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Purification of α-Amylase from Bacillus sp. GHA1 and its partial characterization

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Abstract

Bacillus sp. GHA1 was isolated from water samples and screened for the production of α-amylase. Maximum production of amylase by this strain occurs at 42 °C, pH 6.5 and 72 h after cultivation in production medium. The enzyme was purified through successive applications of ammonium sulfate precipitation, ion exchange and hydrophobic interaction chromatography, resulting in a single band with an apparent molecular weight of 66 kDa, as judged by SDS-PAGE. Calcium analysis of the purified enzyme revealed that it contained three metal ions per molecule. The new extracellular α-amylase is active in a wide range of pH with its maximum activity at pH values 5.5–8.0. The optimum temperature for enzyme activity is 57 °C and the presence of calcium has relatively low influence on its activity and thermostability. The Bacillus sp. GHA1 α-amylase with these properties may be suitable for use in detergent and food industries.

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References

  1. J.E. Nielsen, T.V. Borchert, Biochim. Biophys. Acta 1543 (2000) 253.

    CAS  Google Scholar 

  2. L. Kandra, J. Mol. Struc. 666–667 (2003) 487.

    Google Scholar 

  3. M.J.E.C. van der Maarel, B. van der Veen, J.C.M. Uitdehaag, H. Leemhuis, L. Dijkhuizen, J. Biotechnol. 94 (2002) 137.

    Google Scholar 

  4. A. Pandey, P. Nigam, C.R. Soccol, V.T. Soccol, D. Singh, R. Mohan, Biotechnol. Appl. Biochem. 31 (2000) 135.

    CAS  Google Scholar 

  5. R.H. Sajedi, H. Naderi-Manesh, K. Khajeh, R. Ahmadvand, B. Ranjbar, A. Asoodeh, F. Moradian, Enzyme Microb. Technol. 36 (2005) 666.

    CAS  Google Scholar 

  6. R. Gupta, P. Gigras, H. Mohapatra, V.K. Goswami, B. Chauhan, Process Biochem. 38 (2003) 1599.

    CAS  Google Scholar 

  7. M. Fathi Najafi, Di. Deobagkar, De Deobagkar, Protein Expression Purif. 41 (2005)

    CAS  Google Scholar 

  8. S. Sivaramakrishnan, D. Gangadharan, K.M. Nampoothiri, C.R. Soccol, A. Pandey, Food Technol. Biotechnol. 44 (2006) 173.

    CAS  Google Scholar 

  9. G.B. Manning, L.L. Campbell, J. Biol. Chem. 236 (1961) 2952.

    CAS  Google Scholar 

  10. N. Saito, Arch. Biochem. Biophys. 155 (1973) 290.

    CAS  Google Scholar 

  11. T. Yamamoto, Handbook of Amylases and Related Enzymes, Pergamon Press, Oxford, England, 1988.

    Google Scholar 

  12. S. Ito, Extremophiles 1 (1997) 61.

    Google Scholar 

  13. K. Horikoshi, Agric. Biol. Chem. 35 (1971) 1783.

    CAS  Google Scholar 

  14. N. Wang, Y. Zhang, Q. Wang, J. Liu, H. Wang, Y. Xue, Y. Ma, Biotechnol. J. 1 (2006) 1258.

    CAS  Google Scholar 

  15. K. Igarashi, Y. Hatada, H. Hagihara, K. Saeki, M. Takaiwa, T. Uemura, K. Ara, K. Ozaki, S. Kawai, T. Kobayashi, S. Ito, Appl. Environ. Microbiol. 64 (1998) 3282.

    CAS  Google Scholar 

  16. P.H.A. Sneath, Bergey’s Manual of Systematic Bacteriology, Williams & Wilkins, Baltimore, USA, 1986.

    Google Scholar 

  17. J.M. Parry, P.C.B. Turnbull, J.R. Gibson. A Color Atlas of Bacillus Species, Wolfe Medical, London, 1983.

    Google Scholar 

  18. A. Badoei Dalfard, K. Khajeh, M.R. Soudi, H. Naderi- Manesh, B. Ranjbar, R.H. Sajedi, Enzyme Microb. Technol. 39 (2006) 1409.

