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A Novel α-Amylase from Bacillus mojavensis A21: Purification and Biochemical Characterization

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Abstract

α-Amylase from Bacillus mojavensis A21 (BMA.2) was purified to homogeneity by ultrafiltration, Sephadex G-75 gel filtration and Sepharose mono Q anion exchange chromatography, with a 15.3-fold increase in specific activity and 11% recovery. The molecular weight of the BMA.2 enzyme was estimated to be 58 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration. The optimum temperature and pH were 80 °C and 6.5, respectively. BMA.2 belonged to the EDTA-sensitive α-amylase, but its activity was not stimulated by the presence of Ca2+ ions. The major end-products of starch hydrolysis were maltohexaose, maltopentaose and maltotriose. The N-terminal amino acid sequence of the first ten amino acids of the purified α-amylase was ASVNGTLMQY. Compared to sequences of other amylases, the ten amino acid sequence contains Val at position 3, while amylases from Bacillus licheniformis NH1 and Bacillus sp. SG-1 have Leu and Thr at position 3, respectively.

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Acknowledgements

This work was funded by the Ministry of Higher Education, Scientific Research and Technology-Tunisia. The authors would like to thank Mr A. Hajji from the Faculty of Letters and Human Sciences of Kairouan for his help with English.

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Authors and Affiliations

  1. Laboratoire de Génie Enzymatique et de Microbiologie, Ecole Nationale d’Ingénieurs de Sfax, B.P. “W” 3038, Sfax, Tunisia

    Noomen Hmidet, Hana Maalej, Anissa Haddar & Moncef Nasri

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  1. Noomen Hmidet
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  2. Hana Maalej
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  3. Anissa Haddar
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  4. Moncef Nasri
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Correspondence to Moncef Nasri.

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Hmidet, N., Maalej, H., Haddar, A. et al. A Novel α-Amylase from Bacillus mojavensis A21: Purification and Biochemical Characterization. Appl Biochem Biotechnol 162, 1018–1030 (2010). https://doi.org/10.1007/s12010-009-8902-7

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  • DOI: https://doi.org/10.1007/s12010-009-8902-7

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