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Purification and biochemical characterization of a detergent stable α-amylase from Pseudomonas stutzeri AS22

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Abstract

This study reports the purification and biochemical characterization of a novel maltotetraose-forming-α-amylase from Pseudomonas stutzeri AS22, designated PSA. The P. stutzeri α-amylase (PSA) was purified from the culture supernatant to homogeneity by Sepharose mono Q anion exchange chromatography, ultrafiltration and Sephadex G-100 gel filtration, with a 37.32-fold increase in specific activity, and 31% recovery. PSA showed a molecular weight of approximately 57 kDa by SDS-PAGE. The N-terminal amino acid sequence of the first 7 amino acids was DQAGKSP. This enzyme exhibited maximum activity at pH 8.0 and 55°C, performed stably over a broad range of pH 5.0 ≈ 12.0, but rapidly lost activity above 50°C. Both potato starch and Ca2+ ions have a protective effect on the thermal stability of PSA. The enzyme activity was inhibited by Hg2+, Mn2+, Cd2+, Cu2+, and Co2+, and enhanced by Ba2+. PSA belonged to the EDTA-sensitive α-amylase. The purified enzyme showed high stability towards surfactants (Tween 20, Tween 80 and Triton X-100), and oxidizing agents, such as sodium per borate and H2O2. In addition, PSA showed excellent compatibility with a wide range of commercial solid and liquid detergents at 30°C, suggesting potential application in the detergent industry. Maltotetraose was the specific end product obtained after hydrolysis of starch by the enzyme for an extended period of time, and was not further degraded.

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Authors and Affiliations

  1. Laboratoire de Génie Enzymatique et de Microbiologie, Université de Sfax, Ecole Nationale d’Ingénieurs de Sfax, B.P. 1173-3038, Sfax, Tunisia

    Hana Maalej, Noomen Hmidet, Olfa Ghorbel-Bellaaj & Moncef Nasri

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  1. Hana Maalej
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  2. Noomen Hmidet
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Maalej, H., Hmidet, N., Ghorbel-Bellaaj, O. et al. Purification and biochemical characterization of a detergent stable α-amylase from Pseudomonas stutzeri AS22. Biotechnol Bioproc E 18, 878–887 (2013). https://doi.org/10.1007/s12257-012-0862-z

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  • DOI: https://doi.org/10.1007/s12257-012-0862-z

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