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在vivo中以相分离为媒介的多酶组装

Yao Chen^1,2, Yi Shi¹, Mengyu Li¹

¹State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China.

Journal of agricultural and food chemistry

|March 19, 2025

概括

合成生物学使用RGGRGG凝聚物和ReverseTag/ReverseCatcher系统推进了多酶催化. 这种方法在细胞系统中显著增加了5.4倍的烯产量.

科学领域:

合成生物学合成生物学
生物催化剂是一种生物催化剂.
蛋白质工程是一种蛋白质工程.

背景情况:

多酶组合对于高效的生物催化剂至关重要.
分相蛋白为酶复合体的形成提供了新的平台.
反向标记/反向捕获器系统可以实现特定的蛋白质-蛋白质相互作用.

研究的目的:

研究RGGRGG相隔蛋白质凝聚物的整合与ReverseTag/ReverseCatcher系统,以形成多酶复合体.
为了评估这种策略在增强体内代谢途径性能方面的有效性.

主要方法:

使用粗粒模拟来研究RGGRGG域相互作用和凝结物形成.
使用 ReverseTag/ReverseCatcher 系统将 ReverseTagged 蛋白质固定在 ReverseCatcher_RGGRGG 凝聚剂中.
用 ReverseTag 设计了用于烯生物合成的酶,并将其固定在细胞系统中的 ReverseCatcher_RGGRGG 凝聚物上.

主要成果:

已经证明,RGGRGG域形成了能够与ReverseTag/ReverseCatcher系统接口的凝聚物.
反向标记蛋白在反向捕获器_RGGRGG凝结物中成功自我组装.
与对照组相比,在工程细胞系统中实现了5.4倍的柳科产量增加.

结论:

结合RGGRGG和ReverseTag/ReverseCatcher系统,为构建功能多酶复合体提供了一个有效的策略.
这种方法显著提高了目标代谢物的产生,证明了它在合成生物学应用中的潜力.
这项研究为体内和体外多酶催化和代谢工程开辟了新的途径.

关键词:

RGGRGRGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG 反向Tag/反向Cacter 其他多种酶组合组件

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