食品与发酵工业 >

2020 , Vol. 46 >Issue 1: 43 - 49

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.022753

研究报告

氮离子注入选育高自溶干酪乳杆菌及其肽聚糖水解酶的RT-qPCR分析

  • 崔文明 ,
  • 吕加平 ,
  • 王小鹏 ,
  • 赵莉君 ,
  • 祝超智 ,
  • 张秋会 ,
  • 李苗云 ,
  • 赵改名
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  • 1(河南农业大学 食品科学技术学院,河南 郑州,450002)
    2(农业部农产品加工与质量控制重点开放实验室,中国农业科学院 农产品加工研究所,北京,100193)
博士,讲师(本文通讯作者,E-mail:wenmingcui@126.com)

收稿日期: 2019-07-22

  网络出版日期: 2020-03-27

基金资助

河南省高等学校重点科研项目计划(16A550010)

收起

Real-time quantitative PCR analysis of peptidoglycan hydrolases relative expression of high autolysis Lactobacillus casei mutant induced by nitrogen ion implantation

  • CUI Wenming ,
  • LYU Jiaping ,
  • WANG Xiaopeng ,
  • ZHAO Lijun ,
  • ZHU Chaozhi ,
  • ZHANG Qiuhui ,
  • LI Miaoyun ,
  • ZHAO Gaiming
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  • 1(College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China)
    2 (Key Laboratory of Agro-product Processing and Quality Control, Institute of Agro-food Science and Technology, Chinses Academy of Agricultural Science, Beijing 100193, China)

Received date: 2019-07-22

  Online published: 2020-03-27

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摘要

通过采用低能N+注入对干酪乳杆菌116-a进行诱变,以筛选得到高自溶的干酪乳酸菌突变体,并采用实时荧光定量PCR检测N-乙酰胞壁质酶和N-乙酰氨基葡糖胺糖苷酶在野生菌体和突变体中的表达差异。结果显示:经过离子能量30 keV N+注入诱变,在注入剂量0.5×1015 ions/cm2时,筛选得到遗传稳定性良好的最大正突变菌株,其自溶度为56.3%,最大正突变菌株自溶度比野生型菌株的自溶度提高了25.7%。实时荧光定量PCR检测发现,N-乙酰氨基葡糖苷酶和N-乙酰胞壁质酶在突变体中的表达量比野生菌体中的表达量均有所上调,其中N-乙酰氨基葡糖苷酶表达量上调了约55倍,而N-乙酰胞壁质酶的表达量仅上调2.8倍。因此,N-乙酰氨基葡糖苷酶在干酪乳杆菌116-a自溶中起着重要作用,该酶表达量的上调可以有效提高菌体的自溶。

关键词： 干酪乳酸菌; 自溶; 离子注入; 肽聚糖水解酶

本文引用格式

崔文明 , 吕加平 , 王小鹏 , 赵莉君 , 祝超智 , 张秋会 , 李苗云 , 赵改名 . 氮离子注入选育高自溶干酪乳杆菌及其肽聚糖水解酶的RT-qPCR分析[J]. 食品与发酵工业, 2020 , 46(1) : 43 -49 . DOI: 10.13995/j.cnki.11-1802/ts.022753

Abstract

Lactobacillus casei 116-a was induced by low energy nitrogen ion implantation to screen the high autolysis mutant and investigate differential expression of N-acetylmuramidase and N-acetylglucosaminidase between wild type strain and mutant. The results indicated that a high autolysis mutant with good genetic stability was obtained by N+ implantation with a dose of 0.5×1015 ions/cm2 under 30 keV ion energy. By comparison with the autolysis rate of the wild type strain, the autolysis rate of mutant was increased by 25.7% and up to 56.3%. Quantitative real-time PCR analysis of the expression of N-acetylglucosaminidase and N-acetylmuramidase between mutant and wild type strain revealed the expression of N-acetylglucosaminidase of mutant was up-regulated by about 55 times while the expression of N-acetylmuramidase of mutant was up-regulated by only 2.8 times. Therefore, N-acetylglucosaminidase plays an important role in Lactobacillus casei 116-a autolysis and its gene expression upregulation can effectively improve cell autolysis.

Key words： Lactobacillus casei; autolysis; ion implantation; peptidoglycan hydrolase

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