family GT51 (indirectly involved in the regulation of protein hydrolysis) and the unique peptidase pepX (catabolic peptide), which were closely related to the synthesis of the bacterial cell wall, and therefore it was hypothesized that the presence of peptidases pepX and GT51 was responsible for the highest protease activity of BDⅡ.This study screened a strain of Lacticaseibacillus paracasei BDⅡ with high protease production by a combined phenotypic and genomic approach, elucidated the mechanism of protein hydrolysis and the regulation of protein hydrolysis-related genes, and found that BDⅡ has a great potential for application in functional dairy products.

Key words： protease activity; genome; Lacticaseibacillus paracasei BDII

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