High-yield production of D-1,2,4-butanetriol from lignocellulose-derived xylose by using a synthetic enzyme cascade in a cell-free system.

•A cell-free system was developed for D-1,2,4-butanetriol production in vitro;.•The key factors that influencing BT production efficiency were identified.•A titer of 6.1 g/L BT was achieved from xylose by using the optimized cell-free system.•The feasibility of cell-free system to produce BT in corncob hydrolysates was determined.