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脯氨酸代谢相关基因 CgMCUR1 对甘油假丝酵母和酿酒酵母应激耐受性的影响及应用。

Effect and application of proline metabolism-related gene CgMCUR1 on stress tolerance of Candida glycerinogenes and Saccharomyces cerevisiae.

机构信息

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.

The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.

出版信息

J Appl Microbiol. 2023 Jun 1;134(6). doi: 10.1093/jambio/lxad108.

Abstract

AIMS

To investigate the effect of CgMCUR1 on the phenotype of Candida glycerinogenes and Saccharomyces cerevisiae.

METHODS AND RESULTS

Inhibition of CgMCUR1 expression reduced acetate, H2O2, and high temperature tolerance of C. glycerinogenes. Expression of CgMCUR1 resulted in better acetic acid, H2O2, and high temperature tolerance in recombinant S. cerevisiae. Meanwhile, CgMCUR1 was able to enhance intracellular proline accumulation. The qRT-PCR analysis revealed that overexpression of CgMCUR1 affected proline metabolism in recombinant S. cerevisiae. The overexpression strain also showed reduced levels of cellular lipid peroxidation and an altered ratio of saturated fatty acid (SFA) to unsaturated fatty acid (UFA) in the cell membrane. The ethanol production of recombinant S. cerevisiae at high temperature was 30.9 g l-1, obtaining an increase of 12%, and the conversion rate was increased by 12%. In the undetoxified cellulose hydrolysate, the ethanol yield was 14.7 g l-1 at 30 h with an improvement of 18.5%, and the conversion rate was increased by 15.3%.

CONCLUSIONS

Overexpression of CgMCUR1 rendered the acetic acid, H2O2, and high temperature tolerant of recombinant S. cerevisiae and enhanced the ethanol fermentation performance of recombinant S. cerevisiae under high temperature stress and in undetoxified cellulose hydrolysate by improving intracellular proline accumulation and by changing cellular physiological metabolism.

摘要

目的

研究 CgMCUR1 对 Candida glycerinogenes 和 Saccharomyces cerevisiae 表型的影响。

方法和结果

抑制 CgMCUR1 的表达降低了 C. glycerinogenes 的乙酸盐、H2O2 和高温耐受性。CgMCUR1 的表达导致重组 S. cerevisiae 具有更好的乙酸盐、H2O2 和高温耐受性。同时,CgMCUR1 能够增强细胞内脯氨酸的积累。qRT-PCR 分析显示,CgMCUR1 的过表达影响了重组 S. cerevisiae 中的脯氨酸代谢。过表达菌株还显示出细胞内脂质过氧化水平降低和细胞膜中饱和脂肪酸 (SFA) 与不饱和脂肪酸 (UFA) 比例改变。重组 S. cerevisiae 在高温下的乙醇产量为 30.9 g l-1,增加了 12%,转化率提高了 12%。在未经解毒的纤维素水解物中,30 h 时的乙醇产量为 14.7 g l-1,提高了 18.5%,转化率提高了 15.3%。

结论

CgMCUR1 的过表达使重组 S. cerevisiae 具有耐乙酸盐、H2O2 和高温的能力,并通过提高细胞内脯氨酸积累和改变细胞生理代谢来提高重组 S. cerevisiae 在高温胁迫和未经解毒的纤维素水解物中的乙醇发酵性能。

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