Article ID Journal Published Year Pages File Type
4996734 Bioresource Technology 2017 7 Pages PDF
Abstract

•Arthrobacter chlorophenolicus A6 possesses DNA genes for 4-chlorophenol degradation.•Soluble form CphC-I was expressed, whereas CphB expressed in an insoluble form.•CphC-I exhibited high specific activity when coupled with a flavin reductase, Fre.•vmax = 223.3 μM·min−1, KM = 249.4 μM, and kcat/KM = 0.052 min−1·μM−1.

In this study, cphC-I and cphB, encoding a putative two-component flavin-diffusible monooxygenase (TC-FDM) complex, were cloned from Arthrobacter chlorophenolicus A6. The corresponding enzymes were overexpressed to assess the feasibility of their utilization for the oxidative decomposition of 4-chlorophenol (4-CP). Soluble CphC-I was produced at a high level (∼50%), and subsequently purified. Since CphB was expressed in an insoluble form, a flavin reductase, Fre, cloned from Escherichia coli was used as an alternative reductase. CphC-I utilized cofactor FADH2, which was reduced by Fre for the hydroxylation of 4-CP. This recombinant enzyme complex exhibited a higher specific activity for the oxidation of 4-CP (45.34 U/mg-protein) than that exhibited by CphC-I contained in cells (0.18 U/mg-protein). The Michaelis-Menten kinetic parameters were determined as: vmax = 223.3 μM·min−1, KM = 249.4 μM, and kcat/KM = 0.052 min−1·μM−1. These results could be useful for the development of a new biochemical remediation technique based on enzymatic agents catalyzing the degradation of phenolic contaminants.

Graphical abstractColor change from yellow to orange-red after 4-CP degradation by CphC-I. CphC-I concentration from 0.1 to 25 μM from left to right.Download high-res image (123KB)Download full-size image

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Physical Sciences and Engineering Chemical Engineering Process Chemistry and Technology
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