Intracellular accumulation of hydrophobic 1,3-PDN inhibits methanogenesis in rumen methanogens.

Authors

Bingzi Ouyang, Yurong Cao, Qiushuang Li, Shizhe Zhang, Xiang Zhou, Zhiwu Wu, Xinyi Li, Cancan Wang, Rong Wang, Bo Fu, Xiaojia Liu, Lixiang Yin, Zhiliang Tan, Kui Zhu, Le Luo Guan, Xiumin Zhang, Jie Li, Min Wang

Published in

The ISME journal. Jul 28, 2026. Epub Jul 28, 2026.

Abstract

Methane (CH4) from ruminants is a major source of agricultural greenhouse gas and represents a loss of dietary energy. 3-nitrooxypropanol (3-NOP) is a known methanogenesis inhibitor, but its hydrophilic nature may limit the cellular accessibility to methanogens. Here, we systematically evaluated 1,3-propanediol dinitrate (1,3-PDN), a more hydrophobic derivative of 3-NOP, for its anti-methanogenic potential and underlying mode of action using ruminal fermentation, pure-culture assays, multi-omics analyses, and molecular docking. Intracellular accumulation assays indicated greater cellular accumulation of 1,3-PDN than 3-NOP in rumen-derived methanogen Methanobrevibacter olleyae. Ruminal fermentation assays showed that 1,3-PDN reduced CH4 production by ~55%, and altered hydrogen (H2) metabolism, leading to 11-fold increase in H2 accumulation. Metatranscriptomic profiling revealed that 1,3-PDN significantly altered the active archaeal community, with a notable reduction in Methanobrevibacter_A and suppression of hydrogenotrophic methanogenesis. Molecular docking suggested that 1,3-PDN may bind to the conserved active site of methyl-coenzyme M reductase (MCR), potentially contributing to MCR-associated inhibition. Proteomic analysis further indicated that 1,3-PDN supplementation downregulated key MCR subunits and simultaneously affected other redox-sensitive methanogenesis-related processes, including tetrahydromethanopterin S-methyltransferase (MTR) subunits and proteins involved in the biosynthesis of cofactors F430 and cobalamin. Nitrogen-equivalent assay suggested partial contribution of nitrite to the methanogenesis inhibition and oxidative effects induced by 1,3-PDN. Together, these findings identify 1,3-PDN as an effective inhibitor of ruminal methanogenesis with enhanced cellular enrichment, providing mechanistic insights for the rational development of new CH4 inhibitors.

PMID:
42520218
Bibliographic data and abstract were imported from PubMed on 29 Jul 2026.

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