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Purification and characterization of a novel extracellular halophilic and organic solvent-tolerant amylopullulanase from the haloarchaeon, Halorubrum sp. strain Ha25

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Abstract

A halophilic archaeon, Halorubrum sp. strain Ha25, produced extracellular halophilic organic solvent-tolerant amylopullulanase. The maximum enzyme production was at high salt concentration, 3–4 M NaCl. Optimum pH and temperature for enzyme production were 7.0 and 40 °C, respectively. Molecular mass of purified enzyme was estimated to be about 140 kDa by SDS–PAGE. This enzyme was active on pullulan and starch as substrates. The apparent K m for the enzyme activity on pullulan was 4 mg/ml and for soluble starch was 1.8 mg/ml. Optimum temperature for amylolytic and pullulytic activities was 50 °C. Optimum pH for amylolytic activity was 7 and for pullulytic activity was 7.5. This enzyme was active over a wide range of concentrations (0–4.5 M) of NaCl. The effect of organic solvents on the enzyme activities showed that this enzyme was more stable in the presence of non-polar organic solvents than polar solvents. This study is the first report on amylopullulanase production in halophilic bacteria and archaea.

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Authors and Affiliations

  1. Extremophiles Laboratory, Department of Microbiology, Faculty of Biology, College of Science, School of Biology and Center of Excellence in Phylogeny of Living Organisms, University of Tehran, P. O. Box 14155-6455, Tehran, Iran

    Maryam Siroosi, Mohammad Ali Amoozegar & Mostafa Fazeli

  2. Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

    Khosro Khajeh

  3. Protein Biotechnology Laboratory, Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran

    Mehran Habibi Rezaei

Authors
  1. Maryam Siroosi
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  2. Mohammad Ali Amoozegar
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  3. Khosro Khajeh
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  4. Mostafa Fazeli
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  5. Mehran Habibi Rezaei
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Correspondence to Mohammad Ali Amoozegar.

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Communicated by H. Atomi.

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Siroosi, M., Amoozegar, M.A., Khajeh, K. et al. Purification and characterization of a novel extracellular halophilic and organic solvent-tolerant amylopullulanase from the haloarchaeon, Halorubrum sp. strain Ha25. Extremophiles 18, 25–33 (2014). https://doi.org/10.1007/s00792-013-0589-6

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  • DOI: https://doi.org/10.1007/s00792-013-0589-6

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