Enhancement of nitrous oxide emissions in soil microbial consortia via copper competition between proteobacterial methanotrophs and denitrifiers

Abstract

Unique means of copper scavenging have been identified in proteobacterial methanotrophs, particularly the use of methanobactin, a novel ribosomally synthesized post-translationally modified polypeptide that binds copper with very high affinity. The possibility that copper sequestration strategies of methanotrophs may interfere with copper uptake of denitrifiers in situ and thereby enhance N2O emissions was examined using a suite of laboratory experiments performed with rice paddy microbial consortia. Addition of purified methanobactin from Methylosinus trichosporium OB3b to denitrifying rice paddy soil microbial consortia resulted in substantially increased N2O production, with more pronounced responses observed for soils with lower copper content. The N2O emission-enhancing effect of the soil's native mbnA-expressing Methylocystaceae methanotrophs on the native denitrifiers was then experimentally verified with a Methylocystaceae-dominant chemostat culture prepared from a rice paddy microbial consortium as the inoculum. Lastly, with microcosms amended with varying cell numbers of methanobactin-producing Methylosinus trichosporium OB3b before CH4 enrichment, microbiomes with different ratios of methanobactin-producing Methylocystaceae to gammaproteobacterial methanotrophs incapable of methanobactin production were simulated. Significant enhancement of N2O production from denitrification was evident in both Methylocystaceae-dominant and Methylococcaceae-dominant enrichments, albeit to a greater extent in the former, signifying the comparative potency of methanobactin-mediated copper sequestration while implying the presence of alternative copper abstraction mechanisms for Methylococcaceae. These observations support that copper-mediated methanotrophic enhancement of N2O production from denitrification is plausible where methanotrophs and denitrifiers cohabit.

Agritech to Tame the Nitrogen Cycle

“…For instance, the microbial processes responsible for N 2 O and CH 4 emissions are both sensitive to redox and nutrient balance and are also heavily influenced by the availability of trace metals like copper and iron (Stein 2020). As an example, methanotrophic bacteria produce the copper-chelating chalkophore, metha-nobactin, that prevents access to copper for the expression of nitrous oxide reductase enzymes (NosZ) by denitrifiers (Chang et al 2021). Thus, when copper is at low availability, CH 4 oxidation will proceed at the expense of N 2 O reduction, leading to higher N 2 O emissions with lower CH 4 emissions.…”

Ending the era of Haber–Bosch

“…Denitrification often terminates at N 2 O rather than continuing to dinitrogen due to conditions and processes that slow down or inactivate nitrous oxide reductase (NosZ) enzymes (Zumft & Kroneck, 2007). Among the more interesting mechanisms that inhibit NosZ expression and activity is the competition for copper by microorganisms like methanotrophs that produce the copper-chelating chalkophore, methanobactin (Chang et al, 2021). The conditions, populations, and pathways that produce N 2 O in soils are convoluted and complicated, although clearly, nitrogen excess from fertilizer overuse exacerbates N 2 O production.…”