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异源过氧化氢酶表达和超氧化物歧化酶过表达对增强干酪乳杆菌氧化抗性的影响。

The impact of heterologous catalase expression and superoxide dismutase overexpression on enhancing the oxidative resistance in Lactobacillus casei.

作者信息

Lin Jinzhong, Zou Yexia, Cao Kunlin, Ma Chengjie, Chen Zhengjun

机构信息

State Key Laboratory of Dairy Biotechnology, Technology Center of Bright Dairy and Food Co., Ltd., 1518 Jiangchang Road (W), Shanghai, 200436, China.

Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, China.

出版信息

J Ind Microbiol Biotechnol. 2016 May;43(5):703-11. doi: 10.1007/s10295-016-1752-8. Epub 2016 Feb 27.

Abstract

Two heme-dependent catalase genes were amplified from genomic DNA of Lactobacillus plantarum WCFS1 (KatE1) and Lactobacillus brevis ATCC 367 (KatE2), respectively, and a manganese-containing superoxide dismutase from Lactobacillus casei MCJΔ1 (MnSOD) were cloned into plasmid pELX1, yielding pELX1-KatE1, pELX1-KatE2 and pELX1-MnSOD, then the recombinant plasmids were transferred into L. casei MCJΔ1. The strains of L. casei MCJΔ1/pELX1-KatE1 and L. casei MCJΔ1/pELX1-KatE2 were tolerant at 2 mM H2O2. The survival rates of L. casei MCJΔ1/pELX1-KatE1 and L. casei MCJΔ1/pELX1-KatE2 were 270-fold and 300-fold higher than that of the control strain on a short-term H2O2 exposure, and in aerated condition, the survival cells counts were 146- and 190-fold higher than that of the control strain after 96 h of incubation. Furthermore, L. casei MCJΔ1/pELX1-MnSOD was the best in three recombinants which was superior in the living cell viability during storage when co-storage with Lactobacillus delbrueckii subsp. lactis LBCH-1.

摘要

分别从植物乳杆菌WCFS1(KatE1)和短乳杆菌ATCC 367(KatE2)的基因组DNA中扩增出两个血红素依赖性过氧化氢酶基因,并将干酪乳杆菌MCJΔ1的含锰超氧化物歧化酶(MnSOD)克隆到质粒pELX1中,得到pELX1-KatE1、pELX1-KatE2和pELX1-MnSOD,然后将重组质粒转入干酪乳杆菌MCJΔ1。干酪乳杆菌MCJΔ1/pELX1-KatE1和干酪乳杆菌MCJΔ1/pELX1-KatE2菌株对2 mM H2O2具有耐受性。在短期暴露于H2O2的情况下,干酪乳杆菌MCJΔ1/pELX1-KatE1和干酪乳杆菌MCJΔ1/pELX1-KatE2的存活率分别比对照菌株高270倍和300倍,并且在通气条件下,孵育96小时后存活细胞数分别比对照菌株高146倍和190倍。此外,在与德氏乳杆菌保加利亚亚种LBCH-1共同储存期间,干酪乳杆菌MCJΔ1/pELX1-MnSOD在三种重组体中表现最佳,其活细胞活力更强。

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