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

Abstract

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 °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.

Characterization of Detergent-Compatible Lipases from Candida albicans and Acremonium sclerotigenum under Solid-State Fermentation

“…The results showed that the lipases from both strains could survive the chemical composition of additives in detergent formulations, making them promising options for industrial-level detergent applications. Penicillium crustosum Thom lipase (PCrL) was identified as a novel extracellular lipase from the Ascomycota (a filamentous fungus) strain P22 and exhibited outstanding compatibility and stability with a broad range of commercially available laundry detergents . Efficient functioning of lipase under harsh conditions is crucial for its effectiveness in detergent formulations.…”

“…Additionally, organic solvents have the ability to modulate and enhance the selectivity and activity of certain lipases. PCrL exhibited excellent stability in organic solvents, showing residual activity ranging from 94 to 150% following 24 h of incubation . The use of enzymes, particularly lipases, in organic solvents is becoming increasingly important in industrial applications due to their potential to produce high-value products.…”

“…PCrL exhibited excellent stability in organic solvents, showing residual activity ranging from 94 to 150% following 24 h of incubation. 60 The use of enzymes, particularly lipases, in organic solvents is becoming increasingly important in industrial applications due to their potential to produce high-value products. This is because the use of enzymes in organic solvents can facilitate the synthesis of compounds that may be difficult or impossible to obtain using traditional chemical methods.…”

“…Penicillium crustosum Thom lipase (PCrL) was identified as a novel extracellular lipase from the Ascomycota (a filamentous fungus) strain P22 and exhibited outstanding compatibility and stability with a broad range of commercially available laundry detergents. 60 Efficient functioning of lipase under harsh conditions is crucial for its effectiveness in detergent formulations. To achieve this, the stability and compatibility of the enzyme with all commercially available detergent formulations are important.…”

Alkaliphilic/Alkali-Tolerant Fungi: Molecular, Biochemical, and Biotechnological Aspects

“…These organisms produce alkaline enzymes capable of resisting high pH (proteases, amylases, and lipases) and can be used in the textile, food, pharmaceutical industry, etc. [ 6 , 7 , 8 ]. Even today, the terms alkaliphilic and alkali-tolerant can be perplexing because this classification is highly dependent on the growth conditions and culture medium composition.…”