This is the first report of lipase production from Trichosporon asahii. Here we report 30-fold enhancement (104 U/ml) in lipase production from T asahii MSR 54 after medium optimization by both one variable at - a - time and statistical approach. The strain was identified on the basis of D1/D2 sequences analysis with accession number EU258671. Among seven variables, glucose and CaCl(2) were identified as insignificant variables for lipase production by Plackett Burman (PB) design. Response surface methodology indicated that requirement of malt extract and yeast extract varied with the type of lipase inducer used. Higher malt extract concentrations favored lipase in corn oil and inhibited in Tween-80 while yeast extract (3%) was required in high concentration. On the contrary during kerosene induction both malt extract (0.1%) and yeast extract (0.2%) requirements were minimal. Lipase production corresponded well with biomass. However in kerosene it was a result of induction during starvation accompanied by cell permeabilization. The lipase exhibited S-enantiomeric excess during deacetylation of phenyl ethyl acetate. (C) 2008 Published by Elsevier Ltd.