Project description:Methanotrophs, which help regulate atmospheric levels of methane, are active in diverse natural and man-made environments. This range of habitats and the feast-famine cycles seen by many environmental methanotrophs suggest that methanotrophs dynamically mediate rates of methane oxidation. Global methane budgets require ways to account for this variability in time and space. Functional gene biomarker transcripts are increasingly being studied to inform the dynamics of diverse biogeochemical cycles. Previously, per-cell transcript levels of the methane oxidation biomarker, pmoA, were found to vary quantitatively with respect to methane oxidation rates in model aerobic methanotroph, Methylosinus trichosporium OB3b. In the present study, these trends were explored for two additional aerobic methanotroph pure cultures, Methylocystis parvus OBBP and Methylomicrobium album BG8. At steady-state conditions, per cell pmoA mRNA transcript levels strongly correlated with per cell methane oxidation across the three methanotrophs across many orders of magnitude of activity (R2 = 0.91). Additionally, genome-wide expression data (RNA-seq) were used to explore transcriptomic responses of steady state M. album BG8 cultures to short-term CH4 and O2 limitation. These limitations induced regulation of genes involved in central carbon metabolism (including carbon storage), cell motility, and stress response.
Project description:We report here a methanotroph, Methylotuvimicrobium buryatense 5GB1C, that consumes methane at 500ppm at rates several times higher than any previously published. Analyses of bioreactor-based performance and RNAseq based transcriptomics suggest that this superior ability to utilize low methane is based at least in part on an extremely low non-growth associated maintenance energy and on a 5-fold higher methane specific affinity than previous reports.
Project description:Natural and anthropogenic wetlands are main sources of the atmospheric greenhouse gas methane. Methane emissions from wetlands are mitigated by methanotrophic microorganisms and by processes at the oxic-anoxic interface, such as sulfur cycling, that reduce the activity of methanogens. In this study, we obtained a pure culture (strain HY1) of a versatile wetland methanotroph that oxidizes various organic and inorganic compounds. This strain represents (i) the first isolate that can aerobically oxidize both methane and reduced sulfur compounds and (ii) a new alphapoteobacterial species, named Candidatus Methylovirgula thiovorans. Genomic and proteomic analyses showed that soluble methane monooxygenase and XoxF-type alcohol dehydrogenases are the only enzymes for methane and methanol oxidation, respectively. Unexpectedly, strain HY1 harbors various pathways for respiratory sulfur oxidation and oxidized reduced sulfur compounds to sulfate using the Sox-rDsr pathway (without SoxCD) and the S4I system. It employed the Calvin-Benson-Bassham cycle for CO2 fixation during chemolithoautotrophic growth on the reduced sulfur compounds. Methane and thiosulfate were independently and simultaneously oxidized by strain HY1 for growth. Proteomic and microrespiratory analyses showed that the metabolic pathways for methane and thiosulfate oxidation were induced in the presence of their substrates. The discovery of this versatile methanotroph demonstrates that methanotrophy and thiotrophy is compatible in a single bacterium and adds a new aspect to interactions of methane and sulfur cycles in oxic-anoxic interface environments.
Project description:Lanthanides (Ln) play essential roles in the metabolism of certain bacteria, catalysing key reactions in methane oxidation. This study investigates the diversity and distribution of Ln-dependent proteins, collectively termed the lanthanome, in aerobic methane-oxidizing bacteria (MOB) using genome, plasmid, and metatranscriptome data from methane-rich lake sediments. A custom database of 180 MOB genomes revealed various methanol dehydrogenase (MDH) isoforms, including XoxF variants, distributed across Proteobacteria and Verrucomicrobia phyla. We conducted an experimental study with Methylosinus trichosporium OB3B exposed to CeCl₃ and an ore containing mixed lanthanides, measuring methane oxidation rates and using proteomics to assess shifts in protein expression. Despite differences in adaptation times, methane oxidation rates were consistent across treatments, indicating similar overall metabolic efficiencies after acclimatisation. The genomic analysis uncovered several Ln-binding proteins, including the TonB-dependent receptors LanA and lutH-like, as well as Lanmodulin and LanPepsy, with unique phylogenetic patterns. Metatranscriptomic data showed active lanthanome expression, particularly in Proteobacteria, with the XoxF5 MDH variant prevalent in MOB genomes. The discovery of Ln-binding proteins in plasmids suggests potential horizontal gene transfer, highlighting adaptive mechanisms of MOB to Ln availability and their ecological role in methane cycling. This work expands our understanding of Ln-utilising bacteria, particularly in the context of lanthanide-driven methane oxidation, and offers potential biotechnological applications for Ln-dependent processes.