Analysis of phenol oxidation products by scytalidium thermophilum bifunctional catalase/phenol oxidase (catpo)

Avcı, Gülden
This thesis was aimed to analyze phenol oxidation by the bifunctional catalase/phenol oxidase of the thermophilic fungus Scytalidium thermophilum. Several reactive oxygen species (ROS) are continuously produced in fungi under oxidative stress. Depending on the nature of the ROS species, some are highly toxic and are rapidly detoxified by various cellular enzymatic mechanisms, including the production of catalase. S. thermophilum produces a novel bifunctional catalase-phenol oxidase (CATPO) which is capable of oxidizing phenolics in the absence of hydrogen peroxide. Phenol oxidases convert phenolic compounds to quinones, which are then polymerized mainly by free- radical mediated reactions. In this study, 14 phenolic compounds were selected according to their different chemical structures and functional properties and were analyzed as substrates of CATPO. Among 14 phenolic compounds, only in catechol, chlorogenic acid, catechin and caffeic acid distinct oxidation products were observed by HPLC. This result showed that the range of phenolic compounds oxidized by CATPO is not only limited to catechol, but also larger MW polyphenolic compounds are oxidized. The oxidation products of catechol, caffeic acid, chlorogenic acid and catechin were characterized by LC-ESI-MS. Dimer, trimer, tetramer and oligomer formations were detected. While the maximum conversion efficiency, at 1 hour of reaction, was observed with catechin, minimum conversion efficiency was attained by caffeic acid, under the specified conditions. The oxidation products observed after oxidation of catechol, chlorogenic acid, catechin and caffeic acid by CATPO was compared with the same phenolic compounds oxidation products oxidized by laccase and tyrosinase. CATPO was incapable of oxidizing tyrosinase and laccase-specific substrates tyrosine and ABTS respectively. However, the oxidizing spectrum of substrates indicates that the nature of phenol oxidation by CATPO appears to resemble mainly those of laccase.


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Citation Formats
G. Avcı, “Analysis of phenol oxidation products by scytalidium thermophilum bifunctional catalase/phenol oxidase (catpo),” Ph.D. - Doctoral Program, Middle East Technical University, 2011.