Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Enzymatic Characterization of a Recombinant Levansucrase from Rahnella aquatilis ATCC 15552

  • Kim, Hyun-Jin (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Park, Hae-Eun (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Kim, Min-Jeong (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Lee, Hyeon-Gyu (Department of Food and Nutrition, Hanyang University) ;
  • Yang, Ji-Young (Division of Food Science and Biotechnology, College of Fishery Sciences, Pukyung National University) ;
  • Cha, Jae-Ho (Department of Microbiology, College of Natural Sciences, Pusan National University)
  • Published : 2003.04.01
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Abstract

A 1.25 kb DNA fragment including the lscR gene, which encodes a levansucrase of Rahnella aquatilis ATCC 15552, was subcloned into a high-expression vector, pET-29b, and the recombinant enzyme was overexpressed in Escherichia coli. Most of the levansucrase activity was detected in the cytoplasmic fraction after induction with isopropyl ${\beta}-D-thiogalactoside$. The recombinant enzyme with a tag of six histidine residues at the C-terminus was purified 146-fold by affinity and gel-filtration chromatographies. The molecular mass of the purified LscR was approx. 49 kDa as determined by SDS-PAGE. The optimum pH and temperature of this enzyme for levan formation was pH 6.0 and $30^{\circ}C$, respectively. The optimum substrate concentration for levan formation was 300 mM sucrose. Levan formation was increased by the increase of the enzyme concentrations. Maxium yield of levan formation at optimum substrate concentration, pH, and temperature after 24 h of reaction was approximately 80%.

Keywords

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