BioATRP synthesized ATP-fueled Transient Coacervation

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) (PDMAEMA), 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, glucose oxidase, and urease. ATP-fuelled coacervates also demonstrate the enzymatic catalysis to prove its potential to be exploited as a cellular microreactor. Additionally, we developed biocatalytic polymerization-induced coacervation (BioPIC), improving reaction yield and producing coacervates with distinct characteristics. This method allows in situ and real-time programming over coacervation through BioATRP-controlled polymerization. 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 may lead to the spatial arrangement of multienzyme cascades and offer novel ideas on the architecture of artificial cells with organelles