Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Reaction Characteristics of 4-Methylcatechol 2,3-Dioxygenase from Pseudomonas putida SU10

  • Ha, You-Mee (Department of Biological Science, Sookmyung Women's Universiy) ;
  • Jung, Young-Hee (Department of Biological Science, Sookmyung Women's Universiy) ;
  • Kwon, Dae-Young (Korea Food Research Institute) ;
  • Kim, Young-Soo (Department of Pharmacy, Chungbk National University) ;
  • Kim, Chy-Kyung (Department of Microbiology, Chungbk National University) ;
  • Min, Kyung-Hee (Department of Biological Science, Sookmyung Women's Universiy)
  • Published : 2000.02.01
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Abstract

Reaction characteristics of 4-methylcatechol 2,3-dioxygenase (4MC230) purified from Pseudomonas putida SU10 with a higher activity toward 4-methylcatechol than catechol or 3-cethylcatechol were studied by altering their physical and chemical properties. The enzyme exhibited a maximum activity at pH 7.5 and approximately 40% at pH 6.0 for 4-methylcatechol hydrolysis. The optimum temperature for the enzyme was around $35^{\circ}C$, since the enzyme was unstable at higher temperature. Acetone(10%) stabilized the 4MC230. The effects of solvent and other chemicals (inactivator or reactivator) for the reactivation of the 4MC230 were also investigated. Silver nitrate and hydrogen peroxid severely deactivated the enzyme and the deactivation by hydrogen peroxide severely deactivated the enzyme and the deactivation by hydrogen peroxide was mainly due to the oxidation of ferrous ion to ferric ion. Some solvents acted as an activator and protector for the enzyme from deactivation by hydrogen peroxide. Ascorbate, cysteine, or ferrous ion reactivated the deactivated enzyme by hydrogen peroxide. The addition of ferrous ion together with a reducing agent fully recovered the enzyme activity and increased its activity abut 2 times.

Keywords

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