Pyridine, reactions with—continued hydrolysis

The above-considered calculational data point to high effectiveness of the bifunctional catalysis in hydrolytic reactions and the reactions related to these, due to involvement of molecular chains of water and ammonia, as well as to the preferability in these reactions of a concerted mechanism. This conclusion is fairly general and is corroborated by calculations on other types of nucleophilic reactions, such as hydrolysis of methyl fluoride, tautomerization of pyridine in aqueous solution etc. [110]. An advisable piece of work would apparently, be an analysis, in the light of the conclusions discussed, of mechanisms of the catalytic act in enzymic hydrolysis reactions of the ester and peptide bonds. In the most advanced up-to-date models for, e.g., the reactions with participation of a-chymotrypsin (see Ref. [Ill]), the steps of the base and the acid catalysis are separated. The latter is commonly thought [84, 111] to be operative at the stage of enzymic decomposition of the tetrahedral intermediate. However, taking into account the possibility of realization of the conformationally excited states of the active enzymic center, it would not be hard to think of some realistic schemes of concerted mechanisms, the more so that the fast growing body of calculational material continuously supplies fresh evidence in favor of such mechanisms. 

Work performed in our laboratory over the last several years has systematically addressed many of the problems associated with the thiocyanate chemistry. The use of sodium or potassium trimethylsilanolate for the cleavage reaction provided a method for rapid and specific hydrolysis of the derivatized C-terminal amino acid, which left the shortened peptide with a free C-terminal carboxylate ready for continued rounds of sequencing (3). The use of diphenylphosphoroisothiocyanatidate (DPP-ITC) and pyridine combined the activation and derivatization steps and... 

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