Protein Engineering of Substrate Specificity toward Nitrilases: Strategies and Challenges
Shi-Qian Bian, Zi-Kai Wang, Jin‐Song Gong, Chang Su, Heng Li, Zhenghong Xu, Jin‐Song Shi
Abstract
Nitrilase is extensively applied across diverse sectors owing to its unique catalytic properties. Nevertheless, in industrial production, nitrilases often face issues such as low catalytic efficiency, limited substrate range, suboptimal selectivity, and side reaction products, which have garnered heightened attention. With the widespread recognition that the structure of enzymes has a direct impact on their catalytic properties, an increasing number of researchers are beginning to optimize the functional characteristics of nitrilases by modifying their structures, in order to meet specific industrial or biotechnology application needs. Particularly in the artificial intelligence era, the innovative application of computer-aided design in enzyme engineering offers remarkable opportunities to tailor nitrilases for the widespread production of high-value products. In this discussion, we will briefly examine the structural mechanism of nitrilase. An overview of the protein engineering strategies of substrate preference, regioselectivity and stereoselectivity are explored combined with some representative examples recently in terms of the substrate specificity of enzyme. The future research trends in this field are also prospected.