Purification and properties of tyrosinase isozymes from the gill of Lentinus edodes fruiting body

• Published in: Bioscience, biotechnology, and biochemistry Vol. 60; no. 8; pp. 1273 - 1278
• Main Authors: Kanda, K. (Iwate Biotechnology Research Center, Kitakami (Japan)), Sato, T, Ishii, S, Enei, H, Ejiri, S
• Format: Journal Article
• Language: English
• Published: Tokyo Taylor & Francis 01.08.1996; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; Amino Acids - analysis; Biological and medical sciences; Biotechnology; Enzymes; Food Preservation; Fundamental and applied biological sciences. Psychology; ISOENZIMAS; ISOENZYME; ISOENZYMES; Isoenzymes - chemistry; Isoenzymes - isolation & purification; LENTINUS EDODES; Metabolism; Molecular Sequence Data; Monophenol Monooxygenase - chemistry; Monophenol Monooxygenase - isolation & purification; monophenoloxidase; OXIDOREDUCTASES; OXIDORREDUCTASAS; OXYDOREDUCTASE; Plant physiology and development; Polyporaceae - enzymology; Polyporaceae - ultrastructure; PURIFICACION; PURIFICATION; Sequence Homology, Amino Acid; subunit; tyrosinase; Catalytic subunit; Purification; Enzyme; Isozyme; Alteration; Basidiomycetes; Monophenol monooxygenase; Regulatory subunit; Lentinus edodes; Fungi; Characterization; Browning; Substrate specificity; Oxidoreductases; Edible fungi; Thallophyta
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.60.1273

Six tyrosinase isozymes were purified from the browned gill of the fruiting body of Lentinus edodes by ammonium sulfate fractionation, DEAE-Sephacel and Q-Sepharose column chromatography, and partially denaturing SDS-PAGE. At the step of Q-Sepharose column chromatography, two active fractions (A and B) were obtained. Each fraction was separated to three further fractions A1, A2, and A3, and B1, B2, and B3, respectively, by partially denaturing SDS-PAGE. All these isozymes consisted of two types of polypeptides: alpha polypeptide (A-alpha or B-alpha) and either beta (A-beta or B-beta) or gamma polypeptide (A-gamma or B-gamma). The alpha polypeptide contained the consensus amino acid sequence of the active site of known tyrosinases, which is considered to act as a catalytic subunit. From the results of peptide mapping and the amino acid composition, A-alpha and B-alpha polypeptides were considered to be different proteins. The kinetic properties of the purified tyrosinase isozymes differed greatly according to whether they contained beta or gamma polypeptide, indicating these polypeptides to be a possible regulatory subunit