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Pyranose Dehydrogenase from Agaricus campestris and Agaricus xanthoderma: Characterization and Applications in Carbohydrate Conversions
Food Biotechnology Laboratory, BOKU–University of Natural Resources and Life Sciences Vienna, Muthgasse 11, Vienna 1190, Austria
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 28 June 2013; in revised form: 9 August 2013 / Accepted: 11 August 2013 / Published: 16 August 2013
Abstract: Pyranose dehydrogenase (PDH) is a flavin-dependent sugar oxidoreductase that is limited to a rather small group of litter-degrading basidiomycetes. The enzyme is unable to utilize oxygen as an electron acceptor, using substituted benzoquinones and (organo) metal ions instead. PDH displays a broad substrate specificity and intriguing variations in regioselectivity, depending on substrate, enzyme source and reaction conditions. In contrast to the related enzyme pyranose 2-oxidase (POx), PDHs from several sources are capable of oxidizing α- or β-1→4-linked di- and oligosaccharides, including lactose. PDH from A. xanthoderma is able to perform C-1 and C-2 oxidation, producing, in addition to lactobionic acid, 2-dehydrolactose, an intermediate for the production of lactulose, whereas PDH from A. campestris oxidizes lactose nearly exclusively at the C-1 position. In this work, we present the isolation of PDH-encoding genes from A. campestris (Ac) and A. xanthoderma (Ax) and a comparison of other so far isolated PDH-sequences. Secretory overexpression of both enzymes in Pichia pastoris was successful when using their native signal sequences with yields of 371 U·L−1 for AxPDH and 35 U·L−1 for AcPDH. The pure enzymes were characterized biochemically and tested for applications in carbohydrate conversion reactions of industrial relevance.
Keywords: pyranose dehydrogenase; heterologous expression; agaricus; lactose conversion; lactobionic acid
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Staudigl, P.; Krondorfer, I.; Haltrich, D.; Peterbauer, C.K. Pyranose Dehydrogenase from Agaricus campestris and Agaricus xanthoderma: Characterization and Applications in Carbohydrate Conversions. Biomolecules 2013, 3, 535-552.
Staudigl P, Krondorfer I, Haltrich D, Peterbauer CK. Pyranose Dehydrogenase from Agaricus campestris and Agaricus xanthoderma: Characterization and Applications in Carbohydrate Conversions. Biomolecules. 2013; 3(3):535-552.
Staudigl, Petra; Krondorfer, Iris; Haltrich, Dietmar; Peterbauer, Clemens K. 2013. "Pyranose Dehydrogenase from Agaricus campestris and Agaricus xanthoderma: Characterization and Applications in Carbohydrate Conversions." Biomolecules 3, no. 3: 535-552.