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Identification of a novel Baeyer-Villiger monooxygenase from Acinetobacter radioresistens: close relationship to the Mycobacterium tuberculosis prodrug activator EtaA.

Minerdi D, Zgrablic I, Sadeghi SJ, Gilardi G - Microb Biotechnol (2012)

Bottom Line: Phylogenetic analysis placed the sequence of this novel BVMO in the same clade of the prodrug activator ethionamide monooxygenase (EtaA) and it bears only a distant relation to the other known class I BVMO proteins.In silico analysis of the 3D model of the S13 BVMO generated by homology modelling also supports the similarities with EtaA by binding ethionamide to the active site.In vitro experiments carried out with the purified enzyme confirm that this novel BVMO is indeed capable of typical Baeyer-Villiger reactions as well as oxidation of the prodrug ethionamide.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences and Systems Biology, University of Torino, via Accademia Albertina 13, 10123 Torino, Italy.

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Agarose gel of RT‐PCR products amplified by using A. radioresistens S13 16S rDNA (A: experiment; B: control without RT), alkB (C) and almA genes (D) specific primers. RNA from S13 grown in minimal medium supplemented with C14 (lane 1), C16 (lane 2), C24 (lane 3) and C36 (lane 4) alkane; control experiment of RNA from S13 grown in minimal medium with sodium acetate (lane 5); lane M = DNA mass ladder (XL 1 kb, Eppendorf).
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f6: Agarose gel of RT‐PCR products amplified by using A. radioresistens S13 16S rDNA (A: experiment; B: control without RT), alkB (C) and almA genes (D) specific primers. RNA from S13 grown in minimal medium supplemented with C14 (lane 1), C16 (lane 2), C24 (lane 3) and C36 (lane 4) alkane; control experiment of RNA from S13 grown in minimal medium with sodium acetate (lane 5); lane M = DNA mass ladder (XL 1 kb, Eppendorf).

Mentions: RT‐PCR experiments were performed on the total RNA extracted from A. radioresistens S13 grown in minimal medium supplemented with C14, C16 (medium‐chain‐length alkanes) and C24, C36 (long‐chain‐length alkanes) as the sole energy and carbon source and from bacteria grown in the presence of sodium acetate. Using the two sets of alma‐ and alkB‐specific primers an amplified product of the expected size (499 and 466 bp respectively) was obtained only with RNA extracted from S13 grown with medium‐chain alkanes in the case of ALK primers (Fig. 6), and on RNA extracted from bacteria grown on long‐chain alkanes when BVMO primers were used. No amplified product was obtained from the RNA of bacteria grown with sodium acetate or from the RT‐negative controls. An amplified fragment of the expected size was also obtained using the A. radioresistens S13 16S rDNA‐specific primers on the RNA of the bacterium grown in the presence of medium‐ and long‐chain‐length alkanes and sodium acetate.


Identification of a novel Baeyer-Villiger monooxygenase from Acinetobacter radioresistens: close relationship to the Mycobacterium tuberculosis prodrug activator EtaA.

Minerdi D, Zgrablic I, Sadeghi SJ, Gilardi G - Microb Biotechnol (2012)

Agarose gel of RT‐PCR products amplified by using A. radioresistens S13 16S rDNA (A: experiment; B: control without RT), alkB (C) and almA genes (D) specific primers. RNA from S13 grown in minimal medium supplemented with C14 (lane 1), C16 (lane 2), C24 (lane 3) and C36 (lane 4) alkane; control experiment of RNA from S13 grown in minimal medium with sodium acetate (lane 5); lane M = DNA mass ladder (XL 1 kb, Eppendorf).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815892&req=5

f6: Agarose gel of RT‐PCR products amplified by using A. radioresistens S13 16S rDNA (A: experiment; B: control without RT), alkB (C) and almA genes (D) specific primers. RNA from S13 grown in minimal medium supplemented with C14 (lane 1), C16 (lane 2), C24 (lane 3) and C36 (lane 4) alkane; control experiment of RNA from S13 grown in minimal medium with sodium acetate (lane 5); lane M = DNA mass ladder (XL 1 kb, Eppendorf).
Mentions: RT‐PCR experiments were performed on the total RNA extracted from A. radioresistens S13 grown in minimal medium supplemented with C14, C16 (medium‐chain‐length alkanes) and C24, C36 (long‐chain‐length alkanes) as the sole energy and carbon source and from bacteria grown in the presence of sodium acetate. Using the two sets of alma‐ and alkB‐specific primers an amplified product of the expected size (499 and 466 bp respectively) was obtained only with RNA extracted from S13 grown with medium‐chain alkanes in the case of ALK primers (Fig. 6), and on RNA extracted from bacteria grown on long‐chain alkanes when BVMO primers were used. No amplified product was obtained from the RNA of bacteria grown with sodium acetate or from the RT‐negative controls. An amplified fragment of the expected size was also obtained using the A. radioresistens S13 16S rDNA‐specific primers on the RNA of the bacterium grown in the presence of medium‐ and long‐chain‐length alkanes and sodium acetate.

Bottom Line: Phylogenetic analysis placed the sequence of this novel BVMO in the same clade of the prodrug activator ethionamide monooxygenase (EtaA) and it bears only a distant relation to the other known class I BVMO proteins.In silico analysis of the 3D model of the S13 BVMO generated by homology modelling also supports the similarities with EtaA by binding ethionamide to the active site.In vitro experiments carried out with the purified enzyme confirm that this novel BVMO is indeed capable of typical Baeyer-Villiger reactions as well as oxidation of the prodrug ethionamide.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences and Systems Biology, University of Torino, via Accademia Albertina 13, 10123 Torino, Italy.

Show MeSH
Related in: MedlinePlus