Comprehensive SummaryTo elucidate the relationship between carbohydrate structure and its biological function, glycoenzyme tools are widely applied in a variety of physiological and pathological scenarios. Intricate adjustment via glycoenzyme engineering improved the catalytic efficiency, extended the repertoire of accessible catalytic reactions, and tailored novel substrate specificities. In this review, we introduce the principles of glycoenzyme engineering and summarize the recent advances in the rational design and directed evolution of these enzymes, with an emphasis on screening methods in directed evolution. We also excerpt novel detection methods in glycobiology that can be adapted for the development of next‐generation glycoenzyme tools.