在P. putida中使用基于sacB的新型系统和混合碳来源的新型系统高效地生产拉姆诺脂
Ai-Ping Pang1, Yun Wang1, Teng Zhang1
1National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China; Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
Bioresource technology
|December 18, 2023
概括
研究人员开发了一种新的基因编辑系统,以改善Pseudomonas putida KT2440.0.中的代谢工程. 这提高了宝贵的生物表面活性剂 - - 拉姆诺脂类的产量,在发酵过程中达到创纪录的产量.
科学领域:
- 微生物生物技术 微生物生物技术
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
背景情况:
- 伪omonas putida KT2440是一种用于化学合成的GRAS (一般公认的安全) 菌株.
- 现有的KT2440基因编辑工具效率低,阻碍了代谢工程的努力.
研究的目的:
- 为KT2440.0.开发一种高效的无标记基因破坏系统.
- 为了改造KT2440以提高脂蛋白的产量.
- 通过平衡新陈代谢途径和前体供应来优化脂生物合成.
主要方法:
- 建立了一个基于sacB的新型系统 (pK51mobsacB) 以有效地破坏无标记基因.
- 在KT2440.中引入了脂合成途径,并删除了KT2440.中的竞争途径.
- 利用各种促进剂来微调rhlAB表达,并通过异质rmlBDAC表达增强dTDP-L-rhamnose供应.
- 采用混合碳来源 (葡萄糖/甘油) 来平衡细胞生长和生产.
主要成果:
- 在KT2440.0中实现了高效的无标记基因破坏.
- 成功设计了一种KT2440菌株,用于降低代谢负担的拉姆诺脂类合成.
- 优化了前体供应和基因表达,导致显著的脂产量.
- 在摇瓶中生产了3.64g/L的拉姆诺脂,在5L发酵器中生产了19.77g/L.
结论:
- 新的pK51mobsacB系统显著提高了KT2440.0中的基因编辑效率.
- 通过代谢工程设计的KT2440显示出作为工业脂生产的微生物细胞工厂的高潜力.
- 取得的产量代表了代谢工程KT2440报告的最高产量,验证了该战略.
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