工程糖溶性通路,以改善皮奇亚牧草的Lacto-N-neotetraose生产
Jiao Yang1, Nitesh Kumar Mund1, Lirong Yang1
1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 310027, China.
Enzyme and microbial technology
|January 1, 2025
概括
这项研究将Pichia pastoris改造为产生乳酸-N-新糖 (LNnT),这是一种关键的人类牛奶寡糖. 代谢工程策略显著增加了LNnT产量,证明了其作为微生物细胞工厂的潜力.
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:
- 乳-N-neotetraose (LNnT) 是人乳寡糖化物 (HMO) 的关键成分,为婴儿提供了显著的健康益处.
- 使用普遍公认的安全 (GRAS) 微生物生产LNnT正在引起人们的注意,但Pichia pastoris作为宿主尚未得到充分探索.
- 这项研究解决了对高效微生物生产LNnT的需求.
研究的目的:
- 为了设计Pichia pastoris用于LNnT的新合成.
- 通过代谢工程策略优化LNnT生产.
- 建立P. pastoris作为一个可行的细胞工厂,用于LNnT生物合成.
主要方法:
- 将LNnT生物合成途径基因 (lgtA,lgtB,lac12,gal10) 遗传集成到P. pastoris中.
- 代谢工程方法包括前体路径上调和通过 pfk 基因淘汰的糖溶液流量重定向.
- 在生物反应器中优化发酵,以最大限度地提高LNnT产量.
主要成果:
- 在P. pastoris中初始的LNnT合成产生了0.139μg/L.
- 上调前体供应基因 (UDP-GlcNAc,UDP-Gal) 的产量增加到0.162μg/L.
- 淘汰pfk基因和优化介质导致0.867μg/L,在3L生物反应器中进一步增加到1.24μg/L.
结论:
- 可以成功地为LNnT生产设计Pichia pastoris.
- 代谢工程显著提高了P. pastoris.中的LNnT产量.
- 这项研究验证了P. pastoris作为一个有前途的细胞工厂,用于工业LNnT合成.
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