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食品与发酵工业  2020, Vol. 46 Issue (17): 33-39    DOI: 10.13995/j.cnki.11-1802/ts.023143
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
丁亮亮1, 刘进生1, 顾鹏帅1, 唐蕾1,2*
1(工业生物技术教育部重点实验室(江南大学),江苏 无锡,214122)
2(江南大学 生物工程学院,江苏 无锡,214122)
The role of peroxidase EfeB in Escherichia coli under cell oxidative stress
DING Liangliang1, LIU Jinsheng1, GU Pengshuai1, TANG Lei1,2*
1(Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China)
2(School of Biotechnology, Jiangnan University, Wuxi 214122, China)
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摘要 为了阐明过氧化物酶EfeB对大肠杆菌胞内氧化应激调控的影响,对编码基因efeB进行了同源重组敲除及同源过表达,考察了菌体生长、丙二醛浓度及胞内活性氧水平的变化,并进一步比较了efeB过表达菌株中与氧化胁迫相关基因efeUefeOsodAoxyRkatE和细菌重组修复基因recA在正常和过氧化氢刺激下的表达水平变化。结果表明:过表达efeB对菌体生长有促进作用,过表达菌株Eco/pEE胞内的丙二醛含量相比出发菌株(223.79 nmol/g蛋白)下降了85%,胞内活性氧是出发菌株的1.36倍,同时katE下调了87.62%,efeUefeOsodAoxyRrecA分别上调3.88、1.33、2.48、1.95和1.49倍,过表达菌株Eco/pEE在过氧化氢刺激条件下,katE进一步下调了94.81%,recA进一步上调了8.09倍;反之,菌株缺失efeB生长减缓,胞内丙二醛的含量是出发菌株的1.15倍,胞内活性氧是出发菌株的0.5倍。以上结果提出了大肠杆菌EfeB在细胞氧化应激下新的生理功能。
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关键词:  氧化应激  efeB  大肠杆菌  过氧化氢  丙二醛  活性氧    
Abstract: In order to elucidate the effect of the peroxidase EfeB on the regulation of intracellular oxidative stress in Escherichia coli, Red recombination/disruption and over-expression of the efeB gene were performed. The results showed that after over-expression of efeB, the content of malondialdehyde in the cell was decreased by 85% compared with the parental strain (223.79 nmol/g protein), and the level of intracellular reactive oxygen species was 1.36 folds compared with that of the parental strain. Meanwhile, the expression level of katE was decreased 87.62%, while efeU, efeO, sodA, oxyR and recA up-regulated by 3.88, 1.33, 2.48, 1.95 and 1.49 folds respectively. After exogenous addition of H2O2, katE was further down-regulated by 94.81% while recA was further up-regulated by 8.09 folds in the efeB over-expressing strain Eco/pEE. Conversely, efeB-deficient strain exhibited slower growth rate with higher intracellular malondialdehyde content (1.15-fold of the parental strain). The level of intracellular reactive oxygen species in efeB-deficient strain was 0.5-fold of the parental strain. The results provided new physiological function of EfeB in E.coli under oxidative stress.
Key words:  oxidative stress    efeB    Escherichia coli    hydrogen peroxide    malondialdehyde    reactive oxygen species
收稿日期:  2019-12-18      修回日期:  2020-03-24           出版日期:  2020-09-15      发布日期:  2020-10-14      期的出版日期:  2020-09-15
基金资助: 111引智计划项目(111-2-06);国家轻工技术与工程一流学科自主课题项目(LITE2018-27);江苏省现代工业发酵协同创新中心资助项目(BY2013015-11)
作者简介:  硕士研究生(唐蕾教授为通讯作者,
丁亮亮,刘进生,顾鹏帅,等. 大肠杆菌过氧化物酶EfeB在细胞氧化应激中的作用[J]. 食品与发酵工业, 2020, 46(17): 33-39.
DING Liangliang,LIU Jinsheng,GU Pengshuai,et al. The role of peroxidase EfeB in Escherichia coli under cell oxidative stress[J]. Food and Fermentation Industries, 2020, 46(17): 33-39.
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