0000000000262704
AUTHOR
Even Solem
Tyrosinase versus Catechol Oxidase: One Asparagine Makes the Difference
Tyrosinases mediate the ortho-hydroxylation and two-electron oxidation of monophenols to ortho-quinones. Catechol oxidases only catalyze the oxidation of diphenols. Although it is of significant interest, the origin of the functional discrimination between tyrosinases and catechol oxidases has been unclear. Recently, it has been postulated that a glutamate and an asparagine bind and activate a conserved water molecule towards deprotonation of monophenols. Here we demonstrate for the first time that a polyphenoloxidase, which exhibits only diphenolase activity, can be transformed to a tyrosinase by mutation to introduce an asparagine. The asparagine and a conserved glutamate are necessary to…
Polyphenoloxidase from Riesling and Dornfelder wine grapes (Vitis vinifera) is a tyrosinase.
Abstract Polyphenoloxidases (PPO) of the type-3 copper protein family are considered to be catecholoxidases catalyzing the oxidation of o-diphenols to their corresponding quinones. PPO from Grenache grapes has recently been reported to display only diphenolase activity. In contrast, we have characterized PPOs from Dornfelder and Riesling grapes which display both monophenolase and diphenolase activity. Ultracentrifugation and size exclusion chromatography indicated that both PPOs occur as monomers with Mr of about 38 kDa. Non-reducing SDS–PAGE shows two bands of about 38 kDa exhibiting strong activity. Remarkably, three bands up to 60 kDa displayed only very weak PPO activity, supporting th…
Tyrosinase versus Catecholoxidase: ein Asparagin macht den Unterschied
Tyrosinasen ermoglichen die ortho-Hydroxylierung und Zwei-Elektronen-Oxidation von Monophenolen zu ortho-Chinonen. Catecholoxidasen katalysieren dagegen nur die Oxidation von Diphenolen. Obwohl diese Prozesse von signifikantem Interesse sind, ist die molekulare Basis des funktionellen Unterschiedes zwischen Tyrosinasen und Catecholoxidasen noch unklar. Vor kurzem wurde postuliert, dass eine Glutaminsaure und ein Asparagin ein konserviertes Wassermolekul binden und aktivieren, um dadurch Monophenole zu deprotonieren. Hier zeigen wir, dass eine Polyphenoloxidase, die nur eine Diphenolase-Aktivitat besitzt, durch die Einfuhrung eines Asparagins mittels Mutagenese in eine Tyrosinase umgewandelt…
Are glutamate and asparagine necessary for tyrosinase activity of type-3 copper proteins?
Abstract Type-3 copper proteins (T3CPs) are complex proteins which share similar active sites. Two copper atoms (CuA and CuB) bind dioxygen as a peroxide in a side on coordination. This protein family comprises the enzymes tyrosinase and catechol oxidase as well as the oxygen transporter hemocyanin. T3CPs occur in almost all organisms and exhibit a number of essential functions. In particular, they are involved in all kinds of enzymatic browning reactions and immune defense. The chemical basis of the two catalytic processes, i.e., the o-hydroxylation of monophenols and the two-electron oxidation to o-quinones, is still discussed. Investigations on natural enzymes with known crystal structur…