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A thermoactive glucoamylase with biotechnological relevance from the thermoacidophilic Euryarchaeon Thermoplasma acidophilum

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Abstract

A gene encoding an intracellular glucoamylase was identified in the genome of the extreme thermoacidophilic Archaeon Thermoplasma acidophilum. The gene taGA, consisting of 1,911 bp, was cloned and successfully expressed in Escherichia coli. The recombinant protein was purified 22-fold to homogeneity using heat treatment, anion-exchange chromatography, and gel filtration. Detailed analysis shows that the glucoamylase, with a molecular weight of 66 kDa per subunit, is a homodimer in its active state. Amylolytic activity was measured over a wide range of temperature (40–90°C) and pH (pH 3.5–7) and was maximal at 75°C and at acidic condition (pH 5). The recombinant archaeal glucoamylase uses a variety of polysaccharides as substrate, including glycogen and amylose. Maximal activity was measured towards amylopectin with a specific activity of 4.2 U/mg and increased almost threefold in the presence of manganese. Calcium ions have a pronounced effect on enzyme stability; in the presence of 5 mM CaCl2, the half-life increased from 15 min to 2 h at 80°C.

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Acknowledgments

This work was supported by grant 04-008 202131 from the German Federal Ministry of Education and Research. We would like to thank Moritz Katzer for his support.

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Authors and Affiliations

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

    Christiane Dock, Matthias Hess & Garabed Antranikian

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  1. Christiane Dock
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  2. Matthias Hess
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  3. Garabed Antranikian
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Correspondence to Garabed Antranikian.

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Dock, C., Hess, M. & Antranikian, G. A thermoactive glucoamylase with biotechnological relevance from the thermoacidophilic Euryarchaeon Thermoplasma acidophilum . Appl Microbiol Biotechnol 78, 105–114 (2008). https://doi.org/10.1007/s00253-007-1293-1

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  • DOI: https://doi.org/10.1007/s00253-007-1293-1

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