Molecular analysis of the soluble butane monooxygenase from ‘Pseudomonas butanovora’ a aThe GenBank accession number for the bmoXYBZDC sequence is AY093933.

Sluis, Miriam K.; Sayavedra-Soto, Luis A.; Arp, Daniel J.

doi:10.1099/00221287-148-11-3617

Cited by 87 publications

(79 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…type coxL form II gene sequences and Roseobacter group type coxL form II gene sequences ( Figure 5) Figure 4 Phylogenetic analysis of the derived amino-acid sequences of mmoX genes that encode the active-site subunit of soluble methane monooxygenase, from the Blyth estuary surface microlayer and subsurface water. The dendrogram was derived as described in Materials and methods and rooted with BmoX (soluble butane monooxygenase) from Pseudomonas butanovora (Sluis et al, 2002). The scale bar represents 10% sequence divergence.…”

Section: Resultsmentioning

confidence: 99%

Phylogenetic and functional gene analysis of the bacterial and archaeal communities associated with the surface microlayer of an estuary

et al. 2008

View full text Add to dashboard Cite

The surface microlayer (SML) is the thin biogenic film found at the surface of a water body. The SML is poorly understood but has been shown to be important in biogeochemical cycling and sea-air gas exchange. We sampled the SML of the Blyth estuary at two sites (salinities 21 and 31 psu) using 47 mm polycarbonate membranes. DNA was extracted from the SML and corresponding subsurface water (0.4 m depth) and microbial (bacteria and archaea) community analysis was performed using denaturing gradient gel electrophoresis of 16S rRNA gene PCR amplicons. The diversity of bacterial functional genes that encode enzyme subunits for methane monooxygenase (pmoA and mmoX) and carbon monoxide dehydrogenase (coxL) was assessed using PCR, clone library construction and restriction fragment length polymorphism (RFLP) analysis. Methanotroph genes were present only in low copy numbers and pmoA was detected only in subsurface samples. Diversity of mmoX genes was low and most of the clone sequences detected were similar to those of mmoX from Methylomonas spp. Interestingly, some sequences detected in the SML were different from those detected in the subsurface. RFLP analysis of coxL clone libraries indicated a high diversity of carbon monoxide (CO)-utilizing bacteria in the estuary. The habitats of the closely related coxL sequences suggest that CO-utilizing bacteria in the estuary are recruited from both marine and freshwater/terrestrial inputs. In contrast, methanotroph recruitment appears to occur solely from freshwater input into the estuary.

show abstract

Section: Resultsmentioning

confidence: 99%

Phylogenetic and functional gene analysis of the bacterial and archaeal communities associated with the surface microlayer of an estuary

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Sequence analysis shows BMOH to be most closely related to sMMO (65, 42 and 38 % amino acid identity in the a-, b-and c-subunits of BMOH, respectively) (Sluis et al, 2002). Additionally, sequence alignment and structural modelling show that all residues directly lining the active site cavity of BMOH are the same as those in the methane monooxygenase hydroxylase (MMOH).…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…To initiate growth on C 2 -C 9 alkanes, the Gramnegative b-proteobacterium Thauera butanivorans, formerly called 'Pseudomonas butanovora' (Dubbels et al, 2009), expresses a carboxylate-bridged non-haem di-iron monooxygenase, commonly referred to as soluble butane monooxygenase (sBMO) (Sluis et al, 2002;Takahashi et al, 1980). sBMO belongs to a family of bacterial multicomponent monooxygenases which includes soluble methane monooxygenases (sMMOs), phenol hydroxylases (PHs) and aromatic/alkene monooxygenases (TMOs) (Leahy et al, 2003;Sluis et al, 2002). Due to their unusually large substrate ranges, these powerful oxidizers are of particular interest for their potential in bioremediation (Enzien et al, 1994;Halsey et al, 2007;Parales et al, 2002;Smith & Dalton, 2004) and their ability to serve as industrial biocatalysts (Burton, 2003;Parales et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

