调控乙酰CoA合成酶表达促进乙酸利用及GlcNAc合成

doi:10.3969/j.issn.1673-1689.2020.05.003

首页 > 过刊浏览>2020年第39卷第5期 >16-22. DOI:10.3969/j.issn.1673-1689.2020.05.003

调控乙酰CoA合成酶表达促进乙酸利用及GlcNAc合成
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                        10.3969/j.issn.1673-1689.2020.05.003
                    
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Modulation of Acetyl-CoA Synthetase Expression Promoted Acetate Utilization and N-Acetylglucosamine Production in Bacillus subtilis

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N-乙酰氨基葡萄糖（GlcNAc）是一种重要的功能单糖。在利用枯草芽孢杆菌发酵产GlcNAc的过程中, 乙酸的积累严重抑制了细胞生长和GlcNAc合成。为了获得高效利用乙酸生产GlcNAc的微生物细胞工厂，作者通过突变乙酰CoA（Ac-CoA）合成酶编码基因acsA 5′UTR区域内全局转录调控因子CodY结合序列，调控acsA转录水平，进一步突变AcsA乙酰化调控位点，解除乙酰化修饰，促进乙酸转化为Ac-CoA，最终乙酸积累量由6.3 g/L减少到3.6 g/L，同时细胞干重（DCW）由2.7 g/L增加到5.3 g/L，GlcNAc产量由1.9 g/L提高到5.0 g/L，为进一步获得高产GlcNAc细胞工厂提供了基础。

Abstract:

N-acetyl-D-glucosamine（GlcNAc） is an important functional monosaccharide. In the production of GlcNAc using recombinant Bacillus subtillis（B. subtilis）, accumulation of acetate inhibites cell growth and GlcNAc production, which also decreases GlcNAc yield. To construct a B. subtilis cell factory with efficient acetate utilization and GlcNAc production, expression of acetyl-CoA synthetase AcsA, which catalyzes the conversion of acetate to Ac-CoA, was modulated in this study. Specifically, the global transcriptional regulator CodY binding sites in the 5' UTR region of acsA were mutated to promote transcription of acsA；then, enzyme AcsA was modified by deacetylation to improve its activity. Finally, the accumulated acetate during shake flask fermentation reduced from 6.3 to 3.6 g/L, dry cell weight increased from 2.7 to 5.3 g/L, and the GlcNAc yield was increased by 163% up to 5.0 g/L, which indicated that the developed strain offers improved properties for GlcNAc production.

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马文龙,刘延峰,吕雪芹,李江华,刘龙,堵国成.调控乙酰CoA合成酶表达促进乙酸利用及GlcNAc合成[J].食品与生物技术学报,2020,39(5):16-22.

MA Wenlong, LIU Yanfeng, LYU Xueqin, LI Jianghua, LIU Long, DU Guocheng. Modulation of Acetyl-CoA Synthetase Expression Promoted Acetate Utilization and N-Acetylglucosamine Production in Bacillus subtilis[J]. Journal of Food Science and Biotechnology,2020,39(5):16-22.

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在线发布日期: 2020-06-19
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