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

SYSNO ASEP	0338755
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Pyranose 2-oxidase from Phanerochaete chrysosporium--Expression in E.coli and Biochemical Characterization
Author(s)	Pisanelli, I. (AT) Kujawa, M. (AT) Spadiut, O. (AT) Kittl, R. (AT) Halada, Petr (MBU-M)_{RID, ORCID} Volc, Jindřich (MBU-M)_RID Mozuch, M. D. (US) Kersten, P. (US) Haltrich, D. (AT) Peterbauer, C. (AT)
Source Title	Journal of Biotechnology. - : Elsevier - ISSN 0168-1656 Roč. 142, č. 2 (2009), s. 97-106
Number of pages	10 s.
Language	eng - English
Country	NL - Netherlands
Keywords	Pyranose 2-oxidase ; Phanerochaete chrysosporium ; Lignocellulose degradation
Subject RIV	CE - Biochemistry
CEZ	AV0Z50200510 - MBU-M (2005-2011)
UT WOS	000267567900001
DOI	10.1016/j.jbiotec.2009.03.019
Annotation	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)
Workplace	Institute of Microbiology
Contact	Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
Year of Publishing	2010

Number of the records: 1