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Purification and characterization of a cold-adapted pullulanase from a psychrophilic bacterial isolate

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Abstract

There is a considerable potential of cold-active biocatalysts for versatile industrial applications. A psychrophilic bacterial strain, Shewanella arctica 40-3, has been isolated from arctic sea ice and was shown to exhibit pullulan-degrading activity. Purification of a monomeric, 150-kDa pullulanase was achieved using a five-step purification approach. The native enzyme was purified 50.0-fold to a final specific activity of 3.0 U/mg. The enzyme was active at a broad range of temperature (10–50 °C) and pH (5–9). Optimal activity was determined at 45 °C and pH 7. The presence of various metal ions is tolerated by the pullulanase, while detergents resulted in decreased activity. Complete conversion of pullulan to maltotriose as the sole product and N-terminal amino acid sequence indicated that the enzyme is a type-I pullulanase and belongs to rarely characterized pullulan-degrading enzymes from psychrophiles.

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Acknowledgments

Anke Peters is thanked for excellent technical assistance. FMQ received a scholarship from the KAAD (Katholischer Akademischer Ausländer-Dienst).

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Authors and Affiliations

  1. Institute of Technical Microbiology, Hamburg University of Technology (TUHH), Kasernenstr. 12, 21073, Hamburg, Germany

    Farah Qoura, Skander Elleuche & Garabed Antranikian

  2. Department of Chemistry, Industrial Biocatalysis, Technical University of Munich (TUM), Lichtenbergstr. 4, 85748, Garching, Germany

    Farah Qoura & Thomas Brueck

Authors
  1. Farah Qoura
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  2. Skander Elleuche
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  3. Thomas Brueck
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  4. Garabed Antranikian
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Correspondence to Garabed Antranikian.

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Communicated by M. da Costa.

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792_2014_678_MOESM1_ESM.pdf

Figure S1: Stimulation of pullulanase in the presence of CoCl2. Effect of CoCl2 was tested at different concentrations after pre-incubation at room temperature for 60 min followed by standard activity assays (incubation at 45 °C, pH 7 for 60 min). (PDF 39 kb)

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Qoura, F., Elleuche, S., Brueck, T. et al. Purification and characterization of a cold-adapted pullulanase from a psychrophilic bacterial isolate. Extremophiles 18, 1095–1102 (2014). https://doi.org/10.1007/s00792-014-0678-1

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  • DOI: https://doi.org/10.1007/s00792-014-0678-1

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