Applied Microbiology and Biotechnology

, Volume 82, Issue 4, pp 713–719

Enhancement of display efficiency in yeast display system by vector engineering and gene disruption

  • Kouichi Kuroda
  • Ken Matsui
  • Shinsuke Higuchi
  • Atsushi Kotaka
  • Hiroshi Sahara
  • Yoji Hata
  • Mitsuyoshi Ueda
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-008-1808-4

Cite this article as:
Kuroda, K., Matsui, K., Higuchi, S. et al. Appl Microbiol Biotechnol (2009) 82: 713. doi:10.1007/s00253-008-1808-4

Abstract

Vector engineering and gene disruption in host cells were attempted for the enhancement of α-agglutinin-based display of proteins on the cell surface in yeast. To evaluate the display efficiency by flow cytometric analysis, DsRed-monomer fused with FLAG-tag was displayed and immunostained as a model protein. The use of leu2-d in the expression vector resulted in the enhanced efficiency and ratio of the accessible display of proteins. Moreover, the amount of displayed proteins in SED1-disrupted cells increased particularly during the stationary growth phase. The combination of these improvements resulted in the quantitatively enhanced accessible display of DsRed-monomer on the yeast cell surface. The improved yeast display system would be useful in a wider range of its applications in biotechnology.

Keywords

Cell surface engineering SED1 leu2-d DsRed-monomer Display efficiency 

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kouichi Kuroda
    • 1
  • Ken Matsui
    • 1
  • Shinsuke Higuchi
    • 1
  • Atsushi Kotaka
    • 2
  • Hiroshi Sahara
    • 2
  • Yoji Hata
    • 2
  • Mitsuyoshi Ueda
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Research InstituteGekkeikan Co. Ltd.KyotoJapan