重定向植物的降解途径,用于定向的androst-1,4-diene-3,17-dione微生物生物转化
Xia Ke1,2, Jia-Hao Cui1,2, Qi-Jie Ren1,2
1National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Applied microbiology and biotechnology
|February 1, 2024
概括
这项研究通过基因删除消除副产品并通过基因工程增强酶活性,优化了从植物中产生的类固醇药物的微生物生产. 这导致了更高的产量和更有效的生物转化过程.
科学领域:
- 生物技术是生物技术.
- 微生物工程 微生物工程
- 合成生物学 合成生物学
背景情况:
- 基于类固醇的药物主要是通过植物的微生物转化产生.
- 现有的两步生物工艺实现了高产量,但遭受了副产品的形成.
研究的目的:
- 调查Mycobacterium neoaurum中C19和C22类固醇子途径之间的催化转换.
- 制定策略来消除副产品并提高类固醇中间体的产量.
主要方法:
- 分析了对植物醇诱导的剂量依赖转录反应.
- 利用基因删除 (opccR,sal) 来消除C22类固醇的副产品.
- 建立了一个染色体整合平台,用于使用强大的L2促进剂和特定位点重组的目标基因过度表达.
- 过度表达kstD1基因以增强生物转换.
主要成果:
- 确定了阿尔多酶Sal作为C22类固醇侧链裂变的关键.
- 顺序删除opccR和sal消除了C22副产品,增加了ADD的摩尔产量到71.3%.
- 克服速度限制步骤和优化系统,产生了82.0%的摩尔产量和4.22 g·L−1·day−1的时空产量.
结论:
- 在休息细胞中展示了一种可通过基因操纵控制的代谢开关.
- 为M. neoaurum细胞工厂开发开发了有效的遗传工具.
- 从植物中建立一个有针对性的生物转化路径,以有效地生产C19和C22类类固醇药物中间体.
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