Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity

Abstract Background Bioremediation relying on highly efficient degrading bacteria constitutes a promising and sustainable avenue for controlling and reducing nitrophenol contamination in the environment. A thorough understanding of the bacterial degradation mechanism of nitrophenol is of paramount i...

Full description

Saved in:
Bibliographic Details
Main Authors: Jian Yang, Shanshan Lin, Wei Li, Xianjie Wang, Ru Li
Format: Article
Language:English
Published: BMC 2025-04-01
Series:Microbial Cell Factories
Subjects:
Online Access:https://doi.org/10.1186/s12934-025-02712-1
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850148921787023360
author Jian Yang
Shanshan Lin
Wei Li
Xianjie Wang
Ru Li
author_facet Jian Yang
Shanshan Lin
Wei Li
Xianjie Wang
Ru Li
author_sort Jian Yang
collection DOAJ
description Abstract Background Bioremediation relying on highly efficient degrading bacteria constitutes a promising and sustainable avenue for controlling and reducing nitrophenol contamination in the environment. A thorough understanding of the bacterial degradation mechanism of nitrophenol is of paramount importance for supporting the development of efficient microbial remediation technology. Results In this study, a new bacterium, Rhodococcus sp. 21391, endowed with superior p-nitrophenol (PNP) degradation ability was obtained. Genomic and comparative proteomic analyses revealed that it utilizes the 1,2,4-benzenetriol (BT) pathway for PNP degradation. The catalytic properties of the two-component p-nitrophenol monooxygenase RsNcpAB from the strain were investigated in vitro. The enzyme exhibited a broad substrate selectivity, catalyzing the oxidation of various nitrophenols and halogenated phenols, with significant potential for further research and development. Additionally, the crystal structure of the oxidative component of p-nitrophenol monooxygenase, RsNcpA, was determined. Structural analysis and site-directed mutagenesis revealed that residues Arg100 and His293 in the active site play a crucial role in enzyme catalysis, and a catalytic mechanism model was subsequently proposed. Conclusions This study reports a high-performance nitrophenol-degrading bacterium and enzyme, and reveals their mechanisms at the molecular level. These findings increase the understanding of the bacterial degradation of nitrophenol, thereby providing a crucial foundation for the development of efficient bioremediation technologies. Graphical Abstract
format Article
id doaj-art-faefafa9aa344292bd83ea149f23c45e
institution OA Journals
issn 1475-2859
language English
publishDate 2025-04-01
publisher BMC
record_format Article
series Microbial Cell Factories
spelling doaj-art-faefafa9aa344292bd83ea149f23c45e2025-08-20T02:27:06ZengBMCMicrobial Cell Factories1475-28592025-04-0124111110.1186/s12934-025-02712-1Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificityJian Yang0Shanshan Lin1Wei Li2Xianjie Wang3Ru Li4School of Life and Health Technology, Dongguan University of TechnologyCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of OceanologyCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of OceanologyCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of OceanologyCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of OceanologyAbstract Background Bioremediation relying on highly efficient degrading bacteria constitutes a promising and sustainable avenue for controlling and reducing nitrophenol contamination in the environment. A thorough understanding of the bacterial degradation mechanism of nitrophenol is of paramount importance for supporting the development of efficient microbial remediation technology. Results In this study, a new bacterium, Rhodococcus sp. 21391, endowed with superior p-nitrophenol (PNP) degradation ability was obtained. Genomic and comparative proteomic analyses revealed that it utilizes the 1,2,4-benzenetriol (BT) pathway for PNP degradation. The catalytic properties of the two-component p-nitrophenol monooxygenase RsNcpAB from the strain were investigated in vitro. The enzyme exhibited a broad substrate selectivity, catalyzing the oxidation of various nitrophenols and halogenated phenols, with significant potential for further research and development. Additionally, the crystal structure of the oxidative component of p-nitrophenol monooxygenase, RsNcpA, was determined. Structural analysis and site-directed mutagenesis revealed that residues Arg100 and His293 in the active site play a crucial role in enzyme catalysis, and a catalytic mechanism model was subsequently proposed. Conclusions This study reports a high-performance nitrophenol-degrading bacterium and enzyme, and reveals their mechanisms at the molecular level. These findings increase the understanding of the bacterial degradation of nitrophenol, thereby providing a crucial foundation for the development of efficient bioremediation technologies. Graphical Abstracthttps://doi.org/10.1186/s12934-025-02712-1Nitrophenol biodegradationComparative proteomep-nitrophenol monooxygenaseSubstrate specificity
spellingShingle Jian Yang
Shanshan Lin
Wei Li
Xianjie Wang
Ru Li
Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
Microbial Cell Factories
Nitrophenol biodegradation
Comparative proteome
p-nitrophenol monooxygenase
Substrate specificity
title Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
title_full Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
title_fullStr Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
title_full_unstemmed Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
title_short Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity
title_sort biodegradation of p nitrophenol by rhodococcus sp 21391 unveils a two component p nitrophenol monooxygenase with broad substrate specificity
topic Nitrophenol biodegradation
Comparative proteome
p-nitrophenol monooxygenase
Substrate specificity
url https://doi.org/10.1186/s12934-025-02712-1
work_keys_str_mv AT jianyang biodegradationofpnitrophenolbyrhodococcussp21391unveilsatwocomponentpnitrophenolmonooxygenasewithbroadsubstratespecificity
AT shanshanlin biodegradationofpnitrophenolbyrhodococcussp21391unveilsatwocomponentpnitrophenolmonooxygenasewithbroadsubstratespecificity
AT weili biodegradationofpnitrophenolbyrhodococcussp21391unveilsatwocomponentpnitrophenolmonooxygenasewithbroadsubstratespecificity
AT xianjiewang biodegradationofpnitrophenolbyrhodococcussp21391unveilsatwocomponentpnitrophenolmonooxygenasewithbroadsubstratespecificity
AT ruli biodegradationofpnitrophenolbyrhodococcussp21391unveilsatwocomponentpnitrophenolmonooxygenasewithbroadsubstratespecificity