Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Characterization of the Intact Form of Thermotoga maritima Pectinase TmPecN Expressed in Escherichia coli

  • Kim, Chung Ho (Department of Food and Nutrition, Seowon University) ;
  • Cheong, Jong-Joo (Center for Food and Bioconvergence, Seoul National University)
  • Received : 2014.08.20
  • Accepted : 2014.09.11
  • Published : 2015.06.30
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Abstract

The thermostable pectinase gene TmPec isolated from Thermotoga maritima was introduced into the NdeI site of pRSET-B vector and expressed in its intact form in Escherichia coli BL21. The overexpressed intact form of pectinase (TmPecN protein) was partially purified by heat-denaturation procedure. TmPecN showed the highest activity between 85 and $95^{\circ}C$, and at approximately pH 6.5. Enzyme activity was stably maintained at temperatures below $85^{\circ}C$. In the presence of $Ca^{2+}$, pectinase activity of TmPecN increased to 128.4% of normal level. In contrast, $Ba^{2+}$, $Zn^{2+}$, and $Mn^{2+}$ strongly inhibited TmPecN activity. We conclude that the biochemical properties of the intact form of TmPecN are comparable to those of the recombinant protein TmPec reported previously.

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References

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