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Applied Microbiology and Biotechnology

, Volume 68, Issue 3, pp 327–335 | Cite as

Comparison of cyanide-degrading nitrilases

  • Dakshina M. Jandhyala
  • Richard C. Willson
  • B. Trevor Sewell
  • Michael J. Benedik
Biotechnologically Relevant Enzymes and Proteins

Abstract

Recombinant forms of three cyanide-degrading nitrilases, CynD from Bacillus pumilus C1, CynD from Pseudomonas stutzeri, and CHT from Gloeocercospora sorghi, were prepared after their genes were cloned with C-terminal hexahistidine purification tags and expressed in Escherichia coli, and the enzymes purified using nickel-chelate affinity chromatography. The enzymes were compared with respect to their pH stability, thermostability, metal tolerance, and kinetic constants. The two bacterial genes, both cyanide dihydratases, were similar with respect to pH range, retaining greater than 50% activity between pH 5.2 and pH 8 and kinetic properties, having similar Km (6–7 mM) and Vmax (0.1 mmol min−1 mg−1). They also exhibited similar metal tolerances. However, the fungal CHT enzyme had notably higher Km (90 mM) and Vmax (4 mmol min−1 mg−1) values. Its pH range was slightly more alkaline (retaining nearly full activity above 8.5), but exhibited a lower thermal tolerance. CHT was less sensitive to Hg2+ and more sensitive to Pb2+ than the CynD enzymes. These data describe, in part, the current limits that exist for using nitrilases as agents in the bioremediation of cyanide-containing waste effluent, and may help serve to determine where and under what conditions these nitrilases may be applied.

Keywords

Cyanide Cyanide Hydratase Cyanide Concentration Pseudomonas Stutzeri Start Buffer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We gratefully acknowledge support from the Robert A. Welch Foundation, the University of Houston Environmental Institute, and the Gulf Coast Hazardous Substance Research Center, #069UHH0789, to M.J.B. and from the Wellcome Trust to B.T.S. for support of this project.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Dakshina M. Jandhyala
    • 1
    • 2
  • Richard C. Willson
    • 1
    • 3
  • B. Trevor Sewell
    • 4
  • Michael J. Benedik
    • 1
    • 5
  1. 1.Department of Biology and BiochemistryUniversity of HoustonHoustonUSA
  2. 2.Institute of Veterinary, Animal, and Biomedical SciencesMassey UniversityPalmerston NorthNew Zealand
  3. 3.Department of Chemical EngineeringUniversity of HoustonHoustonUSA
  4. 4.Electron Microscope UnitUniversity of Cape TownCape TownSouth Africa
  5. 5.Department of BiologyTexas A&M UniversityCollege StationUSA

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