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通过重组PgsBCA复合物和发酵优化,在Corynebacterium glutamicum中增强了多-γ-胺酸的合成

Guoqiang Xu¹, Jiyue Wang², Jiancheng Shen¹

¹The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi, 214122, China; Jiangsu Provincial Engineering Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi, 214122, China; Yixing Institute of Food and Biotechnology, Yixing, 214200, China.

Metabolic engineering

|December 31, 2023

概括

这项研究通过优化γ-PGA合成酶复合体,增强了Corynebacterium glutamicum中的聚-γ-胺酸 (γ-PGA) 生产. 获得的最高标位为50.2g/L,这是从葡萄糖中进行 de novo γ-PGA 生物合成的新纪录.

科学领域:

微生物学微生物学
生物技术是生物技术.
合成生物学合成生物学

背景情况:

聚-γ-胺酸 (γ-PGA) 是一种具有多种应用的可生物降解聚合物.
来自Bacillus物种的γ-PGA合成酶复合体 (PgsBCA) 对于γ-PGA生物合成至关重要.
之前的工作确立了一个Corynebacterium glutamicum菌株,用于 de novo γ-PGA生产.

研究的目的:

通过重构和过度表达C. glutamicum中的PgsBCA复合物来增强γ-PGA合成.
研究单个子单位表达水平对γ-PGA生产的影响.
探索来自不同细菌物种的正基因对γ-PGA合成的影响.

主要方法:

在C. glutamicum中复制和过度表达PgsBCA复合物.
调整pgsB,pgsC和pgsA基因的转录水平.
测试来自各种细菌物种的正统PgsB和PgsC基因.
优化发酵条件,包括溶解氧和葡萄糖补充.

主要成果:

这三种成分 (PgsB,PgsC,PgsA) 都对C. glutamicum中的γ-PGA合成至关重要.
优化pgsC和pgsB转录显著增加了γ-PGA产量,而pgsC的影响最大.
将B. licheniformis PgsB替换为B. methylotrophicus PgsB增加了γ-PGA标位,达到17.14 g/L.
在5L发酵器中从葡萄糖中获得了创纪录的de novo γ-PGA标位50.2g/L.

结论:

PgsBCA亚单元的表达水平极大地影响了γ-PGA的产生.
正义的PgsC基因通常抑制了合成,而B. methylotrophicus PgsB显示出有前途的结果.
这项研究提出了一种新且高效的高度新型g-PGA生产策略.

关键词:

溶解的氧气是一种溶解的氧气.料批发发酵的发酵方法在PgsBCA复合体中.重组重组的情况 γ-PGA的 γ-PGA,这是一个很好的方法.

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