Applied Biochemistry and Biotechnology

, Volume 162, Issue 4, pp 1018–1030 | Cite as

A Novel α-Amylase from Bacillus mojavensis A21: Purification and Biochemical Characterization

  • Noomen Hmidet
  • Hana Maalej
  • Anissa Haddar
  • Moncef Nasri


α-Amylase from Bacillus mojavensis A21 (BMA.2) was purified to homogeneity by ultrafiltration, Sephadex G-75 gel filtration and Sepharose mono Q anion exchange chromatography, with a 15.3-fold increase in specific activity and 11% recovery. The molecular weight of the BMA.2 enzyme was estimated to be 58 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration. The optimum temperature and pH were 80 °C and 6.5, respectively. BMA.2 belonged to the EDTA-sensitive α-amylase, but its activity was not stimulated by the presence of Ca2+ ions. The major end-products of starch hydrolysis were maltohexaose, maltopentaose and maltotriose. The N-terminal amino acid sequence of the first ten amino acids of the purified α-amylase was ASVNGTLMQY. Compared to sequences of other amylases, the ten amino acid sequence contains Val at position 3, while amylases from Bacillus licheniformis NH1 and Bacillus sp. SG-1 have Leu and Thr at position 3, respectively.


α-Amylase B. mojavensis A21 Purification Biochemical characterization 



This work was funded by the Ministry of Higher Education, Scientific Research and Technology-Tunisia. The authors would like to thank Mr A. Hajji from the Faculty of Letters and Human Sciences of Kairouan for his help with English.


