The mechanism of tyrosinase-catalysed oxidative decarboxylation of alpha-(3,4-dihydroxyphenyl)-lactic acid.
Mushroom tyrosinase, which is known to catalyse the conversion of o-diphenols into o-benzoquinones, has been shown to catalyse the oxidative decarboxylation of 3,4-dihydroxymandelic acid [Sugumaran (1986) Biochemistry 25, 4489-4492]. To account for this unusual reaction, a quinone methide intermediate has been proposed. Since all attempts to trap this intermediate ended in vain, mechanistic studies were designed to support the formation of this transient product. Replacement of the alpha-proton in 3,4-dihydroxymandelic acid with a methyl group generates alpha-(3,4-dihydroxyphenyl)-lactic acid, the enzymic oxidation of which should produce 3,4-dihydroxyacetophenone as the end product if the oxidative decarboxylation proceeds through the quinone methide intermediate. Accordingly, chemically synthesized alpha-(3,4-dihydroxyphenyl)-lactic acid on enzymic oxidation produced 3,4-dihydroxyacetophenone as the major isolatable product. Non-steady-state kinetic analysis of the enzyme reaction attested to the transient formation of the conventional quinone product. Thus the enzymic oxidation of alpha-(3,4-dihydroxyphenyl)-lactic acid seems to generate the conventional quinone, which, owing to its instability, is rapidly decarboxylated to yield the transient quinone methide. The coupled dieneonephenol re-arrangement and ketol-enol tautomerism transforms the quinone methide into 3,4-dihydroxyacetophenone.
Sugumaran
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M
Department of Biology, University of Massachusetts, Boston 02125.
Dali
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H
Semensi
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V
D001487:Basidiomycota / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D007773:Lactates / Q000138:chemical synthesis / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D011809:Quinones / Q000737:chemistry; D013055:Spectrophotometry, Infrared; D013056:Spectrophotometry, Ultraviolet
D007773:Lactates; D011809:Quinones; C070324:alpha-(3,4-dihydroxyphenyl)lactic acid; D014442:Monophenol Monooxygenase