Sharma Surbhi, Belluati Andrea, Kumar Mohit, Dhiman Shikha
Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10-14, Mainz, 55122, Germany.
Department of Chemistry and Centre for Synthetic Biology, Technical University of Darmstadt, Peter-Grünberg-Straße 4, Darmstadt, 64287, Germany.
Angew Chem Int Ed Engl. 2024 Dec 10;64(11):e202421620. doi: 10.1002/anie.202421620.
A living cell has a highly complex microenvironment whereas numerous enzyme-driven processes are active at once. These procedures are incredibly accurate and efficient, although comparable control has not yet been established in vitro. Here, we design an enzymatic reaction network (ERN) that combines antagonistic and orthogonal enzymatic networks to produce adjustable dynamics of ATP-fueled transient coacervation. Using horseradish peroxidase (HRP)-mediated Biocatalytic Atom Transfer Radical Polymerization (BioATRP), we synthesized poly(dimethylaminoethyl methacrylate), which subsequently formed coacervates with ATP. We rationally explored enzymatic control over coacervation and dissolution, using orthogonal and antagonistic enzyme pairs viz., alkaline phosphatase, Creatine phosphokinase, hexokinase, esterase, and urease. ATP-fuelled coacervates also demonstrate the enzymatic catalysis to prove its potential to be exploited as a cellular microreactor. Additionally, we developed ERN-polymerization-induced transient coacervation (ERN-PIC), with complete control over the system, polymerization, coacervation, and dissolution. Notably, the coacervation process itself determines functional properties, as seen in selective cargo uptake. The strategy offers cutting-edge biomimetic applications, and insights into cellular compartmentalization by bridging the gap between synthetic and biological systems. The development of temporally programmed coacervation is promising for the spatial arrangement of multienzyme cascades, and offers novel ideas on the architecture of artificial cells.
活细胞具有高度复杂的微环境，同时众多酶驱动的过程会同时活跃进行。这些过程极其精确和高效，尽管在体外尚未建立类似的调控机制。在此，我们设计了一个酶促反应网络（ERN），它结合了拮抗和正交的酶网络，以产生由ATP驱动的可调节的瞬时凝聚动力学。利用辣根过氧化物酶（HRP）介导的生物催化原子转移自由基聚合（BioATRP），我们合成了聚甲基丙烯酸二甲氨基乙酯，其随后与ATP形成凝聚物。我们合理地探索了对凝聚和溶解的酶促控制，使用了正交和拮抗的酶对，即碱性磷酸酶、肌酸磷酸激酶、己糖激酶、酯酶和脲酶。由ATP驱动的凝聚物还展示了酶催化作用，以证明其作为细胞微反应器被开发利用的潜力。此外，我们开发了ERN-聚合诱导的瞬时凝聚（ERN-PIC），对系统、聚合、凝聚和溶解具有完全的控制。值得注意的是，凝聚过程本身决定了功能特性，如在选择性货物摄取中所见。该策略提供了前沿的仿生应用，并通过弥合合成系统与生物系统之间的差距，深入了解细胞区室化。时间编程凝聚的发展对于多酶级联反应的空间排列很有前景，并为人工细胞的架构提供了新的思路。
