BOKU - Universität für Bodenkultur Wien - Forschungsinformationssystem
Gewählte Publikation:
Pisanelli, I., Kujawa, M., Spadiut, O., Kittl, R., Halada, P., Volc, J., Mozuch, M.D., Kersten, P., Haltrich, D. and Peterbauer, C..
(2009):
Pyranose 2-oxidase from Phanerochaete chrysosporium—Expression in E. coli and biochemical characterization
J BIOTECHNOL, 142, 97-106; ISSN 0168-1656
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- Abstract:
- The presented work reports the isolation and heterologous expression of the p2ox gene encoding the flavoprotein pyranose 2-oxidase (P2Ox) from the basidiomycete Phanerochaete chrysosporium. The p2ox cDNA was inserted into the bacterial expression vector pET21a(+) and successfully expressed in Escherichia coli. We obtained active, fully flavinylated recombinant P2Ox in yields of approximately 270 mg/l medium. The recombinant enzyme was provided with an N-terminal T7-tag and a C-terminal HiS(6)-tag to facilitate simple one-step purification. We obtained an apparently homogenous enzyme preparation with a specific activity of 16.5 U/mg. Recombinant P2Ox from P. chrysosporium was characterized in some detail with respect to its physical and catalytic properties, both for electron donor (sugar Substrates) and - for the first time - alternative electron acceptors (1,4-benzoquinone, substituted quinones. 2,6-dichloroindophenol and ferricenium ion). As judged from the catalytic efficiencies k(cat)/K-m some of these alternative electron acceptors are better substrates than oxygen, which might have implications for the proposed in vivo function of pyranose 2-oxidase. (C) 2009 Elsevier B.V. All rights reserved.
- Autor*innen der BOKU Wien:
- Haltrich Dietmar
- Kittl Roman
- Kujawa Magdalena
- Peterbauer Clemens Karl
- Find related publications in this database (Keywords)
- Pyranose 2-oxidase
- Phanerochaete chrysosporium
- Lignocellulose degradation
- Heterologous expression
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