• Overexpression and Purification of Monooxygenases Cloned from Arthrobacter chlorophenolicus A6 for Enzymatic Decomposition of 4-Chlorophenol
  • Ryu, Song-Jung;Kang, Christina S.;Kim, Han S.;
  • Department of Environmental Engineering, Konkuk University;Department of Environmental Engineering, Konkuk University;Department of Environmental Engineering, Konkuk University;
  • 4-Chlorophenol 분해박테리아 Arthrobacter chlorophenolicus A6로부터의 monooxygenase의 복제 및 대량발현과 정제 그리고 기질분해활성도 분석
  • 류송정;이소라;김한승;
  • 건국대학교 환경공학과;건국대학교 환경공학과;건국대학교 환경공학과;
Abstract
Arthrobacter chlorophenolicus A6 possesses several monooxygenases (CphC-I, CphC-II, and CphB) that can catalyze the transformation of 4-chlorophenol (4-CP) to hydroxylated intermediates in the initial steps of substrate metabolism. The corresponding genes of the monooxygenases were cloned, and the competent cells were transformed with these recombinant plasmids. Although CphC-II and CphB were expressed as insoluble forms, CphC-I was successfully expressed as a soluble form and isolated by purification. The specific activity of the purified CphC-I was analyzed by using 4-CP, 4-chlorocatechol (4-CC), and catechol (CAT) as substrates. The specific activities for 4-CP, 4-CC, and CAT were determined to be 0.312 U/mg, 0.462 U/mg, 0.246 U/mg, respectively. The results of this study indicated that CphC-I is able to catalyze the degradation of 4-CC and CAT in addition to 4-CP, which is a primary substrate. This research is expected to provide the fundamental information for the development of an eco-friendly biochemical degradation of aromatic hydrocarbons.

Keywords: 4-Chlorophenol;Arthrobacter chlorophenolicus A6;Monooxygenase;Enzyme cloning and expression;

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This Article

  • 2014; 19(3): 47-55

    Published on Jun 30, 2014

  • 10.7857/JSGE.2014.19.3.047
  • Received on Mar 13, 2014
  • Revised on Mar 27, 2014
  • Accepted on Mar 30, 2014