The influence of water on protease-catalyzed peptide synthesis in acetonitrile/water mixtures.
Protease-catalyzed peptide synthesis in acetonitrile/water mixtures, containing 0-90% water, was investigated. alpha-Chymotrypsin, as well as thermolysin, were deposited on solid supports, prior to exposure to the reaction media. Peptide syntheses were performed using both a kinetically controlled process (chymotrypsin) and an equilibrium-controlled synthesis (thermolysin). The activity of chymotrypsin decreased at low water contents. However, at low water contents (1-10%) hydrolytic side reactions were suppressed and high yields of dipeptides were obtained. Optimal water content for the thermolysin-catalyzed reaction was 4-8%. The dipeptides produced were fully soluble in the reaction mixtures. High operational stability for alpha-chymotrypsin was obtained during 216 h of reaction.
Reslow
M
M
Department of Biotechnology, Chemical Center, University of Lund, Sweden.
Adlercreutz
P
P
Mattiasson
B
B
D000097:Acetonitriles; D000327:Adsorption; D002384:Catalysis; D002918:Chymotrypsin; D004151:Dipeptides / Q000138:chemical synthesis; D004355:Drug Stability; D006868:Hydrolysis; D007202:Indicators and Reagents; D007700:Kinetics; D010447:Peptide Hydrolases / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis*; D012995:Solubility; D012996:Solutions; D013820:Thermolysin / Q000378:metabolism; D014867:Water
D000097:Acetonitriles; D004151:Dipeptides; D007202:Indicators and Reagents; D010455:Peptides; D012996:Solutions; D014867:Water; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D013820:Thermolysin; C032159:acetonitrile