et al. 2009

Self Cite

View full text Add to dashboard Cite

Soluble butane monooxygenase (sBMO), a three-component di-iron monooxygenase complex expressed by the C 2 -C 9 alkane-utilizing bacterium Thauera butanivorans, was kinetically characterized by measuring substrate specificities for C 1 -C 5 alkanes and product inhibition profiles. sBMO has high sequence homology with soluble methane monooxygenase (sMMO) and shares a similar substrate range, including gaseous and liquid alkanes, aromatics, alkenes and halogenated xenobiotics. Results indicated that butane was the preferred substrate (defined by k cat : K m ratios). Relative rates of oxidation for C 1 -C 5 alkanes differed minimally, implying that substrate specificity is heavily influenced by differences in substrate K m values. The low micromolar K m for linear C 2 -C 5 alkanes and the millimolar K m for methane demonstrate that sBMO is two to three orders of magnitude more specific for physiologically relevant substrates of T. butanivorans. Methanol, the product of methane oxidation and also a substrate itself, was found to have similar K m and k cat values to those of methane. This inability to kinetically discriminate between the C 1 alkane and C 1 alcohol is observed as a steady-state concentration of methanol during the two-step oxidation of methane to formaldehyde by sBMO. Unlike methanol, alcohols with chain length C 2 -C 5 do not compete effectively with their respective alkane substrates. Results from product inhibition experiments suggest that the geometry of the active site is optimized for linear molecules four to five carbons in length and is influenced by the regulatory protein component B (butane monooxygenase regulatory component; BMOB). The data suggest that alkane oxidation by sBMO is highly specialized for the turnover of C 3 -C 5 alkanes and the release of their respective alcohol products. Additionally, sBMO is particularly efficient at preventing methane oxidation during growth on linear alkanes ¢C 2, despite its high sequence homology with sMMO. These results represent, to the best of our knowledge, the first kinetic in vitro characterization of the closest known homologue of sMMO.

show abstract

“…The two main clades separating the Type I and Type II methanotrophs are indicated. The related butane monooxygenase (bmo) from Pseudomonas butanovora (Sluis et al, 2002), belonging to the c-subdivision of the Proteobacteria, was used as an outgroup. The accession numbers of the reference mmoX sequences are shown in parentheses.…”

Section: Phenotypic Characterization Of Dmmox1 and Dmmox2 Mutationsmentioning

confidence: 99%

Duplication of the mmoX gene in Methylosinus sporium: cloning, sequencing and mutational analysis

et al. 2006

View full text Add to dashboard Cite

The soluble methane monooxygenase (sMMO) is a key enzyme for methane oxidation, and is found in only some methanotrophs, including Methylosinus sporium 5. sMMO expression is regulated at the level of transcription from a σ 54 promoter by a copper-switch, and is only expressed when the copper-to-biomass ratio during growth is low. Extensive phylogenetic and genetic analyses of sMMOs and other soluble di-iron monooxygenases reveal that these enzymes have only been acquired relatively recently through horizontal gene transfer. In this study, further evidence of horizontal gene transfer was obtained, through cloning and sequencing of the genes encoding the sMMO enzyme complex plus the regulatory genes mmoG and mmoR, and identification of a duplicate copy of the mmoX gene in Ms. sporium. mmoX encodes the α subunit of the hydroxylase of the sMMO enzyme, which constitutes the active site (Prior & Dalton, 1985). The mmoX genes were characterized at the molecular and biochemical levels. Although both copies were transcribed, only mmoX copy 1 was essential for sMMO activity. Construction of an sMMO− mutant by marker-exchange mutagenesis gave some possible insights into the role of the water-soluble pigment in siderophore-mediated iron acquisition. Finally, the amenability of Ms. sporium to genetic manipulation was demonstrated by complementing the sMMO− mutant by heterologous expression of sMMO genes from Methylosinus trichosporium OB3b and Methylococcus capsulatus (Bath), and it was shown that Ms. sporium could be used as an alternative model organism for molecular analysis of MMO regulation.

show abstract

Molecular analysis of the soluble butane monooxygenase from ‘Pseudomonas butanovora’ a aThe GenBank accession number for the bmoXYBZDC sequence is AY093933.

Cited by 87 publications

References 79 publications

Phylogenetic and functional gene analysis of the bacterial and archaeal communities associated with the surface microlayer of an estuary

Phylogenetic and functional gene analysis of the bacterial and archaeal communities associated with the surface microlayer of an estuary

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

Duplication of the mmoX gene in Methylosinus sporium: cloning, sequencing and mutational analysis

Contact Info

Product

Resources

About