Skip to Main Content
U.S. Forest Service
Caring for the land and serving people

United States Department of Agriculture

Home > Search > Publication Information

  1. Share via EmailShare on FacebookShare on LinkedInShare on Twitter
    Dislike this pubLike this pub
    Author(s): Ines Pisanelli; Magdalena Kujawa; Oliver Spadiut; Roman Kittl; Petr Halada; Jindrich Volc; Michael D. MozuchPhilip Kersten; Dietmar Haltrich; Clemens Peterbauer
    Date: 2009
    Source: Journal of biotechnology. Vol. 142, no. 2 (June 15, 2009): pages 97-106.
    Publication Series: Miscellaneous Publication
    PDF: View PDF  (726.34 KB)

    Description

    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/lmedium. The recombinant enzyme was provided with an N-terminal T7-tag and a C-terminal His6-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 kcat/Km, 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.

    Publication Notes

    • We recommend that you also print this page and attach it to the printout of the article, to retain the full citation information.
    • This article was written and prepared by U.S. Government employees on official time, and is therefore in the public domain.

    Citation

    Pisanelli, Ines; Kujawa, Magdalena; Spadiut, Oliver; Kittl, Roman; Halada, Petr; Volc, Jindrich; Mozuch, Michael D.; Kersten, Philip; Haltrich, Dietmar; Peterbauer, Clemens. 2009. Pyranose 2-oxidase from Phanerochaete chrysosporium : expression in E. coli and biochemical characterization. Journal of biotechnology. Vol. 142, no. 2 (June 15, 2009): pages 97-106.

    Cited

    Google Scholar

    Keywords

    Pyranose 2-oxidase, Phanerochaete chrysosporium, lignocellulose degradation, heterologous expression, lignocellulose, biodegradation, enzymes, Basidiomycetes, molecular genetics, fungi, genetics, wood-decaying fung, industrial applications, gene expression, biotechnology, pyranose, Escherichia coli, Bacteria, flavoproteins, recombinant DNA, sugars, microbial metabolism, chemical kinetics, oxygen

    Related Search


    XML: View XML
Show More
Show Fewer
Jump to Top of Page
https://www.fs.usda.gov/treesearch/pubs/35193