This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Murai, T. Articles by Tanaka, A. Search for Related Content PubMed PubMed Citation Articles by Murai, T. Articles by Tanaka, A. Agricola Articles by Murai, T. Articles by Tanaka, A.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Apr 1997, 1362-1366, Vol 63, No. 4
Copyright © 1997, American Society for Microbiology

Construction of a starch-utilizing yeast by cell surface engineering

T Murai, M Ueda, M Yamamura, H Atomi, Y Shibasaki, N Kamasawa, M Osumi, T Amachi and A Tanaka
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Japan.

We have engineered the cell surface of the yeast Saccharomyces cerevisiae by anchoring active glucoamylase protein on the cell wall, and we have endowed the yeast cells with the ability to utilize starch directly as the sole carbon source. The gene encoding Rhizopus oryzae glucoamylase with its secretion signal peptide was fused with the gene encoding the C-terminal half (320 amino acid residues from the C terminus) of yeast alpha-agglutinin, a protein involved in mating and covalently anchored to the cell wall. The constructed plasmid containing this fusion gene was introduced into S. cerevisiae and expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter from S. cerevisiae. The glucoamylase activity as not detected in the culture medium, but it was detected in the cell pellet fraction. The glucoamylase protein transferred to the soluble fraction from the cell wall fraction after glucanase treatment but not after sodium dodecyl sulfate treatment, indicating the covalent binding of the fusion protein to the cell wall. Display of the fused protein was further confirmed by immunofluorescence microscopy and immunoelectron microscopy. The transformant cells could surely grow on starch as the sole carbon source. These results showed that the glucoamylase was anchored on the cell wall and displayed as its active form. This is the first example of an application of cell surface engineering to utilize and improve the metabolic ability of cells.

This article has been cited by other articles:

• Shimma, Y.-i., Saito, F., Oosawa, F., Jigami, Y. (2006). Construction of a Library of Human Glycosyltransferases Immobilized in the Cell Wall of Saccharomyces cerevisiae. Appl. Environ. Microbiol. 72: 7003-7012 [Abstract] [Full Text]  
• Shiraga, S., Kawakami, M., Ishiguro, M., Ueda, M. (2005). Enhanced Reactivity of Rhizopus oryzae Lipase Displayed on Yeast Cell Surfaces in Organic Solvents: Potential as a Whole-Cell Biocatalyst in Organic Solvents. Appl. Environ. Microbiol. 71: 4335-4338 [Abstract] [Full Text]  
• Shigechi, H., Koh, J., Fujita, Y., Matsumoto, T., Bito, Y., Ueda, M., Satoh, E., Fukuda, H., Kondo, A. (2004). Direct Production of Ethanol from Raw Corn Starch via Fermentation by Use of a Novel Surface-Engineered Yeast Strain Codisplaying Glucoamylase and {alpha}-Amylase. Appl. Environ. Microbiol. 70: 5037-5040 [Abstract] [Full Text]  
• Abe, H., Shimma, Y.-i., Jigami, Y. (2003). In vitro oligosaccharide synthesis using intact yeast cells that display glycosyltransferases at the cell surface through cell wall-anchored protein Pir. Glycobiology 13: 87-95 [Abstract] [Full Text]  
• Matsumoto, T., Fukuda, H., Ueda, M., Tanaka, A., Kondo, A. (2002). Construction of Yeast Strains with High Cell Surface Lipase Activity by Using Novel Display Systems Based on the Flo1p Flocculation Functional Domain. Appl. Environ. Microbiol. 68: 4517-4522 [Abstract] [Full Text]  
• Lynd, L. R., Weimer, P. J., van Zyl, W. H., Pretorius, I. S. (2002). Microbial Cellulose Utilization: Fundamentals and Biotechnology. Microbiol. Mol. Biol. Rev. 66: 506-577 [Abstract] [Full Text]  
• Moukadiri, I., Jaafar, L., Zueco, J. (1999). Identification of Two Mannoproteins Released from Cell Walls of a Saccharomyces cerevisiae mnn1 mnn9 Double Mutant by Reducing Agents. J. Bacteriol. 181: 4741-4745 [Abstract] [Full Text]  
• Murai, T., Ueda, M., Kawaguchi, T., Arai, M., Tanaka, A. (1998). Assimilation of Cellooligosaccharides by a Cell Surface-Engineered Yeast Expressing beta -Glucosidase and Carboxymethylcellulase from Aspergillus aculeatus. Appl. Environ. Microbiol. 64: 4857-4861 [Abstract] [Full Text]