Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Cloning and Characterization of an Esterase from Xanthomonas oryzae pv. oryzae

  • Kang, Han-Chul (Department of Molecular Physiology, National Institute of Agricultural Biotechnology, Rural Development Administration) ;
  • Kim, Jong-Bum (Department of Molecular Physiology, National Institute of Agricultural Biotechnology, Rural Development Administration) ;
  • Lee, Hak-Sun (Department of Molecular Physiology, National Institute of Agricultural Biotechnology, Rural Development Administration) ;
  • Cho, Kang-Jin (Department of Molecular Physiology, National Institute of Agricultural Biotechnology, Rural Development Administration)
  • Published : 2008.06.30
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Abstract

The gene encoding a putative esterase of Xanthomonas oryzae pv. oryzae was cloned using PCR technique. The gene was expressed with His6 tag in E. coli. One-step purification of the recombinant esterase with Ni-NTA resin resulted in one band by SDS-PAGE analysis. The purified enzyme showed a molecular weight of 30 kDa, as expected, therefore the enzyme was a mononer. The enzyme was the most active toward p-nitrophenyl (p-NP) acetate and p-NP-butyrate to a lesser extent. However, the enzyme could not hydrolyze p-NP-myristate, palmitate, and stearate. Therefore, the enzyme is considered as an esterase, very different from lipase. The purified esterase had optimal pH at around 8.0 and was stable in a broad range of pH values. The optimal temperature ranged from 30 to $40^{\circ}C$, and the residual activity observed after heat treatment at $55^{\circ}C$ for 20 min was 72 % of the initial activity. The activity was inhibited by the presence of copper and cobalt ions.

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