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通过表型分类进行体内持续进化,以缓解β-氨酸生产中的代谢瓶

Fuqiang Song¹, Heng Zhang², Ke Wang²

¹Science Center for Future Foods, Jiangnan University, Wuxi, Jiangsu 214122, China; Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology, Jiangnan University, Wuxi 214122, China.

Trends in biotechnology

|September 12, 2025

概括

我们通过进化酶L-阿斯巴酸-α-decarboxylase来增强工程大肠杆菌中的β-氨酸 (β-Alanine) 生产. 这种生物传感器引导的进化方法提高了62.45%的特定产量.

科学领域:

合成生物学合成生物学
代谢工程是代谢工程.
酶工程是什么?酶工程是什么?

背景情况:

β-氨酸 (β-Alanine) 是一个关键的平台化学物质,由于路径瓶,其生物合成效率有限.
酶的限制,特别是L-阿斯巴酸-α-脱碳酶 (PanD_bsu),阻碍了有效的β-氨酸生产.

研究的目的:

通过工程Escherichia coli.克服β-氨酸生物合成的局限性.
增强L-酸盐-α-脱碳酶 (PanD_bsu) 的活性和稳定性,以改善β-氨酸的产生.

主要方法:

使用模块化路径优化和组合调节的Escherichia coli MG1655的系统工程.
开发一个 in vivo 进化平台,将基础编辑和生物传感器指导结合起来,用于高通量查.
通过位点和和代突变进行PanD_bsu的蛋白质工程,然后进行结构和功能分析.

主要成果:

使用生物传感器引导进化平台生成具有增强活动的PanD_bsu变体.
鉴定了一种PanD_bsu^T4E突变,该突变在工程菌株MA31.1.中增加了62.45%的特定β-氨酸产量.
发现PanD_bsu^T4E突变体通过Glu-Lys盐桥表现出稳定的四级结构.

结论:

开发了一个可扩展的战略,以解决微生物细胞工厂的途径瓶.
将蛋白质工程与生物传感器引导进化的整合是优化代谢途径和增强化学品生产的强大方法.

关键词:

L-阿斯巴甜酸-α-脱碳酶酶的作用基础编辑的基础编辑生物传感器引导的进化.路径工程工程的路径工程.这是一种β-氨酸 (β-alanine).

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