The invention provides a carbonyl reductase mutant and its application in catalytically synthesizing 17β-hydroxy steroid compounds. The mutant has a mutation in a core amino acid of the wild-type carbonyl reductase. The mutants provided by the invention can significantly improve the ability of carbonyl reductase to convert a series of 17-carbonyl steroid compounds, and have high stereoselectivity. The recombinant cell provided by the invention is used as a biocatalyst, which can obtain a high yield target product with a high substrate feeding concentration, has few by-products, and the yield is not lower than 95%, and the conversion time is short, and the amount of the biocatalyst used is small, The space-time yield is much higher than the current reported level, the preparation method is simple and convenient, the conditions are mild, and the environment is friendly.