期刊
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
卷 37, 期 1-6, 页码 9-15出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.molcatb.2005.08.008
关键词
RGD; peptide synthesis; alcalase; organic solvents; enzyme
The tripeptide Bz-Arg-Gly-Asp(-NH2)-OH was synthesized by a combination of chemical and enzymatic methods in this study. Firstly, Gly-Asp-(NH2)(2) was synthesized by a novel chemical method in three steps including chloroacetylation of L-aspartic acid, esterification of chloroacetyl L-aspartic acid and ammonolysis of chloroacetyl L-aspartic acid diethyl ester. Secondly, the linkage of the third amino acid (Bz-Arg-OEt) to Gly-Asp-(NH2)(2) was completed by enzymatic method under kinetic control condition. An industrial alkaline protease alcalase was used in water-organic cosolvents systems. The synthesis reaction conditions were optimized by examining the effects of several factors including water content, temperature, pH and reaction time on the yield of the synthesis product Bz-Arg-Gly-Asp(-NH2)-OH. The optimum conditions are pH 8.0, 35 degrees C, in ethanol/Tris-HCI buffer system (85:15, v/v), 8 h with the tripeptide yield of 73.6%. (c) 2005 Elsevier B.V. All rights reserved.
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