Hydrolysis of N — Benzoyl — L — Tyrosine Ethyl Ester Anchored in CD by α-Chymotrypsin and Subtilisin

  • K. Sandeep Prabhu
  • C. S. Ramadoss

Abstract

N-Benzoyl-L-tyrosine ethyl ester (BTEE), a water insoluble substrate, was rendered soluble upon complexation with β-cyclodextrin (or its methyl derivative) and but not with γ-CD. Kinetics of the hydrolysis of such CD anchored BTEE by subtilisin Carlsberg showed a 2 to 4 fold increase in both kcat and Km values, over the methanol solubilised substrate. Whereas, α-chymotrypsin hydrolyzed the CD anchored BTEE with only a marginally higher reaction rate than the methanol solubilised BTEE.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. [1]
    Zaks, A. and Klibanov, A. M., Enzymatic catalysis in nonaqueous solvents. J. Biol. Chem., 263(7), 3194–3201 (1986)Google Scholar
  2. [2]
    Cook, H. W. and Lands, W. E. M., Further studies of the kinetics of oxgenation of arachidonic acid by soybean lipoxygenase. Can. J. Biochem., 53, 1220–1231 (1975)PubMedCrossRefGoogle Scholar
  3. [3]
    Sandeep Prabhu, K. and Ramadoss, C. S., α-Chymotcypsin catalyzed hydrolysis of N-benzoyl-L-tyrosine ethyl ester anchored in β-cyclodextrin. Biocatal. Biotransform., 12, 281–291 (1995)CrossRefGoogle Scholar
  4. [4]
    Jyh-Ping Chen., Enhancement of enzymatic hydrolysis rate of olive oil in water by dime?hyl-β-cyclodextrin. Biotechnol. Lett., 2(9), 633–636 (1989)Google Scholar
  5. [5]
    Otero, C., Cruzado, C and Ballesteros, A., Use of cyclodextrins in enzymology to enhance me solubility of hydrophobic compounds in water. Appl. Biochem. Biotechnol, 27, 185–194(1991)CrossRefGoogle Scholar
  6. [6]
    Jyothirmayi, N. and Ramadoss, C.S., Soybean lipoxygenase catalyzed oxygenation of unsaturated fatty acid encapsulated in cyclodextrin. Biochim. Biophys. Acta., 1083, 193–200 (1991)PubMedCrossRefGoogle Scholar
  7. [7]
    Lowry, O.H., Rosenbrough, N.J., Farr, A.L. and Randall, R.J., Protein measurement with the folin phenol reagent J. Biol. Chem., 193, 265–275(1951)PubMedGoogle Scholar
  8. [8]
    Corey, D.R. and Margaret, P.A., Cyclic peptides as proteases: a reevaruation. Proc. Natl. Acad. Sci. (USA), 91, 4106–4109 (1994)CrossRefGoogle Scholar
  9. [9]
    Ezure, Y., Maruo, S., Kojima, M., Yamashita, H. and Sugiyama, M. Effect of α-cydodextrin on thermostabüity of glucoamylase. Agric. Biol. Chem., 52(4), 1073–1074 (1988)CrossRefGoogle Scholar
  10. [10]
    Karrupiah, N and Sharma, A., Cyclodextrins as protein folding aids. Biochem. Biophys. Res. Commun., 211(1), 60–66 (1995)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 1996

Authors and Affiliations

  • K. Sandeep Prabhu
    • 1
  • C. S. Ramadoss
    • 1
  1. 1.Division of Biological SciencesVittal Mallya Scientific Research FoundationBangaloreIndia

Personalised recommendations