Select your language

ISSN 1995-5537

"Biotechnology" journal V. 5, No. 2, 2012
Р. 92-97, Bibliography 20, Russian.
Universal Decimal classification: 665.58.002.39 (088.8)


Yu. A. Shesterenko, O. V. Sevastyanov, I. I. Romanovskaya

Bogatsky Physico-Chemical Institute of National Academy of Sciences of Ukraine, Odesa

According to modified method, partially purified tyrosinase preparation from Agaricus bisporus mushroom was  isolated. Using the native electrophoresis, 17 protein fractions were found, 12 of which demonstrated well_marked phenoloxidase activity, accounting for 92.5% of total protein. By the SDS-PAGE method, 9 protein fractions were revealed; the main of them were fractions with molecular masses of 12 and 41–48 kDa. The kinetic parameters of phenol and catechol oxidation catalyzed by free and immobilized tyrosinase were studied. Et was shown that absence of enzyme entrapment in matrix significantly influences on Km and Vmax of catechol oxidation, but at phenol bioconversion the Km increased 6.3-fold along with appreciable decreasing of Vmax. During the investigation of catechol oxidation, catalyzed by free and immobilized tyrosinase, as well as phenol by free enzyme, the substrate inhibition was revealed.

Key words: tyrosinase, electrophoresis, phenol oxidation, kinetic parameters, immobilization.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008