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 dilu

The hydrolytic efficacy of D-Lac and its various iterations on D-PL was evaluated in 25 mL triangular vials utilizing a 14 mL reaction system comprising 768 mM DL-PL, Tris-HCl buffer (500 mM, pH 7.5), and 200 uL of diluted purified enzyme. The reaction was allowed to proceed for 20 minutes at 30°C, after which the enzyme was inactivated through boiling and the resulting product was analyzed through reversed-phase high-performance liquid chromatography.