Assembly of a Rieske non-heme iron oxygenase multicomponent system from Phenylobacterium immobile E DSM 1986 enables pyrazon cis-dihydroxylation in E. coli
Autor: | Daniela Resende, Per-Olof Syrén, Wendy Escobedo-Hinojosa, Elsa Potoudis, Theresa Farr, Bernhard Hauer, Andreas Hunold |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: |
Cis-dihydroxylation
Oxygenase Iron 030303 biophysics medicine.disease_cause Applied Microbiology and Biotechnology 03 medical and health sciences chemistry.chemical_compound Rieske non-heme iron oxygenases medicine Escherichia coli Gene Ferredoxin 030304 developmental biology Phenylobacterium immobile Applied Genetics and Molecular Biotechnology 0303 health sciences General Medicine Caulobacteraceae Pyridazines chemistry Biochemistry Dihydroxylation Biodegradation Biocatalysis Oxygenases Heterologous expression Pyrazon oxygenase DNA Biotechnology |
Zdroj: | Applied Microbiology and Biotechnology |
Popis: | Phenylobacterium immobile strain E is a soil bacterium with a striking metabolism relying on xenobiotics, such as the herbicide pyrazon, as sole carbon source instead of more bioavailable molecules. Pyrazon is a heterocyclic aromatic compound of environmental concern and its biodegradation pathway has only been reported in P. immobile. The multicomponent pyrazon oxygenase (PPO), a Rieske non-heme iron oxygenase, incorporates molecular oxygen at the 2,3 position of the pyrazon phenyl moiety as first step of degradation, generating a cis-dihydrodiendiol. The aim of this work was to identify the genes encoding for each one of the PPO components and enable their functional assembly in Escherichia coli. P. immobile strain E genome sequencing revealed genes encoding for RO components, such as ferredoxin-, reductase-, α- and β-subunits of an oxygenase. Though, P. immobile E displays three prominent differences with respect to the ROs currently characterized: (1) an operon-like organization for PPO is absent, (2) all the elements are randomly scattered in its DNA, (3) not only one, but 19 different α-subunits are encoded in its genome. Herein, we report the identification of the PPO components involved in pyrazon cis-dihydroxylation in P. immobile, its appropriate assembly, and its functional reconstitution in E. coli. Our results contributes with the essential missing pieces to complete the overall elucidation of the PPO from P. immobile. Bundesministerium für Bildung und Forschung Consejo Nacional de Ciencia y Tecnología Projekt DEAL |
Databáze: | OpenAIRE |
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