在Saccharomyces cerevisiae中有效生产7-脱水胆固醇的途径重构
Yuchen Han1,2, Huayi Gao3, Shuo Wang3
1Key Laboratory of Engineering Biology for Low-carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
ACS synthetic biology
|June 27, 2025
概括
这项研究将酵母改造为产生7-脱胆固醇 (7-DHC),这是维生素D的前体. 优化策略导致7-DHC产量大幅增加,在生物反应堆中达到3.26g L-1以实现可持续生产.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
背景情况:
- 7-脱水胆固醇 (7-DHC) 是一种至关重要的固醇,也是维生素D3的主要前体.
- 对于各种应用来说,7-DHC的高效和可持续生产至关重要.
- 目前用于7-DHC生产的方法可能面临可扩展性和可持续性的限制.
研究的目的:
- 构建一种能够对7-脱胆固醇进行de novo生物合成的Saccharomyces cerevisiae菌株.
- 通过代谢工程策略,优化工程酵母菌株以提高7-DHC的产量.
- 为了实现7-DHC的高度,可扩展和可持续的生产.
主要方法:
- 在Saccharomyces cerevisiae中构建一个新的7-DHC生物合成途径.
- 引入异构的DHCR24和关键酶的过度表达.
- 优化厄戈斯特醇通路,使用有机溶剂,表面活性剂和e-聚氨酸.
- 该途径在过氧体和氧化还原水平再平衡中的亚细胞局部化.
- 在5升生物反应器中进行大规模发酵.
主要成果:
- 最初的7-DHC标位为109.0毫克L-1通过途径构建和酶修改实现.
- 通过动态调节厄戈斯特醇通路和DHCR24多拷贝表达,7-DHC标位增加了86.3%.
- 增加了99.1%的7-DHC标位,添加了e-polylysine.
- 在路径重构后,摇动瓶中的最终7-DHC标位为517.4毫克L-1.
- 在5L生物反应器中成功扩大到3.26g L-1.
结论:
- 经过工程设计的Saccharomyces cerevisiae菌株通过新的生物合成途径有效地产生7-脱胆固醇.
- 代谢工程策略,包括途径优化和辅因子平衡,显著提高7-DHC产量.
- 开发的路径重构策略为7-DHC生产提供了一种可持续和可扩展的方法.
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