Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris.

cDNA ofAureobasidium melanogenumlipase comprises 1254 bp encoding 417 amino acids, whereas genomic DNA of lipase comprises 1311 bp with one intron (57 bp). The lipase gene contains a putative signal peptide encoding 26 amino acids. TheA. melanogenumlipase gene was successfully expressed inPichia pastoris. Recombinant lipase in an inducible expression system showed the highest lipase activity of 3.8 U/mL after six days of 2% v/v methanol induction. The molecular mass of purified recombinant lipase was estimated as 39 kDa using SDS-PAGE. Optimal lipase activity was observed at 35–37 °C and pH 7.0 usingp-nitrophenyl laurate as the substrate. Lipase activity was enhanced by Mg2+, Mn2+, Li+, Ca2+, Ni2+, CHAPS, DTT, and EDTA and inhibited by Hg2+, Ag+, SDS, Tween 20, and Triton X-100. The addition of 10% v/v acetone, DMSO,p-xylene, and octanol increased lipase activity, whereas that of propanol and butanol strongly inhibited it. Lipase activity ofAureobasidium melanogenumlipase expressed inPichia pastorisusing inducible and constitutive expression system. [ABSTRACT FROM PUBLISHER]