The lipase production of Burkholderia sp. GXU56 was influenced by carbon and nitrogen sources, inorganic salts, initial pH of the medium and cultivation temperature. The maximum lipase production was 580.52 U/mL and reached 5 times the level of the basic medium in the optimum medium at pH 8.0, 32 degrees C, 200 rpm and 40-48 h. The lipase was purified 53.6 fold to homogeneity and the molecular weight was 35 KDa on SDS-PAGE. The optimum pH and temperature of the lipase were 8.0 and 40 degrees C, respectively, and it was stable in the range of pH 7-8.5 and at temperatures below 45 degrees C. The lipase activity was strongly inhibited by Zn2+, Cu2+, Co2+, Fe2+, Fe3+ ions and SDS, while it was stimulated by Li+ and Ca2+ ions and in presence of 0.1 % CTAB, 0.1 % Triton X-100 and 10 % DMSO. K-m and V-max of the lipase were calculated to be 0.038 mmol/L, and 0.029 mmol/L min(-1), respectively, with PNPB as the substrate. The GXU56 lipase showed enantioselective hydrolysis of (R,S)-methyl mandelate to (R)-mandelic acid, which is an important intermediate in the pharmaceutical industry.