Skip to main content
SpringerLink
Log in

Purification and Characterization of NAD+-Dependent Salicylaldehyde Dehydrogenase from Carbaryl-Degrading Pseudomonas sp. Strain C6

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

NAD+-dependent salicylaldehyde dehydrogenase (SALDH) which catalyzes the oxidation of salicylaldehyde to salicylate was purified form carbaryl-degrading Pseudomonas sp. strain C6. The enzyme was found to be a functional homotrimer (150 kDa) with subunit molecular mass of 50 kDa and contained calcium (1.8 mol/mol of enzyme). These properties were found to be unique. External addition of metal ions showed no effect on the activity and addition of chelators showed moderate inhibition of the activity. Potassium ions were found to enhance the activity significantly. SALDH showed higher affinity for salicylaldehyde (K m = 4.5 μM) and accepts mono- as well as di-aromatic aldehydes; however it showed poor activity on aliphatic aldehydes. Chloro-/nitro-substituted benzaldehydes were potent substrate inhibitors as compared to benzaldehyde and 3-hydroxybenzaldehyde, while 2-naphthaldehyde and salicylaldehyde were moderate. The kinetic data revealed that SALDH, though having broad specificity, is more efficient for the oxidation of salicylaldehyde as compared to other aromatic aldehyde dehydrogenases which gives an advantage for Pseudomonas sp. strain C6 to bioremediate carbaryl and other aromatic aldehydes efficiently.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

MSM:

Minimal salt medium

SALDH:

Salicylaldehyde dehydrogenase

TLC:

Thin layer chromatography

DMSO:

Dimethyl sulfoxide

References

  1. Andersen, A. (2006). International Journal of Toxicology, 25(Suppl 1), 11–27.

    CAS  Google Scholar 

  2. Caboni, P., Aissani, N., Cabras, T., Falqui, A., Marotta, R., Liori, B., et al. (2013). Journal of Agricultural and Food Chemistry, 61, 1794–1803.

    Article  CAS  Google Scholar 

  3. Deneer, J. W., Seinen, W., & Hermens, J. L. M. (1988). Aquatic Toxicology, 12, 185–192.

    Article  CAS  Google Scholar 

  4. Iwasawa, A., Niwano, Y., Kohno, M., & Ayaki, M. (2011). Biocontrol Science, 16, 165–170.

    Article  CAS  Google Scholar 

  5. Mazza, M., Montanari, L., & Pavanetto, F. (1976). Farmaco Science, 31, 334–344.

    CAS  Google Scholar 

  6. Niknahad, H., Shuhendler, A., Galati, G., Siraki, A. G., Easson, E., Poon, R., et al. (2003). Chemico-Biological Interactions, 143–144, 119–128.

    Article  Google Scholar 

  7. Poirier, M., Fournier, M., Brousseau, P., & Morin, A. (2002). Journal of Toxicology and Environmental Health. Part A, 65, 1437–1451.

    Article  CAS  Google Scholar 

  8. Smiesko, M., & Benfenati, E. (2004). Journal of Chemical Information and Computer Sciences, 44, 976–984.

    CAS  Google Scholar 

  9. Becker, T. W., Krieger, G., & Witte, I. (1996). Free Radical Research, 24, 325–332.

    Article  CAS  Google Scholar 

  10. Lindahl, R. (1992). Critical Reviews in Biochemistry and Molecular Biology, 27, 283–335.

    Article  CAS  Google Scholar 

  11. Netzeva, T. I., & Schultz, T. W. (2005). Chemosphere, 61, 1632–1643.

    Article  CAS  Google Scholar 

  12. Roy, K., & Das, R. N. (2010). Journal of Hazardous Materials, 183, 913–922.

    Article  CAS  Google Scholar 

  13. Vasiliou, V., Pappa, A., & Petersen, D. R. (2000). Chemico-Biological Interactions, 129, 1–19.

    Article  CAS  Google Scholar 

  14. Patnaik, P. (2007). A comprehensive guide to the hazardous properties of chemical substances (3rd ed.). New Jersey: Wiley.

