PRODUCTION, PARTIAL PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF POLYPHENOL OXIDASE FROM TRICOTHECIUMSP (ENDOPHYTIC FUNGUS OF COCOA PODHUSK)SARTINI, ABD. RAUF PATONG, TJODI HARLIM AND PIRMAN
Fungal strains are considered to be excellent sources for industrial polyphenol oxidase production. Polyphenol oxidase could be produced by endophytic fungus Trichotheciumsp from podhusk of Theobroma cacaoL. The aim of this research is to partial purify and characterize of the enzyme. The enzyme was produced by submerged fermentation using media with composition, i.e: 1 % yeast extract, 25 ppm CuSO 4.5H 2O, 0.034 % gallic acid, 0.1 % KH2PO 4, 0.05 % MgSO 4.7H 2 0. The steps of purification were done by 60 - 90 % (NH 4 ) 2SO 4 precipitation, dialyzed, then partial purified by gel filtration chromatography using sephadex G-100, respectively. Polyacrylamide gel electrophoresis of the polyphenol oxidase showed 2 bands. Biochemical characteristics of the polyphenol oxidase are laccase and tyrosinase category. Optimum temperature and pH are 30 o C and 7.0, respectively.. The enzymes was inhibited by ascorbic acid, sodium metabisulphite, and disodium EDTA. The enzymes obeys Michealis-Menten kinetic with Kmand V max values being 0.625 mM and 2083 U/mL, catechol as substrate.
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