Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24 h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 mu moles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10 mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non-aqueous environment. (C) 2009 Elsevier Ltd. All rights reserved.