Identification of a novel self-sufficient styrene monooxygenase from Rhodococcus opacus 1CP

Sequence analysis of a 9-kb genomic fragment of the actinobacterium Rhodococcus opacus 1CP led to identification of an open reading frame encoding a novel fusion protein, StyA2B, with a putative function in styrene metabolism via styrene oxide and phenylacetic acid. Gene cluster analysis indicated t...

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Veröffentlicht in:Journal of bacteriology Jg. 191; H. 15; S. 4996
Hauptverfasser: Tischler, Dirk, Eulberg, Dirk, Lakner, Silvia, Kaschabek, Stefan R, van Berkel, Willem J H, Schlömann, Michael
Format: Journal Article
Sprache:Englisch
Veröffentlicht: United States 01.08.2009
Schlagworte:
Bacterial Proteins - classification
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacterial Proteins - physiology
Chromatography, High Pressure Liquid
Epoxy Compounds - chemistry
Epoxy Compounds - metabolism
Gas Chromatography-Mass Spectrometry
Genome, Bacterial - genetics
Genome, Bacterial - physiology
Models, Biological
Molecular Sequence Data
Oxygenases - classification
Oxygenases - genetics
Oxygenases - metabolism
Oxygenases - physiology
Phylogeny
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Rhodococcus - enzymology
Rhodococcus - genetics
Rhodococcus - metabolism
Styrene - chemistry
Styrene - metabolism
ISSN:1098-5530, 1098-5530
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Zusammenfassung:Sequence analysis of a 9-kb genomic fragment of the actinobacterium Rhodococcus opacus 1CP led to identification of an open reading frame encoding a novel fusion protein, StyA2B, with a putative function in styrene metabolism via styrene oxide and phenylacetic acid. Gene cluster analysis indicated that the highly related fusion proteins of Nocardia farcinica IFM10152 and Arthrobacter aurescens TC1 are involved in a similar physiological process. Whereas 413 amino acids of the N terminus of StyA2B are highly similar to those of the oxygenases of two-component styrene monooxygenases (SMOs) from pseudomonads, the residual 160 amino acids of the C terminus show significant homology to the flavin reductases of these systems. Cloning and functional expression of His(10)-StyA2B revealed for the first time that the fusion protein does in fact catalyze two separate reactions. Strictly NADH-dependent reduction of flavins and highly enantioselective oxygenation of styrene to (S)-styrene oxide were shown. Inhibition studies and photometric analysis of recombinant StyA2B indicated the absence of tightly bound heme and flavin cofactors in this self-sufficient monooxygenase. StyA2B oxygenates a spectrum of aromatic compounds similar to those of two-component SMOs. However, the specific activities of the flavin-reducing and styrene-oxidizing functions of StyA2B are one to two orders of magnitude lower than those of StyA/StyB from Pseudomonas sp. strain VLB120.
Bibliographie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:1098-5530
1098-5530
DOI:10.1128/JB.00307-09