Polymer-engineered condensates for enzyme activation

Abstract

Enzyme condensates are powerful tools for controlling enzymatic reactions in living cells. Recent advances in polymer science have enabled the design of artificial enzyme condensates in vitro, providing a promising approach to enhance enzymatic activity and stability for various biotechnological applications. In this review, we describe a systematic approach to engineering enzyme condensates through polymer-based strategies. First, we consider the design principles for tailoring the state of the enzyme condensates using charged polymers, including approaches that utilize enzymes as scaffolds or clients, and compare these condensates with other enzyme activation methods, highlighting the advantages and potential limitations of enzyme condensates. Second, we review the major factors that affect enzyme performance within the condensates, including size-dependent effects and local environmental changes. These data are supported by recent mechanistic studies using various enzyme systems, including oxidoreductases. Finally, we focus on possible applications and outline the key challenges in expanding the utility of enzyme condensates from single-enzyme to multienzyme systems and from solution-based to surface-bound architectures. Our comprehensive overview of enzyme condensate engineering provides a new perspective to bridge cellular organization principles and innovations in enzyme catalysis. This review highlights recent advances in engineering artificial enzyme condensates in vitro using charged polymers. Based on our recent findings, we describe strategies for designing condensates through interactions between polymers and enzymes or coenzymes. We then summarize enzyme activation mechanisms triggered by enzyme condensates, including size-dependent effects and conformational changes in enzymes. We also discuss potential applications and future directions, including multienzyme systems, integration with solid surfaces, and combination with rational enzyme design.