As a valuable chemical, 1,3-propanediol (1,3-PD) could be biosynthesized by glycerol fermentation. However, no natural microorganisms that could directly convert glucose into 1,3-PD have been found so far. In this work, genes coding for two enzymes, glycerol-3-phosphate dehydrogenase (GPD, EC 1.1.1.8) and glycerol-3-phosphatase (GPP, EC 3.1.3.21), which were responsible for glycerol production, were organized into the plasmid pUC18K under control of the respective lac promoters. Two recombinant proteins were expressed successfully in wild-type Klebsiella pneumoniae. A glycerol concentration of 6.8 g l(-1) was obtained in flask culture. When glucose was exhausted, dihydroxyacetone was added and medium pH was adjusted to 7.0, and then a 1,3-PD concentration of 0.58 g l(-1) was achieved with engineered K. pneumoniae from glucose.