Chinese Journal of Biochemistry and Molecular Biology 2008;24(5):426-431

Enzymatic Synthesis of Endomorphin by Immobilized Papain on Sodium Alginate-Chitosan

Ping HE ; Zhuolie HUANG ; Chunyi LI ; Weijia CHEN ; Guanghong WU ; Zhizhan CHU

Keywords

endomorphin; immobilized papain; organic solvent

Country

China

Language

Chinese

Abstract

The enzymatic synthesis of endomorphins were carried out by immobilized papain on sodium alginate-chitosan (IPSAC). The reaction has been carried out in two steps. First, Trp-Phe-NH2 was obtained by Boc-Trp-OH coupling with Phe-NH2 using IPSAC catalyst in microaqueous acetonitrile system with a 27.8 % yielding as determined by high performance liquid chromatography (HPLC). The catalytic properties of IPSAC were examined by studying the dependence of pH, ionic strength, solution content, reaction temperature,enzyme loading and reaction time over the yields. The results of orthogonal experiments indicated that pH was the most significant factor that influenced this synthesis. Second, Boc-Tyr-Pro-OMe and Trp-Phe-NH2 were suspended into the microaqueous acetonitrile to produce Tyr-Pro-Trp-Phe-NH2 (endomorphin-1) with a 35.2% yield.