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Purification, biochemical characterization and gene sequencing of a thermostable raw starch digesting α-amylase from Geobacillus thermoleovorans subsp. stromboliensis subsp. nov.

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Abstract

This study reports the purification and biochemical characterization of a raw starch-digesting α-amylase from Geobacillus thermoleovorans subsp. stromboliensis subsp. nov. (strain PizzoT). The molecular weight was estimated to be 58 kDa by SDS–PAGE. The enzyme was highly active over a wide range of pH from 4.0–10.0. The optimum temperature of the enzyme was 70°C. It showed extreme thermostability in the presence of Ca2+, retaining 50% of its initial activity after 90 h at 70°C. The enzyme efficiently hydrolyzed 20% (w/v) of raw starches, concentration normally used in starch industries. The α-amylase showed an high stability in presence of many organic solvents. In particular the residual activity was of 73% in presence of 15% (v/v) ethyl alcohol, which corresponds to ethanol yield in yeast fermentation process. By analyzing its complete amyA gene sequence (1,542 bp), the enzyme was proposed to be a new α-amylase.

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Acknowledgments

This study has been financed partially under the bilateral agreement of Italian and Turkish governments through project no: TBAG-U/192(106T756) and Ministry of Foreign Affairs-Italian MFA.

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Authors and Affiliations

  1. Institute of Biomolecular Chemistry, CNR-National Research Council of Italy, Via Campi Flegrei 34, 80078, Pozzuoli, NA, Italy

    Ilaria Finore, Annarita Poli, Ida Romano, Laura Dipasquale, Barbara Nicolaus & Licia Lama

  2. Department of Environmental Sciences, University of Naples “Parthenope”, Centro Direzionale (Isola C4), 80143, Naples, Italy

    Ilaria Finore

  3. Department of Chemical Engineering, Bogazici University, Bebek, 34342, Istanbul, Turkey

    Ceyda Kasavi & Betul Kirdar

  4. Department of Bioengineering, Marmara University, Goztepe Campus, 34722, Istanbul, Turkey

    Ebru Toksoy Oner

Authors
  1. Ilaria Finore
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  2. Ceyda Kasavi
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  3. Annarita Poli
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  4. Ida Romano
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  5. Ebru Toksoy Oner
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  6. Betul Kirdar
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  7. Laura Dipasquale
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  8. Barbara Nicolaus
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  9. Licia Lama
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Correspondence to Licia Lama.

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Finore, I., Kasavi, C., Poli, A. et al. Purification, biochemical characterization and gene sequencing of a thermostable raw starch digesting α-amylase from Geobacillus thermoleovorans subsp. stromboliensis subsp. nov.. World J Microbiol Biotechnol 27, 2425–2433 (2011). https://doi.org/10.1007/s11274-011-0715-5

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  • DOI: https://doi.org/10.1007/s11274-011-0715-5

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