用大肠杆菌MG1655进行多维工程,以有效生物合成二可酸乳糖
Yihan Chen1, Chunhua Zhao2, Ruiyan Wang2
1State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
Journal of agricultural and food chemistry
|February 24, 2025
概括
这项研究开发了一种新的无等离子体大肠杆菌菌株,用于高效的 difucosyllactose (DFL) 生产. 这种改造品种取得了高的DFL产量,克服了这种有价值的母乳寡糖的先前方法的局限性.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 代谢工程是代谢工程.
背景情况:
- 二糖乳糖 (DFL) 是母乳的关键成分,具有显著的健康益处.
- 现有的使用工程菌株的DFL合成方法患有等离子体不稳定性和抗生素依赖性,阻碍了工业应用.
- 开发强大的,基于细胞工厂的生产系统对于可扩展的DFL制造至关重要.
研究的目的:
- 设计一种稳定,无等离子体的大肠杆菌菌株,用于高产量的DFL生产.
- 为了优化参与DFL生物合成的关键酶的表达.
- 在大型生物反应堆中证明开发的菌株的商业可行性.
主要方法:
- 多复制SAMT和fut3Bc基因的基因组集成进入大肠杆菌MG1655.5.
- 在GDP-l-fucose生物合成途径中过度表达基因.
- 使用AlphaFold 3对Fut3Bc进行酶工程,以创建一个有益的突变 (F24Y).
- 在7L生物反应器中,优化菌株 (MGA2S-5) 的料批量培养.
主要成果:
- 一个优化的无等离子体大肠杆菌菌株 (MGA2S-5) 已成功地用集成的SAMT和突变的fut3Bc (F24Y) 构建.
- 该菌株在7L生物反应器中在料批量条件下达到35.04g/L的高DFL度.
- 观察到完全消耗中间产品,表明有效的代谢流向DFL.
结论:
- 没有等离子体的大肠杆菌菌株可以有效地被设计为高水平的DFL生产.
- 酶工程和基因组整合策略显著提高了DFL产量和生产效率.
- 开发的MGA2S-5菌株为工业规模制造二氧化硫酸乳糖提供了一个有前途的平台.
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