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20 minutes acid analyses active components active fractions alcchol aliquots amino acid ammonium sulfate aqueous mushroom extract assayed photometrically Biochem Biol centrifugation Chem chloride chlorogenic acid chromatography of mushroom column of Dowex-2 concentration contained approximately continuous-flow paper cupric ion cuprous ion curtain cyanide D,L-DOPA desalted determined dialysis diphenols distilled water DOPA dopachrome coloration Dowex-2 ion exchange electrophoretic elution enzymatic enzymatic activity enzyme preparations equilibrated flask freeze-dried gradient hydrochloric acid hydrolysates illustrated in Figure investigated ion exchange resin material melanin method micromole molecular species molecules naphthoresorcinol ninhydrin ninhydrin-amino acid reaction nitrogen nitrogen content non-protein obtained optical densities oxidation oxidise para-cresol phenol phenylalanine phosphate buffer photometric assay Pigment present protein purification quantities reacted resolution of mushroom results are illustrated room temperature sample Sephadex sodium substances substrate specificity patterns thermostabilities Tris-chloride buffer tubes tyrosinase tyrosine vanillic acid yielded Рн ф ф