Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of an Extradiol Dioxygenase Which Preferentially Acts on 4-Methylcatechol

  • Ha, You-Mee (Department of Biological Science, Sookmyung Women′s University) ;
  • Jung, Young-Hee (Department of Biological Science, Sookmyung Women's University) ;
  • Kwon, Dae-Young (Korea Food Research Institute) ;
  • Kim, Young-Chang (Department of Microbiology, Chungbuk National University) ;
  • Kim, Young-Soo (Department of Pharmacy, Chungbuk National University) ;
  • Kim, Chy-Kyung (Department of Microbiology, Chungbuk National University) ;
  • Min, Kyung-Hee (Department of Biological Science, Sookmyung Women′s University)
  • Published : 1999.06.01
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Abstract

A catechol 2,3-dioxygenase (C23O) was purified to apparent homogeneity from Pseudomonas putida SU10 through several purification steps consisting of ammonium sulfate precipitation and chromatographies on DEAE 5PW, Superdex S-200, and Resource-Q. Gel filtration indicated a molecular mass under nondenaturing conditions of about 130 kDa. The enzyme has a subunit of 34 kDa as was determined by SDS-PAGE. These results suggest that the native enzyme is composed of four identical subunits. The N-terminal amino acid sequence (30 residues) of the enzyme has been determined and exhibits high identity with other extradiol dioxygenases. The reactivity of this enzyme towards catechol and methyl-substituted catechols is somewhat different from that seen for other catechol 2,3-dioxygenases, with 4-methylcatechol cleaved at a higher rate than catechol or 3-methylcatechol. $K_m$ values of the enzyme for these substrates are between 3.5 and 5.7 M.

Keywords

References

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