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通过重编程pyruvate合成来进行前体平衡,以便在工程Escherichia coli中产生高产量的l-isoleucine

Xiaojing Huo¹, Xuesen Xia², Nan Xue¹

¹Department of Microbial Physiological & Metabolic Engineering, State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.

Bioresource technology

|February 25, 2026

概括

研究人员通过平衡其前体,增强了微生物生产必不可少的氨基酸l-isoleucine (l-Ile). 这种代谢工程策略显著增加了葡萄糖的l-Ile产量,提高了其工业可行性.

科学领域:

代谢工程是代谢工程.
合成生物学合成生物学
生物化学生产生化生产

背景情况:

氨基氨酸 (l-Ile) 是一种必不可少的分支链氨基酸,对于食品,料和制药应用至关重要.
目前l-Ile的微生物发酵产量不足于最佳,限制了其工业规模的生产.
有效的合成需要精确控制前体分子,2-氧丁酸盐和酸盐.

研究的目的:

通过代谢工程来增强从葡萄糖中获得的l-异黄素 (l-Ile) 产量.
为了优化2-oxobutyrate和pyruvate前体之间的平衡,以增加l-Ile的生产.
开发一个强大的微生物平台,以实现高效和成本效益的l-Ile合成.

主要方法:

构建了一个无等离子体的l-threonine (l-Thr) 合成模块,以提高2-oxobutyrate的供应.
通过修改运输系统和酶表达,优化了l-Ile合成途径.
通过删除pykFA并利用葡萄糖PTS运输系统来分割基酸盐的工程前体平衡.
减弱了TCA循环流量,并引入了白氨酸脱酶,以最大限度地减少损失和解决辅因子失衡.

主要成果:

达到0.67mol/mol葡萄糖的高l-异黄素产量和1.72g/L/h的生产率.
通过平衡2-氧丁酸盐/酸盐的比率,显著降低了副产品l-氨酸 (l-Val).
证明了一种可行的策略,以促进l-isoleucine的下游净化.

结论:

综合代谢工程策略成功增强了l-异黄素的生产.
均衡前体比对于l-Ile的高产微生物合成至关重要.
开发的工程菌株作为一个有前途的细胞工厂,用于l-isoleucine和衍生品生产.

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