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A Simple and Efficient Method for the Purification of Membrane-Bound Levansucrase from Zymomonas mobilis

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Abstract

A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilisATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 m MnCl2 as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg−1. The native enzyme formed large aggregates with an apparent molecular mass of more than 106 Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits.

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Authors and Affiliations

  1. Institute of Microbiology and Biotechnology, University of Latvia, Kronvalda boulevard 4, 1586 Riga, Latvia

    Armands Vigants

  2. Institute of Chemistry, GKSS Research Center Geesthacht GmbH, Kantstr. 55, 14513 Teltow, Germany, DE

    Hans-Georg Hicke

  3. Department of Biotechnology, RWTH Aachen, Sammelbau Biologie, Worringer Weg 1, 52056 Aachen, Germany, DE

    Stefan Peter Marx

Authors
  1. Armands Vigants
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  2. Hans-Georg Hicke
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  3. Stefan Peter Marx
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Received: 10 October 2000 / Accepted: 17 November 2000

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Vigants, A., Hicke, HG. & Marx, S. A Simple and Efficient Method for the Purification of Membrane-Bound Levansucrase from Zymomonas mobilis . Curr Microbiol 42, 415–418 (2001). https://doi.org/10.1007/s002840010239

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  • DOI: https://doi.org/10.1007/s002840010239

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