过表达谷氧还蛋白基因GRX5提高酿酒酵母乙酸耐性

doi:10.11949/j.issn.0438-1157.20141174

化工学报 ›› 2015, Vol. 66 ›› Issue (4): 1434-1439.DOI: 10.11949/j.issn.0438-1157.20141174

过表达谷氧还蛋白基因GRX5提高酿酒酵母乙酸耐性

方青¹, 张明明¹, 陈洪奇¹, 熊亮¹, 赵心清², 白凤武^1,2

1. 大连理工大学生命科学与技术学院, 辽宁大连 116023;
2. 上海交通大学生命科学技术学院, 上海 200240

收稿日期:2014-08-05 修回日期:2015-01-18 出版日期:2015-04-05 发布日期:2015-04-05
通讯作者: 赵心清
作者简介:方青(1989-),男,硕士研究生。
基金资助:
国家高技术研究发展计划项目(2012AA021205, 2012AA101805);国家自然科学基金项目(21376043)。

Improvement of acetic acid tolerance of Saccharomyces cerevisiae by overexpressing glutaredoxin encoding gene GRX5

FANG Qing¹, ZHANG Mingming¹, CHEN Hongqi¹, XIONG Liang¹, ZHAO Xinqing², BAI Fengwu^1,2

1. School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116023, Liaoning, China;
2. School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China

Received:2014-08-05 Revised:2015-01-18 Online:2015-04-05 Published:2015-04-05
Supported by:
supported by the National High Technology Research and Development Program of China (2012AA021205, 2012AA101805) and the National Natural Science Foundation of China (21376043).

摘要/Abstract

摘要：

利用可再生的纤维素原料生产燃料乙醇是国内外研究的热点。但纤维素原料一些预处理过程产生的乙酸对酿酒酵母细胞生长和乙醇发酵产生强烈抑制,因此,提高酿酒酵母细胞的乙酸耐受性是提高纤维素乙醇发酵效率的重要手段。本文研究了谷氧还蛋白家族中GRX5p的编码基因的过表达对酿酒酵母在乙酸胁迫条件下细胞生长和发酵性能的影响。结果表明,过表达GRX5的重组菌株在含有5 g·L^-1乙酸的平板中生长优于对照菌株;在含有5 g·L^-1乙酸的培养基中进行乙醇发酵,过表达GRX5的重组菌株可在48 h基本消耗培养基中所有的葡萄糖,发酵周期比对照菌株缩短了12 h。过表达GRX5菌株的乙醇生产强度为0.897 g·L^-1·h^-1,比对照提高了28.5%。代谢物分析结果表明,过表达GRX5的重组菌株可产生更多的保护性物质海藻糖和甘油,有利于增强菌株胁迫耐受性。

关键词: 谷氧还蛋白, GRX5, 燃料乙醇, 发酵效率, 乙酸胁迫, 酿酒酵母

Abstract:

Fuel ethanol production using renewable cellulosic materials has attracted widespread attention by researchers. However, acetic acid released from the pretreatment process exerts severe inhibition on yeast cell growth and ethanol fermentation. Therefore, improvement of acid tolerance of industrial Saccharomyces cerevisiae benefits efficient cellulosic ethanol production. In this work, glutaredoxin encoding gene GRX5 was overexpressed in S. cerevisiae, and it was found that the GRX5 overexpression strain grew better than that of the control strain in the plates containing 5 g·L^-1 acetic acid. When ethanol fermentation in the presence of 5 g·L^-1 acetic acid was evaluated, it was found that the GRX5 overexpression strain consumed all glucose in the broth within 48 h, 12 h shorter than that of the control strain. Ethanol productivity of the GRX5 overexpressed strain was determined to be 0.897 g·L^-1·h^-1, with a 28.5% increase than that of the control strain. Analysis of the key metabolites showed that the GRX5 overexpression strain produced higher concentration of trehalose and glycerol, which might explain its improved cell viability in the presence of acetic acid stress.

Key words: glutaredoxin, GRX5, fuel ethanol, fermentation efficiency, acetic acid stress, Saccharomyces cerevisiae

中图分类号:

Q819

方青, 张明明, 陈洪奇, 熊亮, 赵心清, 白凤武. 过表达谷氧还蛋白基因GRX5提高酿酒酵母乙酸耐性[J]. 化工学报, 2015, 66(4): 1434-1439.

FANG Qing, ZHANG Mingming, CHEN Hongqi, XIONG Liang, ZHAO Xinqing, BAI Fengwu. Improvement of acetic acid tolerance of Saccharomyces cerevisiae by overexpressing glutaredoxin encoding gene GRX5[J]. CIESC Journal, 2015, 66(4): 1434-1439.

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过表达谷氧还蛋白基因GRX5提高酿酒酵母乙酸耐性

Improvement of acetic acid tolerance of Saccharomyces cerevisiae by overexpressing glutaredoxin encoding gene GRX5

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