Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Stable Expression of Aspercillus Awamori Glucoamylase in Distiller's Yeast

Bio/Technology volume 6, pages 417–421 (1988)Cite this article

Abstract

A strain of distiller's yeast stably secreting high levels of Aspergillus awamori glucoamylase was constructed by introducing an expression cassette containing glucoamylase cDNA and a dominant selectable marker into the yeast chromosome. Glucoamylase expression levels were increased by modifying the yeast promoter and the sequences upstream of the start of translation. Fermentation studies with the improved strain showed that production of glucoamylase was constant for over 800 generations of continuous culture. This strain utilizes about 95% of the carbohydrates in soluble starch at 25% (w/v) substrate concentration and produces high yields of ethanol.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. De Mot, R., Van Dijck, K., Donkers, A. and Verachtert, H. 1985. Potentialities and limitations of direct alcohol fermentation of starchy material with amylolytic yeasts. Appl. Microbiol. Biotechnol. 22:222–226.

    Article  CAS  Google Scholar 

  2. Innis, M.A., Holland, M.J., McCabe, P.C., Cole, G.E., Wittman, V.P., Tal, R., Watt, K.W.K., Gelfand, D.H., Holland, J.P., and Meade, J.H. 1985. Expression, glycosylation, and secretion of an Aspergillus glucoamylase by Saccharomyces cerevisiae. Science 228:21–26.

    Article  CAS  Google Scholar 

  3. Holland, M.J., Holland, J.P., Thill, G.P., and Jackson, K.A. 1981. The primary structures of two yeast enoiase genes. J. Biol. Chem. 256:1385–1395.

    CAS  Google Scholar 

  4. McAlister, L. and Holland, M.J. 1982. Targeted deletion of a yeast enolase gene. J. Biol. Chem. 257:7181–7188.

    CAS  PubMed  Google Scholar 

  5. Cohen, R., Yokoi, T., Holland, J.P., Pepper, A.E., and Holland, M.J. 1987. Transcription of the constitutively expressed yeast enolase gene ENOl is mediated by positive and negative cis-acting regulatory sequences. Mol. Cell. Biol. 7:2753–2761.

    Article  CAS  Google Scholar 

  6. Innis, M.A. and McCormick, F. 1986. Procedures for expression, modification, and analysis of human fibroblast interferon (IFN-β) genes in heterologous cells. Methods Enzymol. 119:397–403.

    Article  CAS  Google Scholar 

  7. Gelfand, D.H. et al. Cetus Corporation, Emeryville, CA, 1983, unpublished data.

    Google Scholar 

  8. Tamaki, H. 1978. Genetic studies of ability to ferment starch in Saccharomyces: gene polymorphism. Mol. Gen. Genet. 164:205–209.

    Article  Google Scholar 

  9. Kniskern, P.J. 1986. Unusually high-level expression of a foreign gene (hepatitis B virus core antigen) in Saccharomyces cerevmae. Gene 46:135–145.

    Article  CAS  Google Scholar 

  10. Smith, R.A., Duncan, M.J., and Moir, D.T. 1985. Heterologous protein secretion from yeast. Science 229:1219–1224.

    Article  CAS  Google Scholar 

  11. Inlow, D., McRae, J., and Ben-Bassat, A. 1988. Fermentation of cornstarch to ethanol with genetically engineered yeast. Biotechnology and Bioengineering In press.

  12. Hanahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166:557–580.

    Article  CAS  Google Scholar 

  13. Klebe, R.J., Harriss, J.V., Sharp, Z.D., and Douglas, M.G. 1983. A general method for poiyethylene-glycol-induced genetic transformation of bacteria and yeast. Gene 25:333–341.

    Article  CAS  Google Scholar 

  14. Imanaka, T., and Aiba, S. 1981. A perspective on the application of genetic engineering: Stability of recombinant plasmids. Annals of the NY Acad. of Science 369:1–14.

    Article  CAS  Google Scholar 

  15. Gerhardt, P. (ed.). 1981. Manual of methods for general bacteriology, p.333–334. American Society for Microbiology, Washington, DC.

    Google Scholar 

  16. Oka, A., Sugisaki, H., and Takanami, M. 1980. Nucleotide sequence of the kanamycin resistance transposon Tn903. J. Mol. Biol. 147:217–226.

    Article  Google Scholar 

  17. Southern, E. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98:503–517.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Duane Inlow and Arie Ben-Bassat: Fermentation Research and Development, Cetus Corporation, 1400 53rd St., Emeryville, CA 94608.

  2. Michael A. Innis: Corresponding author.

Authors and Affiliations

  1. Departments of Microbial Genetics, Cetus Corporation, 1400 53rd St., Emeryville, CA, 94608

    Georgette E. Cole, Peter C. McCabe, Duane Inlow, David H. Gelfand, Arie Ben-Bassat & Michael A. Innis

Authors
  1. Georgette E. Cole
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter C. McCabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Duane Inlow
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David H. Gelfand
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Arie Ben-Bassat
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michael A. Innis
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cole, G., McCabe, P., Inlow, D. et al. Stable Expression of Aspercillus Awamori Glucoamylase in Distiller's Yeast. Nat Biotechnol 6, 417–421 (1988). https://doi.org/10.1038/nbt0488-417

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0488-417

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing