The hydrolytic activity of D-Lac and its variants on D-PL was determined in 25 mL triangular vials using a 14 mL reaction system containing 768 mM DL-PL Tris-HCl buffer 500 mM pH 75 and 200 uL of puri

The enzymatic hydrolysis of D-Lac and its derivatives on D-PL was assessed using a 14 mL reaction system in 25 mL triangular vials. The reaction system comprised of 768 mM DL-PL, Tris-HCl buffer (500 mM, pH 7.5), and 200 uL of purified enzyme. The mixture was incubated at 30°C for 20 minutes, followed by enzyme inactivation using boiling. Subsequently, the reaction products were analyzed using reversed-phase high-performance liquid chromatography.