铁蛋白DrfE对耐辐射异常球菌抗氧化酶活性的影响

doi:10.13560/j.cnki.biotech.bull.1985.2017.02.024

生物技术通报 ›› 2017, Vol. 33 ›› Issue (2): 164-171.doi: 10.13560/j.cnki.biotech.bull.1985.2017.02.024

铁蛋白DrfE对耐辐射异常球菌抗氧化酶活性的影响

吴小丽^1,2, 刘盈盈², 江世杰^2,3, 陈云², 刘小利^1,2, 汪雨舟⁴, 平淑珍², 王劲^1,2

1. 西南科技大学生命科学与工程学院,绵阳 621000;
2. 中国农业科学院生物技术研究所,北京 100081;
3. 四川大学生命科学学院,成都 610065;
4. 中国人民大学附属中学,北京 100080

收稿日期:2016-11-07 出版日期:2017-02-26 发布日期:2017-02-08
作者简介:吴小丽,女,硕士研究生,研究方向：特殊环境微生物功能基因资源利用;E-mail：wuxiaoli513@126.com
基金资助:
国家重点基础研究发展计划（“973计划”）（2013CB733903）

Effect of Ferritin DrfE on Antioxidant Enzyme Activity in Deinococcus radiodurans

WU Xiao-li^1,2, LIU Ying-ying², JIANG Shi-jie^2,3, CHEN Yun², LIU Xiao-li^1,2, WANG Yu-zhou⁴, PING Shu-zhen², WANG Jin^1,2

1. College of Life Science and Engineering,Southwest University of Science and Technology,Mianyang 621000;
2. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081;
3. College of Life Sciences,Sichuan University,Chengdu 610065;
4. The High School Affiliated to Renmin University of China,Beijing 100080

Received:2016-11-07 Published:2017-02-26 Online:2017-02-08

摘要/Abstract

摘要： 为研究耐辐射异常球菌（Deinococcus radiodurans）中drfE编码的铁蛋白DrfE的功能,通过基因回补试验构建drfE基因的回补菌株（pdrfE∷Δ）,对野生型WT、突变株ΔdrfE及回补菌株pdrfE∷Δ进行不同浓度H₂O₂和NaCl胁迫,分别测定野生型WT、突变株ΔdrfE和回补菌株pdrfE∷Δ体内Fe²⁺含量及抗氧化酶类活性。结果显示,drfE基因缺失导致菌株对氧化和盐胁迫敏感,该基因的回补能够恢复菌株对氧化和盐胁迫的抗性。drfE基因缺失导致耐辐射异常球菌体内Fe²⁺浓度由232 μmol/L增加至293 μmol/L;80 mmol/L H₂O₂处理30 min后,突变株ΔdrfE的CAT活性下降32.74%,SOD下降41.3%。以上结果表明DrfE蛋白可能参与耐辐射异常球菌铁代谢途径,并且在细胞抗氧化体系中发挥重要作用。

关键词: 耐辐射异常球菌, 铁蛋白DrfE, 非生物胁迫, 抗氧化酶, 铁离子

Abstract: To investigate the function of the ferritin DrfE encoded by gene drfE in Deinococcus radiodurans,we constructed the complemented strain pdrfE∷Δ,and compared the stress resistances of the wild-type WT,mutant ΔdrfE and complemented strain pdrfE∷Δ under different doses of hydrogen peroxide（H₂O₂）and NaCl. Also,the Fe²⁺concentration and antioxidant enzyme activities of three strains were measured under oxidative stress. The results showed that the deletion of gene drfE led the strain to be more sensitive to oxidative and high-salt stresses and the complementation of gene drfE restored the resistance of the strain to oxidative and salt stress. Furthermore,the deletion of drfEresulted in the Fe²⁺ concentration in D. radiodurans cell increased from 232 μmol/L to 293 μmol/L in vivo. After treated with 80 mmol/L H₂O₂for 30 min,the CAT and SOD activities of the mutant ΔdrfE decreased by 32.74% and 41.3%,respectively,than that of wild-type. The above findings implied that the ferritin DrfE might be involved in iron metabolic pathway of D. radiodurans and played a key role in antioxidant system.

Key words: Deinococcus radiodurans, ferritin DrfE, abiotic stress, antioxidant enzyme, iron ion

吴小丽, 刘盈盈, 江世杰, 陈云, 刘小利, 汪雨舟, 平淑珍, 王劲. 铁蛋白DrfE对耐辐射异常球菌抗氧化酶活性的影响[J]. 生物技术通报, 2017, 33(2): 164-171.

WU Xiao-li, LIU Ying-ying, JIANG Shi-jie, CHEN Yun, LIU Xiao-li, WANG Yu-zhou, PING Shu-zhen, WANG Jin. Effect of Ferritin DrfE on Antioxidant Enzyme Activity in Deinococcus radiodurans[J]. Biotechnology Bulletin, 2017, 33(2): 164-171.

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铁蛋白DrfE对耐辐射异常球菌抗氧化酶活性的影响

Effect of Ferritin DrfE on Antioxidant Enzyme Activity in Deinococcus radiodurans

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