Počet záznamů: 1

Pyranose 2-oxidase from Phanerochaete chrysosporium--Expression in E.coli and Biochemical Characterization

  1. 1.
    0338755 - MBU-M 2010 RIV NL eng J - Článek v odborném periodiku
    Pisanelli, I. - Kujawa, M. - Spadiut, O. - Kittl, R. - Halada, Petr - Volc, Jindřich - Mozuch, M. D. - Kersten, P. - Haltrich, D. - Peterbauer, C.
    Pyranose 2-oxidase from Phanerochaete chrysosporium--Expression in E.coli and Biochemical Characterization.
    Journal of Biotechnology. Roč. 142, č. 2 (2009), s. 97-106 ISSN 0168-1656
    Výzkumný záměr: CEZ:AV0Z50200510
    Klíčová slova: Pyranose 2-oxidase * Phanerochaete chrysosporium * Lignocellulose degradation
    Kód oboru RIV: CE - Biochemie
    Impakt faktor: 2.881, rok: 2009

    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)
    Trvalý link: http://hdl.handle.net/11104/0182446