Theoretical Research on Substrate Specificity Differences within the Feruloyl Esterase Family

Abstract

This study comprehensively investigated the substrate specificity differences among four feruloyl esterases using computational biology methods, offering novel insights into enzymatic functions, structural factors influencing substrate specificity, and the optimization of enzyme performance. The research aimed to elucidate the molecular mechanisms underlying the substrate specificity of feruloyl esterases (FAEs) and to assess the impact of disulfide bonds on their activity. FAEs are crucial in bioenergy and biocatalysis, yet the understanding of their substrate specificity remains limited. The study comprised three parts. First, the analysis of substrate specificity differences among four FAEs; second, the evaluation of the effects of disrupting disulfide bonds on FAE activity; and finally, the proposal of computational strategies for improving FAE applications. By employing homology modeling, active site analysis, molecular docking, and dynamic simulations, this study constructed FAE structural models and simulated FAE-substrate interactions. The results revealed that specific amino acid residues play a key role in substrate recognition, and the disruption of disulfide bonds can affect the enzyme's substrate affinity. The discussion section explores the molecular mechanisms of FAEs' substrate specificity and the theoretical and practical contributions of computational predictions to enzyme engineering. The study suggests future research directions to deepen the understanding of FAE substrate specificity and offers new strategies for synthetic biology and biocatalysis. In conclusion, this research provides new insights into the functions of FAEs, the structural basis of substrate specificity, and how to optimize enzyme performance through computational methods. performance through computational methods.