Purification, Biochemical and Kinetic Characterization of a Novel Alkaline sn-1,3-Regioselective Triacylglycerol Lipase from Penicillium crustosum Thom Strain P22 Isolated from Moroccan Olive Mill Wastewater

A novel extracellular lipase from a filamentous fungus Ascomycota strain, P22, was isolated from olive mill wastewater, then purified and characterized. This strain was identified as Penicillium crustosum Thom based on sequencing analyses. Penicillium crustosum Thom strain P22 lipase (PCrL) was purified 63-fold to homogeneity using ammonium sulfate precipitation and chromatography on a Q-Sepharose Fast Flow column, with a total yield of 34%. The purified PCrL had a molecular mass of 28 kDa, estimated by SDS-PAGE. The 20 NH2-terminal amino-acid residues showed a high degree of homology with those of other Penicillium lipases. The specific activity of PCrL at pH 9 and 37 degrees C were found to be 5000 and 10,000 U/mg on olive oil and trioctanoin emulsions, respectively. PCrL exhibited clear regioselectivity toward the sn-1 position of the surface-coated triglycerides which were esterified with alpha-eleostearic acid at the sn-1/3 position. PCrL was completely inhibited by 53 mu M of Orlistat, 5 mM of phenylmethylsulfonylfluoride, and 2 mM of diiodopropyl fluorophosphate, suggesting that it belonged to the serine lipase family. PCrL showed high activity and stability in the presence of water-immiscible organic solvents, surfactant, and oxidizing agents, and showed considerable compatibility with commercial laundry detergents. Washing performance analysis revealed that it could effectively remove oil stains. Hence, PCrL has several attractive properties that make it a promising potential candidate for detergent formulations.

Automated summary

A novel extracellular lipase P22 from Penicillium crustosum Thom was isolated and characterized. The specific activities of P22 lipase were found to be 5000 and 10,000 U/mg on olive oil and trioctanoin emulsions, respectively. It exhibited clear regioselectivity toward the sn-1 position of surface-coated triglycerides and showed high activity and stability in the presence of water-immiscible organic solvents, surfactants, and oxidizing agents. It also showed compatibility with commercial laundry detergents and effectively removed oil stains.