通过在大肠杆菌中构建酸化-脱酸化反应,从葡萄糖中生物合成曼诺
Yuyao Wang1, Enhui Chen2, Yanfei Wang1
1Tianjin University of Science and Technology, National Engineering Laboratory for Industrial Enzymes, Tianjin 300457, China; National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
Enzyme and microbial technology
|March 22, 2024
概括
研究人员开发了一种新的方法,利用工程化大肠杆菌菌株从葡萄糖中合成d-mannose. 这种新途径实现了63%的转化率,大大改善了以前用于d-曼诺斯生产的酶方法.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 生物化学生产 生化生产
背景情况:
- D-曼诺是一种有价值的单糖,用于食品,制药和化品行业.
- 现有的d-mannose生产方法,依赖于化学合成或酶性表皮化,面临诸如低转化率和复杂的净化过程等局限性,因为反应平衡.
- 需要有效和可扩展的d-曼诺斯合成路径.
研究的目的:
- 开发一种新的,高效的微生物发酵策略,从葡萄糖中生产d-mannose.
- 设计一种代谢修饰的大肠杆菌菌株,能够产生高产量的d-曼诺斯合成.
- 为了克服传统的d-曼诺斯生产方法的局限性.
主要方法:
- 通过删除果酸酶 (PfkA) 基因以积累果糖-6-酸盐 (F6P) 来设计大肠杆菌菌株.
- 鉴定并利用了内源性酸酶YniC,它对-6-酸盐具有很高的特异性.
- 构建了一条涉及曼-6-酸盐异构酶和YniC的重组合成途径,以将F6P转化为d-曼.
主要成果:
- 这种改造品种在48小时内从葡萄糖中产生了25.2g/L的d-曼诺斯.
- 获得了63%的高转化率,显著优于传统的2-epimerase方法的15%转化率.
- 证明了指导F6P向d-曼诺斯合成的有效途径.
结论:
- 这项研究提出了一种高效且可扩展的方法,用于从易于获得的葡萄糖中合成d-曼诺斯.
- 在工程E. coli中开发的化-脱化途径为现有的生产技术提供了更好的替代方案.
- 工程菌株为工业规模的d-mannose生产提供了一个具有成本效益的平台.
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