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Updated: Jun 24, 2025

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通过代谢工程和生物催化剂生产高产氨酸

Haihong Chen¹, Yaohong Wang¹, Weishan Wang²

¹State Key Laboratory of Bioreactor Engineering and School of Biotechnology, East China University of Science and Technology, Shanghai, China.

Nature biotechnology

|June 5, 2024

概括

在这项研究中,Rhodobacter sphaeroides被改造为产生coproporphyrin III (CPIII),达到16.5g/L的高产量. 进一步的酶催化产生了和氨酸,为有价值的氨酸提供了可持续的生物生产方法.

科学领域:

生物技术和合成生物学
微生物工程微生物工程
生物化学生物化学

背景情况:

氨酸是多功能化合物,在医学,能源和材料领域都有应用.
能有效且可扩展的氨酸生产,如氨酸III (CPIII) 对于它们的广泛使用至关重要.
当前的生产方法可能会面临产量和可持续性的限制.

研究的目的:

开发一种高产量的微生物平台,使用Rhodobacter sphaeroides生产甲酸III (CPIII).
为提高CPIII产量设计生物生产途径.
利用CPIII作为合成有价值的金属氨酸的前体,包括和氨酸.

主要方法:

在Rhodobacter sphaeroides中进行全基因组的CRISPRi查,以确定CPIII过度生产的关键遗传标.
代谢工程策略,包括利用PrrA酸化,以提高CPIII的产量.
酶催化使用工程金属酸酶和甲基脱碳酶用于金属氨酸的下游合成.
料批发发酵和试点规模生物反应器研究 (200L) 进行过程优化和扩展.
开发高纯度 (>95%) 的CPIII净化方案.

主要成果:

通过CRISPRi查识别hemN作为改善CPIII生产的关键目标.
通过优化料批发发酵,通过优化料批发发酵实现了最大CPIII产量16.5g/L.
从CPIII中通过酶催化成功合成了血红素和抗瘤剂菲林.
在5L生物反应器中,证明了高的酶转化率 (63%的,98%的) 和产量 (10.8 g/L,21.3 g/L).
为CPIII.建立了一个可扩展的净化过程.

结论:

工程化Rhodobacter球状体作为高产量CPIII生物生产的高效细胞工厂.
开发的战略使得可扩展和可持续的和氨酸的生产成为可能.
这种方法为有价值的氨酸的工业和医疗应用提供了可行的解决方案.

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