A simple and efficient method for the purification of membrane-bound levansucrase from Zymomonas mobilis.
A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 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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Institute of Microbiology and Biotechnology, University of Latvia, Kronvalda boulevard 4, 1586 Riga, Latvia.
Hicke
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Marx
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SP
D002462:Cell Membrane / Q000201:enzymology; D002845:Chromatography / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006602:Hexosyltransferases / Q000302:isolation & purification*; D007700:Kinetics; D017355:Zymomonas / Q000201:enzymology* / Q000502:physiology
D006602:Hexosyltransferases; C021513:levansucrase