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

• Published in: International journal of molecular sciences Vol. 23; no. 19
• Main Authors: Hasnaoui, Ismail, Dab, Ahlem, Mechri, Sondes, Abouloifa, Houssam, Saalaoui, Ennouamane, Jaouadi, Bassem, Noiriel, Alexandre, Asehraou, Abdeslam, Abousalham, Abdelkarim
• Format: Journal Article
• Language: English
• Published: MDPI 07-10-2022
• ISSN: 1422-0067
• DOI: 10.3390/ijms231911920

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 NH
2
-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 °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 α-eleostearic acid at the
sn
-1/3 position. PCrL was completely inhibited by 53 µM 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.