Further characterization of lipase B from Ustilago maydis expressed in Pichia pastoris: a member of the Candida antarctica lipase B-like superfamily

Abstract Lipases from the basidiomycete fungus Ustilago maydis are promising but underexplored biocatalysts due to their high homology with Candida antarctica lipases. This study provides a comprehensive characterization of a recombinant CALB-like lipase from U. maydis expressed in Pichia pastoris (rUMLB), and compares its properties with those of the well-studied recombinant lipase B from C. antarctica (rCALB). Biochemical analyses included evaluations of optimal pH, temperature, triglyceride (TG) preference for short and medium acyl chains, phospholipase and amidase activities, enantiopreference, thermostability, stability in organic solvents, and responses to NaCl concentration. rUMLB, a glycosylated enzyme with a molecular weight of 38.6 kDa, exhibited cold-active behavior at 0°C and preferred hydrolysis on short-chain fatty acid TGs, like rCALB. Both enzymes displayed strong (R)-enantiopreference and increased activity in non-polar solvents; however, rUMLB was more sensitive to polar solvents. Notably, rUMLB, and was activated at high salt concentrations as previously reported for rCALB. rUMLB uniquely demonstrated significant phospholipase activity towards natural phospholipids, a feature not observed in rCALB. These comparative insights highlight the functional diversity within the CALB-type lipase family, underscoring UMLB’s potential as a versatile biocatalyst and providing valuable information for biotechnological applications and the understanding of enzyme structure-function relationships of CALB superfamily.