Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation of a Novel Freshwater Agarolytic Cellvibrio sp. KY-YJ-3 and Characterization of Its Extracellular ${\beta}$-Agarase

  • Rhee, Young-Joon (School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Han, Cho-Rong (School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Kim, Won-Chan (School of Applied Biological Chemistry, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Jun, Do-Youn (School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Rhee, In-Ku (School of Applied Biological Chemistry, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Kim, Young-Ho (School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University)
  • Received : 2010.07.06
  • Accepted : 2010.07.21
  • Published : 2010.10.28
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Abstract

A novel agarolytic bacterium, KY-YJ-3, producing extracellular agarase, was isolated from the freshwater sediment of the Sincheon River in Daegu, Korea. On the basis of Gram-staining data, morphology, and phylogenetic analysis of the 16S rDNA sequence, the isolate was identified as Cellvibrio sp. By ammonium sulfate precipitation followed by Toyopearl QAE-550C, Toyopearl HW-55F, and MonoQ column chromatographies, the extracellular agarase in the culture fluid could be purified 120.2-fold with a yield of 8.1%. The specific activity of the purified agarase was 84.2 U/mg. The molecular mass of the purified agarase was 70 kDa as determined by dodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified agarase were $35^{\circ}C$ and pH 7.0, respectively. The purified agarase failed to hydrolyze the other polysaccharide substrates, including carboxymethyl-cellulose, dextran, soluble starch, pectin, and polygalacturonic acid. Kinetic analysis of the agarose hydrolysis catalyzed by the purified agarase using thin-layer chromatography showed that the main products were neoagarobiose, neoagarotetraose, and neoagarohexaose. These results demonstrated that the newly isolated freshwater agarolytic bacterium KY-YJ-3 was a Cellvibrio sp., and could produce an extracellular ${\beta}$-agarase, which hydrolyzed agarose to yield neoagarobiose, neoagarotetraose, and neoagarohexaose as the main products.

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References

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