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Welte, C.

Publications and source records attributed to Welte, C..

5 recordsLinked to original sources

Adaptation of the freshwater anaerobic methanotroph 'Ca. Methanoperedens vercellensis' to low pH levels reveals membrane lipid remodelling

Anaerobic methanotrophic archaea are key members of the biological methane filter, thereby preventing emissions of this strong greenhouse gas into the atmosphere. Previous studies on freshwater anaerobic methanotrophs targeted the activity of these microorganisms at circumneutral pH whereas molecular ecology studies identified this phylotype also in acidic environments such as peatlands; it is currently unknown whether they can adapt to low pH and remain effective in the biological methane filter in low pH environments. Here we show that a granular enrichment culture of the freshwater methanotroph Ca. M. vercellensis loses activity when experiencing pH stress but remains metabolically active down to pH 5.65 with appropriate adaptation time, indicating that adaptive changes are necessary to accommodate anaerobic methane oxidation at lower pH. Analyses of archaeal lipids revealed an increase in zwitterionic intact polar lipids over anionic lipids as an adaptation. This coincided with a change in granule structure while methane oxidation rate and enrichment state of Ca. M. vercellensis remained stable. We show that Ca. M. vercellensis remains metabolically active at lower pH values, despite increased maintenance energy demands and the need for cytoplasmic pH homeostasis. Our study demonstrates that adaptations to stress by slow-growing microorganisms may require long-term observation and is thereby instrumental for a better understanding of methane cycling in acidic ecosystems.

microbiology↗

Atomic resolution structures of key enzyme MCR in anaerobic methanotrophy reveal novel and extensive post-translational modifications.

Anaerobic methanotrophic archaea (ANME) are crucial to planetary carbon cycling. They oxidise methane in anoxic niches by transferring electrons directly to nitrate or metal oxides and alternatively to sulfate-reducing bacteria. Due to their physiological complexity, no ANME species have been isolated, hampering the biochemical investigation of the enzymatic processes involved in anaerobic methane oxidation. To study the methane-capturing enzyme of these microorganisms, we circumvented the isolation barrier by exploiting microbial enrichments of freshwater nitrate-reducing ANME-2d grown in bioreactors, and marine ANME-2c in syntrophy with bacterial partners. The crystal structures of their Methyl-Coenzyme M Reductases (MCRs), refined to true atomic resolution, provided the most precise image of the enzyme to date. Despite their physiological differences, these ANMEs have extremely conserved MCR structures, similar to homologs from methanogenic Methanosarcinales, rather than the phylogenetically distant MCR of ANME-1 isolated from Black Sea mats. The three studied MCRs are highly modified, with seven post-translational modifications. Among them was a novel 3(S)-methylhistidine on the {gamma}-chain of both ANME-2d MCRs. Labelling with gaseous krypton did not reveal any internal channels that would facilitate alkane diffusion to the active site as observed in the ethane-specialized enzyme. Based on our data, the methanotrophic MCRs should follow the same radical reaction mechanism proposed for the methane-generating homologues. The described pattern of post-translational modifications underscores the importance of native purification as a powerful approach to discovering intrinsic enzymatic features in uncultivated microorganisms existing in nature.

microbiology↗

The clumped isotope signatures of multiple methanogenesis metabolisms

Methane is a potent greenhouse gas, an important energy source, and a potential biosignature on extraterrestrial planetary bodies. The relative abundances of doubly substituted ("clumped") methane isotopologues (13CH3D and 12CH2D2) offer important information on the sources and sinks of methane. However, the clumped isotope signatures of microbially produced methane from different methanogenic pathways lack a systematic investigation. In this study, we provide a dataset encompassing the relative isotopologue abundances produced by hydrogenotrophic, methylotrophic, acetoclastic, and methoxydotrophic methanogenesis. We find that a statistical "combinatorial effect" generates significant differences in 12CH2D2 compositions between hydrogenotrophic methanogenesis and other pathways. The thermodynamic drive of methanogenic reactions and phylogenetic affiliation may also influence the isotope compositions of methane. Our study provides new experimental constraints on the isotope signatures of different microbial methanogenic pathways, and evidence of the mechanisms responsible for the observed differences. TeaserA novel stable isotope tool to track and differentiate sources of biological methane.

microbiology↗

Physiological stress response to sulfide exposure of freshwater anaerobic methanotrophic archaea

Freshwater wetlands and coastal sediments are becoming hotspots for the emission of the greenhouse gas methane. Eutrophication-induced deposition of organic matter leads to elevated methanogenesis and sulfate reduction, thereby increasing the concentrations of methane and toxic sulfide, respectively. However, the effects of sulfide stress on the anaerobic methanotrophic biofilter have not been well explored. Here, we show how an enrichment culture dominated by the freshwater anaerobic methane-oxidizing archaeon Candidatus (Ca.) Methanoperedens responds to short-term and long-term exposure to sulfide in a bioreactor. The methane-oxidizing activity decreased to 45% and 20% but partially recovered to 70% and 30% within 5 days after short- and long-term sulfide exposure, respectively. Metagenomics indicated that Ca. Methanoperedens remained dominant in the enrichment throughout the entire experiment. The first short-term sulfide pulse led to increased expression of genes encoding for sulfide detoxification by low abundant community members, whereas long-term exposure resulted in upregulation of Ca. Methanoperedens genes encoding sulfite reductases of Group III (Dsr-LP). Ca. Methanoperedens consumed Polyhydroxyalkanoates during long-term sulfide exposure, possibly to aid in stress adaptation. Together, these results provide a valuable baseline for understanding fundamental ecophysiological adaptations in sulfate- and nitrate-rich aquatic ecosystems. Short synopsis statementThis study investigated how freshwater anaerobic methanotrophic archaea responded to sulfide exposure, revealing a transient inhibition and physiological adaptation mechanisms.

microbiology↗

Mechanisms of extracellular electron transfer in anaerobic methanotrophic archaea

Anaerobic methanotrophic (ANME) archaea are environmentally important uncultivated microorganisms mitigating the release of the potent greenhouse gas methane. During methane oxidation ANME archaea engage in extracellular electron transfer (EET) with other microorganisms, metal oxides, and electrodes, through a currently unknown mechanism. To shed light on this mechanism, we cultivated ANME-2d archaea (Ca. Methanoperedens) in bioelectrochemical systems and observed strong methane-dependent current (91-93% of total current) associated with high enrichment of Ca. Methanoperedens on the anode (up to 82% of the community) determined by metagenomics and transmission electron microscopy. Electrochemistry and metatranscriptomics indicated that the EET mechanism was similar at various electrode potentials pointing to the involvement of an so far uncharacterized short-range electron transport protein complex and OmcZ nanowires, suggesting a unique EET pathway in all ANME-2 archaea. Our findings furthermore indicate that bioelectrochemical cells might be powerful tools for the cultivation, and possibly isolation, of uncultured electroactive microorganisms.

microbiology↗