Deletion of glycerol-3-phosphate dehydrogenase genes improved 2,3-butanediol production by reducing glycerol production in pyruvate decarboxylase-deficient Saccharomyces cerevisiae.

•Glycerol-free production of 2,3-BD production by the cofactor engineered yeast is feasible.•108.6 g/L 2,3-BD was produced with a high yield (0.462 g 2,3-BDO/g glucose) in the YP medium based fed-batch fermentation.•A batch fermentation with a cassava hydrolysate resulted in 47.44 g/L 2,3-BD titer.