Tyrosinase-catalyzed hydroxylation of hydroquinone, a depigmenting agent, to hydroxyhydroquinone: A kinetic study

• Published in: Bioorganic & medicinal chemistry 2014-07, Vol.22 (13), p.3360-3369
• Main Authors: García-Molina, María del Mar, Muñoz Muñoz, Jose Luis, Martinez-Ortiz, Francisco, Martinez, José Rodriguez, García-Ruiz, Pedro Antonio, Rodriguez-López, José Neptuno, García-Cánovas, Francisco
• Format: Article
• Language: English
• Subjects: 2-Hydroxy-p-benzoquinone; Ascorbic acid; Ascorbic Acid - chemistry; Biocatalysis; Hydrogen peroxide; Hydrogen Peroxide - chemistry; Hydroquinone; Hydroquinones - chemistry; Hydroquinones - metabolism; Hydroxylation; Kinetic; Kinetics; Molecular Structure; Monophenol Monooxygenase - metabolism; Skin Lightening Preparations - chemistry; Skin Lightening Preparations - metabolism; Tyrosinase
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2014.04.048

Hydroquinone (HQ) is used as a depigmenting agent. In this work we demonstrate that tyrosinase hydroxylates HQ to 2-hydroxyhydroquinone (HHQ). Oxy-tyrosinase hydroxylates HQ to HHQ forming the complex met-tyrosinase-HHQ, which can evolve in two different ways, forming deoxy-tyrosinase and p-hydroxy-o-quinone, which rapidly isomerizes to 2-hydroxy-p-benzoquinone or on the other way generating met-tyrosinase and HHQ. In the latter case, HHQ is rapidly oxidized by oxygen to generate 2-hydroxy-p-benzoquinone, and therefore, it cannot close the enzyme catalytic cycle for the lack of reductant (HHQ). However, in the presence of hydrogen peroxide, met-tyrosinase (inactive on hydroquinone) is transformed into oxy-tyrosinase, which is active on HQ. Similarly, in the presence of ascorbic acid, HQ is transformed into 2-hydroxy-p-benzoquinone by the action of tyrosinase; however, in this case, ascorbic acid reduces met-tyrosinase to deoxy-tyrosinase, which after binding to oxygen, originates oxy-tyrosinase. This enzymatic form is now capable of reacting with HQ to generate p-hydroxy-o-quinone, which rapidly isomerizes to 2-hydroxy-p-benzoquinone. The formation of HHQ during the action of tyrosinase on HQ is demonstrated by means of high performance liquid chromatography mass spectrometry (HPLC–MS) by using hydrogen peroxide and high ascorbic acid concentrations. We propose a kinetic mechanism for the tyrosinase oxidation of HQ which allows us the kinetic characterization of the process. A possible explanation of the cytotoxic effect of HQ is discussed.