工程氧化辅因子平衡,以改善大肠杆菌中的5-甲基四基叶酸盐生产
Jinning Yang1,2,3,4, Yaokang Wu1,2,3,4, Xueqin Lv1,2,3,4
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Journal of agricultural and food chemistry
|April 16, 2024
概括
绿色生产的5-甲基四基酸盐 (5-MTHF) 使用工程E. 大肠杆菌实现了创纪录的产量. 代谢工程策略显著提高了5-MTHF的积累,为潜在的工业应用.
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:
- 5-甲基四基叶酸 (5-MTHF) 是叶酸的活性形式,对人类健康至关重要.
- 5-MTHF的化学合成引发了环境问题.
- 微生物合成为5-MTHF生产提供了一个可持续的替代方案.
研究的目的:
- 开发一种代谢工程化的大肠杆菌菌株,用于高产量的5-MTHF微生物生产.
- 为了优化代谢途径,增强叶酸的减少和辅因子的再生.
- 通过发酵实现5-MTHF的工业相关标位.
主要方法:
- 工程设计的E.E. 大肠杆菌BL21 (DE3) 通过修改糖解和酸路径.
- 引入了形式脱酶,用于NAD(P) H再生.
- 过度表达的跨酶基因 (pntAB,sthA) 来平衡辅因子池.
- 在叶酸到5-MTHF通路中的过度表达的关键酶.
- 在3L发酵器中使用料批发发酵.
主要成果:
- 最初的菌株修改增加了5-MTHF,达到44.57μg/L.
- 形式脱酶的表达增强了5-MTHF的产生,达到247.36μg/L.
- 优化的Z13菌株达到3009.03μg/L的5MTHF峰值标位.
- 这代表了在E.E.中报告的最高5-MTHF产量. 大肠杆菌.
结论:
- 代谢工程的E. 代谢工程. 大肠杆菌对可持续的5-MTHF生产是有效的.
- 优化辅因子再生和通路流量对于高产量至关重要.
- 取得的标位是迈向工业规模微生物5-MTHF制造的重大进展.
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