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Alcalase®‐catalysed synthesis of the precursor tetrapeptide N ‐benzoylarginylglycylaspartylserinamide (Bz‐RGDS‐NH 2 ) of the cell‐adhesion peptide arginylglycylaspartylserine (RGDS)
Author(s) -
Huang YiBing,
Yang Sen,
Cai Yu,
Wang Hua,
Gao Gui,
Wu XiaoXia,
Xu Li,
Zhang XueZhong
Publication year - 2008
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20070123
Subject(s) - tetrapeptide , chemistry , yield (engineering) , tripeptide , substrate (aquarium) , catalysis , acetonitrile , peptide , peptide synthesis , hydrolysis , organic chemistry , stereochemistry , biochemistry , materials science , oceanography , metallurgy , geology
In the present study, a precursor tetrapeptide Bz‐RGDS‐NH 2 ( N ‐benzoylarginylglycylaspartylserinamide) of cell‐adhesion peptide RGDS (arginylglycylaspartylserine) was synthesized by a novel route. First of all, the precursor tripeptide GDS‐NH 2 (glycylaspartylserinamide) was synthesized by a chemical method only using aspartic acid and serine at gram scale in four steps. The linkage of the fourth amino acid Bz‐Arg‐OEt ( N ‐benzoyl‐ L ‐arginine ethyl ester) to GDS‐NH 2 was completed by an enzymatic method under kinetic control in water‐miscible organic media. An industrial alkaline protease, Alcalase®, with a wide substrate specificity, was used as the catalyst. The effects of organic solvents, pH value, reaction temperature, water content and molar ratio of substrates on the yield of Bz‐RGDS‐NH 2 synthesis were examined. The optimum reaction conditions were found to be pH 10.0, 35 °C, 8 h, in an acetonitrile/(Na 2 CO 3 /NaHCO 3 ) buffer system (93:7, v/v) with a maximal yield of 65.2%. We found that secondary hydrolysis of the peptide product did not take place in these water‐miscible organic solvents.