Fig. 1

Recombinant SFAR4 proteins for lipase/esterase activity analysis. a YPD agar plates supplemented with 1.5 % tributyrin emulsion and secreted recombinant SFAR4 that hydrolyzes tributyrin to produce clear zones. b YPD supplemented with 0. 5 % CaCl₂ and 2 % Tween-20, with hydrolytic activity indicated the formation of a white calcium precipitate. Transformed SFAR4 yeast colonies were grown on agar plates for 5 days. c Substrate specificity was determined by measuring activities for acyl p-nitrophenyl esters at 5 mM concentrations and monitoring the release of p-nitrophenol photometrically at 348 nm. These assays were performed using 50 mM Good’s buffer (pH 7.0) at 30 °C. One unit of enzyme is defined as the amount of enzyme that hydrolyzes 1 μmol of the substrate per minute