在有氧和氧气有限的疗法下,蛋白质减少细胞中的重组蛋白质表达
Alvaro R Lara1, Jose Utrilla2, Luz María Martínez3
1Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark.
Biotechnology and bioengineering
|January 5, 2024
概括
这项研究使用遗传装置对减少蛋白质组的大肠杆菌 (Escherichia coli,PR) 进行了基因工程,以提高工业蛋白质生产. 该PR菌株显著增加了基于flavin mononucleotide的光蛋白 (FbFP) 的产量,为生物制造提供了一个有希望的替代品.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 生物技术是生物技术.
背景情况:
- 工业细胞培养面临来自宿主复杂性和有限的生物反应器质量转移的挑战.
- 最小细胞方法和基因工程为克服这些局限性提供了解决方案.
研究的目的:
- 开发和评估工程最小细胞系统,以增强大肠杆菌中蛋白质的生产.
- 优化表达策略,以增加基于黄素单核酸的光蛋白 (FbFP) 的产量.
主要方法:
- 使用了减少蛋白质组 (PR) 的大肠杆菌菌株.
- 工程构成性蛋白质生成器 (CPG),微空气蛋白质生成器 (MPG) 和微空气转录级联 (MTC) 用于FbFP表达.
- 实现了代谢和表达模型.
- 通过删除pyruvate dehydrogenase复合调节剂和表达维特里奥斯基拉血红蛋白来设计菌株.
主要成果:
- 使用CPG的PR菌株产生的FbFP比使用野生类型 (WT) 多47% (有氧) 和35% (氧气有限).
- 在氧气限制下,PR菌株的MTC产生的FbFP光度比WT高出9%.
- 使用MTC的工程菌株在氧气限制下,与WT相比,FbFP表达的增加超过50%.
结论:
- 工程最小细胞系统与优化的遗传装置显著提高蛋白质的生产在大肠杆菌.
- 蛋白质组减少与定制表达策略相结合,为工业生物工艺提供了可行的替代方案.
- 进一步的菌株工程,包括代谢修饰和异质基因表达,在氧气有限的条件下提高生产力.
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