Cui Huanqing, Zhang Yage, Liu Sihan, Cao Yang, Ma Qingming, Liu Yuan, Lin Haisong, Li Chang, Xiao Yang, Hassan Sammer Ul, Shum Ho Cheung
Department of Mechanical Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong (SAR), China.
Advanced Biomedical Instrumentation Centre, Hong Kong Science Park, Shatin, New Territories, Hong Kong (SAR), China.
Nat Commun. 2024 Aug 8;15(1):6771. doi: 10.1038/s41467-024-51043-z.
Hierarchical compartmentalization responding to changes in intracellular and extracellular environments is ubiquitous in living eukaryotic cells but remains a formidable task in synthetic systems. Here we report a two-level compartmentalization approach based on a thermo-responsive aqueous two-phase system (TR-ATPS) comprising poly(N-isopropylacrylamide) (PNIPAM) and dextran (DEX). Liquid membraneless compartments enriched in PNIPAM are phase-separated from the continuous DEX solution via liquid-liquid phase separation at 25 °C and shrink dramatically with small second-level compartments generated at the interface, resembling the structure of colloidosome, by increasing the temperature to 35 °C. The TR-ATPS can store biomolecules, program the spatial distribution of enzymes, and accelerate the overall biochemical reaction efficiency by nearly 7-fold. The TR-ATPS inspires on-demand, stimulus-triggered spatiotemporal enrichment of biomolecules via two-level compartmentalization, creating opportunities in synthetic biology and biochemical engineering.
响应细胞内和细胞外环境变化的分层区室化在活的真核细胞中普遍存在，但在合成系统中仍然是一项艰巨的任务。在此，我们报告了一种基于由聚（N-异丙基丙烯酰胺）（PNIPAM）和葡聚糖（DEX）组成的热响应性双水相系统（TR-ATPS）的两级区室化方法。富含PNIPAM的无液膜区室在25℃下通过液-液相分离与连续的DEX溶液相分离，并通过将温度升高至35℃，在界面处产生小的二级区室，从而显著收缩，类似于胶体体的结构。TR-ATPS可以储存生物分子，对酶的空间分布进行编程，并将整体生化反应效率提高近7倍。TR-ATPS通过两级区室化激发了生物分子按需、刺激触发的时空富集，为合成生物学和生化工程创造了机会。
