Customizing biocatalysts by reducing ΔG‡: Integrating ground-state destabilization and transition-state stabilization

Enzymatic catalysts are increasingly recognized for their potential to revolutionize the chemical industry by enabling precise control of reaction pathways, reducing energy consumption, and minimizing waste, thereby offering exceptional selectivity and sustainability. Enzyme engineering, which focuses on modifying natural enzymes to meet industrial needs, plays a pivotal role in this transformation. A key objective of enzyme engineering is to lower the free energy barrier (ΔG^‡). This review delves into computationally driven strategies designed to accomplish this objective. These strategies are classified into two distinct approaches: ground-state destabilization (GSD) and transition-state stabilization (TSS). For example, GSD may involve reshaping the hydrogen bonding network to elevate ground state energy, while TSS can stabilize the transition state by modulating local electric fields. GSD strategies involve modulating substrate-binding free energy to destabilize the ground state, reshaping hydrogen bonding networks, and refining binding conformations. TSS methods involve modulating proton and electron transfers, optimizing local electric fields, and modifying active sites based on transition-state models. Furthermore, this review contrasts GSD and TSS approaches, discussing their computational underpinnings, respective advantages, and limitations throughout the text to provide a comprehensive understanding of their applications. Finally, we explore technical challenges, the impact of emerging technologies, and the directions and trends of future research.