The bioconversion of glycerol was carried out using Lactobacillus strain. Lactobacillus was grown on nutrient broth. The pre-culture was added to MRS (De Man, Rogosa and Sharpe) broth medium where glycerol was used as carbon source to acquire 1,3-propanediol (1,3-PDO). Bioconversion of glycerol was carried out at 200C, 250C, 300C, 350C and 400C, using Lactobacillus strain. The atmospheric pressure and temperature were maintained for 1, 2, 3 and 4days. Optimum result was obtained at 350C and 3rd day where 0.59mole of 1,3-PDO produced per mole of glycerol. Resulted 1,3-PDO was characterized by Gas chromatography (GC), Fourier Transform Infrared Spectroscopy (FTIR) and Proton Nuclear Magnetic Resonance (1H NMR) techniques.