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Applied Biochemistry and Biotechnology

, Volume 162, Issue 4, pp 1018–1030 | Cite as

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

  • Noomen Hmidet
  • Hana Maalej
  • Anissa Haddar
  • Moncef Nasri
Article

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.

Keywords

α-Amylase B. mojavensis A21 Purification Biochemical characterization 

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Noomen Hmidet
    • 1
  • Hana Maalej
    • 1
  • Anissa Haddar
    • 1
  • Moncef Nasri
    • 1
  1. 1.Laboratoire de Génie Enzymatique et de MicrobiologieEcole Nationale d’Ingénieurs de SfaxSfaxTunisia

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