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在非传统的酵母和细菌中选广泛宿主表达促进体的穿表达载体
Liyun Ji1, Shuo Xu1, Yue Zhang1
1State Key Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
Microbial cell factories
|August 16, 2024
概括
这项研究开发了来自Yarrowia lipolytica和Kluyveromyces marxianus的广谱促进器库,用于合成生物学应用. 主要酵母促进剂在大肠杆菌等细菌中显示出跨领域的实用性,使生物产品合成能够得到增强.
科学领域:
- 合成生物学 合成生物学
- 微生物生物技术 微生物生物技术
- 代谢工程是代谢工程.
背景情况:
- 非传统的酵母和细菌在合成生物学中至关重要,用于将可再生原料转化为有价值的产品.
- 优化代谢途径和生物产品产量需要在微生物系统中使用战略性促进剂.
- 对于生物合成工程师来说,为各种主机开发广泛的促进器库是必不可少的.
研究的目的:
- 使用来自Yarrowia lipolytica和Kluyveromyces marxianus的构成性促进体构建和描述促进体-报告体载体.
- 系统地评估这些促进剂在各种微生物宿主 (包括酵母和细菌) 的强度和跨领域适用性.
- 确定适合增强合成生物制品合成在合成生物学应用中的促进剂.
主要方法:
- 从Y. lipolytica (yl.hp4d, yl.FBA1in, yl.TEF1, yl.TDH1, yl.EXP1) 中选择了五个组成促进体,以及从K. marxianus中选择了五个 (km.PDC1, km.FBA1, km.TEF1, km.TDH3, km.ENO1).
- 使用α-氨酶和红色光蛋白 (RFP) 作为记者基因构建的促进体-记者载体.
- 在Y. lipolytica,K. marxianus,Pichia pastoris,Escherichia coli和Corynebacterium glutamicum中系统地描述了促进剂的强度.
主要成果:
- 所有五种K. marxianus促销者都在Y. lipolytica中表达了基因,反之亦然,强度各不相同.
- 在Y. lipolytica TEF1 (yl.TEF1) 和K. marxianus TEF1 (km.TEF1) 促销者证明了在P. pastoris,大肠杆菌和C. glutamicum中适应性的表现.
- 在大肠杆菌中,km.TEF1促进体达到T7促进体RFP表达强度的约20%,突出显示其强大的细菌表达.
结论:
- 非传统的酵母促进剂在合成生物学中对受监管系统具有多样化,跨领域的适用性.
- 这些促进体可以有效地管理碳流量,并在各种微生物宿主中增强目标生物产物合成.
- 开发的促进者库为动态调节和改进的生物生产在工程微生物中提供了创新的工具.
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