A Comparative Study of the Performance of Solid Supported and Soluble α-Chymotrypsin for the Enzymatic Synthesis of Kyotorphin

Abstract

The performance of both free and solid-supported α-chymotrypsin for the synthesis of ZTyr-ArgNH₂ (Z-kyotorphin amide) has been studied. The effect on the reaction yield of cosolvent/buffer proportions and reaction temperatures has been examined by a Box-Wilson method. Such factors influenced both preparations to a different extent. A kinetic analysis of reaction rate data has shown that the kinetic data are consistent with a proposed sequential mechanism. Model equations and numerically estimated kinetic parameters are presented. Statistical tests validate the model. In addition, the performance of the reaction has also been assessed by instantaneous and overall fractional yield calculations. Despite the high byproduct formation, the immobilized enzyme could be the choice for a technological process aiming at the large scale synthesis of such a peptide.

Key Words:

• Kinetically controlled enzymatic peptide synthesis
• α-chymotrypsin
• CPG-supported
• α-chymotrypsin
• Instantaneous and overall fractional yields
• Reaction rate analysis
• Non-linear regression methods