Advertisement

Biochemistry (Moscow)

, Volume 74, Issue 1, pp 81–84 | Cite as

Expression and functional analysis of aminotransferase involved in indole-3-acetic acid biosynthesis in Azospirillum brasilense Yu62

  • Shi-Mei Ge
  • Li Tao
  • San-Feng ChenEmail author
Article

Abstract

In this study, atrC (a novel gene from Azospirillum brasilense identified in our laboratory) was expressed in Escherichia coli, and SDS-PAGE analysis of the expressed AtrC revealed the apparent molecular weight of 45 kD. When analyzed under non-denaturing PAGE conditions and using L-tryptophan as a substrate, the purified AtrC protein exhibited aminotransferase activity, while crude protein extracts from A. brasilense Yu62 showed two activity bands with molecular masses estimated as 44 and 66 kD. Thus, we deduced that AtrC protein is identical to the 44 kD band of crude protein extracts. The optimal temperature and pH for the catalytic activity of the purified AtrC are 30°C and pH 7.0, respectively.

Key words

function analysis aminotransferase Azospirillum brasilense indole-3-acetic acid 

Abbreviations

IAA

indole-3-acetic acid

IPA

indole-3-pyruvic acid

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Okon, Y. (1985) Trends Biotechnol., 3, 223–228.CrossRefGoogle Scholar
  2. 2.
    Zimmer, W., Wesche, M., and Timmermans, L. (1998) Curr. Microbiol., 36, 327–331.PubMedCrossRefGoogle Scholar
  3. 3.
    Carreno-Lopez, R., Campos-Reales, N., and Elmerich, C. (2000) Mol. Gen. Genet., 264, 521–530.PubMedCrossRefGoogle Scholar
  4. 4.
    Abdel-Salam, M. S., and Klingmuller, W. (1987) Mol. Gen. Genet., 210, 165–170.CrossRefGoogle Scholar
  5. 5.
    Baca, B. E., Soto-Urzua, L., Xochihua-Corona, Y. G., and Cuervo-Garcia, A. (1996) Soil Biol. Biochem., 26, 57–63.CrossRefGoogle Scholar
  6. 6.
    Pedraza, R. O., Ramirez-Mata, A., Xiqui, M. L., and Baca, B. E. (2004) FEMS Microbiol. Lett., 233, 15–21.PubMedCrossRefGoogle Scholar
  7. 7.
    Prinsen, E., Costacurta, A., Michiels, K., Vanderleyden, J., and van Onckelen, H. (1993) Mol. Plant-Microbe Interact., 6, 609–615.Google Scholar
  8. 8.
    Costacurta, A., Keijers, V., and Vanderleyden, J. (1994) Mol. Gen. Genet., 243, 463–472.PubMedGoogle Scholar
  9. 9.
    Hartmann, A., and Zimmer, W. (1994) in Azospirillum-Plant Associations (Okon, Y., ed.) CRC Press, Boca Raton, pp. 15–39.Google Scholar
  10. 10.
    Patten, C. L., and Glick, B. R. (1996) Can. J. Microbiol., 42, 207–220.PubMedCrossRefGoogle Scholar
  11. 11.
    Soto-Urzua, L., Xochinua-Corona, Y. G., Flores-Encarnacion, M., and Baca, B. E. (1996) Can. J. Microbiol., 42, 294–298.PubMedGoogle Scholar
  12. 12.
    Xie, B., Xu, K., Zhao, H. X., and Chen, S. F. (2005) FEMS Microbiol. Lett., 248, 57–63.PubMedCrossRefGoogle Scholar
  13. 13.
    Xie, B., and Chen, S. F. (2006) Prog. Nat. Sci., 16, 366–372.CrossRefGoogle Scholar
  14. 14.
    Yang, J. B., Cao, Z. L., and Li, J. L. (1984) J. China Agric. Univ., 10, 321–329 (Chinese).Google Scholar
  15. 15.
    Kittell, B. L., Helinski, D. R., and Ditta, G. S. (1989) J. Bacteriol., 171, 5458–5466.PubMedGoogle Scholar
  16. 16.
    Sambrook, J., and Russell, D. W. (2001) Molecular Cloning: a Laboratory Manual, 3rd Edn., Cold Spring Harbor Laboratory, NY.Google Scholar
  17. 17.
    Matsui, I., Matsui, E., Sakai, Y., Kikuchi, H., Kawarabayasi, Y., Ura, H., Kawaguchi, S. I., Kuramitsu, H., and Harata, K. (2000) J. Biol. Chem., 275, 4871–4879.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  1. 1.Department of Microbiology and Immunology, College of Biological SciencesChina Agricultural UniversityBeijingP. R. China
  2. 2.College of Biological Sciences and National Key Laboratory for AgrobiotechnologyChina Agricultural UniversityBeijingP. R. China

Personalised recommendations