Identification of the upstream 4-chlorophenol biodegradation pathway using a recombinant monooxygenase from Arthrobacter chlorophenolicus A6

Bioresource Technology
Suyeon ChoHan S Kim

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 Ni2+-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 63.22U/mg-protein, and the Michaelis-Menten kinetic parameters were vmax=0.21mM/min, KM=0.19mM, and kcat/KM=0.04mM-1min-1. These results would be useful for the development of a novel biochemical treatment technology for 4-CP and phenolic hydrocarbons.

Citations

Oct 5, 2018·The Journal of Biological Chemistry·Panu Pimviriyakul, Pimchai Chaiyen
Nov 30, 2018·Journal of Applied Microbiology·M Sánchez-GonzálezR Rojas-Herrera
Oct 1, 2019·Microbial Biotechnology·Panu PimviriyakulPimchai Chaiyen
Jun 27, 2019·Biodegradation·Zhi-Qing ZhaoYan-Mei Li
Sep 24, 2018·Environmental Science and Pollution Research International·Jun Won YangHan S Kim
Mar 17, 2020·Environmental Science and Pollution Research International·José Andrés CorderoSadahiko Itoh
Oct 19, 2021·The Science of the Total Environment·Hefei ShiJinyou Shen

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