Enzymes greatly enhance the rate of reactions by a variety of physical organic mechanisms. One of the more contentious of these has been desolvation. To get a quantitative assessment of this contribution, we examined acetyl transfer to oxydianions. This is a model reaction for enzymes in which a high-energy acyl phosphate is formed. The aqueous reaction of phosphate or phosphonates with p-nitrophenyl acetate (pNPA) shows a substantial negative deviation from the Brønsted correlation obtained with monoanionic nucleophiles and from the reactivity of other, larger, oxydianions (molybdate, arsenate and vanadate). This, and other data, suggests a significant contribution of desolvation to the activation energy. To further investigate this we studied the effect of various DMSO (dimethyl sulfoxide)/H_2O mixtures on the reaction of chloromethylphosphonate, and of molybdate, on the reaction with pNPA. Increasing the DMSO concentration from 1% to 90% (v/v) increases the second-order rate constant for each of these reactions by over a factor of 5000. This is over a 1000 times greater than the enhancing effect on the reaction of phenoxides and over 105 times the (inhibiting) effect on the reaction with neutral nucleophiles (imidazole). Extrapolation to pure DMSO yields a rate enhancement of ∼10^5, relative to the reaction in water, for the oxydianions. This enhancement is over 3 orders of magnitude greater than that seen with monoanionic phenoxide nucleophiles. This suggests a significant role for desolvation in the reactions in the enzyme-catalyzed nucleophilic reactions of inorganic phosphate but a modest role in reactions with less highly charged nucleophiles
