Purification and characterization of tyrosinase from Streptomyces vinceudrauppus DSV 5

Abstract

Streptomyces vinceudrauppus DSV 5 is the first investigated report for tyrosinase activity. The studies presented in this research show that this organism may be a future source for larger production of tyrosinase. The enzyme was purified initially with 140 ml of culture filtrate. The crude enzyme was primarily purified by centrifugation, followed by ammonium sulfate precipitation and ultrafiltration and employed to ion exchange chromatography. Thereafter, the enzyme was loaded on a Sephadex G-75 column and, after ultra filtration, 0.54 mg of purified tyrosinase were obtained and confirmed results from sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Tyrosinase kinetics was determined with L-DOPA as substrate, the kinetic parameters are Km - 0.17 mM and Vmax - 227 IU/ml were determined.