Phosphates stepwise hydrolysis

Phosphates. Insoluble orthophosphates may be precipitated with phosphate ion derived from trimethyl or triethyl phosphate by stepwise hydrolysis. Thus 1.8M sulphuric acid containing zirconyl ions and trimethyl phosphate on... 

Stepwise Hydrolysis of a Cobalt (III) Phosphate Complex (X = CXi6H4(N02)2)... 

Methyl-p-nitrophenyl phosphate coordinated to the two metal centers in 37 undergoes hydrolysis by a two-step addition-elimination mechanism [73]. The free phosphate hydrolyzes by a concerted mechanism. In both phosphate monoester and diester hydrolysis, the two Co(m) centers in 32 and 37 stabilize the five-coordinate phosphate species (transition state or intermediate) by bringing the phosphate and nucleophile together. This stabilization leads to a change in mechanism from dissociative to concerted for a phosphate monoester hydrolysis [96] and from concerted to stepwise for phosphate diester hydrolysis [73]. 

SCHEME 10.2 Common pathways of QM formation in biological systems, (a) Stepwise two-electron oxidation by cytochrome P450 or a peroxidase, (b) Enzymatic oxidation of a catechol followed by spontaneous isomerization of the resulting n-quinone. (c) Enzymatic hydrolysis of a phosphate ester followed by base-catalyzed elimination of a leaving group from the benzylic position. 

Recently Benkovic and Schrayl28b and Clark and Kirby,26c have investigated the hydrolysis of dibenzylphosphoenolpyruvic acid and mono-benzylphospho-enolpyruvic acid which proceed via stepwise loss of benzyl alcohol (90%) and the concomitant formation of minor amounts (10%) of dibenzylphosphate and monobenzylphosphate, respectively. The pH-rate profiles for release of benzyl alcohol reveal that the hydrolytically reactive species must involve a protonated carboxyl group or its kinetic equivalent. In the presence of hydroxylamine the course of the reaction for the dibenzyl ester is diverted to the formation of dibenzyl phosphate (98%) and pyruvic acid oxime hydroxamate but remains unchanged for the monobenzyl ester except for production of pyruvic acid oxime hydroxamate. The latter presumably arises from phosphoenolpyruvate hydroxamate. These facts were rationalized according to scheme (44) for the dibenzyl ester, viz. 

It should be recalled here that the alcoholic hydroxyl of serine does not possess a dissociation constant within the pH range, accessible to enzymic reactions. Therefore, this amino acid cannot influence the pH-activity curve. On the other hand, it is well known that DFP inhibition is initially reversible and becomes only slowly irreversible. This has been demonstrated for true ChE from electric eel by Nachmansohn and associates (46) and for plasma ChE by Mackworth and Webb (47). Similarly, a stepwise reaction with inhibitors, containing the diethyl phosphoryl moiety, has been made probable by Hobbiger (34)- Therefore, it appears possible that phosphates are first attacked by the imidazol moiety of the esteratic site, in conformity with the catalytic influence of free imidazol on phosphate hydrolysis (48). This step is followed by transfer to serine. The final product is a trialkyl phosphate XV, which is not split by imidazol (scheme F). 

Hydrolysis of phosphate monoesters may proceed by either the associative or dissociative pathway depending on the type of phosphate ester (including the pK values of stepwise deprotonation of the phosphate ester), the properties of the leaving group and the pH of the solution. The neutral diprotonated form of the phosphate monoesters (and other neutral phosphoramidates and polyphosphates) appears to react via the associative pathway, while the mono-and di-anionic forms react solely by the dissociative pathway (Shen and Morgan, 1973 Wijesekera, 1992). 

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