Formation of amide bond catalyzed by lipase in aqueous phase for peptide synthesis

Publication date: Available online 2 April 2016 Source:Journal of Molecular Catalysis B: Enzymatic Author(s): Chia-Hung Kuo, Lin Jer-An, Ching-Ming Chien, Chang-Han Tsai, Yung-Chuan Liu, Chwen-Jen Shieh A dipeptide N-acetyl-L-phenylalanyl-L-tyrosinamide (N-Ac-Phe-Tyr-NH2), with angiotensin I converting enzyme (ACE) inhibitor activatity, 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-NH2 with a reaction time of 3.8min, a temperature of 20.9°C, an enzyme amount of 6.5U, 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. Graphical abstract

Source: Journal of Molecular Catalysis B: Enzymatic - April 2, 2016 Category: Biochemistry Source Type: research