Applied Microbiology and Biotechnology

, Volume 51, Issue 1, pp 65–70

Development of an arming yeast strain for efficient utilization of starch by co-display of sequential amylolytic enzymes on the cell surface

  • T. Murai
  • M. Ueda
  • Y. Shibasaki
  • N. Kamasawa
  • M. Osumi
  • T. Imanaka
  • A. Tanaka
ORIGINAL PAPER

DOI: 10.1007/s002530051364

Cite this article as:
Murai, T., Ueda, M., Shibasaki, Y. et al. Appl Microbiol Biotechnol (1999) 51: 65. doi:10.1007/s002530051364

Abstract

The construction of a whole-cell biocatalyst with its sequential reaction has been performed by the genetic immobilization of two amylolytic enzymes on the yeast cell surface. A recombinant strain of Saccharomyces cerevisiae that displays glucoamylase and α-amylase on its cell surface was constructed and its starch-utilizing ability was evaluated. The gene encoding Rhizopus oryzae glucoamylase, with its own secretion signal peptide, and a truncated fragment of the α-amylase gene from Bacillus stearothermophilus with the prepro secretion signal sequence of the yeast α factor, respectively, were fused with the gene encoding the C-terminal half of the yeast α-agglutinin. The constructed fusion genes were introduced into the different loci of chromosomes of S. cerevisiae and expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. The glucoamylase and α-amylase activities were not detected in the culture medium, but in the cell pellet fraction. The transformant strain co-displaying glucoamylase and α-amylase could grow faster on starch as the sole carbon source than the transformant strain displaying only glucoamylase.

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • T. Murai
    • 1
  • M. Ueda
    • 1
  • Y. Shibasaki
    • 2
  • N. Kamasawa
    • 2
  • M. Osumi
    • 2
  • T. Imanaka
    • 1
  • A. Tanaka
    • 1
  1. 1.Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan Tel.: +81-75-753-5524 Fax: +81-75-753-5534JP
  2. 2.Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo 112-8681, JapanJP