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Impact of copper on the physiology and transcriptome of Methylosinus trichosporium OB3b grown on either methane or methanol

ABSTRACT: This study investigates the combined effects of copper availability and carbon source (methane versus methanol) on the physiology and transcriptome of Methylosinus trichosporium OB3b, a model Type II methanotroph. Methanotrophs oxidize methane to carbon dioxide through intermediates such as methanol, formaldehyde, and formate, using two forms of methane monooxygenase (MMO): the copper-dependent particulate MMO (pMMO) and the soluble MMO (sMMO). Although Msn. trichosporium OB3b can utilize both methane and methanol as sole carbon sources, growth on methanol is significantly affected by copper. In the presence of copper, methanol-grown cells exhibited impaired growth, formate accumulation, and decreased pH, while buffering with MOPS restored growth. NADH/NAD ratios increased nearly tenfold in methanol-grown cultures with copper, indicating redox imbalance. Transmission electron microscopy revealed disrupted intracytoplasmic membranes under these conditions. Transcriptomic analysis showed that genes involved in methanol oxidation were differentially expressed in response to copper and carbon source, and two siderophore biosynthetic gene clusters were repressed during methanol growth with copper. Overall, the findings suggest that Msn. trichosporium OB3b modulates metallophore expression and carbon metabolism to manage oxidative and redox stress under methanol growth conditions. Understanding these regulatory mechanisms is critical for optimizing methanotroph-based biotechnological applications, including methanol-driven biosynthesis of valuable compounds such as methanobactin.

ORGANISM(S): Methylosinus trichosporium OB3b

PROVIDER: GSE310588 | GEO | 2026/01/03

REPOSITORIES: GEO

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Project description:Metabolic flexibility in aerobic methane oxidising bacteria (methanotrophs) enhances cell growth and survival in instances where resources are variable or limiting. Examples include the production of intracellular compounds (such as glycogen or polyhydroxyalkanoates) in response to unbalanced growth conditions and the use of some energy substrates, besides methane, when available. Indeed, recent studies show that verrucomicrobial methanotrophs can grow mixotrophically through oxidation of hydrogen and methane gases via respiratory membrane-bound group 1d [NiFe] hydrogenases and methane monooxygenases respectively. Hydrogen metabolism is particularly important for adaptation to methane and oxygen limitation, suggesting this metabolic flexibility may confer growth and survival advantages. In this work, we provide evidence that, in adopting a mixotrophic growth strategy, the thermoacidophilic methanotroph, Methylacidiphilum sp. RTK17.1 changes its growth rate, biomass yields and the production of intracellular glycogen reservoirs. Under nitrogen-fixing conditions, removal of hydrogen from the feed-gas resulted in a 14 % reduction in observed growth rates and a 144% increase in cellular glycogen content. Concomitant with increases in glycogen content, the total protein content of biomass decreased following the removal of hydrogen. Transcriptome analysis of Methylacidiphilum sp. RTK17.1 revealed a 3.5-fold upregulation of the Group 1d [NiFe] hydrogenase in response to oxygen limitation and a 4-fold upregulation of nitrogenase encoding genes (nifHDKENX) in response to nitrogen limitation. Genes associated with glycogen synthesis and degradation were expressed constitutively and did not display evidence of transcriptional regulation. Collectively these data further challenge the belief that hydrogen metabolism in methanotrophic bacteria is primarily associated with energy conservation during nitrogen fixation and suggests its utilisation provides a competitive growth advantage within hypoxic habitats.

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Impact of copper on the physiology and transcriptome of Methylosinus trichosporium OB3b grown on either methane or methanol

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