在大肠杆菌中对3 - 福科西拉克托斯过度生产的组合优化策略
Zhihui Du1, Yingying Zhu1, Zhen Lu2
1State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
Journal of agricultural and food chemistry
|June 15, 2024
概括
这项研究利用大肠杆菌 (Escherichia coli) 来增强3-糖糖糖 (3-FL) 的产生,这是一种有益的母乳寡糖. 优化的菌株取得了高产量,证明了工业规模3FL制造的潜力.
科学领域:
- 微生物工程 微生物工程
- 代谢工程是代谢工程.
- 生物技术是生物技术.
背景情况:
- 3-糖乳糖 (3-FL) 是一种关键的母乳寡糖,对婴儿健康有显著的益处.
- 之前的工作确立了 *Escherichia coli* BL21(DE3) 对于 *in vivo* 3-FL生物合成.
- 为了高效和可扩展的3FL生产,需要进一步优化.
研究的目的:
- 为了增强代谢工程 * Escherichia coli * * 的 3-FL 生产.
- 通过基因和蛋白质工程来改善细胞外3-FL产量.
- 评估工程微生物菌株的工业适用性.
主要方法:
- 优化培养条件以增加3-FL产量.
- 竞争性通路基因的失活,以重定向代谢流量.
- 评估用于增强细胞外分泌的传送系统.
- 在α1,3-fucosyltransferase (FutM2) 活性位点 (Q126) 的位点和突变发生.
- 对FutM2突变的结构分析和分子动力学模拟.
主要成果:
- 培养条件优化使3-FL产量翻了一番.
- 工程 *E. coli* 菌株显示显著改善了3-FL生物合成.
- 这种FutM2-Q126A突变体表现出增强的酶活性.
- 最好的菌株BD3-A在摇瓶中达到6.91g/L,在食批量种植中达到52.1g/L.
- 结构分析证实了残留物修饰对酶结构的积极影响.
结论:
- 代谢和蛋白质工程策略成功增强了大肠杆菌中的3-FL产量.
- 开发的BD3-A菌株对具有成本效益的,大规模的工业3-FL制造具有很高的潜力.
- 这项工作有助于可持续生产功能性寡糖.
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