    Book  Google Scholar 

  15. Connors, M. A., & Barnsley, E. A. (1982). Journal of Bacteriology, 149, 1096–1101.

    CAS  Google Scholar 

  16. Eaton, R. W., & Chapman, P. J. (1992). Journal of Bacteriology, 174, 7542–7554.

    CAS  Google Scholar 

  17. Fuenmayor, S. L., Wild, M., Boyes, A. L., & Williams, P. A. (1998). Journal of Bacteriology, 180, 2522–2530.

    CAS  Google Scholar 

  18. Grund, E., Denecke, B., & Eichenlaub, R. (1992). Applied and Environmental Microbiology, 58, 1874–1877.

    CAS  Google Scholar 

  19. Hayatsu, M., Hirano, M., & Nagata, T. (1999). Applied and Environmental Microbiology, 65, 1015–1019.

    CAS  Google Scholar 

  20. Kuhm, A. E., Stolz, A., Ngai, K. L., & Knackmuss, H. J. (1991). Journal of Bacteriology, 173, 3795–3802.

    CAS  Google Scholar 

  21. Shamsuzzaman, K. M., & Barnsley, E. A. (1974). Biochemical and Biophysical Research Communications, 60, 582–589.

    Article  CAS  Google Scholar 

  22. Swetha, V. P., & Phale, P. S. (2005). Applied and Environmental Microbiology, 71, 5951–5956.

    Article  CAS  Google Scholar 

  23. Williams, P. A., Catterall, F. A., & Murray, K. (1975). Journal of Bacteriology, 124, 679–685.

    CAS  Google Scholar 

  24. Barnsley, E. A. (1976). Journal of Bacteriology, 125, 404–408.

    CAS  Google Scholar 

  25. Chapalamadugu, S., & Chaudhry, G. R. (1991). Applied and Environmental Microbiology, 57, 744–750.

    CAS  Google Scholar 

  26. Seo, J. S., Keum, Y. S., & Li, Q. X. (2013). Biodegradation. doi:10.1007/s10532-013-9629-2.

    Google Scholar 

  27. Eggeling, L., & Sahm, H. (1985). European Journal of Biochemistry, 150, 129–134.

    Article  CAS  Google Scholar 

  28. Li, X., Li, Y., Wei, D., Li, P., Wang, L., & Feng, L. (2010). Microbiological Research, 165, 706–712.

    Article  CAS  Google Scholar 

  29. MacKintosh, R. W., & Fewson, C. A. (1988). The Biochemical Journal, 255, 653–661.

    CAS  Google Scholar 

  30. Shrivastava, R., Basu, A., & Phale, P. S. (2011). Archives of Microbiology, 193, 553–563.

    Article  CAS  Google Scholar 

  31. van Ophem, P. W., Van, B. J., & Duine, J. A. (1992). European Journal of Biochemistry, 206, 511–518.

    Article  Google Scholar 

  32. Yeung, C. K., Yep, A., Kenyon, G. L., & McLeish, M. J. (2008). Biochimica et Biophysica Acta, 1784, 1248–1255.

    Article  CAS  Google Scholar 

  33. Zhao, H., Li, Y., Chen, W., & Cai, B. (2007). Chinese Science Bulletin, 52, 1942–1948.

    Article  CAS  Google Scholar 

  34. Davies, J. I., & Evans, W. C. (1964). The Biochemical Journal, 91, 251–261.

    CAS  Google Scholar 

  35. Rossello-Mora, R. A., Lalucat, J., & Garcia-Valdes, E. (1994). Applied and Environmental Microbiology, 60, 966–972.

    CAS  Google Scholar 

  36. Schell, M. A. (1983). Journal of Bacteriology, 153, 822–829.

    CAS  Google Scholar 

  37. Li, S., Li, X., Zhao, H., & Cai, B. (2011). Microbiological Research, 166, 643–653.

    Article  CAS  Google Scholar 

  38. Coitinho, J. B., Costa, D. M., Guimaraes, S. L., de Goes, A. M., & Nagem, R. A. (2012). Acta Crystallographica, Section F: Structural Biology and Crystallization Communications, 68, 93–97.

    CAS  Google Scholar 

  39. Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  40. Laemmli, U. K. (1970). Nature, 227, 680–685.

