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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Purification and Characterization of an Indican-hydrolyzing β-glucosidase from Agrobacterium tumefaciens

Agrobacterium tumefaciens 유래 인디칸 분해활성을 갖는 β-glucosidase의 분리와 특성분석

  • Hwang, Chang-Sun (Department of Biological Sciences, College of Natural Sciences, Chonnam National University) ;
  • Lee, Jin-Young (Department of Biological Sciences, College of Natural Sciences, Chonnam National University) ;
  • Kim, Geun-Joong (Department of Biological Sciences, College of Natural Sciences, Chonnam National University)
  • 황창선 (전남대학교 자연과학대학 생물학과) ;
  • 이진영 (전남대학교 자연과학대학 생물학과) ;
  • 김근중 (전남대학교 자연과학대학 생물학과)
  • Received : 2012.12.05
  • Accepted : 2012.12.24
  • Published : 2012.12.31
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Abstract

Indican (indoxyl-${\beta}$-D-glucoside) is a colorless natural compound and can be used as a precursor for the production of indigo. This production step only require an enzyme, ${\beta}$-glucosidase, that readily screened from microbial resource by using selective media supplemented with indican as a sole carbon source. Agrobacterium tumefaciens was well grown in this media and thus presumed to produce a related enzyme. The corresponding gene, encoding a protein with a calculated molecular mass of 51 kDa, was cloned and overexpressed as MBP fusion proteins. The purified enzyme was determined to be a dimer and showed the maximum activity for indican at pH 7.0 and $40^{\circ}C$. The kinetic parameters for indican, Km and Vmax, were determined to be 1.4 mM and 373.8 ${\mu}M/min/mg$, respectively. The conversion yield of indican into indigo using this enzyme was about 1.7-1.8 folds higher than that of previously isolated enzyme from Sinorhizobium meliloti. Additionally, this enzyme was able to hydrolyze various ${\beta}$-1,4 glycoside substrates.

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References

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