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Biochemical characterization of an extracellular polyextremophilic α-amylase from the halophilic archaeon Halorubrum xinjiangense

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Abstract

An extracellular haloalkaliphilic thermostable α-amylase producing archaeon was isolated from the saltwater Lake Urmia and identified as Halorubrum xinjiangense on the basis of morphological, biochemical, and molecular properties. The enzyme was purified to an electrophoretically homogenous state by 80 % cold ethanol precipitation, followed by affinity chromatography. The concentrated pure amylase was eluted as a single peak on fast protein liquid chromatography. The molecular mass of the purified enzyme was about 60 kDa, with a pI value of 4.5. Maximum amylase activity was at 4 M NaCl or 4.5 M KCl, 70 °C, and pH 8.5. The K m and V max of the enzyme were determined as 3.8 mg ml−1 and 12.4 U mg−1, respectively. The pure amylase was stable in the presence of SDS, detergents, and organic solvents. In addition, the enzyme (20 U) hydrolyzed 69 % of the wheat starch after a 2-h incubation at 70 °C in an aqueous/hexadecane two-phase system.

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Acknowledgments

This work was supported financially by Grant Number 88-04-33-9700 from Tehran University of Medical Sciences, Tehran, Iran.

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Authors and Affiliations

  1. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy and Biotechnology Research Center, Tehran University of Medical Sciences, P.O. Box 14155-6451, 14174, Tehran, Iran

    Mahsa Moshfegh, Ahmad Reza Shahverdi & Mohammad Ali Faramarzi

  2. Department of Animal Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran

    Gholamreza Zarrini

Authors
  1. Mahsa Moshfegh
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  2. Ahmad Reza Shahverdi
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  3. Gholamreza Zarrini
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  4. Mohammad Ali Faramarzi
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Correspondence to Mohammad Ali Faramarzi.

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Communicated by S. Albers.

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Moshfegh, M., Shahverdi, A.R., Zarrini, G. et al. Biochemical characterization of an extracellular polyextremophilic α-amylase from the halophilic archaeon Halorubrum xinjiangense . Extremophiles 17, 677–687 (2013). https://doi.org/10.1007/s00792-013-0551-7

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  • DOI: https://doi.org/10.1007/s00792-013-0551-7

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