Cohn’s
            <i>Crenothrix</i>
            is a filamentous methane oxidizer with an unusual methane monooxygenase

Stoecker, Kilian; Bendinger, Bernd; Schöning, Björn; Nielsen, Per Halkjær; Nielsen, Jeppe Lund; Baranyi, Christian; Toenshoff, Elena R.; Daims, Holger; Wagner, Michael

doi:10.1073/pnas.0506361103

Cited by 232 publications

(183 citation statements)

References 27 publications

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“…The clone library RFLP analysis revealed one major OTU (88% of clone library) with closest sequence similarity to a known Methylobacter pmoA gene sequence (AAF08210) (Costello and Lidstrom, 1999). The two remaining low-abundance OTUs, both with a relative abundance of 6%, were similar to pmoA clones from uncultivated bacteria collected from a river plain aquifer (AAZ06139) (Erwin et al, 2005) and a sand water filter (ABC59829) (Stoecker et al, 2006).…”

Section: Resultsmentioning

confidence: 83%

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

et al. 2008

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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.

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Section: Resultsmentioning

confidence: 83%

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

et al. 2008

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“…The group of MOB comprises the three families Methylococcaceae, Methylocystaceae, and Beijerinckiaceae [5,[9][10][11][12]. The only exception is Crenothrix polyspora, a filamentous, sheathed microorganism recently discovered to be methanotrophic [52]. Members of the Methylococcaceae are termed type I MOB and belong to the γ-subdivision of the Proteobacteria phylum.…”

Section: Introductionmentioning

confidence: 99%

Diversity of Aerobic Methanotrophic Bacteria in a Permafrost Active Layer Soil of the Lena Delta, Siberia

et al. 2008

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With this study, we present first data on the diversity of aerobic methanotrophic bacteria (MOB) in an Arctic permafrost active layer soil of the Lena Delta, Siberia. Applying denaturing gradient gel electrophoresis and cloning of 16S ribosomal ribonucleic acid (rRNA) and pmoA gene fragments of active layer samples, we found a general restriction of the methanotrophic diversity to sequences closely related to the genera Methylobacter and Methylosarcina, both type I MOB. In contrast, we revealed a distinct species-level diversity. Based on phylogenetic analysis of the 16S rRNA gene, two new clusters of MOB specific for the permafrost active layer soil of this study were found. In total, 8 out of 13 operational taxonomic units detected belong to these clusters. Members of these clusters were closely related to Methylobacter psychrophilus and Methylobacter tundripaludum, both isolated from Arctic environments. A dominance of MOB closely related to M. psychrophilus and M. tundripaludum was confirmed by an additional pmoA gene analysis. We used diversity indices such as the Shannon diversity index or the Chao1 richness estimator in order to compare the MOB community near the surface and near the permafrost table. We determined a similar diversity of the MOB community in both depths and suggest that it is not influenced by the extreme physical and geochemical gradients in the active layer.

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“…Known CuMMOs are restricted to methanotrophs and autotrophic ammonia-oxidizers, nearly all of which grow solely on C1 compounds (Table 1) (Konneke et al, 2005;Stoecker et al, 2006;Dunfield et al, 2007;Baani and Liesack, 2008;MartensHabbena et al, 2009;Ettwig et al, 2010;Semrau et al, 2010). The resulting 'restricted role' paradigm has implications for applications involving CuMMOs.…”

Section: Introductionmentioning

confidence: 99%

Hydrocarbon monooxygenase in Mycobacterium: recombinant expression of a member of the ammonia monooxygenase superfamily

Coleman

et al. 2011

The ISME Journal

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The copper membrane monooxygenases (CuMMOs) are an important group of enzymes in environmental science and biotechnology. Areas of relevance include the development of green chemistry for sustainable exploitation of methane (CH 4 ) reserves, remediation of chlorinated hydrocarbon contamination and monitoring human impact in the biogeochemical cycles of CH 4 and nitrogen. Challenges for all these applications are that many aspects of the ecology, physiology and structure-function relationships in the CuMMOs are inadequately understood. Here, we describe genetic and physiological characterization of a novel member of the CuMMO family that has an unusual physiological substrate range (C 2 -C 4 alkanes) and a distinctive bacterial host (Mycobacterium). The Mycobacterial CuMMO genes (designated hmoCAB) were amenable to heterologous expression in M. smegmatis-this is the first example of recombinant expression of a complete and highly active CuMMO enzyme. The apparent specific activity of recombinant cells containing hmoCAB ranged from 2 to 3 nmol min -1 per mg protein on ethane, propane and butane as substrates, and the recombinants could also attack ethene, cis-dichloroethene and 1,2-dichloroethane. No detectable activity of recombinants or wild-type strains was seen with methane. The specific inhibitor allylthiourea strongly inhibited growth of wild-type cells on C 2 -C 4 alkanes, and omission of copper from the medium had a similar effect, confirming the physiological role of the CuMMO for growth on alkanes. The hydrocarbon monooxygenase provides a new model for studying this important enzyme family, and the recombinant expression system will enable biochemical and molecular biological experiments (for example, site-directed mutagenesis) that were previously not possible.

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Cohn’s Crenothrix is a filamentous methane oxidizer with an unusual methane monooxygenase

Cited by 232 publications

References 27 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

Diversity of Aerobic Methanotrophic Bacteria in a Permafrost Active Layer Soil of the Lena Delta, Siberia

Hydrocarbon monooxygenase in Mycobacterium: recombinant expression of a member of the ammonia monooxygenase superfamily

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