This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Alves, A. M. C. R.
Right arrow Articles by Wösten, H. A. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Alves, A. M. C. R.
Right arrow Articles by Wösten, H. A. B.
Agricola
Right arrow Articles by Alves, A. M. C. R.
Right arrow Articles by Wösten, H. A. B.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2004, p. 6379-6384, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6379-6384.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Highly Efficient Production of Laccase by the Basidiomycete Pycnoporus cinnabarinus

Alexandra M. C. R. Alves,1,2 Eric Record,3 Anne Lomascolo,3 Karin Scholtmeijer,4 Marcel Asther,3 Joseph G. H. Wessels,1 and Han A. B. Wösten1,2*

Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Haren,1 BioMaDe, Groningen,4 Microbiology, Institute of Biomembranes, University of Utrecht, Utrecht, The Netherlands,2 UMR 1163 de Biotechnologie des Champignons Filamenteux, INRA/Université de Provence, IFR-BAIM, Marseille, France3

Received 24 February 2004/ Accepted 24 June 2004

An efficient transformation and expression system was developed for the industrially relevant basidiomycete Pycnoporus cinnabarinus. This was used to transform a laccase-deficient monokaryotic strain with the homologous lac1 laccase gene placed under the regulation of its own promoter or that of the SC3 hydrophobin gene or the glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Schizophyllum commune. SC3-driven expression resulted in a maximal laccase activity of 107 nkat ml–1 in liquid shaken cultures. This value was about 1.4 and 1.6 times higher in the cases of the GPD and lac1 promoters, respectively. lac1-driven expression strongly increased when 25 g of ethanol liter–1 was added to the medium. Accordingly, laccase activity increased to 1,223 nkat ml–1. These findings agree with the fact that ethanol induces laccase gene expression in some fungi. Remarkably, lac1 mRNA accumulation and laccase activity also strongly increased in the presence of 25 g of ethanol liter–1 when lac1 was expressed behind the SC3 or GPD promoter. In the latter case, a maximal laccase activity of 1,393 nkat ml–1 (i.e., 360 mg liter–1) was obtained. Laccase production was further increased in transformants expressing lac1 behind its own promoter or that of GPD by growth in the presence of 40 g of ethanol liter–1. In this case, maximal activities were 3,900 and 4,660 nkat ml–1, respectively, corresponding to 1 and 1.2 g of laccase per liter and thus representing the highest laccase activities reported for recombinant fungal strains. These results suggest that P. cinnabarinus may be a host of choice for the production of other proteins as well.

* Corresponding author. Mailing address: Microbiology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Phone: 31 30 2533448. Fax: 31 30 2513655. E-mail: h.a.b.wosten{at}bio.uu.nl.

Applied and Environmental Microbiology, November 2004, p. 6379-6384, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6379-6384.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»