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Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
The acid hydrolysis mechanism of acetals catalyzed by a supramolecular assembly in basic solution
Michael D Pluth1, Robert G Bergman, Kenneth N Raymond
1Department of Chemistry, University of California, Berkeley, and Division of Chemical Sciences, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Abstract:
A self-assembled supramolecular host catalyzes the hydrolysis of acetals in basic aqueous solution. The mechanism of hydrolysis is consistent with the Michaelis-Menten kinetic model. Further investigation of the rate-limiting step of the reaction revealed a negative entropy of activation (DeltaS(double dagger) = -9 cal mol(-1) K(-1)) and an inverse solvent isotope effect (k(H(2)O)/k(D(2)O) = 0.62). These data suggest that the mechanism of hydrolysis that takes place inside the assembly proceeds through an A-2 mechanism, in contrast to the A-1 mechanism operating in the uncatalyzed reaction. Comparison of the rates of acetal hydrolysis in the assembly with the rate of the reaction of unencapsulated substrates reveals rate accelerations of up to 980 over the background reaction for the substrate 1,1-diethoxyethane.
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