Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Surface Immobilizntion on Silica of Endoxylanase Produced from Recombinant Bacillus subtilis

  • Kang, Su-Cheol (Department of Bioscience and Biotechnology, Sejong University) ;
  • Kim, Hye-Jeong (Department of Bioscience and Biotechnology, Sejong University) ;
  • Nam, Soo-Wan (Department of Biotechnology and Bioengineering, Dong-Eui University) ;
  • Oh, Deok-Kun (Department of Biotechnology and Bioengineering, Dong-Eui University)
  • Published : 2002.10.01
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Abstract

The plasmid, pJHKJ4, containing the endoxylanase gene, was introduced into Bacillus subtilis DB 104. The recombinant cells produced 587 unit/ml of endoxylanase at 33 h. The endoxylanase was immobilized covalently on the surface of silica fur effective xylan hydrolysis. The activities of the immobilized and free endoxylanases were optimal at pH 6.5 and 10 mM $MnSO_4$. The optimal temperature of the immobilized endoxylanase was $60^{\circ}C$, whereas that of the free endoxylanase was $65^{\circ}C$. Under these optimal conditions, the activity of the immobilized endoxylanase was 1.7 times higher than that of the fee endoxylanase. From microscope photographs, the immobilized endoxylanase was found to be bounded and evenly distributed on the surface of silica, a nonporous solid support. The enzyme kinetics between the immobilized and free endoxylanases was estimated to be uncompetitive, when plotting double-reciprocal plots against xylan concentrations and endoxylanase activities. These results suggest that the higher activity of the immobilized endoxylanase may be due to increased formation of enzyme-substrate complex, because of the easy accessibility of the immobilized enzyme to the polysaccharide-xylan as a high molecular weight substrate.

Keywords

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