A pH-Indicator-Based Screen for Hydrolytic Haloalkane Dehalogenase

  • Huimin Zhao
Part of the Methods in Molecular Biology™ book series (MIMB, volume 230)

Abstract

Microbial hydrolytic haloalkane dehalogenases catalyze the cleavage of halogen-carbon bonds of a variety of aliphatic halogenated compounds, including a broad range of chlorinated (C2–C6) and brominated (C2–C8) alkanes, with water as the sole co-substrate, resulting in the production of halide ions, protons, and alcohols (1,2). Based primarily on substrate specificity and sequence homology, these enzymes have been classified into two general classes that are represented by the enzymes from Xanthobacter autotrophicus GJ10 and Rhodococcus rhodochrous (3). The study of these enzymes has been motivated largely by their potential use in waste treatment, bioremediation and industrial biocatalysis (4,5). A substantial amount of mechanistic and structural information is available for these enzymes (6, 7, 8, 9).

Keywords

Codon Lipase Chlorinate Alkane Gelatin 

References

  1. 1.
    Fetzner, S. and Lingens, F. (1994) Bacterial dehalogenases: biochemistry, genetics, and biotechnological applications. Microbiol. Rev. 58, 641–685.PubMedGoogle Scholar
  2. 2.
    Slater, J. H., Bull, A. T., and Hardman, D. J. (1997) Microbial dehalogenation of halogenated alkanoic acids, alcohols and alkanes. Adv. Microb. Physiol. 38, 133–176.PubMedCrossRefGoogle Scholar
  3. 3.
    Damborsky, J., Nyandoroh, M. G., Nemec, M., Holoubek, I., Bull, A. T., and Hardman, D. J. (1997) Some biochemical properties and the classification of a range of bacterial haloalkane dehalogenase. Biotechnol. Appl. Biochem. 26, 19–25.PubMedGoogle Scholar
  4. 4.
    Swanson, P. E. (1999) Dehalogenases applied to industrial-scale biocatalysis. Curr. Opin. Biotechnol. 10, 365–369.PubMedCrossRefGoogle Scholar
  5. 5.
    Copley, S. D. (1998) Microbial dehalogenases: enzymes recruited to convert xenobiotic substrates. Curr. Opin. Chem. Bio. 2, 613–617.CrossRefGoogle Scholar
  6. 6.
    Schindler, J. F., Naranjo, P. A., Honaberger, D. A., et al. (1999) Haloalkane dehalogenase: steady-state kinetics and halide inhibition. Biochemistry 38, 5772–5778.PubMedCrossRefGoogle Scholar
  7. 7.
    Newman, J., Peat, T. S., Richard, R., et al. (1999) Haloalkane dehalogenase: structure of a Rhodococcus enzyme. Biochemistry 38, 16,105–16,114.PubMedCrossRefGoogle Scholar
  8. 8.
    Verschueren, K. H. G., Seljee, F., Rozeboom, H. J., Kalk, K. H., and Dijkstra, B. W. (1993) Crystallographic analysis of the catalytic mechanism of haloalkane dehalogenase. Nature 363, 693–698.PubMedCrossRefGoogle Scholar
  9. 9.
    Schanstra, J. P., Kingma, J., and Janssen, D. (1996) Specificity and kinetics of haloalkane dehalogenase. J. Biol. Chem. 271, 14,747–14,753.PubMedCrossRefGoogle Scholar
  10. 10.
    Arnold, F. H. (1998) Design by directed evolution. Acc. Chem. Res. 31, 125–131.CrossRefGoogle Scholar
  11. 11.
    Kuchner, O. and Arnold, F. H. (1997) Directed evolution of enzyme catalysts. Trends Biotechnol. 15, 523–530.PubMedCrossRefGoogle Scholar
  12. 12.
    Janes, L. E., Lowendahl, C., and Kazlauskas, R. J. (1998) Quantitative screening of hydrolase libraries using pH indicators: identifying active and enantioselective hydrolases. Chem. Eur. J. 4, 2324–2331.CrossRefGoogle Scholar
  13. 13.
    Zhao, H. (1998) Improved recombinant haloaliphatic dehalogenases. PCT/US 00/06132.Google Scholar
  14. 14.
    Affholter, J. A., Swanson, P. E., Kan, H. L., and Richard, R.A. (1998) Recombinant haloaliphatic dehalogenases. PCT/US 98/36080.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2003

Authors and Affiliations

  • Huimin Zhao
    • 1
  1. 1.Department of Chemical and Biomolecular EngineeringUniversity of IllinoisUrbana

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