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α-Amylase activity from the halophilic archaeon Haloferax mediterranei

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Abstract

The halophilic archaeon Haloferax mediterranei is able to grow in a minimal medium containing ammonium acetate as a carbon and nitrogen source. When this medium is enriched with starch, α-amylase activity is excreted to the medium in low concentration. Here we report methods to concentrate and purify the enzyme. The relative molecular mass of the enzyme, determined by gel filtration, is 50±4 kDa, and on SDS-PAGE analysis a single band appeared at 58 kDa. These results indicated that the halophilic α-amylase is a monomeric enzyme. The enzyme showed a salt requirement for both stability and activity, being stable from 2 to 4 M NaCl, with maximal activity at 3 M NaCl. The enzyme displayed maximal activity at pHs from 7 to 8, and its optimal temperature was in a range from 50 °C to 60 °C. The results also implicated several prototropic groups in the catalytic reaction.

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Acknowledgements

We thank Dr. J.M. Sansano Gil (Department of Organic Chemistry, University of Alicante) for collaboration in the polarimetry studies. RMN experiments and HPLC were carried out at the Servicios Técnicos de Investigación de la Universidad de Alicante (Spain). This work was supported by funds from CICYT-PB98-0969 and GV01-120.

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Authors and Affiliations

  1. División de Bioquímica, Facultad de Ciencias, Universidad de Alicante, Apartado 99, 03080 , Alicante, Spain

    F. Pérez-Pomares, V. Bautista, J. Ferrer, C. Pire, F. C. Marhuenda-Egea & M. J. Bonete

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  1. F. Pérez-Pomares
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  2. V. Bautista
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  3. J. Ferrer
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  4. C. Pire
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  5. F. C. Marhuenda-Egea
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  6. M. J. Bonete
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Correspondence to M. J. Bonete.

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Communicated by W.D. Grant

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Pérez-Pomares, F., Bautista, V., Ferrer, J. et al. α-Amylase activity from the halophilic archaeon Haloferax mediterranei . Extremophiles 7, 299–306 (2003). https://doi.org/10.1007/s00792-003-0327-6

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  • DOI: https://doi.org/10.1007/s00792-003-0327-6

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