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

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Enzymes in Nonaqueous Solvents

Part of the book series: Methods in Biotechnology ((MIBT,volume 15))

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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.

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References

  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.

    Article  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  4. Teale, F. W. J. (1960) The ultraviolet fluorescence of proteins in neutral solution. Biochem.J. 76, 381–388.

    CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  7. Woody, R. W. (1995) Circular dichroism. Methods Enzymol. 246, 34–71.

    Article  CAS  Google Scholar 

  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, 187–240.6

    Article  CAS  Google Scholar 

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© 2001 Humana Press Inc.

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Kise, H. (2001). Detection of Structural Changes of Enzymes in Nonaqueous Media by Fluorescence and CD Spectroscopy. In: Vulfson, E.N., Halling, P.J., Holland, H.L. (eds) Enzymes in Nonaqueous Solvents. Methods in Biotechnology, vol 15. Humana Press. https://doi.org/10.1385/1-59259-112-4:203

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  • DOI: https://doi.org/10.1385/1-59259-112-4:203

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-929-2

  • Online ISBN: 978-1-59259-112-1

  • eBook Packages: Springer Protocols

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