Abstract Global climate deterioration intensifies the demand for exploiting efficient CO 2 utilization approaches. Converting CO 2 to biorefinery feedstock affords an alternative strategy for third‐generation biorefineries. However, upcycling CO 2 into complex chiral carbohydrates remains a major challenge. Previous attempts at sugar synthesis from CO 2 either produce mixtures with poor stereoselectivity or require ATP as a cofactor. Here, by redesigning glycolaldehyde synthase, the authors constructed a synthetic pathway for biorefinery feedstock D‐xylulose from CO 2 that does not require ATP as a cofactor. The artificial D‐xylulose pathway only requires a three‐step enzyme cascade reaction to achieve the stereoselective synthesis of D‐xylulose at a concentration of 1.2 g L −1 . Our research opens up an alternative route toward future production of chemicals and fuels from CO 2 .