The application of ketoreductase-based biocatalytic reduction to access optically pure Prelog or anti-Prelog alcohols offers a valuable approach for asymmetric synthesis. Despite this, control of the stereopreferences of ketoreductases as desired remains challenging, since natural ketoreductases usually display Prelog preference and it is difficult to transfer the knowledge from engineered anti-Prelog ketoreductases to the others. Here, we present the discovery of a switch between Prelog and anti-Prelog reduction toward halogen-substituted acetophenones in six short-chain dehydrogenase/reductases (SDRs). Through carefully analysis of the structural information and multiple-sequence alignment of several reported SDRs with Prelog or anti-Prelog stereopreference, the key residues that might control their stereopreferences were identified using Lactobacillus fermentum short-chain dehydrogenase/reductase 1 (LfSDR1) as the starting enzyme. Protein engineering at these positions of LfSDR1 could improve its anti-...