Detection of Laccase, Peroxidase, and Polyphenol Oxidase on a Single Polyacrylamide Gel Electrophoresis

Abstract Laccase, polyphenol oxidase and peroxidase are widely distributed in both plants and micro-organisms. Among these enzymes, the identification of one particular activity in the presence of the others is often difficult as enzymes can often oxidise the same substrates. A method is described, that is suitable to differentiate the three activities on the same polyacrylamide gel electrophoresis. The method is based on the use of two substrates, 4-aminoN,N-diethylaniline and 4-tert-butyl-catechol, so that coloured spots appear corresponding to the different activities. Moreover, a comparison among different mushroom species belonging to the same genus is reported using the described method.

[1]  A. Rinaldi,et al.  Polyphenol oxidase activity staining in polyacrylamide electrophoresis gels. , 1997, Journal of biochemical and biophysical methods.

[2]  A. Rescigno,et al.  EFFECTS OF SOME SUBSTITUTED ANTHRAQUINONES AND ANTHRONES ON LACCASE PRODUCTION IN PZ''UÀ OTUS SAJOR CAJU , 1996 .

[3]  Professor Dr. Gunter M. Rothe Electrophoresis of Enzymes , 1994, Springer Labmanual.

[4]  R. Dawley,et al.  Differentiation of tyrosinase and laccase using 4-hexyl-resorcinol, a tyrosinase inhibitor , 1993 .

[5]  A. Rescigno,et al.  Improved chromatographic purification of peroxidase and beta-glucosidase from Hordeum vulgare seedlings. , 1993, Preparative biochemistry.

[6]  S. Koga,et al.  Generation of superoxide during the enzymatic action of tyrosinase. , 1992, Archives of biochemistry and biophysics.

[7]  V. Kahn Tiron as a substrate for horseradish peroxidase , 1989 .

[8]  Richard W. Miller,et al.  Tiron as a substrate for mushroom tyrosinase , 1987 .

[9]  V. Kahn Tropolone—a compound that can aid in differentiating between tyrosinase and peroxidase , 1985 .

[10]  W. Peczyńska-Czoch,et al.  Analysis, Production, and Isolation of an Extracellular Laccase from Polyporus anceps , 1980, Applied and environmental microbiology.

[11]  A. Leonowicz,et al.  Quantitative estimation of laccase forms in some white-rot fungi using syringaldazine as a substrate , 1981 .

[12]  M. Hayn,et al.  A rapid assay for catechol oxidase and lactase using 2-nitro-5-thiobenzoic acid. , 1977, Analytical biochemistry.

[13]  L. C. Loon Tobacco polyphenoloxidases : a specific staining method indicating non - identity with peroxidases , 1971 .

[14]  S. Patil,et al.  Potato phenolases. Purification and properties. , 1965, The Journal of biological chemistry.