Protein engineering approaches for enhanced catalytic efficiency

Protein engineering has emerged as a straightforward and efficient strategy to expand the application fields of natural enzymes as biocatalysts. It has developed rapidly depending on the structure–function analysis, computational modeling, and comprehending of catalytic processes to guide the modification and application of protein biocatalyst. Furthermore, advanced technologies such as high-throughput screening were recently developed for protein engineering and these significantly facilitate the rational design and collection of improved enzymes to further overcome the potential limitations of enzyme catalyst in the industry. In this review, we first summarize current effective approaches including rational protein design, directed evolution, and semi-rational design and highlight the progress in applying these approaches in the design of enhanced enzymes or de novo enzymes with a rich variety of properties. In addition to these approaches, other advanced strategies responsible for enhanced activities of enzymes, including displaying system, high-throughput screening, spatial organization of enzymes, and electron cryomicroscopy, are also discussed. In general, this chapter would help us understand the mechanism and approaches of protein design, screening, and evaluation tools for enhanced catalytic efficiency and it may offer new insights and guidance to obtain excellent enzymes for future biocatalysis and cell factories research.