适应性实验室进化和Cupriavidus necator的代谢工程,以改善挥发性脂肪酸的代谢
Eric C Holmes1, Alissa C Bleem1, Christopher W Johnson1
1Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory, Golden, CO, 80401, USA.
Metabolic engineering
|November 3, 2024
概括
删除多基酸盐 (PHB) 基因会影响挥发性脂肪酸 (VFA) 的生长. 适应性进化发现了调节基因中的关键突变,使得改造的Cupriavidus necator菌株能够改善VFA生物转化.
科学领域:
- 微生物生物技术 微生物生物技术
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
背景情况:
- 将废物流转化为增值产品的生物转化对生物经济至关重要.
- 挥发性脂肪酸 (VFA) 是微生物碳上循环的有希望的原料.
- 库普里亚维杜斯 (Cupriavidus necator H16) 是一个适合于VFA生物转换的工业宿主.
研究的目的:
- 通过使用VFA,研究聚基酸盐 (PHB) 操作子删除对C. necator生长的影响.
- 识别促进微生物在VFA基质上生长的基因修饰.
- 开发改进的C.necator平台菌株,以实现高效的VFA生物转化.
主要方法:
- 使用适应性实验室进化 (ALE) 进行选择,以改善VFA的生长.
- 在基因分析中发现了法R,H16_A1372/H16_A1373和H16_A2296-A2298.8的突变.
- 进行了代谢分析 (NADH 丰度) 和基因表达分析.
主要成果:
- 删除PHB操作子导致VFA基板的显著生长缺陷.
- 发R和H16_A1373的突变恢复并改善了VFA的生长.
- 工程系菌株呈现了降低的NADH水平和与新陈代谢和应激耐受性相关的改变基因表达.
结论:
- 在VFA上的C. necator的碳和能量代谢中,PHB生物合成起着至关重要的作用.
- PhaR是影响全球代谢网络和VFA利用的关键调节器.
- 开发了具有增强VFA增长能力的工程C. necator菌株,为改进废物流的价值化铺平了道路.
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