在大肠杆菌中有效合成维生素B5,通过工程基托班托酸氧甲基转移酶和辅因子供应
Kun Qiu1,2,3,4, Fuqiang Song2,3,4, Ke Wang2,3
1Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
研究人员设计了一个微生物工厂,以提高d-pantothenic acid (维生素B5) 的生产. 这涉及优化关键的酶和途径,从而显著提高d-pantothenic酸产量和生产力.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 酶工程是什么? 酶工程是什么?
背景情况:
- d-Pantothenic acid (d-PA) 或维生素B5对于生理功能至关重要,作为辅酶A的前体.
- 松酸甲基转移酶 (PanB) 是d-PA合成中的速度限制酶,需要辅因子5,10-甲基甲基酸 (5,10-CH2-THF).
- 优化d-PA生产需要对酶和辅因子供应通路进行工程.
研究的目的:
- 通过代谢工程策略来增强d-PA的产生.
- 为了提高番酸甲基转移酶 (PanB) 的效率,并优化辅因子供应.
- 开发一个强大的微生物细胞工厂,用于高产量的d-PA合成.
主要方法:
- 识别并设计了转录因子 *bhsA* 以减轻d-PA生产的限制.
- 使用酶工程,与PanC共同表达PanB并产生一种高活性PanBM突变体 (V123I/K124W).
- 增强了 de novo 合成并抑制了 5,10-CH2-THF 的降解,并引入了 d-PA 载体 PanT.
主要成果:
- 改造的PanBM酶显示d-PA产量增加了2.5倍.
- 优化辅因子供应和细胞外运输显著提高了d-PA产量.
- 开发的无等离子体菌株DPA23在5L生物反应器中达到78.48g/Ld-PA,生产率为2.69g/L/h.
结论:
- 代谢和酶工程策略成功增强了d-PA的生产.
- 优化的微生物细胞工厂证明了d-PA的高效率和生产率.
- 这项研究为建造d-PA及其衍生物的微生物工厂提供了有价值的框架.
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