Banner for LabNotes
From Lab Notes
Contact Information
Forest Products Laboratory
One Gifford Pinchot Drive
Madison, WI 53726-2398
Phone: (608) 231-9200
Fax: (608) 231-9592
Email

 

You are here: FPL Home  / Information Products & Services  / Publications

Requested Product

Title: Pyranose 2-oxidase from Phanerochaete chrysosporium-Expression in E. coli and biochemical characterization

Source: Journal of Biotechnology 142(2009)97-106; 2009

Author(s)Pisanelli, Ines; Kujawa, Magdalena; Spadiut, Oliver; Kittl, Roman; Halada, Petr; Volc, Jindrich; Mozuch, Michael D.; Kersten, Philip; Haltrich, dietmar, Peterbauer, Clemens

Publication Year: 2009  View PDF »

Category: Journal Articles
Associated Research Project(s):   FPL-4712-3A

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 cDNAwas insertedinto 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.

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

Publication Review Process: Informally Refereed (Peer-Reviewed)

File size: 727 kb(s)

Date posted: 02/09/2010

This publication is also viewable on Treesearch:  view
RITS Product ID: 31951
Current FPL Scientist associated with this product
Kersten, Philip J.
Research Microbiologist
  

Additional items that might interest you
View the video celebrating FPL's 100 years of public service in 2010, from the producers of the Greatest Good....view

Research Highlights from FPL....view

Termite Eradication: A search for the Holy Grail.... view

Moisture Management in Residential Construction Series videos...view

Wood Floor Systems in Residential Construction Series videos....view
- FPL's Mission and Strategic Plan -

FPL's mission is to identify and conduct innovative wood and fiber utilization research that contributes to conservation and productivity of the forest resource, thereby sustaining forests, the economy, and quality of life. ... ..more »

- FPL Research Emphasis Areas -
Advanced Composites

As an integral part of the FPL mission, we improve the long-term sustainability of our Nation's forests by creating valuable composite products from biobased materials ... ..more »


Advanced Structures

The FPL has been in the forefront of wood-frame housing research since 1910 and has long been recognized as a world leader in such housing-related areas as engineered wood ... ..more »


Forest Biorefinery

We all know the compelling reasons that the United States needs to reduce its dependence on fossil fuels. Historically, the greatest increases in energy demand have been for transportation fuels ... ..more »


Nanotechnology

A leader in wood products research for over a century, the FPL is positioning itself to become the lead Federal research facility for the application of nanotechnology in forest products ... more »


Woody Biomass Utilization

Forests in the United States contain a substantial amount of small-diameter, overstocked, and underutilized material.FPL research projects are exploring the potential of the small-diameter ... ..more »