Protease-catalyzed peptide synthesis in frozen aqueous systems: the "freeze-concentration model".

Freezing of the aqueous reaction mixtures can cause dramatically increased peptide yields in protease-catalyzed kinetically controlled peptide synthesis reactions. The "Freeze-concentration model" which implies that the enzymic reaction within a macroscopically frozen system takes place in an unfrozen liquid phase is presented as a reasonable basis for the understanding of the effect of freezing. It is shown that L-amino acid esters are effective nucleophiles in alpha-chymotrypsin-catalyzed acyl transfer reactions when these are performed in frozen state.