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To investigate the production and characteristicsof thermostable levansucrase from Rahnella aquatilis ATCC33071, the levansucrase gene from R. aquatilis was clonedand expressed in Escherichia coli without induction system.Expression of levansucrase gene in E. coli had no notable ordetrimental effect on the growth of host strain, and therecombinant levansucrase exhibited levan synthesis activity.Levansucrase was secreted to the periplasm in E. coli, andaddition of 0.5% glycine yielded further secretion of levansucraseto the growth medium and resulted in an increase of totallevansucrase activity. Furthermore, the cellular levansucrasewas evaluated for the production of levan by using toluenepermeabilizedwhole-cells. The levansucrase was thermostableat 37oC. The molecular size of levan was 1×106 Da, as determinedby HPLC, and the degree of polymerization of levan variedwith incubation temperatures: Low incubation temperaturewas preferable for the production of high-molecular size levan.The present study demonstrated that the mass production oflevan and levan oligosaccharides can be achieved by glycinesupplementation to the growth medium or by toluenepermeabilizedwhole-cells.

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