The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1

Abstract Transmembrane alkane monooxygenase (AlkB)‐type monooxygenases, especially AlkB2 monooxygenases, are crucial for aerobic degradation of the medium‐to‐long‐chain n‐alkanes in hydrocarbon‐utilizing microorganisms. In this study, we identified a GntR/VanR transcription regulator AlkR of Pseudom...

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Main Authors: Wanli Peng, Xiuli Wang, Qinchen Liu, Zhihong Xiao, Fulin Li, Nannan Ji, Zhuo Chen, Jiaying He, Junhao Wang, Zixin Deng, Shuangjun Lin, Rubing Liang
Format: Article
Language:English
Published: Wiley 2025-04-01
Series:mLife
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Online Access:https://doi.org/10.1002/mlf2.70004
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author Wanli Peng
Xiuli Wang
Qinchen Liu
Zhihong Xiao
Fulin Li
Nannan Ji
Zhuo Chen
Jiaying He
Junhao Wang
Zixin Deng
Shuangjun Lin
Rubing Liang
author_facet Wanli Peng
Xiuli Wang
Qinchen Liu
Zhihong Xiao
Fulin Li
Nannan Ji
Zhuo Chen
Jiaying He
Junhao Wang
Zixin Deng
Shuangjun Lin
Rubing Liang
author_sort Wanli Peng
collection DOAJ
description Abstract Transmembrane alkane monooxygenase (AlkB)‐type monooxygenases, especially AlkB2 monooxygenases, are crucial for aerobic degradation of the medium‐to‐long‐chain n‐alkanes in hydrocarbon‐utilizing microorganisms. In this study, we identified a GntR/VanR transcription regulator AlkR of Pseudomonas aeruginosa SJTD‐1 involved in the negative regulation of AlkB2 and deciphered its nature of DNA binding and ligand release. The deletion of alkR enhanced the transcription levels of the alkB2 gene and the utilization efficiency of the medium‐to‐long‐chain n‐alkanes by strain SJTD‐1. The dimer of AlkR recognizes and binds to a conserved palindromic motif in the promoter of the alkB2 gene, and structural symmetry is vital for DNA binding and transcription repression. The long‐chain fatty acyl coenzyme A compounds can release AlkR and stimulate transcription of alkB2, reflecting the effect of alkane catabolic metabolites. Structural insights unveiled that the arginine residues and scaffold residues of AlkR are critical for DNA binding. Further bioinformatics analysis of AlkR revealed the widespread VanR–AlkB couples distributed in Pseudomonadaceae with high conservation in the sequences of functional genes and intergenic regions, highlighting a conserved regulatory pattern for n‐alkane utilization across this family. These findings demonstrate the regulatory mechanism and structural basis of GntR/VanR transcription regulators in modulating n‐alkane biodegradation and provide valuable insights in improving the bioremediation efficiency of hydrocarbon pollution.
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spelling doaj-art-58289ebab80f4faba0d44ad79b0606ae2025-08-20T03:53:39ZengWileymLife2770-100X2025-04-014212614210.1002/mlf2.70004The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1Wanli Peng0Xiuli Wang1Qinchen Liu2Zhihong Xiao3Fulin Li4Nannan Ji5Zhuo Chen6Jiaying He7Junhao Wang8Zixin Deng9Shuangjun Lin10Rubing Liang11State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaState Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology Shanghai Jiao Tong University Shanghai ChinaAbstract Transmembrane alkane monooxygenase (AlkB)‐type monooxygenases, especially AlkB2 monooxygenases, are crucial for aerobic degradation of the medium‐to‐long‐chain n‐alkanes in hydrocarbon‐utilizing microorganisms. In this study, we identified a GntR/VanR transcription regulator AlkR of Pseudomonas aeruginosa SJTD‐1 involved in the negative regulation of AlkB2 and deciphered its nature of DNA binding and ligand release. The deletion of alkR enhanced the transcription levels of the alkB2 gene and the utilization efficiency of the medium‐to‐long‐chain n‐alkanes by strain SJTD‐1. The dimer of AlkR recognizes and binds to a conserved palindromic motif in the promoter of the alkB2 gene, and structural symmetry is vital for DNA binding and transcription repression. The long‐chain fatty acyl coenzyme A compounds can release AlkR and stimulate transcription of alkB2, reflecting the effect of alkane catabolic metabolites. Structural insights unveiled that the arginine residues and scaffold residues of AlkR are critical for DNA binding. Further bioinformatics analysis of AlkR revealed the widespread VanR–AlkB couples distributed in Pseudomonadaceae with high conservation in the sequences of functional genes and intergenic regions, highlighting a conserved regulatory pattern for n‐alkane utilization across this family. These findings demonstrate the regulatory mechanism and structural basis of GntR/VanR transcription regulators in modulating n‐alkane biodegradation and provide valuable insights in improving the bioremediation efficiency of hydrocarbon pollution.https://doi.org/10.1002/mlf2.70004AlkB2 monooxygenaseAlkR transcription regulatorregulatory mechanismstructure featuresVanR–AlkB couples
spellingShingle Wanli Peng
Xiuli Wang
Qinchen Liu
Zhihong Xiao
Fulin Li
Nannan Ji
Zhuo Chen
Jiaying He
Junhao Wang
Zixin Deng
Shuangjun Lin
Rubing Liang
The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
mLife
AlkB2 monooxygenase
AlkR transcription regulator
regulatory mechanism
structure features
VanR–AlkB couples
title The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
title_full The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
title_fullStr The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
title_full_unstemmed The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
title_short The GntR/VanR transcription regulator AlkR represses AlkB2 monooxygenase expression and regulates n‐alkane degradation in Pseudomonas aeruginosa SJTD‐1
title_sort gntr vanr transcription regulator alkr represses alkb2 monooxygenase expression and regulates n alkane degradation in pseudomonas aeruginosa sjtd 1
topic AlkB2 monooxygenase
AlkR transcription regulator
regulatory mechanism
structure features
VanR–AlkB couples
url https://doi.org/10.1002/mlf2.70004
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