Detection of Structural Changes of Enzymes in Nonaqueous Media by Fluorescence and CD Spectroscopy

  • Hideo Kise
Part of the Methods in Biotechnology book series (MIBT, volume 15)

Abstract

The ability of enzymes to catalyze synthetic reactions in mainly organic solvents has been well documented. In general, enzymes are insoluble in organic solvents. Therefore, dispersions of hydrated enzymes or immobilized enzymes have been most extensively used for synthetic reactions in organic solvents. In these reaction systems, the nature of organic solvent and water-organic ratio strongly influence the activity and specificity of enzymes. It is reasonably assumed that changes in activity and specificity of enzymes are mainly ascribed to changes in the higher structures of enzymes. Therefore, analysis of enzyme structure is important for the selection of the solvent, but methods to analyze dispersed or immobilized enzymes in organic solvents are rather limited.

Keywords

Quartz Hydrated Acetone DMSO Tyrosine 

References

  1. 1.
    Kijima, T., Yamamoto, S., and Kise, H. (1994) Fluorescence spectroscopic study of subtilisins as relevant to their catalytic activity in aqueous-organic media. Bull. Chem. Soc. Jpn. 67, 2819–2824.CrossRefGoogle Scholar
  2. 2.
    Kijima, T., Yamamoto, S., and Kise, H. (1996) Study on tryptophan fluorescence and catalytic activity of α-chymotrypsin in aqueous-organic media. Enzyme Microb. Technol. 18, 2–6.CrossRefGoogle Scholar
  3. 3.
    Sasaki, T., Kobayashi, M., and Kise, H. (1997) Active conformation of α-chymotrypsin in organic solvents as studied by circular dichroism. Biotechnol. Tech. 11, 387–390.CrossRefGoogle Scholar
  4. 4.
    Teale, F. W. J. (1960) The ultraviolet fluorescence of proteins in neutral solution. Biochem.J. 76, 381–388.Google Scholar
  5. 5.
    Genov, N., Nicolov, P., Betzel, C., Wilson, K., and Dolashka, P. (1993) Fluorescence properties of subtilisins and related proteinases (subtilases): relation to X-ray models. J. Photochem. Photobiol. B 18, 265–272.CrossRefGoogle Scholar
  6. 6.
    Yang, J. T., Wu, C.-S. C., and Martinez, H. M. (1986) Calculation of protein conformation from circular dichroism. Methods Enzymol. 130, 208–290.CrossRefGoogle Scholar
  7. 7.
    Woody, R. W. (1995) Circular dichroism. Methods Enzymol. 246, 34–71.CrossRefGoogle Scholar
  8. 8.
    Birktoft, J. J. and Blow, D. M. (1972) Structure of crystalline α-chymotrypsin. V. The atomic structure of tosyl-α-chymotrypsin at 2 Å resolution. J. Mol. Biol. 68, 187–240.6CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Hideo Kise
    • 1
  1. 1.Institute of Materials ScienceUniversity of TsukubaIbarakiJapan

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