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Purification and Characterization of a Thermostable ${\beta}-1$,3-1,4-Glucanase from Laetiporus sulphureus var. miniatus

  • Hong, Mi-Ri (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Kim, Yeong-Su (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Joo, Ah-Reum (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Lee, Jung-Kul (Department of Chemical Engineering, Konkuk University) ;
  • Kim, Yeong-Suk (Department of Forest Products, Kookmin University) ;
  • Oh, Deok-Kun (Department of Bioscience and Biotechnology, Konkuk University)
  • Published : 2009.08.31

Abstract

A ${\beta}-1$,3-1,4-glucanase from the fungus Laetiporus sulphureus var. miniatus was purified as a single 26 kDa band by ammonium sulfate precipitation, HiTrap Q HP, and UNO Q ion-exchange chromatography, with a specific activity of 29 U/mg. The molecular mass of the native enzyme was 52 kDa as a dimer by gel filtration. ${\beta}-1$,3-1,4-Glucanase showed optimum activity at pH 4.0 and $75^{\circ}C$. The half-lives of the enzyme at $70^{\circ}C$ and $75^{\circ}C$ were 152 h and 22 h, respectively. The enzyme showed the highest activity for barley ${\beta}$-glucan as ${\beta}-1$,3-1,4-glucan among the tested polysaccharides and p-nitrophenyl-${\beta}$-D-glycosides with a $K_m$, of 0.67 mg/ml, a $k_{cat}$ of 13.5 $S^{-1}$ and a $k_{cat}/K_m$ of 20 mg/ml/s.

Keywords

References

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