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在工程化大肠杆菌中生物合成Sialyllacto-N-tetraose c

Chenchen Li¹, Mengli Li¹, Wei Gao¹

¹State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, Jiangsu 214122, China.

Journal of agricultural and food chemistry

|November 7, 2024

概括

研究人员设计了一种高产的大肠杆菌菌株,用于生产甲酸-N-四糖c (LSTc),这是一种关键的人类牛奶寡糖. 这种代谢工程方法实现了显著的LST c标位,提供了一个有前途的微生物细胞工厂战略.

科学领域:

微生物学微生物学
代谢工程是代谢工程.
生物化学生物化学

背景情况:

人类牛奶寡糖 (HMO) 对婴儿健康至关重要.
酸-N-四糖c (LST c) 是一种复杂的HMO,具有重要的开发和应用重要性.
高效的LSTc生物合成是一个挑战.

研究的目的:

开发一个微生物细胞工厂,用于高效的LSTc生产.
使用多变量模块化代谢工程 (MMME) 来优化LSTc生物合成的代谢途径.
提高前体供应和辅助因子再生,以增加LST的产量.

主要方法:

利用多变量模块化代谢工程 (MMME) 来选基转移酶和平衡代谢流.
通过阻断竞争性途径并增强前体 (UDP-GlcNAc,UDP-Gal) 供应,设计了一种大肠杆菌菌株 (W15).
建立了一个CTP辅因子再生系统.

主要成果:

构建了能够进行LSTc生物合成的工程菌株.
在振动瓶培养中达到220.9 mg/L的LST c标位,其中有W15菌株.
在3L料批发发酵中达到922.2 mg/L LST c,生产率为10.25 mg/L/h,DCW特异性产量为38.70 mg/g.

结论:

MMME是开发微生物细胞工厂的有效策略,用于LSTc等复杂的HMO.
工程化的大肠杆菌菌株W15显示出高水平的LSTc生产.
这项研究为LSTc的工业生产提供了一种可行的方法.

关键词:

在 CTP 辅助因子再生过程中.埃舍里希亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.代谢工程是代谢工程.sialyllacto-N-tetraose c 的使用情况.

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