模块化路径工程的迪特尔类合成酶和美酸路径的密尔蒂拉迪因生产
Yongjin J Zhou1, Wei Gao, Qixian Rong
1Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian 116023, People's Republic of China.
Journal of the American Chemical Society
|January 28, 2012
概括
使用在酵母中的模块化通路工程策略,增强了微生物的毫特二烯生产. 关键酶的蛋白质融合改善了新陈代谢的流动,从而提高了这种重要的植物类前体的产量.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 微生物生物技术 微生物生物技术
背景情况:
- 微生物生产的植物类药物比植物提取具有优势,但在途径优化方面面临挑战.
- 之前的研究集中在前体通路 (MVA/MEP) 上,忽视了二类合成酶之间的相互作用.
- 米尔蒂拉迪因是Salvia miltiorrhiza中发现的tanshionones的关键前体.
研究的目的:
- 开发一个模块化通路工程 (MOPE) 战略,用于在酵母中快速组装合成毫拉迪因通路.
- 设计二烯合成酶 (SmCPS,SmKSL) 和前体供应酶的蛋白质融合,以提高代谢流量.
- 系统地评估有效的微生物生产的线路变异.
主要方法:
- 应用MOPE策略,在Saccharomyces cerevisiae中组装合成毫拉迪因通路.
- 预测和分析SmCPS和SmKSL之间的分子相互作用.
- 构建蛋白质融合 (SmCPS-SmKSL,BTS1-ERG20) 以增强代谢流量的道化.
- 评估多个路径变体和优化在双胞胎酵母菌株.
主要成果:
- SmCPS-SmKSL和BTS1-ERG20的蛋白质融合显著改善了密尔蒂拉迪因的产生.
- 在工程融合中观察到,副产品的积累减少.
- 在菌株YJ2X中的优化途径在15L生物反应器中实现了365 mg/L的毫蒂拉标位.
结论:
- MOPE策略使微生物类路径的快速和合理工程成为可能.
- 对二类合成酶和前体酶的系统工程对于高效的微生物植物类生产至关重要.
- 这项工作展示了成功的方法,用于高滴度的微生物合成的米尔蒂拉迪因.
相关概念视频
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