Bacillus sp. SCSIO 15029中一种耐高温Mn-超氧化物歧化酶的克隆、表达及鉴定

doi:10.11978/2015125

热带海洋学报 ›› 2016, Vol. 35 ›› Issue (4): 63-70.doi: 10.11978/2015125CSTR: 32234.14.2015125

Bacillus sp. SCSIO 15029中一种耐高温Mn-超氧化物歧化酶的克隆、表达及鉴定

邓盾^{1, 2, 3}, 张云^{1, 3}, 孙爱君^{1, 3}, 胡云峰^{1, 2, 3}

1. 中国科学院南海海洋研究所, 中国科学院热带海洋生物资源与生态重点实验室, 广东广州510301;
2. 中国科学院大学, 北京 100049;
3. 中国科学院南海海洋研究所, 广东省海洋药物重点实验室, 广东广州510301

收稿日期:2015-10-13 出版日期:2016-07-29 发布日期:2016-08-04
通讯作者: 胡云峰。E-mail: yunfeng.hu@scsio.ac.cn
作者简介:邓盾(1985—), 男,安徽省滁州市人, 博士研究生, 研究方向为生物催化与酶工程。E-mail: dengdun2008@126.com
基金资助:
中国科学院战略性先导科技专项(XDA11030404)

Cloning, expression and characterization of a thermo-tolerant manganese superoxide dismutase from Bacillus sp. SCSIO 15029

DENG Dun^{1, 2, 3}, ZHANG Yun^{1, 3}, SUN Aijun^{1, 3}, HU Yunfeng^{1, 2, 3}

1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China;
3. Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China

Received:2015-10-13 Online:2016-07-29 Published:2016-08-04
Contact: HU Yunfeng. E-mail: yunfeng.hu@scsio.ac.cn
Supported by:
Strategic Priority Research Program of the Chinese Academy of Sciences (XDA11030404)

摘要/Abstract

摘要： 本研究从南海沉积物中分离到一株耐高温菌Bacillus sp. SCSIO 15029, 从其基因组中克隆了一个编码超氧化物歧化酶的基因BaSOD, 并在大肠杆菌BL21(DE3)中实现了可溶性异源表达。随后, 对重组BaSOD进行了酶学性质鉴定, BaSOD的最适pH为7.5, 最适反应温度为40℃, 酶活为5511.46U·mg^-1。经鉴定, BaSOD是一种锰离子特异的超氧化物歧化酶(superoxide dismutase, SOD), 该酶具有良好的热稳定性, 在60℃处理30min酶活基本没有变化, 在70℃下处理30min剩余酶活为71%, 具有较好的工业应用前景。

关键词: 枯草芽孢杆菌, 深海高温酶, 超氧化物歧化酶

Abstract: A putative superoxide dismutase gene (BaSOD) was cloned from Bacillus sp. SCSIO 15029, which was isolated from the sediment of the South China Sea. The BaSOD gene was expressed in Escherichia coli BL21(DE3) by using pET-28a(+) expression system. The recombinant enzyme was soluble in E. coli, and its biochemical properties were studied. BaSOD exhibits maximum activity at pH 7.5 and 40℃ with a specific activity of BaSOD at 5511.46 U·mg^-1. BaSOD is a manganese-dependent superoxide dismutase. The enzyme activity was basically not affected after incubation at 60℃ for 30 min. The residual activity was 71% after incubation at 70℃ for 30 min. BaSOD shows good thermo-stability and possesses very good potential for industrial utilization.

Key words: Bacillus, deep-sea thermophilic enzyme, superoxide dismutase

中图分类号:

Q939

邓盾, 张云, 孙爱君, 胡云峰. Bacillus sp. SCSIO 15029中一种耐高温Mn-超氧化物歧化酶的克隆、表达及鉴定[J]. 热带海洋学报, 2016, 35(4): 63-70.

DENG Dun, ZHANG Yun, SUN Aijun, HU Yunfeng. Cloning, expression and characterization of a thermo-tolerant manganese superoxide dismutase from Bacillus sp. SCSIO 15029[J]. Journal of Tropical Oceanography, 2016, 35(4): 63-70.

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Bacillus sp. SCSIO 15029中一种耐高温Mn-超氧化物歧化酶的克隆、表达及鉴定

Cloning, expression and characterization of a thermo-tolerant manganese superoxide dismutase from Bacillus sp. SCSIO 15029

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