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Identification of several intracellular carbohydrate-degrading activities from the halophilic archaeon Haloferax mediterranei

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Abstract

Three different amylolytic activities, designated AMY1, AMY2, and AMY3 were detected in the cytoplasm of the extreme halophilic archaeon Haloferax mediterranei grown in a starch containing medium. This organism had also been reported to excrete an α-amylase into the external medium in such conditions. The presence of these different enzymes which are also able to degrade starch may be related to the use of the available carbohydrates and maltodextrins, including the products obtained by the action of the extracellular amylase on starch that may be transported to the cytoplasm of the organism. The behavior of these intracellular hydrolytic enzymes on starch is reported here and compared with their extracellular counterpart. Two of these glycosidic activities (AMY1, AMY3) have also been purified and further characterized. As with other halophilic enzymes, they were salt dependent and displayed maximal activity at 3 M NaCl, and 50°C. The purification steps and molecular masses have also been reported. The other activity (AMY2) was also detected in extracts from cells grown in media with glycerol instead of starch and in a yeast extract medium. This enzyme was able to degrade starch yielding small oligosaccharides and displayed similar halophilic behavior with salt requirement in the range 1.5–3 M NaCl.

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Acknowledgments

This work was supported by funds from BIO2005-08991-C02-01.

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Authors and Affiliations

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

    F. Pérez-Pomares, S. Díaz, V. Bautista, C. Pire, G. Bravo, J. Esclapez, B. Zafrilla & María-José Bonete

Authors
  1. F. Pérez-Pomares
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  2. S. Díaz
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  3. V. Bautista
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  4. C. Pire
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  5. G. Bravo
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  6. J. Esclapez
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  7. B. Zafrilla
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  8. María-José Bonete
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Corresponding author

Correspondence to María-José Bonete.

Additional information

Communicated by F. Robb.

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792_2009_246_MOESM1_ESM.pdf

Fig. 1 Supplemental Thin layer chromatography for the degradation reaction of AMY2 and AMY3 on starch, maltotriose and maltohexose. S, G, M, T y H: starch, glucose, maltose, maltotriose and maltohexaose standars. S2 and H2: degradation of starch and maltohexaose by AMY2. S3 and H3: degradation of starch and maltohexaose by AMY3. S3a, S3b, S3c, S3d: degradation of starch by non pooled fractions from the last purification step of AMY3 (a: next to AMY3 pool, before AMY3 pool, b: previous to fraction a, c: next to the AMY3 pool, after it, and d: fraction after b) (PDF 78 kb)

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Pérez-Pomares, F., Díaz, S., Bautista, V. et al. Identification of several intracellular carbohydrate-degrading activities from the halophilic archaeon Haloferax mediterranei . Extremophiles 13, 633–641 (2009). https://doi.org/10.1007/s00792-009-0246-2

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  • DOI: https://doi.org/10.1007/s00792-009-0246-2

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