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Purification and biochemical characterization of an acidophilic amylase from a newly isolated Bacillus sp. DR90

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Abstract

An acidophilic and Ca2+-independent amylase was purified from a newly isolated Bacillus sp. DR90 by ion-exchange chromatography, and exhibited a molecular weight of 68.9 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme were found to be 4.0 and 45 °C, respectively. The enzyme activity was increased by Ba2+, Fe2+ and Mg2+, and decreased by Hg2+ and Zn2+, while it was not affected by Na+, K+, phenylmethylsulfonyl fluoride and β-mercaptoethanol. Ca2+ and EDTA did not have significant effect on the enzyme activity and thermal stability. The values of K m and V max for starch as substrate were 4.5 ± 0.13 mg/ml and 307 ± 12 μM/min/mg, respectively. N,N-dialkylimidazolium-based ionic liquids such as 1-hexyl-3-methylimidazolium bromide [HMIM][Br] have inhibitory effect on the enzyme activity. Thin layer chromatography analyses displayed that maltose and glucose are the main products of the enzyme reaction on starch. Regarding the features of the enzyme, it may be utilized as a novel candidate for industrial applications.

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Acknowledgments

We gratefully thank the Institute of Biotechnology and research council of Ferdowsi University of Mashhad for their financial support (Grant number: 3/17458; 06-02-1390 and 4065; 06-02-1389).

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Authors and Affiliations

  1. Department of Chemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

    Ahmad Asoodeh & Ashraf Alemi

  2. Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

    Ahmad Asoodeh

  3. Department of Chemistry, Tarbiat Modares University, P.O. Box 14155, 4938, Tehran, Iran

    Akbar Heydari & Jafar Akbari

Authors
  1. Ahmad Asoodeh
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  2. Ashraf Alemi
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  3. Akbar Heydari
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  4. Jafar Akbari
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Correspondence to Ahmad Asoodeh.

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Communicated by H. Atomi.

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Asoodeh, A., Alemi, A., Heydari, A. et al. Purification and biochemical characterization of an acidophilic amylase from a newly isolated Bacillus sp. DR90. Extremophiles 17, 339–348 (2013). https://doi.org/10.1007/s00792-013-0520-1

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  • DOI: https://doi.org/10.1007/s00792-013-0520-1

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