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Isolation and initial characterization of a novel type of Baeyer-Villiger monooxygenase activity from a marine microorganism.

Willetts A, Joint I, Gilbert JA, Trimble W, Mühling M - Microb Biotechnol (2012)

Bottom Line: The enzyme is able to use NADH as the source of reducing power necessary to accept the atom of diatomic oxygen not incorporated into the oxyfunctionalized substrate.A particular feature of this BVMO lies in its truncated C-terminal domain, which results in a relatively small protein (357 amino acids; 38.4 kDa).Although low, abundance was detected in samples from many marine provinces, confirming the potential for biodiscovery in marine microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth, UK. andrewj.willetts@btconnect.com

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Related in: MedlinePlus

Phylogenetic analysis of the BVMO amino acid sequence of Stenotrophomonas isolate PML 168 and 67 similar sequences identified by BLASTp search. Evolutionary history was inferred using the neighbour‐joining method (Saitou and Nei, 1987). The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Poisson correction method (Zuckerkandl and Pauling, 1965) and are in the units of the number of amino acid substitutions per site. The analysis involved those 66 amino acid sequences which showed greatest similarity to the BVMO sequence of isolate 168 as judged by a BLASTp search within the NCBI database. All positions containing gaps and missing data were eliminated. There were a total of 265 positions in the final data set. Evolutionary analyses were conducted in MEGA5 (Tamura et al., 2007). The cluster containing the Stenotrophomonas sequences is indicated by a bar.
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f2: Phylogenetic analysis of the BVMO amino acid sequence of Stenotrophomonas isolate PML 168 and 67 similar sequences identified by BLASTp search. Evolutionary history was inferred using the neighbour‐joining method (Saitou and Nei, 1987). The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Poisson correction method (Zuckerkandl and Pauling, 1965) and are in the units of the number of amino acid substitutions per site. The analysis involved those 66 amino acid sequences which showed greatest similarity to the BVMO sequence of isolate 168 as judged by a BLASTp search within the NCBI database. All positions containing gaps and missing data were eliminated. There were a total of 265 positions in the final data set. Evolutionary analyses were conducted in MEGA5 (Tamura et al., 2007). The cluster containing the Stenotrophomonas sequences is indicated by a bar.

Mentions: A BLASTp search within the NCBI database revealed a large number of similar sequences of which the 67 most similar sequences were selected for a subsequent phylogenetic analysis (Fig. 2). This analysis showed that a gene from S. maltophila strain K279a, annotated as a putative monooxygenase (Crossman et al., 2008), was closely related to that of isolate PML 168 and that these two ORFs together with the only other available Class B flavoprotein from a strain of Stenotrophomonas (S. maltophila R551‐3) formed a separate cluster (Fig. 2). The reliability of the branching order in the phylogenetic tree was evaluated by analysing the consensus between the branching order of this neighbour‐joining (NJ) based tree and that determined using two further algorithms for phylogenetic clustering similar to the approach presented by Mühling and colleagues (2008) – the minimum evolution (ME) and the maximum likelihood (ML) methods. The similarity of the three trees was assessed by calculating the branch score distances, using the programme treedist, part of the Phylip software package (http://evolution.gs.washington.edu/phylip.html; Felsenstein, 2004). This analysis revealed that the branching order within the ME‐ and NJ‐based trees was identical (score = 0), while that of the ML‐based tree was very similar to the ME and the NJ tree (score = 0.41), thus supporting the robustness of the phylogenetic clusters. Importantly, the cluster containing the Class B flavoprotein sequences of the three Stenotrophomonas strains was identical in all three trees.


Isolation and initial characterization of a novel type of Baeyer-Villiger monooxygenase activity from a marine microorganism.

Willetts A, Joint I, Gilbert JA, Trimble W, Mühling M - Microb Biotechnol (2012)

Phylogenetic analysis of the BVMO amino acid sequence of Stenotrophomonas isolate PML 168 and 67 similar sequences identified by BLASTp search. Evolutionary history was inferred using the neighbour‐joining method (Saitou and Nei, 1987). The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Poisson correction method (Zuckerkandl and Pauling, 1965) and are in the units of the number of amino acid substitutions per site. The analysis involved those 66 amino acid sequences which showed greatest similarity to the BVMO sequence of isolate 168 as judged by a BLASTp search within the NCBI database. All positions containing gaps and missing data were eliminated. There were a total of 265 positions in the final data set. Evolutionary analyses were conducted in MEGA5 (Tamura et al., 2007). The cluster containing the Stenotrophomonas sequences is indicated by a bar.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815331&req=5

f2: Phylogenetic analysis of the BVMO amino acid sequence of Stenotrophomonas isolate PML 168 and 67 similar sequences identified by BLASTp search. Evolutionary history was inferred using the neighbour‐joining method (Saitou and Nei, 1987). The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Poisson correction method (Zuckerkandl and Pauling, 1965) and are in the units of the number of amino acid substitutions per site. The analysis involved those 66 amino acid sequences which showed greatest similarity to the BVMO sequence of isolate 168 as judged by a BLASTp search within the NCBI database. All positions containing gaps and missing data were eliminated. There were a total of 265 positions in the final data set. Evolutionary analyses were conducted in MEGA5 (Tamura et al., 2007). The cluster containing the Stenotrophomonas sequences is indicated by a bar.
Mentions: A BLASTp search within the NCBI database revealed a large number of similar sequences of which the 67 most similar sequences were selected for a subsequent phylogenetic analysis (Fig. 2). This analysis showed that a gene from S. maltophila strain K279a, annotated as a putative monooxygenase (Crossman et al., 2008), was closely related to that of isolate PML 168 and that these two ORFs together with the only other available Class B flavoprotein from a strain of Stenotrophomonas (S. maltophila R551‐3) formed a separate cluster (Fig. 2). The reliability of the branching order in the phylogenetic tree was evaluated by analysing the consensus between the branching order of this neighbour‐joining (NJ) based tree and that determined using two further algorithms for phylogenetic clustering similar to the approach presented by Mühling and colleagues (2008) – the minimum evolution (ME) and the maximum likelihood (ML) methods. The similarity of the three trees was assessed by calculating the branch score distances, using the programme treedist, part of the Phylip software package (http://evolution.gs.washington.edu/phylip.html; Felsenstein, 2004). This analysis revealed that the branching order within the ME‐ and NJ‐based trees was identical (score = 0), while that of the ML‐based tree was very similar to the ME and the NJ tree (score = 0.41), thus supporting the robustness of the phylogenetic clusters. Importantly, the cluster containing the Class B flavoprotein sequences of the three Stenotrophomonas strains was identical in all three trees.

Bottom Line: The enzyme is able to use NADH as the source of reducing power necessary to accept the atom of diatomic oxygen not incorporated into the oxyfunctionalized substrate.A particular feature of this BVMO lies in its truncated C-terminal domain, which results in a relatively small protein (357 amino acids; 38.4 kDa).Although low, abundance was detected in samples from many marine provinces, confirming the potential for biodiscovery in marine microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth, UK. andrewj.willetts@btconnect.com

Show MeSH
Related in: MedlinePlus