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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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Multiple sequence alignment of different flavoprotein monooxygenases including several Baeyer‐Villiger monoxygenases (BVMO)s proteins. FAD‐ and NADPH‐binding domains are highlighted in yellow (Rosmann fold), the fingerprint consensus sequence is highlighted in green. Identical residues between two sequences are indicated by an asterisk (*); similar residues are indicated by two dots (high similarity) or one dot (low similarity). PAMO = phenylacetone monooxygenase from Thermobifida fusca, 1W4X; STMO = steroid monooxygenase from Rhodococcus rhodochrous, BAA24454; CPMO = cyclopentanone monooxygenase from Comamonas testosteronii NCIMB 9872, CAD10798; CHMO = cyclohexanone monooxygenase from Acinetobacter calcoaceticus NCIMB 9871, AAG10021; EtaA = ethionamide‐activating monooxygenase from Mycobacterium tuberculosis H37Rv, CAB06212; HAPMO = 4‐hydroxyacetophenone monooxygenase from Pseudomonas fluorescens ACB, AF355751; BVMO = Baeyer‐Villiger monooxygenase from Acinetobacter radioresistens strain S13, HQ685899 (this work).
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f3: Multiple sequence alignment of different flavoprotein monooxygenases including several Baeyer‐Villiger monoxygenases (BVMO)s proteins. FAD‐ and NADPH‐binding domains are highlighted in yellow (Rosmann fold), the fingerprint consensus sequence is highlighted in green. Identical residues between two sequences are indicated by an asterisk (*); similar residues are indicated by two dots (high similarity) or one dot (low similarity). PAMO = phenylacetone monooxygenase from Thermobifida fusca, 1W4X; STMO = steroid monooxygenase from Rhodococcus rhodochrous, BAA24454; CPMO = cyclopentanone monooxygenase from Comamonas testosteronii NCIMB 9872, CAD10798; CHMO = cyclohexanone monooxygenase from Acinetobacter calcoaceticus NCIMB 9871, AAG10021; EtaA = ethionamide‐activating monooxygenase from Mycobacterium tuberculosis H37Rv, CAB06212; HAPMO = 4‐hydroxyacetophenone monooxygenase from Pseudomonas fluorescens ACB, AF355751; BVMO = Baeyer‐Villiger monooxygenase from Acinetobacter radioresistens strain S13, HQ685899 (this work).

Mentions: The sequence homology search using the amino acid sequence coded by ORF1491 revealed significant homology with several multifunctional flavin‐containing monooxygenases, the N‐hydroxylating monooxygenases and BVMO. The presence of two Rossmann folds, as evidenced by two GXGXX(G/A) motifs (Fig. > 3), which is a common sequence among FAD and NAD(P)H‐dependent oxidoreductases, discriminates the enzyme from the mechanistically related flavoprotein hydroxylases (Eppink et al., 1997). Furthermore, the presence of an identified sequence fingerprint (FXGXXXHXXXW(P/D) (Fig. 3) suggests that the protein is a flavin‐containing Baeyer‐Villiger monooxygenase (Fraaije et al., 2002). Indeed a search in the protein sequence database revealed that the closest homologue with known activity is ethionamide‐activating monooxygenase from Mycobacterium tuberculosis, sharing a 49.7% sequence identity and 71.0% sequence similarity.


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)

Multiple sequence alignment of different flavoprotein monooxygenases including several Baeyer‐Villiger monoxygenases (BVMO)s proteins. FAD‐ and NADPH‐binding domains are highlighted in yellow (Rosmann fold), the fingerprint consensus sequence is highlighted in green. Identical residues between two sequences are indicated by an asterisk (*); similar residues are indicated by two dots (high similarity) or one dot (low similarity). PAMO = phenylacetone monooxygenase from Thermobifida fusca, 1W4X; STMO = steroid monooxygenase from Rhodococcus rhodochrous, BAA24454; CPMO = cyclopentanone monooxygenase from Comamonas testosteronii NCIMB 9872, CAD10798; CHMO = cyclohexanone monooxygenase from Acinetobacter calcoaceticus NCIMB 9871, AAG10021; EtaA = ethionamide‐activating monooxygenase from Mycobacterium tuberculosis H37Rv, CAB06212; HAPMO = 4‐hydroxyacetophenone monooxygenase from Pseudomonas fluorescens ACB, AF355751; BVMO = Baeyer‐Villiger monooxygenase from Acinetobacter radioresistens strain S13, HQ685899 (this work).
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Related In: Results  -  Collection

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f3: Multiple sequence alignment of different flavoprotein monooxygenases including several Baeyer‐Villiger monoxygenases (BVMO)s proteins. FAD‐ and NADPH‐binding domains are highlighted in yellow (Rosmann fold), the fingerprint consensus sequence is highlighted in green. Identical residues between two sequences are indicated by an asterisk (*); similar residues are indicated by two dots (high similarity) or one dot (low similarity). PAMO = phenylacetone monooxygenase from Thermobifida fusca, 1W4X; STMO = steroid monooxygenase from Rhodococcus rhodochrous, BAA24454; CPMO = cyclopentanone monooxygenase from Comamonas testosteronii NCIMB 9872, CAD10798; CHMO = cyclohexanone monooxygenase from Acinetobacter calcoaceticus NCIMB 9871, AAG10021; EtaA = ethionamide‐activating monooxygenase from Mycobacterium tuberculosis H37Rv, CAB06212; HAPMO = 4‐hydroxyacetophenone monooxygenase from Pseudomonas fluorescens ACB, AF355751; BVMO = Baeyer‐Villiger monooxygenase from Acinetobacter radioresistens strain S13, HQ685899 (this work).
Mentions: The sequence homology search using the amino acid sequence coded by ORF1491 revealed significant homology with several multifunctional flavin‐containing monooxygenases, the N‐hydroxylating monooxygenases and BVMO. The presence of two Rossmann folds, as evidenced by two GXGXX(G/A) motifs (Fig. > 3), which is a common sequence among FAD and NAD(P)H‐dependent oxidoreductases, discriminates the enzyme from the mechanistically related flavoprotein hydroxylases (Eppink et al., 1997). Furthermore, the presence of an identified sequence fingerprint (FXGXXXHXXXW(P/D) (Fig. 3) suggests that the protein is a flavin‐containing Baeyer‐Villiger monooxygenase (Fraaije et al., 2002). Indeed a search in the protein sequence database revealed that the closest homologue with known activity is ethionamide‐activating monooxygenase from Mycobacterium tuberculosis, sharing a 49.7% sequence identity and 71.0% sequence similarity.

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