Continuous Production ofN-(Benzyloxycarbonyl)-L-Glycyl-L-Phenylalanine Methyl Ester Utilizing Extractive Reaction in Aqueous/Organic Biphasic Medium

Abstract

N-(Benzyloxycarbonyl)-L-glycyl-L-phenylalanine methyl ester was continuously synthesized, enzymatically, utilizing an extractive reaction in an aqueous/organic biphasic system. The extremely high yield, ca. 100%, was obtained continuously in a water/butyl acetate biphasic medium.

Novel kinetic analysis of enzymatic dipeptide synthesis: Effect of pH and substrates on thermolysin catalysis

“…Thermolysin shows specificity against hydrophobic or bulky amino residues at the amino side of the splitting point in peptide or protein substrates. These properties (a high thermal stability, a tolerance to organic solvents, and a specificity against hydrophobic residues) should have permitted thermolysin a wide applications in a number of catalytic processes, including the continuous synthesis of peptide derivatives containing hydrophobic residues such as Z-AspPheOMe (Hirata et al, 1997;Miyanaga et al, 1995;Murakami et al, 1996;Hirata, 1997a, 1997b;Nagayasu et al, 1994aNagayasu et al, , 1994b, F-AspPheOMe Murakami, Hayashi et al, 1999;Murakami, Yoshida et al, 2000) and Z-GlyPheOMe (Murakami, Oda, et al, 1999;Murakami, Oda et al, 2000) mainly in reaction media containing organic solvents. It is obvious that the major specificity site of thermolysin is at S1Ј position, which accepts large hydrophobic residues, thereby the enzyme preferentially cleaves peptides at the N-terminal side of hydrophobic or bulky amino residues, such as Ile, Leu, Val, and Phe.…”