    Article  CAS  Google Scholar 

  41. Britton, H. T. S., & Robinson, R. A. (1931). Journal of the Chemical Society, 0, 1456–1462.

    Article  CAS  Google Scholar 

  42. Singh, R., Trivedi, V., & Phale, P. (2013). Archives of Microbiology, 195, 521–535.

    Article  CAS  Google Scholar 

  43. Monod, J., Wyman, J., & Changeux, J. P. (1965). Journal of Molecular Biology, 12, 88–118.

    Article  CAS  Google Scholar 

  44. Wood, W. A. (1977). Trends in Biochemical Sciences, 2, 223–226.

    Article  CAS  Google Scholar 

  45. Brabson, J. S., & Switzer, R. L. (1975). The Journal of Biological Chemistry, 250, 8664–8669.

    CAS  Google Scholar 

  46. Callewaert, D. M., Rosemblatt, M. S., & Tchen, T. T. (1974). The Journal of Biological Chemistry, 249, 1737–1741.

    CAS  Google Scholar 

  47. Edwards, Y. H., Edwards, P. A., & Hopkinson, D. A. (1973). FEBS Letters, 32, 235–237.

    Article  CAS  Google Scholar 

  48. Kuhm, A. E., Knackmuss, H. J., & Stolz, A. (1993). The Journal of Biological Chemistry, 268, 9484–9489.

    CAS  Google Scholar 

  49. Legrain, C., & Stalon, V. (1976). European Journal of Biochemistry, 63, 289–301.

    Article  CAS  Google Scholar 

  50. Marshall, M., & Cohen, P. P. (1972). The Journal of Biological Chemistry, 247, 1641–1653.

    CAS  Google Scholar 

  51. O’Toole, N., Barbosa, J. A., Li, Y., Hung, L. W., Matte, A., & Cygler, M. (2003). Protein Sciences, 12, 327–336.

    Article  Google Scholar 

  52. Penninckx, M., Simon, J. P., & Wiame, J. M. (1974). European Journal of Biochemistry, 49, 429–442.

    Article  CAS  Google Scholar 

  53. Rosemblatt, M. S., Callewaert, D. M., & Tchen, T. T. (1973). The Journal of Biological Chemistry, 248, 6014–6018.

    CAS  Google Scholar 

  54. Gomez-Manzo, S., Chavez-Pacheco, J. L., Contreras-Zentella, M., Sosa-Torres, M. E., Arreguin-Espinosa, R., Perez, d. l. M., et al. (2010). Journal of Bacteriology, 192, 5718–5724.

    Article  CAS  Google Scholar 

  55. Zellner, G., & Jargon, A. (1997). Archives of Microbiology, 168, 480–485.

    Article  CAS  Google Scholar 

  56. Bradbury, S. L., & Jakoby, W. B. (1972). Proceedings of the National Academy of Sciences of the United States of America, 69, 2373–2376.

    Article  CAS  Google Scholar 

  57. Guerrillot, L., & Vandecasteele, J. P. (1977). European Journal of Biochemistry, 81, 185–192.

    Article  CAS  Google Scholar 

  58. Richardson, I. W., & Anthony, C. (1992). The Biochemical Journal, 287(Pt 3), 709–715.

    CAS  Google Scholar 

  59. Oubrie, A., Rozeboom, H. J., Kalk, K. H., Duine, J. A., & Dijkstra, B. W. (1999). Journal of Molecular Biology, 289, 319–333.

    Article  CAS  Google Scholar 

  60. Arfman, N., Van, B. J., De Vries, G. E., Harder, W., & Dijkhuizen, L. (1991). The Journal of Biological Chemistry, 266, 3955–3960.

    CAS  Google Scholar 

  61. Vonck, J., Arfman, N., De Vries, G. E., Van, B. J., Van Bruggen, E. F., & Dijkhuizen, L. (1991). The Journal of Biological Chemistry, 266, 3949–3954.

    CAS  Google Scholar 

  62. Klinman, J. P. (1972). The Journal of Biological Chemistry, 247, 7977–7987.

    CAS  Google Scholar 

  63. Rietveld, E. C., de Zwart, M., Cox, P. G., & Seutter-Berlage, F. (1987). Biochimica et Biophysica Acta, 914, 162–169.

    Article  CAS  Google Scholar 

  64. Hansch, C. & Leo, A. (1979). Substituent constants for correlation analysis in chemistry and biology Wiley.

Download references

Acknowledgments

We acknowledge the senior research fellowship to RS and VDT from CSIR, Govt. of India and the research grant to PP from DBT, Govt. of India. Metal analysis at SAIF-IITB is gratefully acknowledged.

Author information

Authors and Affiliations

  1. Department of Biosciences and Bioengineering, Indian Institute of Technology—Bombay, Powai, Mumbai, 400 076, India

    Randhir Singh, Vikas D. Trivedi & Prashant S. Phale

Authors
  1. Randhir Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vikas D. Trivedi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Prashant S. Phale
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Prashant S. Phale.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOC 8654 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Singh, R., Trivedi, V.D. & Phale, P.S. Purification and Characterization of NAD+-Dependent Salicylaldehyde Dehydrogenase from Carbaryl-Degrading Pseudomonas sp. Strain C6. Appl Biochem Biotechnol 172, 806–819 (2014). https://doi.org/10.1007/s12010-013-0581-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-013-0581-8

Keywords

Search

Navigation