Newlase F is a rude enzyme which contains triacylglycerol lipase and acid protease. Hydrolysis of dipeptide heptyl esters with Newlase F was studied in phosphate buffer-organic solvent by HPLC. When the Newlase F’s level reached 5 mg/mL under mild condition (pH 7.0, 30(C), the lipase had the highest activity. The reaction was also affected greatly by organic solvents and their concentrations. It is found that protease in Newlase F does not hydrolyze amide bond under this condition (pH 7.0, r.t.). Newlase F is a rude enzyme which contains triacylglycerol lipase and acid protease. Hydrolysis of dipeptide heptyl esters with Newlase F was studied in a phosphate buffer-organic solvent by HPLC. When the Newlase F’s level reached 5 mg / mL under mild condition (pH 7.0, 30 (C), the lipase had the highest activity. The reaction was also affected greatly by organic solvents and their concentrations. It is found that protease in Newlase F does not hydrolyze amide bond under this condition (pH 7.0, rt).