Identification of the upstream 4-chlorophenol biodegradation pathway using a recombinant monooxygenase from Arthrobacter chlorophenolicus A6. (December 2017)

• Record Type: Journal Article
• Authors: Cho, Su Yeon; Kwean, Oh Sung; Yang, Jun Won; Cho, Wooyoun; Kwak, Seonyeong; Park, Sungyoon; Lim, Yejee; Kim, Han S.
• Is Part Of: Bioresource technology. Volume 245:Part B(2017)
• Page Start: 1800
• Page End: 1807
• Publication Date: 2017-12
• Subjects: 4-Chlorophenol -- Arthrobacter chlorophenolicus A6 -- Two-component flavin-diffusible monooxygenase -- CphC-I -- Initial biodegradation pathway; Biomass -- Periodicals; Biomass energy -- Periodicals; Bioremediation -- Periodicals; Agricultural wastes -- Periodicals; Factory and trade waste -- Periodicals; Organic wastes -- Periodicals; Bioénergie -- Périodiques; Déchets agricoles -- Périodiques; Déchets industriels -- Périodiques; Déchets organiques -- Périodiques; Déchets (Combustible) -- Périodiques; 662.88
• DOI: 10.1016/j.biortech.2017.05.006
• Languages: English
• ISSNs: 0960-8524

Abstract

Graphical abstract: Highlights: TC-FDM catalyzes initial 4-chlorophenol biodegradation pathway. 4-CP is converted sequentially to BQN → HQN → HQL by CphC-I. The activity of CphC-I for 4-CP was 63.22 U/mg-protein. vmax = 0.21 mM/min, KM = 0.19 mM, and kcat / KM = 0.04 mM −1 min −1 . Abstract: This study aimed to clarify the initial 4-chlorophenol (4-CP) biodegradation pathway promoted by a two-component flavin-diffusible monooxygenase (TC-FDM) consisting of CphC-I and CphB contained in Arthrobacter chlorophenolicus A6 and the decomposition function of CphC-I. The TC-FDM genes were cloned from A. chlorophenolicus A6, and the corresponding enzymes were overexpressed. Since CphB was expressed in an insoluble form, Fre, a flavin reductase obtained from Escherichia coli, was used. These enzymes were purified using Ni 2+ -NTA resin. It was confirmed that TC-FDM catalyzes the oxidation of 4-CP and the sequential conversion of 4-CP to benzoquinone (BQN) → hydroquinone (HQN) → HQL. This indicated that CphC-I exhibits substrate specificity for 4-CP, BQN, and HQN. The activity of CphC-I for 4-CP was 63.22 U/mg-protein, and the Michaelis-Menten kinetic parameters were vmax = 0.21 mM/min, KM = 0.19 mM, and kcat / KM = 0.04 mM −1 min −1 . These results would be useful for the development of a novel biochemical treatment technology for 4-CP and phenolic hydrocarbons.