Applied Microbiology and Biotechnology

, Volume 75, Issue 3, pp 549–555

Preparation of a whole-cell biocatalyst of mutated Candida antarctica lipase B (mCALB) by a yeast molecular display system and its practical properties

  • Michiko Kato
  • Jun Fuchimoto
  • Takanori Tanino
  • Akihiko Kondo
  • Hideki Fukuda
  • Mitsuyoshi Ueda
Biotechnologically Relevant Enzymes and Proteins

DOI: 10.1007/s00253-006-0835-2

Cite this article as:
Kato, M., Fuchimoto, J., Tanino, T. et al. Appl Microbiol Biotechnol (2007) 75: 549. doi:10.1007/s00253-006-0835-2

Abstract

To prepare a whole-cell biocatalyst of a stable lipase at a low price, mutated Candida antarctica lipase B (mCALB) constructed on the basis of the primary sequences of CALBs from C. antarctica CBS 6678 strain and from C. antarctica LF 058 strain was displayed on a yeast cell surface by α-agglutinin as the anchor protein for easy handling and stability of the enzyme. When mCALB was displayed on the yeast cell surface, it showed a preference for short chain fatty acids, an advantage for producing flavors; although when Rhizopus oryzae lipase (ROL) was displayed, the substrate specificity was for middle chain lengths. When the thermal stability of mCALB on the cell surface was compared with that of ROL on a cell surface, T1/2, the temperature required to give a residual activity of 50% for heat treatment of 30 min, was 60°C for mCALB and 44°C for ROL indicating that mCALB displayed on cell surface has a higher thermal stability. Furthermore, the activity of the displayed mCALB against p-nitrophenyl butyrate was 25-fold higher than that of soluble CALB, as reported previously. These findings suggest that mCALB-displaying yeast is more practical for industrial use as the whole-cell biocatalyst.

Keywords

Candida antarctica lipase BMutationYeast cell surface engineeringSubstrate specificity

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Michiko Kato
    • 1
  • Jun Fuchimoto
    • 1
  • Takanori Tanino
    • 2
  • Akihiko Kondo
    • 2
  • Hideki Fukuda
    • 3
  • Mitsuyoshi Ueda
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Department of Chemical Science and Engineering, Faculty of EngineeringKobe UniversityKobeJapan
  3. 3.Division of Molecular Science, Graduate School of Science and TechnologyKobe UniversityKobeJapan