De novo engineering of programmable and multi-functional biomolecular condensates for controlled biosynthesis

作者：Wenwen Yu, Ke Jin, Dandan Wang, Nan‐Kai Wang, Yangyang Li, Yanfeng Liu, Jianghua Li, Guocheng Du, Xueqin Lv, Jian Chen, Rodrigo Ledesma‐Amaro, Long Liu · 发表于：Nature Communications · 年份：2024 · DOI：10.1038/s41467-024-52411-5 · 被引用次数：43 · 研究领域：RNA Research and Splicing、RNA modifications and cancer、RNA and protein synthesis mechanisms

There is a growing interest in the creation of engineered condensates formed via liquid–liquid phase separation (LLPS) to exert precise cellular control in prokaryotes. However, de novo design of cellular condensates to control metabolic flux or protein translation remains a challenge. Here, we present a synthetic condensate platform, generated through the incorporation of artificial, disordered proteins to realize specific functions in Bacillus subtilis. To achieve this, the “stacking blocks” strategy is developed to rationally design a series of LLPS-promoting proteins for programming condensates. Through the targeted recruitment of biomolecules, our investigation demonstrates that cellular condensates effectively sequester biosynthetic pathways. We successfully harness this capability to enhance the biosynthesis of 2’-fucosyllactose by 123.3%. Furthermore, we find that condensates can enhance the translation specificity of tailored enzyme fourfold, and can increase N-acetylmannosamine titer by 75.0%. Collectively, these results lay the foundation for the design of engineered condensates endowed with multifunctional capacities. Biomolecular condensates can serve as synthetic organelle-like compartments within prokaryotes. Here, the authors report a synthetic condensate platform, generated through the incorporation of artificial, disordered proteins, and demonstrate control of metabolic flux and protein translation in Bacillus subtilis.