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http://dx.doi.org/10.7857/JSGE.2014.19.3.047

Overexpression and Purification of Monooxygenases Cloned from Arthrobacter chlorophenolicus A6 for Enzymatic Decomposition of 4-Chlorophenol  

Ryu, Song-Jung (Department of Environmental Engineering, Konkuk University)
Kang, Christina S. (Department of Environmental Engineering, Konkuk University)
Kim, Han S. (Department of Environmental Engineering, Konkuk University)
Publication Information
Journal of Soil and Groundwater Environment / v.19, no.3, 2014 , pp. 47-55 More about this Journal
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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