结合的合成途径可以改善复合大肠杆菌菌株中3-基酸的产生
Dafeng Zhou1, Diego Leandro Quiroga-Sánchez1, Xuan Zhang2
1Department of Biological Science and Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Biotechnology notes (Amsterdam, Netherlands)
|October 17, 2024
概括
在工程E. coli*中合马洛尼尔-CoA和propionyl-CoA通路显著增强了3-基酸 (3-HP) 的生产. 删除特定的基因进一步提高了3-HP的产量,证明了一个有前途的代谢工程策略.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 生物技术是生物技术.
背景情况:
- 3-基酸 (3-HP) 是一种有价值的平台化学物质.
- 从葡萄糖中有效的微生物生产3-HP仍然是一个挑战.
- 优化辅因子再生对于改善3HP生物合成至关重要.
研究的目的:
- 通过合马洛尼尔-CoA和propionyl-CoA通路,增强复合大肠杆菌中的3-HP生产.
- 研究NADP+/NADPH再生在3-HP生物合成中的作用.
- 通过代谢工程策略来改善3HP标位.
主要方法:
- 工程 *E. coli* 菌株过度表达了来自马洛尼尔-CoA (乙-CoA 碳氧化酶 - ACC,马洛尼尔-CoA 减少酶 - MCR) 和-CoA (propionyl-CoA 脱酶 - PACD) 途径的关键酶.
- 在用葡萄糖作为基质的摇瓶中进行培养.
- 基因修饰包括基因删除 (*ygfH*, *prpC*) 以优化代谢流量.
- 补充生物和二碳酸盐.
主要成果:
- 单一途径的过度表达产生了低的3HP标位 (0.26g/L对于马洛尼尔-CoA,0.11g/L对于普罗尼尔-CoA).
- 结合两种途径 (菌株*Ec*-PAM) 增加了3HP的产量,达到1.29g/L.
- 用生物和碳酸盐补充剂进一步改善了产量.
- 在*Ec*-ΔY-ΔP-PAM菌株中删除了*ygfH*和*prpC*基因,导致3HP标位增加1.5倍至1.94g/L.
结论:
- 马洛尼尔-CoA和propionyl-CoA通路的同时表达有效地提高了大肠杆菌中的3-HP产量.
- 代谢工程策略,包括基因删除和辅因子再生优化,对于高位数3HP生物合成至关重要.
- 这项研究提出了一种有效的方法来改善从葡萄糖中产生3-HP的微生物产量.
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