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Volume 159, Issue Pt_3

Genetic and chemical characterization of ibuprofen degradation by Ibu-2 Free

Abstract

Ibu-2 has the unusual ability to cleave the acid side chain from the pharmaceutical ibuprofen and related arylacetic acid derivatives to yield corresponding catechols under aerobic conditions via a previously uncharacterized mechanism. Screening a chromosomal library of Ibu-2 DNA in EPI300 allowed us to identify one fosmid clone (pFOS3G7) that conferred the ability to metabolize ibuprofen to isobutylcatechol. Characterization of pFOS3G7 loss-of-function transposon mutants permitted identification of five ORFs, , whose predicted amino acid sequences bore similarity to the large and small units of an aromatic dioxygenase (), a sterol carrier protein X (SCPx) thiolase (), a domain of unknown function 35 (DUF35) protein () and an aromatic CoA ligase (). Two additional ORFs, and , which encode putative ferredoxin reductase and ferredoxin components of an aromatic dioxygenase system, respectively, were also identified on pFOS3G7. Complementation of a markerless loss-of-function deletion mutant restored catechol production as did complementation of the Tn mutant. Expression of subcloned alone in did not impart full metabolic activity unless coexpressed with . CoA ligation followed by ring oxidation is common to phenylacetic acid pathways. However, the need for a putative SCPx thiolase (IpfD) and DUF35 protein (IpfE) in aerobic arylacetic acid degradation is unprecedented. This work provides preliminary insights into the mechanism behind this novel arylacetic acid-deacylating, catechol-generating activity.

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2013-03-01
2024-04-20
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Genetic and chemical characterization of ibuprofen degradation by Sphingomonas Ibu-2
Microbiology 159, 621 (2013); https://doi.org/10.1099/mic.0.062273-0
/content/journal/micro/10.1099/mic.0.062273-0
/content/journal/micro/10.1099/mic.0.062273-0
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