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Applied Biochemistry and Biotechnology

, Volume 113, Issue 1–3, pp 161–171 | Cite as

Effects of carbon source on expression of alcohol oxidase activity and on morphologic pattern of YR-1 strain, a filamentous fungus isolated from petroleum-contaminated soils

  • Carmen Rodríguez Robelo
  • Vanesa Zazueta Novoa
  • Roberto Zazueta-SandovalEmail author
Article

Abstract

Soluble alcohol oxidase (AO) activity was detected in the supernatant fraction of a high-speed centrifugation procedure after ballistic cellular homogenization to break the mycelium from a filamentous fungus strain named YR-1, isolated from petroleum-contaminated soils. AO activity from aerobically grown mycelium was detected in growth media containing different carbon sources, including alcohols and hydrocarbons but not in glucose. In previous work, zymogram analysis conducted with crude extracts from aerobic mycelium of YR-1 strain indicated the existence of two AO enzymes originally named AO-1 and AO-2. In the present study, we were able to separate the AO-1 band into two bands depending on culture conditions, carbon source, and polyacrylamide gel electrophoresis (PAGE) separation conditions; the enzyme activity pattern in zymograms from cell-free extracts exhibited three different bands after native PAGE. New nomenclature was used for upper bands AO-1 and AO-2 and lower band AO-3, respectively. The expression of AO activity was studied in the absence of glucose in the culture media and in the presence of hydrocarbons or petroleum as sole carbon source, suggesting that AO expression could be subjected to two regulatory possibilities: carbon catabolite regulation by glucose and induction by hydrocarbons. The possibility of catabolic inhibition of AO by glucose in the active enzyme was also tested, and the results confirm that this kind of regulatory mechanism is not present in AO activity.

Index Entries

Alcohol oxidase filamentous fungi hydrocarbon biodegradation petroleum contamination YR-1 strain 

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References

  1. 1.
    Atlas, R. M. (1995), Chem. Eng. News 73(14), 32–42.Google Scholar
  2. 2.
    Alper, J. (1993), Biotechnology, 11, 973–975.CrossRefGoogle Scholar
  3. 3.
    Guenguerich, F. P. and Macdonald, T. L. (1990), FASEB J. 4, 2453–2459.Google Scholar
  4. 4.
    Kellner, D. G., Maves, S. A., and Sligar, S. P. (1997), Curr. Opin. Biotechnol. 8, 274–278.PubMedCrossRefGoogle Scholar
  5. 5.
    Jones, J. G. and Bellion, E. (1991), Biochem. J. 280, 475–481.PubMedGoogle Scholar
  6. 6.
    Evers, M. E., Titorenko, V., Harder, W., ven der Hlei, Y., and Veehhuis, M. (1996), Yeast 12, 917–923.PubMedCrossRefGoogle Scholar
  7. 7.
    Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951), J. Biol. Chem. 193, 265–275.PubMedGoogle Scholar
  8. 8.
    Gleason, M. A., Ortori, S., and Sudberry, P. E. (1986), J. Gen. Microbiol. 132, 3459–3465.Google Scholar
  9. 9.
    Bartnicki-Garcia, S. and Nickerson, W. J. (1962), J. Bacteriol. 84, 841–858.PubMedGoogle Scholar
  10. 10.
    Torres-Guzman, J. C., Arreola-Garcia, G. A., Zazueta-Sandoval, R., Carrillo-Rayas, T., Martínez-Cadena, G., and Gutiérrez-Corona F. (1994), Curr. Genet. 26, 166–171.PubMedCrossRefGoogle Scholar
  11. 11.
    Janssen, F. W., Kerwin, R. M., and Ruelius, H. W. (1975), in Methods in Enzymology, vol. XLI, Academic, London, UK, pp. 364–369.Google Scholar
  12. 12.
    Laemmli, U. K. (1971), Nature 227, 680–685.CrossRefADSGoogle Scholar
  13. 13.
    Alvarado-Caudillo, Y., Bravo-Torres, J. C., Zazueta-Novoa, V., Silva-Jiménez, H., Torres-Guzmán, J. C., Gutiérrez-Corona, J. F., and Zazueta-Sandoval, R. (2002), Appl. Biochem. Biotechnol. 98–100, 243–255.PubMedCrossRefGoogle Scholar
  14. 14.
    Sakai, Y. and Tani, Y. (1992), Gene 114, 67–73.PubMedCrossRefGoogle Scholar
  15. 15.
    Benen, J. A. E., Sánchez-Torres, P., Wagemaker, M. J. M., Fraaije, M. W., van Berkel, W. J. H., and Visser, J. (1998), J. Biol. Chem. 273, 7865–7872.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Carmen Rodríguez Robelo
    • 1
  • Vanesa Zazueta Novoa
    • 1
  • Roberto Zazueta-Sandoval
    • 1
    Email author
  1. 1.Instituto de Investigación en Biología Experimental, Facultad de QuímicaUniversidad de GuanajuatoGuanajuatoMéxico

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