A Metarhizium anisopliae spore surface lipase (MASSL) strongly bound to the fungal spore surface has been purified by ion exchange chromatography on DEAE sepharose followed by ultrafiltration and hydrophobic interaction chromatography on phenyl sepharose. Electrophoretic analyses showed that the molecular weight of this lipase is similar to 66 kDa and pI is 5.6. Protein sequencing revealed that identified peptides in MASSL shared identity with several lipases or lipase-related sequences. The enzyme was able to hydrolyze triolein, the animal lipid cholesteryl stearate and all pNP ester substrates tested with some preference for esters with a short acyl chain. The values of K(m) and V(max) for the substrates pNP palmitate and pNP laurate were respectively 0.474 mM and 1.093 mMol min(-1) mg(-1) and 0.712 mM and 5.696 mMol min(-1) mg-1. The optimum temperature of the purified lipase was 30 degrees C and the enzyme was most stable within the most acid pH range (pH 3-6). Triton X-100 increased and SDS reduced enzyme lipolytic activity. MASSL activity was stimulated by Ca(2+), Mg(2+) and Co(2+) and inhibited by Mn(2+). The inhibitory effect on activity exerted by EDTA and ECTA was limited, while the lipase inhibitor Ebelactone B completely inhibited MASSL activity as well as PMSF. Methanol 0.5% apparently did not affect MASSL activity while beta-mercaptoethanol activated the enzyme. (C) 2009 Elsevier Ltd. All rights reserved.