A Novel and Simple Method for the Purification of Extracellular Levansucrase from Zymomonas mobilis |
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| Authors: | Armands Vigants Stefan Peter Marx Raimonds Linde Solvita Ore Martins Bekers Ilmara Vina Hans-Georg Hicke |
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| Affiliation: | (1) Institute of Microbiology and Biotechnology, University of Latvia, Kronvalda Boulevard 4, 1586 Riga, Latvia, LV;(2) Department of Biotechnology, RWTH Aachen, Sammelbau Biologie, Worringer Weg 1, 52056 Aachen, Germany, DE;(3) Institute of Chemistry, GKSS Research Center Geesthacht GmbH, Kantstrasse 55, 14513 Teltow, Germany, DE |
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| Abstract: | A new and simple method for the purification of extracellular levansucrase from Zymomonas mobilis from highly viscous fermentation broth was developed. After incubation of the fermentation broth with a fructose-polymer cleaving enzyme preparation (Fructozyme, Novozymes, DK) for 48 h, levansucrase precipitated as aggregates and was redissolved in a 3 M urea solution. By ongoing size-exclusion chromatography on Sephacryl S-300 the final levansucrase preparation was purified 100-fold and exhibited a specific activity of 25–35 U/mgprotein. The levansucrase was stable in 3 M urea solution for at least four months without inactivation. To maximize the enzyme yield the dynamic changes of extracellular levansucrase activity during fermentation were investigated. The highest levansucrase activity was observed during the logarithmic phase of growth (15–19 h of fermentation).Received: 26 September 2002 / Accepted: 24 October 2002 |
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