Chia-Hung Kuo, Jer-An Lin, Ching-Ming Chien
Jul 1, 2016
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Influential Citations
5
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Journal
Journal of Molecular Catalysis B-enzymatic
Abstract
Abstract A dipeptide N -acetyl- l -phenylalanyl- l -tyrosinamide ( N -Ac-Phe-Tyr-NH 2 ), with angiotensin I converting enzyme (ACE) inhibitor activity, was synthesized via porcine pancreatic lipase catalyzed amidation of N -acetyl-phenylalanine ethyl ester with l -tyrosinamide in an aqueous phase. Response surface methodology was employed to evaluate the effects of synthesis parameters. The optimum synthesis conditions obtained an 84.45% yield of N -Ac-Phe-Tyr-NH 2 with a reaction time of 3.8 min, a temperature of 20.9 °C, an enzyme amount of 6.5 U, and a substrate molar ratio of 2.5:1 (Tyr:Phe). The kinetics of lipase and α-chymotrypsin catalyzed amidation was compared using the Ping-Pong mechanism. The lipase showed a lower apparent kinetic constant than α-chymotrypsin indicating that the acyl lipase intermediate had a higher affinity toward tyrosinamide in the amidation. In addition, because the lipase can avoid the secondary hydrolysis of synthesized peptide, it is expected to be an effective method for obtaining a good yield of dipeptide.