Biochemical characterization of an alkaline and detergent-stable Lipase from Fusarium annulatum Bugnicourt strain CBS associated with olive tree dieback

Abstract

This work describes a novel extracellular lipolytic carboxylester hydrolase named FAL, with lipase and phospholipase A1 (PLA1) activity, from a newly isolated filamentous fungus Ascomycota CBS strain, identified as Fusarium annulatum Bunigcourt. FAL was purified to about 62-fold using ammonium sulphate precipitation, Superdex® 200 Increase gel filtration and Q-Sepharose Fast Flow columns, with a total yield of 21%. The specific activity of FAL was found to be 3500 U/mg at pH 9 and 40°C and 5000 U/mg at pH 11 and 45°C, on emulsions of triocanoin and egg yolk phosphatidylcholine, respectively. SDS-PAGE and zymography analysis estimated the molecular weight of FAL to be 33 kDa. FAL was shown to be a PLA1 with a regioselectivity to the sn-1 position of surface-coated phospholipids esterified with α-eleostearic acid. FAL is a serine enzyme since its activity on triglycerides and phospholipids was completely inhibited by the lipase inhibitor Orlistat (40 μM). Interestingly, compared to Fusarium graminearum lipase (GZEL) and the Thermomyces lanuginosus lipase (Lipolase®), this novel fungal (phospho)lipase showed extreme tolerance to the presence of non-polar organic solvents, non-ionic and anionic surfactants, and oxidants, in addition to significant compatibility and stability with some available laundry detergents. The analysis of washing performance showed that it has the capability to efficiently eliminate oil-stains. Overall, FAL could be an ideal choice for application in detergents.

Heterologous expression and characterization of a novel thermostable and alkali stable recombinant lipase enzyme from Bacillus thuringensis into E. coliBL21(DE3) for detergent formulation

“…Saraswat et al (2017) used a lipase enzyme for detergent formulation and found 15% increased wash efficiency from commercial detergent. Similarly, Dab et al (2023) reported a lipase from Fusarium annulatum which showed 50% increase in wash potential when applied in combination with commercial detergents. Li et al (2014) also reported a lipase from Pseudomonas stutzeri for detergent formulation with considerable increase in stain removal efficiency.…”

Optimization of Lipase Production from Pseudomonas fluorescens

“…20 Lipase enzymes break down greasy stains, and surfactants in detergents stick to the residue and remove stains from clothing. 21 Lipase is used along with other enzymes to remove the glue from the spools of yarn that aid in dyeing. 22 Lipases significantly increase oil yield during vegetable oil processing and improve the appearance of the final product.…”