Biochemical characterization of lipase from Cryptococcus albidus (D24) and its application in sugar fatty acid ester synthesis.

Abstract

Microbial lipases are widely used in industrial applications. The lipase enzyme from Cryptococcus albidus (D24) strain was biochemically characterized and evaluated for its potential in catalyzing esterification reactions, particularly in the synthesis of sugar fatty acid esters. The enzyme demonstrated enhanced activity in solvents like acetone, isopropanol, and dimethylformamide. The effects of different metal ions on the stability of the enzyme were also evaluated, and the results revealed an increased activity with Mn²⁺, K⁺ and Co²⁺. The molecular weight of the D24 lipase was determined to be 36.31 kDa, placing it within the range of other yeast-originated lipases. The kinetic parameters, including Km and Vmax, were calculated to be 1.58 × 10^-4 mM p-nitrophenyl palmitate (pNPP) and 26.31 U/min respectively, according to the Lineweaver-Burk plot. The enzyme exhibited promising results in catalyzing the esterification reaction to yield L-proline-glucose ester, as well as the synthesis of fructose monopalmitate. Thin layer chromatography (TLC) analysis of the lipase-catalyzed synthesis of fructose monopalmitate showed that, from 24 to 40 h, fructose monopalmitate concentration increased from 4.5 to 8.4% (w/w of the reaction mixture). These findings suggest the potential of D24 lipase for applications in synthesis of industrial products. This study is the first to report on the biochemical properties of the D24 lipase, offering a novel and sustainable enzymatic alternative for the future biotechnological applications.