用NADH再生策略在E.中生物合成聚3-基酸-同乳酸盐). 大肠杆菌
Ju Wu1, Xuan Gong1, Pengye Guo1
1State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
Biodesign research
|December 19, 2025
概括
一种基于酸盐脱酶 (PtxD) 的新型NADH再生系统有效地增强了Escherichia coli中的基于乳酸的共聚合物生物合成. 这一策略可以增强乳酸的产生,而不会对细菌生长或P3HB-co-LA) 合成产生负面影响.
科学领域:
- 生物技术和代谢工程 生物技术和代谢工程
- 合成生物学 合成生物学
- 聚合物科学 聚合物科学
背景情况:
- 基于乳酸盐的共聚合物的有效生物合成需要强大的辅因子NADH的供应.
- 传统方法经常将NADH再生与中央碳代谢联系起来,可能会限制产量.
- 开发脱的NADH再生系统对于优化共聚合物合成至关重要.
研究的目的:
- 使用基于酸盐脱酶 (PtxD) 的NADH再生策略,构建一个高效的乳酸盐基共聚物生物合成平台.
- 为了研究PtxD表达水平对乳酸合并到共聚物的影响.
- 评估工程菌株在摇瓶和生物反应器培养物中的性能.
主要方法:
- 工程 *Escherichia coli* 菌株 WJPCTP-01 带有集成的 *ptxD* 基因用于NADH再生.
- 使用不同度的异烯β-d-1-thiogalactopyranoside (IPTG) 来控制PtxD表达.
- 在摇瓶和5L生物反应器中使用葡萄糖和树脂糖作为碳来源进行培养.
- 量化P(3HB-co-LA) 共聚合物生产和乳酸分数.
主要成果:
- 基于PtxD的策略成功地将NADH再生与碳代谢脱,增强乳酸合成.
- 合成的共聚合物中的乳酸盐分数在6.2至16.7mol%之间,取决于IPTG度.
- 使用葡萄糖和西洛斯,P(3HB-co-LA) 的产量分别达到3.24 g/L (23.0 mol% LA) 和2.23 g/L (39.0 mol% LA).
- 在生物反应器发酵过程中,使用西洛斯的合成速率为0.12g/L/h,获得了8.57g/L P(3HB-co-41.3 mol% LA) 的标位.
结论:
- 基于PtxD的NADH再生系统是一种有效的策略,用于增强大肠杆菌中的乳酸基共聚合物生物合成.
- 这种方法为乳酸合成提供足够的NADH,而不会影响细菌生长或整体聚合物生产.
- 该工程菌株显示了工业规模生产P(3HB-co-LA) 共聚合物的巨大潜力.
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