Methanobactin from Methylosinus trichosporium OB3b inhibits N2O reduction in denitrifiers

Methanotrophs synthesize methanobactin, a secondary metabolite that binds copper with an unprecedentedly high affinity. Such a strategy may provide methanotrophs a “copper monopoly” that can inhibit the activity of copper-containing enzymes of other microbes, e.g., copper-dependent N 2 O reductases. Here, we show that methanobactin from Methylosinus trichosporium OB3b inhibited N 2 O reduction in denitrifiers. When Pseudomonas stutzeri DCP-Ps1 was incubated in cocultures with M. trichosporium OB3b or with purified methanobactin from M. trichosporium OB3b, stoichiometric N 2 O production was observed from NO 3 − reduction, whereas no significant N 2 O accumulation was observed in cocultures with a mutant defective in methanobactin production. Copper uptake by P. stutzeri DCP-Ps1 was inhibited by the presence of purified methanobactin, leading to a significant downregulation of nosZ transcription. Similar findings were observed with three other denitrifier strains. These results suggest that in situ stimulation of methanotrophs can inadvertently increase N 2 O emissions, with the potential for increasing net greenhouse gas emissions.