Reaction engineering for consecutive enzymatic reactions in peptide synthesis: application to the synthesis of a pentapeptide.
A single-pot enzymatic synthesis of Z-CCK5 (4-8) is presented in this work, employing Z-Gly-Trp-OBzl as acyl donor, under kinetic control. The first goal of the work is the development of a synthetic strategy allowing the use of the same medium for two reactions catalyzed by immobilized alpha-chymotrypsin, discriminating between simultaneous and consecutive addition systems. The second goal is the maximization of the pentapeptide yield as a function of the molar excess of both nucleophiles employed. A maximum yield of 36% was obtained, and the addition strategy as well as the optimal initial concentrations of substrates have been determined.
Ruiz
S
S
Departament of d'Enginyeria Química, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Feliu
J A
JA
Caminal
G
G
Alvaro
G
G
López-Santín
J
J
D000097:Acetonitriles; D000327:Adsorption; D000595:Amino Acid Sequence; D002766:Cholecystokinin / Q000031:analogs & derivatives* / Q000096:biosynthesis; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004800:Enzymes, Immobilized; D007700:Kinetics; D010446:Peptide Fragments / Q000096:biosynthesis*; D015202:Protein Engineering; D012844:Sincalide / Q000737:chemistry
D000097:Acetonitriles; D004151:Dipeptides; D004800:Enzymes, Immobilized; D010446:Peptide Fragments; C110473:benzyloxycarbonyl-glycyl-tryptophyl-methionyl-aspartyl(OBu-t)-phenylalaninamide; D002766:Cholecystokinin; D002918:Chymotrypsin; D012844:Sincalide; C032159:acetonitrile