Jw. Keillor et al., CATALYSIS OF AMIDE HYDROLYSIS AND FORMATION UNDER NEUTRAL CONDITIONS BY A ZWITTERIONIC IMIDAZOLIUM THIOLATE, Journal of the American Chemical Society, 116(11), 1994, pp. 4669-4673
As a simple model for the hydrolysis of peptides mediated by the cyste
ine proteases, 2-(mercaptomethyl)1-methylimidazole (1) was used to cat
alyze the hydrolysis of four nonactivated amides, namely, formamide, d
imethylformamide, N-formylmorpholine, and formanilide (3a-d), at 98 de
grees C, pD 7.6-8.0, mu = 1.0 (KCl). Progress of the hydrolysis reacti
ons was followed by H-1 NMR, and the kinetics as a function of added I
I] were used to determine the second-order catalytic rate constants (k
(cat)). A putative intermediate S-formyl thiolester of 1 (4) was not o
bserved to build up during the course of the hydrolysis: the partition
ing of authentic 4 between H2O and morpholine and between H2O and anil
ine was determined (98 degrees C, pD 8.0). The hydrolysis of N-formylm
orphline was observed to be catalyzed by added phosphate buffer under
the same conditions. When the hydrolysis of a 200 mM D2O solution of N
-formylmorpholine was allowed to proceed to completion, an equilibrium
position of 33 mM amide, 167 mM HCO2H, and 167 mM morpholine was atta
ined: that same equilibrium position was obtained starting with a solu
tion 200 mM in each of HCO2H and morpholine. The conditional equilibri
um constant, K'(eq) = [NFM]/([HCO2H](tot)-[morpholine](tot)), was foun
d to be 1.2 M(-1).