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 HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fujita, Y.
Right arrow Articles by Kondo, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fujita, Y.
Right arrow Articles by Kondo, A.
Agricola
Right arrow Articles by Fujita, Y.
Right arrow Articles by Kondo, A.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2004, p. 1207-1212, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1207-1212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Synergistic Saccharification, and Direct Fermentation to Ethanol, of Amorphous Cellulose by Use of an Engineered Yeast Strain Codisplaying Three Types of Cellulolytic Enzyme

Yasuya Fujita,1 Junji Ito,2 Mitsuyoshi Ueda,3 Hideki Fukuda,1 and Akihiko Kondo2*

Division of Molecular Science, Graduate School of Science and Technology,1 Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501,2 Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan3

Received 2 September 2003/ Accepted 3 November 2003

A whole-cell biocatalyst with the ability to induce synergistic and sequential cellulose-degradation reaction was constructed through codisplay of three types of cellulolytic enzyme on the cell surface of the yeast Saccharomyces cerevisiae. When a cell surface display system based on {alpha}-agglutinin was used, Trichoderma reesei endoglucanase II and cellobiohydrolase II and Aspergillus aculeatus ß-glucosidase 1 were simultaneously codisplayed as individual fusion proteins with the C-terminal-half region of {alpha}-agglutinin. Codisplay of the three enzymes on the cell surface was confirmed by observation of immunofluorescence-labeled cells with a fluorescence microscope. A yeast strain codisplaying endoglucanase II and cellobiohydrolase II showed significantly higher hydrolytic activity with amorphous cellulose (phosphoric acid-swollen cellulose) than one displaying only endoglucanase II, and its main product was cellobiose; codisplay of ß-glucosidase 1, endoglucanase II, and cellobiohydrolase II enabled the yeast strain to directly produce ethanol from the amorphous cellulose (which a yeast strain codisplaying ß-glucosidase 1 and endoglucanase II could not), with a yield of approximately 3 g per liter from 10 g per liter within 40 h. The yield (in grams of ethanol produced per gram of carbohydrate consumed) was 0.45 g/g, which corresponds to 88.5% of the theoretical yield. This indicates that simultaneous and synergistic saccharification and fermentation of amorphous cellulose to ethanol can be efficiently accomplished using a yeast strain codisplaying the three cellulolytic enzymes.

* Corresponding author. Mailing address: Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan. Phone: 81-78-803-6196. Fax: 81-78-803-6196. E-mail: kondo{at}cx.kobe-u.ac.jp.

Applied and Environmental Microbiology, February 2004, p. 1207-1212, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1207-1212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tsai, S.-L., Oh, J., Singh, S., Chen, R., Chen, W. (2009). Functional Assembly of Minicellulosomes on the Saccharomyces cerevisiae Cell Surface for Cellulose Hydrolysis and Ethanol Production. Appl. Environ. Microbiol. 75: 6087-6093 [Abstract] [Full Text]  
  • French, C. E. (2009). Synthetic biology and biomass conversion: a match made in heaven?. J R Soc Interface 6: S547-S558 [Abstract] [Full Text]  
  • Ito, J., Kosugi, A., Tanaka, T., Kuroda, K., Shibasaki, S., Ogino, C., Ueda, M., Fukuda, H., Doi, R. H., Kondo, A. (2009). Regulation of the Display Ratio of Enzymes on the Saccharomyces cerevisiae Cell Surface by the Immunoglobulin G and Cellulosomal Enzyme Binding Domains. Appl. Environ. Microbiol. 75: 4149-4154 [Abstract] [Full Text]  
  • Nevoigt, E. (2008). Progress in Metabolic Engineering of Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 72: 379-412 [Abstract] [Full Text]  
  • Okano, K., Zhang, Q., Kimura, S., Narita, J., Tanaka, T., Fukuda, H., Kondo, A. (2008). System Using Tandem Repeats of the cA Peptidoglycan-Binding Domain from Lactococcus lactis for Display of both N- and C-Terminal Fusions on Cell Surfaces of Lactic Acid Bacteria. Appl. Environ. Microbiol. 74: 1117-1123 [Abstract] [Full Text]  
  • Katahira, S., Fujita, Y., Mizuike, A., Fukuda, H., Kondo, A. (2004). Construction of a Xylan-Fermenting Yeast Strain through Codisplay of Xylanolytic Enzymes on the Surface of Xylose-Utilizing Saccharomyces cerevisiae Cells. Appl. Environ. Microbiol. 70: 5407-5414 [Abstract] [Full Text]  


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