Cloning of two gene clusters involved in the catabolism of 2,4-dinitrophenol by Paraburkholderia sp. strain KU-46 and characterization of the initial DnpAB enzymes and a two-component monooxygenases DnpC1C2
Copyright © 2023 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved..
Little is currently known about the metabolism of the industrial pollutant 2,4-dinitrophenol (DNP), particularly among gram-negative bacteria. In this study, we identified two non-contiguous genetic loci spanning 22 kb of Paraburkholderia (formerly Burkholderia) sp. strain KU-46. Additionally, we characterized four key initial genes (dnpA, dnpB, and dnpC1C2) responsible for DNP degradation, providing molecular and biochemical evidence for the degradation of DNP via the formation of 4-nitrophenol (NP), a pathway that is unique among DNP utilizing bacteria. Reverse transcription polymerase chain reaction (PCR) analysis indicated that dnpA, which encodes the initial hydride transferase, and dnpB which encodes a nitrite-eliminating enzyme, were induced by DNP and organized in an operon. Moreover, we purified DnpA and DnpB from recombinant Escherichia coli to demonstrate their effect on the transformation of DNP to NP through the formation of a hydride-Meisenheimer complex of DNP, designated as H--DNP. The function of DnpB appears new since all homologs of the DnpB sequences in the protein database are annotated as putative nitrate ABC transporter substrate-binding proteins. The gene cluster responsible for the degradation of DNP after NP formation was designated dnpC1C2DXFER, and DnpC1 and DnpC2 were functionally characterized as the FAD reductase and oxygenase components of the two-component DNP monooxygenase, respectively. By elucidating the hqdA1A2BCD gene cluster, we are now able to delineate the final degradation pathway of hydroquinone to β-ketoadipate before it enters the tricarboxylic acid cycle.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:136 |
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| Enthalten in: |
Journal of bioscience and bioengineering - 136(2023), 3 vom: 19. Sept., Seite 223-231 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Liu, Yaxuan [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 22.08.2023 Date Revised 22.08.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jbiosc.2023.05.013 |
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| Förderinstitution / Projekttitel: |
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NLM35845736X |
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| 100 | 1 | |a Liu, Yaxuan |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Cloning of two gene clusters involved in the catabolism of 2,4-dinitrophenol by Paraburkholderia sp. strain KU-46 and characterization of the initial DnpAB enzymes and a two-component monooxygenases DnpC1C2 |
| 264 | 1 | |c 2023 | |
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| 500 | |a Date Completed 22.08.2023 | ||
| 500 | |a Date Revised 22.08.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2023 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved. | ||
| 520 | |a Little is currently known about the metabolism of the industrial pollutant 2,4-dinitrophenol (DNP), particularly among gram-negative bacteria. In this study, we identified two non-contiguous genetic loci spanning 22 kb of Paraburkholderia (formerly Burkholderia) sp. strain KU-46. Additionally, we characterized four key initial genes (dnpA, dnpB, and dnpC1C2) responsible for DNP degradation, providing molecular and biochemical evidence for the degradation of DNP via the formation of 4-nitrophenol (NP), a pathway that is unique among DNP utilizing bacteria. Reverse transcription polymerase chain reaction (PCR) analysis indicated that dnpA, which encodes the initial hydride transferase, and dnpB which encodes a nitrite-eliminating enzyme, were induced by DNP and organized in an operon. Moreover, we purified DnpA and DnpB from recombinant Escherichia coli to demonstrate their effect on the transformation of DNP to NP through the formation of a hydride-Meisenheimer complex of DNP, designated as H--DNP. The function of DnpB appears new since all homologs of the DnpB sequences in the protein database are annotated as putative nitrate ABC transporter substrate-binding proteins. The gene cluster responsible for the degradation of DNP after NP formation was designated dnpC1C2DXFER, and DnpC1 and DnpC2 were functionally characterized as the FAD reductase and oxygenase components of the two-component DNP monooxygenase, respectively. By elucidating the hqdA1A2BCD gene cluster, we are now able to delineate the final degradation pathway of hydroquinone to β-ketoadipate before it enters the tricarboxylic acid cycle | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a 2,4-Dinitrophenol | |
| 650 | 4 | |a 4-Nitrophenol | |
| 650 | 4 | |a Hydride transferase | |
| 650 | 4 | |a Nitrite-eliminating enzyme | |
| 650 | 4 | |a Nitroaromatics | |
| 650 | 4 | |a Paraburkholderia | |
| 650 | 4 | |a Two-component monooxygenase | |
| 650 | 7 | |a Mixed Function Oxygenases |2 NLM | |
| 650 | 7 | |a EC 1.- |2 NLM | |
| 650 | 7 | |a 2,4-Dinitrophenol |2 NLM | |
| 650 | 7 | |a Q13SKS21MN |2 NLM | |
| 650 | 7 | |a Oxygenases |2 NLM | |
| 650 | 7 | |a EC 1.13.- |2 NLM | |
| 700 | 1 | |a Yamamoto, Taisei |e verfasserin |4 aut | |
| 700 | 1 | |a Kohaya, Nozomi |e verfasserin |4 aut | |
| 700 | 1 | |a Yamamoto, Kota |e verfasserin |4 aut | |
| 700 | 1 | |a Okano, Kenji |e verfasserin |4 aut | |
| 700 | 1 | |a Sumiyoshi, Takaaki |e verfasserin |4 aut | |
| 700 | 1 | |a Hasegawa, Yoshie |e verfasserin |4 aut | |
| 700 | 1 | |a Lau, Peter C K |e verfasserin |4 aut | |
| 700 | 1 | |a Iwaki, Hiroaki |e verfasserin |4 aut | |
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