Applied Microbiology and Biotechnology

, Volume 86, Issue 6, pp 1895–1902

Synthesis of functional dipeptide carnosine from nonprotected amino acids using carnosinase-displaying yeast cells

  • Chiaki Inaba
  • Shinsuke Higuchi
  • Hironobu Morisaka
  • Kouichi Kuroda
  • Mitsuyoshi Ueda
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-009-2396-7

Cite this article as:
Inaba, C., Higuchi, S., Morisaka, H. et al. Appl Microbiol Biotechnol (2010) 86: 1895. doi:10.1007/s00253-009-2396-7

Abstract

Carnosine (β-alanyl-l-histidine) is one of the bioactive dipeptides and has antioxidant, antiglycation, and cytoplasmic buffering properties. In this study, to synthesize carnosine from nonprotected amino acids as substrates, we cloned the carnosinase (CN1) gene and constructed a whole-cell biocatalyst displaying CN1 on the yeast cell surface with α-agglutinin as the anchor protein. The display of CN1 was confirmed by immunofluorescent labeling, and CN1-displaying yeast cells showed hydrolytic activity for carnosine. When carnosine was synthesized by the reverse reaction of CN1, organic solvents were added to the reaction mixture to reduce the water content. The CN1-displaying yeast cells were lyophilized and examined for organic solvent tolerance. Results showed that the CN1-displaying yeast cells retained their original hydrolytic activity in hydrophobic organic solvents. In the hydrophobic organic solvents and hydrophobic ionic liquids, the CN1-displaying yeast cells catalyzed carnosine synthesis, and carnosine was synthesized from nonprotected amino acids in only one step. The results of this research suggest that the whole-cell biocatalyst displaying CN1 on the yeast cell surface can be used to synthesize carnosine with ease and convenience.

Keywords

Carnosine Whole-cell biocatalyst Carnosinase (CN1) Functional dipeptide Cell surface engineering Cell surface display 

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Chiaki Inaba
    • 1
  • Shinsuke Higuchi
    • 1
  • Hironobu Morisaka
    • 1
  • Kouichi Kuroda
    • 1
  • Mitsuyoshi Ueda
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan