• M. Solís-Oba Centro de Investigación en Biotecnología Aplicada del Instituto Politécnico Nacional
  • E. Bárzana Facultad de Química, Universidad Nacional Autónoma de México
  • M. García-Garibay Departamento de Biotecnología, Universidad Autónoma Metropolitana Unidad Iztapalapa
  • G. Viniegra-González Departamento de Biotecnología, Universidad Autónoma Metropolitana Unidad Iztapalapa
Keywords: ABTS, recycling, oxidation, dyes, lacction


The commercial enzyme laccase cloned from Myceliophtera termophila and expressed in Aspergillus oryzae was used to produce an active, stable and oxidized form of the chemical mediator ABTS (2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) in order to test the feasibility of using the monocation ABTS + as a recyclable intermediate for the oxidation of a variety of aromatic chemicals in the absence of the enzyme. It was shown that ABTS +, by itself, oxidized, indigo blue, tannic and galic acids, brilliant blue G, Coomassie blue,
orange 7 and p-cresol. Oxidation was assessed by important changes in their visible absorption spectra. It was also shown that oxidation of these compounds by a mixture of ABTS and laccase can be done in a cyclic manner, such a way the oxidation rate was 94 times higher for indigo, 17 times for brilliant blue G, 34 times for orange 7 and 5 times higher for p-cresol compared with using the mediator or the laccase alone. Those results show the feasibility of a recycling scheme for the final oxidation of a wide variety of aromatic compounds in which the enzyme is not directly in contact with the compound but through the intermediate ABTS +.


Bourbonnais R., Paice M. (1990). Oxidation of nonphenolic substrates. An expanded role for laccase in lignin biodegradation. FEBS Letters 267, 99–102.

Bourbonnais R., Paice M., Freiermuth B., Bodie E., Borneman S. (1997). Reactivities of various mediators and laccases with Kraft pulp and lignin model compounds, Applied and Environmental Microbiology, 4627-4632.

Collins P., Dobson A., Field J. (1998), Reduction of the 2,2 -Azino-bis (3-Ethylbenzthiazoline-6- Sulfonate) Cation Radical by Physiological Organic Acids in the Absence and Presence of Manganese. Applied and Environmental Microbiology 64 (6), 2026-2031.

Greca M., Costa-Ferreira M., Pessoa M. (2001). Decolorization of an anthraquinone-type dye using a laccase formulation. Bioresource Technology 79, 171-177. Ho G., Liao Ch. (1983). Activation of a siliceous carrier for enzyme immobilization. United States Patent 4,384,045.

Johannes Ch., Majcherczyk A. (2000a). Natural mediators in the oxidation of polycyclic aromatic hydrocarbons by laccase mediator system. Applied and Environmental Microbiology 66, 524-528.

Johannes Ch., Majcherczyk A. (2000b). Laccase activity tests and laccase inhibitors. Journal of Biotechnology 78, 193-199.

Li K., Helm R., Eriksson K. (1998). Mechanistic studies of the oxidation of a non-phenolic lignin model compound by the laccase/Ihidroxibenzotriazole redox system. Biotechnology and Applied Biochemistry 27, 239-243.

Lonergan G., Mew E., Schliephake K., Baker W., (1997). Phenolic substrate for fluorometric detection of laccase activity. FEMS Microbiology Letters 153, 485-490.

Majcherczyk A., Johannes C., (2000). Radical mediated indirect oxidation of a PEG-coupled polycyclic aromatic hydrocarbon (PAH) model compound by fungal laccase. Biochimica et Biophysica Acta-General Subjects 1474 (2), 157-162.

Novo Nordisk, Deni Lite II S Product Sheet, Enzyme Business, Denmark, 1999.

Potthast A., Rosenau T., Chen C., Gratzl J. (1996). A novel method for the conversion of benzyl alcohols to benzaldehydes by laccasecatalysed oxidation. Journal of Molecular Catalysis A. 108, 5-9.

Rodríguez E., Pickard M., Vázquez-Duhalt R. (1999). Industrial dye decolorization by laccase from ligninolytic fungi. Current Microbiology 38, 27-32.

Solís-Oba M., Ugalde-Saldivar V., Gonzalez I., Viniegra-Gonzalez G. (2005). An electrochemical-spectrophotometrical study of the oxidized forms of the mediator 2,2’- azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) produced by immobilized laccase. Journal of Electroanalytical Chemistry 579, 59-66.

Thurston Ch. (1994). The structure and function of fungal laccase, review article. Microbiology 140, 19-26.

Xu F. (1996). Oxidation of phenols, anilines and benzenethiols by fungal laccase: correlation between activity and redox potentials as well as halide inhibition. Biochemistry 35, 7608- 7614.
How to Cite
Solís-Oba, M., Bárzana, E., García-Garibay, M., & Viniegra-González, G. (2020). THE ABTS●+ AN OXIDANT AGENT OF DIFFERENT CHEMICAL COMPOUNDS AND ITS RECYCLING PROCESS BETWEEN LACCASE AND SUBSTRATE. Revista Mexicana De Ingeniería Química, 6(3), 275-281. Retrieved from