  1. 1.
    Van der Maarel, M. J. E. C., Van der Veen, B., Uitdehaag, J. C. M., Leemhuis, H., & Dijkhuizen, L. (2002). Journal of Biotechnology, 94, 137–155.CrossRefGoogle Scholar
  2. 2.
    Gupta, R., Gigras, P., Mohapatra, H., Goswami, V. K., & Chauhan, B. (2003). Process Biochemistry, 38, 1599–1616.CrossRefGoogle Scholar
  3. 3.
    McTigue, M. A., Kelly, C. T., Doyle, E. M., & Fogarty, W. M. (1995). Enzyme and Microbial Technology, 17, 570–573.CrossRefGoogle Scholar
  4. 4.
    Burhan, A., Nisa, U., Gokhan, C., Omer, C., Ashabil, A., & Osman, G. (2003). Process Biochemistry, 38, 1397–1403.CrossRefGoogle Scholar
  5. 5.
    Asgher, M., Javaid, A. M., Rahman, S. U., & Legge, R. L. (2007). Journal of Food Engineering, 79, 950–955.CrossRefGoogle Scholar
  6. 6.
    Saxena, R. K., Dutt, K., Agarwal, L., & Nayyar, P. (2007). Bioresource Technology, 98, 260–265.CrossRefGoogle Scholar
  7. 7.
    Hashim, S. O., Delgado, O. D., Martinez, M. A., Kaul, R. H., Mulaa, F. J., & Mattiasson, B. (2005). Enzyme and Microbial Technology, 36, 139–146.CrossRefGoogle Scholar
  8. 8.
    Arikan, B. (2008). Bioresource Technology, 99, 3071–3076.CrossRefGoogle Scholar
  9. 9.
    Hmidet, N., Bayoudh, A., Berrin, J. G., Kanoun, S., Juge, N., & Nasri, M. (2008). Process Biochemistry, 43, 499–510.CrossRefGoogle Scholar
  10. 10.
    Najafi, M. F., Deobagkar, D., & Deobagkar, D. (2005). Protein Expression and Purification, 41, 349–354.CrossRefGoogle Scholar
  11. 11.
    Bolton, D. J., Kelly, C. T., & Fogarty, W. M. (1997). Enzyme and Microbial Technology, 20, 340–343.CrossRefGoogle Scholar
  12. 12.
    Hamilton, L. M., Kelly, C. T., & Fogarty, W. M. (1999). Process Biochemistry, 35, 27–31.CrossRefGoogle Scholar
  13. 13.
    Haddar, A., Bougatef, A., Agrebi, R., Sellami-Kamoun, A., & Nasri, M. (2009). Process Biochemistry, 44, 29–35.CrossRefGoogle Scholar
  14. 14.
    Haddar, A., Agrebi, R., Bougatef, A., Hmidet, N., Sellami-Kamoun, A., & Nasri, M. (2009). Bioresource Technology, 100, 3366–3373.CrossRefGoogle Scholar
  15. 15.
    Miller, J. H. (1972). Experiments in Moleculer Genetics (pp. 431–435). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.Google Scholar
  16. 16.
    Miller, G. L. (1959). Analytical Chemistry, 31, 426–428.CrossRefGoogle Scholar
  17. 17.
    Bradford, M. (1976). Analytical Biochemistry, 72, 248–254.CrossRefGoogle Scholar
  18. 18.
    Laemmli, U. K. (1970). Nature, 227, 680–685.CrossRefGoogle Scholar
  19. 19.
    Ivanova, V. N., Dobreva, E. P., & Emanuilova, E. I. (1993). Journal of Biotechnology, 28, 277–289.CrossRefGoogle Scholar
  20. 20.
    Tebo, B., Ferriera, S., Johnson, J., Kravitz, S., Beeson, K., Sutton, G., Rogers, Y.H., Friedman, R., Frazier, M. & Venter, J.C Accession NZ_ABCF01000001, EMBL/GenBank/DDBJ databases (2007). Bacillus sp. SG-1, whole genome sequence.Google Scholar
  21. 21.
    Gray, G. L., Mainzer, S. E., Rey, M. W., Lamsa, M. H., Kindle, K. L., Carmona, C., et al. (1986). Journal of Bacteriology, 166, 635–643.Google Scholar
  22. 22.
    Kuhn, H., Fietzek, P. P., & Lampen, J. O. (1982). Journal of Bacteriology, 149, 372–373.Google Scholar
  23. 23.
    Hayashida, S., Teramoto, Y., & Inoue, T. (1988). Applied and Environmental Microbiology, 54, 1516–1522.Google Scholar
  24. 24.
    Machius, M., Wiegand, G., & Huber, R. (1995). Journal of Molecular Biology, 246, 545–559.CrossRefGoogle Scholar
  25. 25.
    Buisson, G., Duee, D., Haser, R., & Payan, F. (1987). EMBO Journal, 6, 3909–3916.Google Scholar
  26. 26.
    Goyal, N., Gupta, J. K., & Soni, S. K. (2005). Enzyme and Microbial Technology, 37, 723–734.CrossRefGoogle Scholar
  27. 27.
    Hagihara, H., Igarashi, K., Hayashi, Y., Endo, K., Ikawa-Kitayama, K., Ozaki, K., et al. (2001). Applied and Environmental Microbiology, 67, 1744–1750.CrossRefGoogle Scholar
  28. 28.
    Kelly, C. T., McTigue, M. A., Doyle, E. M., & Fogarty, W. M. (1995). Journal of Industrial Microbiology and Biotechnology, 15, 446–448.CrossRefGoogle Scholar
  29. 29.
    Morgan, F. J., & Priest, F. G. (1981). Journal of Applied Bacteriology, 50, 107–114.Google Scholar
  30. 30.
    Igarashi, K., Hatada, Y., Hagihara, H., Saeki, K., Takaiwa, M., Uemura, T., et al. (1998). Applied and Environmental Microbiology, 64, 3282–3289.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Noomen Hmidet
    • 1
  • Hana Maalej
    • 1
  • Anissa Haddar
    • 1
  • Moncef Nasri
    • 1
  1. 1.Laboratoire de Génie Enzymatique et de MicrobiologieEcole Nationale d’Ingénieurs de SfaxSfaxTunisia

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