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Over half of the culture\u2019s bacterial community consists of two closely related\n                      <jats:italic toggle=\"yes\">Desulfobacterota<\/jats:italic>\n                      strains, designated ORM2a and ORM2b, whose mechanisms of benzene activation are unknown. Three proteomics data sets were integrated and analyzed using high-quality OR metagenomes and metagenome-assembled genomes (MAGs), including a complete circularized ORM2a MAG, to identify active metabolic pathways and proteins expressed during methanogenic benzene degradation. Among the proteins identified were Bam-like subunits of an ATP-independent benzoyl-CoA degradation pathway, as well as downstream \u03b2-oxidation proteins yielding acetate. The most abundant proteins identified mapped to two ORM2a gene clusters of unknown function. Homologous and syntenic gene clusters were identified in the MAGs of ORM2b and a sulfate-reducing\n                      <jats:italic toggle=\"yes\">Pelotomaculum<\/jats:italic>\n                      that also degrades benzene, as well as in nine contigs assembled from hydrothermal vent metagenomes. Extensive homology and structural predictions suggest that the first cluster\u2014termed the \u201cMagic\u201d gene cluster\u2014encodes for enzymes catalyzing the chemically challenging activation of benzene and subsequent transformation steps yielding benzoyl-CoA. The second (\u201cNanopod\u201d) gene cluster encodes a transmembrane complex that may facilitate benzene transport across the cell membrane. Phylogenomic analyses place ORM2a and ORM2b within a novel genus of strict anaerobes specialized for benzene degradation, which we propose naming \u201c\n                      <jats:italic toggle=\"yes\">Candidatus<\/jats:italic>\n                      Anaerobenzenivorax.\u201d\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>Benzene is a widespread, persistent, and toxic pollutant that can accumulate in anoxic environments such as groundwater and sediments. Benzene can be metabolized in the absence of oxygen; however, despite decades of research, the biochemical mechanisms for benzene activation under anaerobic conditions remain unproven. This study provides strong genetic and proteomic evidence for a new suite of enzymes that initiate anaerobic benzene activation. These findings lay a foundation for future biochemical studies and expand our understanding of how microbes carry out difficult chemical reactions in the absence of oxygen.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aem.02083-25","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T13:00:36Z","timestamp":1774616436000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Identification of a highly expressed gene cluster likely coding for benzene activation enzymes in a methanogenic enrichment culture"],"prefix":"10.1128","volume":"92","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3430-0792","authenticated-orcid":true,"given":"Courtney R. 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