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通过Corynebacterium glutamicum表达Rhizopus oryzae氧化系统生产11α-氧类固醇衍生物

Beatriz Galán¹, Carmen Felpeto-Santero², José Luis García²

¹Department of Microbial and Plant Biotechnology, Margarita Salas Centre for Biological Research-CSIC, Madrid, Spain. bgalan@cib.csic.es.

Methods in molecular biology (Clifton, N.J.)

|August 29, 2023

概括

这项研究证明了Corynebacterium glutamicum作为微生物宿主成功地用于类固醇11α-氧化,这是一个关键的制药过程. 这种工程菌株为传统的真菌生物转化方法提供了更清洁,更有效的替代方案.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
微生物工程微生物工程

背景情况:

类固醇11α-氧化对于生产避孕药和葡萄糖皮质激素等药品至关重要.
目前使用真菌的工业方法由于多个细胞染色体P450 (CYPs) 产生基化类固醇的混合物.
开发更高效和更具体的生物转化过程对于制药应用是必不可少的.

研究的目的:

为了设计一种细菌菌株,以实现高效和特定的类固醇11α-氧化.
确定Corynebacterium glutamicum作为表达真核体类固醇氧酶的合适宿主.
为在制药类固醇生物转化中使用复合C. glutamicum提供概念证明.

主要方法:

化学合成了Rhizopus oryzae 11α-hydroxylase (CYP509C12) 和其氧化还原合作伙伴 (RoCPR1) 的基因,将其合成为合成操作子 (FUN).
将FUN操作子克隆到pECXK-99E矢量中以创建pXKFUN等离子体.
用 pXKFUN 转化了强大的,GRAS (通常被认为是安全的) 细菌菌株Corynebacterium glutamicum R31,以产生重组菌株C. glutamicum R31 (pXKFUN).

主要成果:

改造的C. glutamicum R31 (pXKFUN) 菌株有效地进行了类固醇11α-氧化.
C. glutamicum证明了有效地运输C19和C21类固醇.
细菌宿主没有降解类固醇或引入不必要的副作用,导致更纯净的产品.

结论:

科里尼细菌 (Corynebacterium glutamicum) 作为表达真核体类固醇生物转化酶的有效和干净的底盘.
这种工程细菌系统为工业生产基化类固醇提供了一个有前途的替代方案.
开发的协议促进使用重组C. glutamicum的类固醇11α-氧化,为改善制药制造铺平了道路.

关键词:

通过11α-氧化.生物转化生物转化在Corynebacterium glutamicum的研究中,我们发现在真菌CYP450中.类固醇是一种类固醇.

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