Assembly of a Rieske non-heme iron oxygenase multicomponent system from Phenylobacterium immobile E DSM 1986 enables pyrazon cis-dihydroxylation in E. coli

• Abstract: 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. Key points: • Phenylobacterium immobile E DSM 1986 harbors the only described pyrazon oxygenase (PPO). • We elucidated the genes encoding for all PPO components. • Heterologous expression of PPO enabled pyrazon dihydroxylation in E. coli JW5510. 

Ämnesord

NATURVETENSKAP  -- Biologi -- Mikrobiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Microbiology (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Microbiology in the medical area (hsv//eng)

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

Biocatalysis

Biodegradation

Cis-dihydroxylation

Phenylobacterium immobile

Pyrazon oxygenase

Rieske non-heme iron oxygenases

Aromatic compounds

Biochemistry

Encoding (symbols)

Genes

Hydroxylation

Iron

Molecular oxygen

Porphyrins

Signal encoding

Biodegradation pathways

Environmental concerns

Genome sequencing

Heterocyclic aromatic compounds

Heterologous expression

Multi-component systems

Multicomponents

Sole carbon source

Escherichia coli

ampicillin

bacterial DNA

chloridazon

ferredoxin

oxidoreductase

oxygenase

rieske non heme iron oxygenase

unclassified drug

pyridazine derivative

bacterium

bioavailability

chemical compound

chemical reaction

coliform bacterium

detection method

enzyme

enzyme activity

gene expression

genome

microbial activity

alpha chain

Article

bacterial genome

bacterial strain

beta chain

controlled study

degradation

dihydroxylation

gene identification

gene sequence

genetic code

nonhuman

Phenylobacterium immobile e dsm 1986

soil microflora

Caulobacteraceae

genetics

Bacteria (microorganisms)

Oxygenases

Pyridazines

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)