    CAS  Google Scholar 

  19. P. Bernfeld, Methods Enzymol. 1 (1955) 149.

    CAS  Google Scholar 

  20. E.O. Hägele, E. Schaich, P. Lehmann, H. Bürk, A.W. Wahlefeld, Clin. Chem. 28 (1982) 2201.

    Google Scholar 

  21. H. Sheehan, B.V. McCleary, Biotechnol. Tech. 2 (1988) 289.

    CAS  Google Scholar 

  22. M.M. Bradford, Anal. Biochem. 72 (1976) 248.

    CAS  Google Scholar 

  23. U.K. Laemmli, Nature 227 (1970) 680.

    CAS  Google Scholar 

  24. C.N. Pace, F. Vajdos, L. Fee, G. Grimsley, T. Gray, Protein Sci. 4 (1995) 2411.

    CAS  Google Scholar 

  25. E.S. Demirkan, B. Mikami, M. Adachi, T. Higasa, S. Utsumi, Process Biochem. 40 (2005) 2629.

    CAS  Google Scholar 

  26. H. Hagihara, K. Igarashi, Y. Hayashi, K. Endo, K. Ikawa-Kitayama, K. Ozaki, Appl. Environ. Microbiol. 67 (2001) 1744.

    CAS  Google Scholar 

  27. N.J. Shih, R.G. Labbe, Appl. Environ. Microbiol. 61 (1995) 1776.

    CAS  Google Scholar 

  28. V. Paquet, C. Croux, G. Goma, P. Soucaille, Appl. Environ. Microbiol. 57 (1991) 212.

    CAS  Google Scholar 

  29. G. Mamo, B.A. Gashe, A. Gessesse, J. Appl. Microbiol. 86 (1999) 557.

    CAS  Google Scholar 

  30. M. Asgher, M. Javaid Asad, S.U. Rahman, R.L. Legge, J. Food Eng. 79 (2007) 950.

    CAS  Google Scholar 

  31. H. Iefuji, M. Chino, M. Kato, Y. Iimura, Biochem. J. 318 (1996) 989.

    CAS  Google Scholar 

  32. A. Burhan, U. Nisa, C. Gokhan, C. Omer, A. Ashabil, G. Osman, Process Biochem. 38 (2003) 1397.

    CAS  Google Scholar 

  33. C. Vieille, G.J. Zeikus, Microbiol. Mol. Biol. Rev. 65 (2001) 1.

    CAS  Google Scholar 

  34. R.D. Wind, R.M. Buitelaar, G. Eggink, H.J. Huizing, L. Dijkhuizen, Appl. Microbiol. Biotechnol. 41 (1994) 155.

    CAS  Google Scholar 

  35. M. Hashida, H. Bisgaard-Frantzen, Trends Glycosci. Glycotechnol. 68 (2000) 389.

    Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Razi University, 67149-67346, Kermanshah, Iran

    A. Ahmadi & S. Ghobadi

  2. Department of Biochemistry, Faculty of Science, Tarbiat Modarres University, P.O. Box 14115-175, Tehran, Iran

    K. Khajeh & A. Badoei Dalfard

  3. Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    B. Nomanpour

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  1. A. Ahmadi
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  2. S. Ghobadi
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  3. K. Khajeh
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  4. B. Nomanpour
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  5. A. Badoei Dalfard
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Correspondence to S. Ghobadi.

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Ahmadi, A., Ghobadi, S., Khajeh, K. et al. Purification of α-Amylase from Bacillus sp. GHA1 and its partial characterization. JICS 7, 432–440 (2010). https://doi.org/10.1007/BF03246029

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  • DOI: https://doi.org/10.1007/BF03246029

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