{"doi":"10.1155/2017/5793620","title":"Expression, Purification, and Characterization of (<i>R</i>)-Sulfolactate Dehydrogenase (ComC) from the Rumen Methanogen<i>Methanobrevibacter millerae</i>SM9","abstract":"<jats:p>(<jats:italic>R</jats:italic>)-Sulfolactate dehydrogenase (EC 1.1.1.337), termed ComC, is a member of an NADH/NADPH-dependent oxidoreductase family of enzymes that catalyze the interconversion of 2-hydroxyacids into their corresponding 2-oxoacids. The ComC reaction is reversible and in the biosynthetic direction causes the conversion of (<jats:italic>R</jats:italic>)-sulfolactate to sulfopyruvate in the production of coenzyme M (2-mercaptoethanesulfonic acid). Coenzyme M is an essential cofactor required for the production of methane by the methyl-coenzyme M reductase complex. ComC catalyzes the third step in the first established biosynthetic pathway of coenzyme M and is also involved in methanopterin biosynthesis. In this study, ComC from<jats:italic>Methanobrevibacter millerae</jats:italic>SM9 was cloned and expressed in<jats:italic>Escherichia coli</jats:italic>and biochemically characterized. Sulfopyruvate was the preferred substrate using the reduction reaction, with 31% activity seen for oxaloacetate and 0.2% seen for<jats:italic>α</jats:italic>-ketoglutarate. Optimal activity was observed at pH 6.5. The apparent<jats:italic>K</jats:italic><jats:sub>M</jats:sub>for coenzyme (NADH) was 55.1 <jats:italic>μ</jats:italic>M, and for sulfopyruvate, it was 196 <jats:italic>μ</jats:italic>M (for sulfopyruvate the<jats:italic>V</jats:italic><jats:sub>max</jats:sub>was 93.9 <jats:italic>μ</jats:italic>mol min<jats:sup>−1</jats:sup> mg<jats:sup>−1</jats:sup>and<jats:italic>k</jats:italic><jats:sub>cat</jats:sub>was 62.8 s<jats:sup>−1</jats:sup>). The critical role of ComC in two separate cofactor pathways makes this enzyme a potential means of developing methanogen-specific inhibitors for controlling ruminant methane emissions which are increasingly being recognized as contributing to climate change.</jats:p>","journal":"Archaea","year":2017,"id":656824,"datarank":0.3596842909197557,"base_score":2.3978952727983707,"endowment":2.3978952727983707,"self_citation_contribution":0.3596842909197557,"citation_network_contribution":0.0,"self_endowment_contribution":0.3596842909197557,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":10,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1714526,"name":"Linley R. Schofield","orcid":"0000-0001-7288-2792","position":1,"is_corresponding":false},{"id":1714527,"name":"Carrie Sang","orcid":"0000-0003-4368-1558","position":2,"is_corresponding":false},{"id":1714528,"name":"Debjit Dey","orcid":null,"position":3,"is_corresponding":false},{"id":1714529,"name":"Ron S. Ronimus","orcid":"0000-0001-8669-5840","position":4,"is_corresponding":false},{"id":181780,"name":"Yanli Zhang","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Expression, Purification, and Characterization of (<i>R</i>)-Sulfolactate Dehydrogenase (ComC) from the Rumen Methanogen<i>Methanobrevibacter millerae</i>SM9","abstract":"<jats:p>(<jats:italic>R</jats:italic>)-Sulfolactate dehydrogenase (EC 1.1.1.337), termed ComC, is a member of an NADH/NADPH-dependent oxidoreductase family of enzymes that catalyze the interconversion of 2-hydroxyacids into their corresponding 2-oxoacids. The ComC reaction is reversible and in the biosynthetic direction causes the conversion of (<jats:italic>R</jats:italic>)-sulfolactate to sulfopyruvate in the production of coenzyme M (2-mercaptoethanesulfonic acid). Coenzyme M is an essential cofactor required for the production of methane by the methyl-coenzyme M reductase complex. ComC catalyzes the third step in the first established biosynthetic pathway of coenzyme M and is also involved in methanopterin biosynthesis. In this study, ComC from<jats:italic>Methanobrevibacter millerae</jats:italic>SM9 was cloned and expressed in<jats:italic>Escherichia coli</jats:italic>and biochemically characterized. Sulfopyruvate was the preferred substrate using the reduction reaction, with 31% activity seen for oxaloacetate and 0.2% seen for<jats:italic>α</jats:italic>-ketoglutarate. Optimal activity was observed at pH 6.5. The apparent<jats:italic>K</jats:italic><jats:sub>M</jats:sub>for coenzyme (NADH) was 55.1 <jats:italic>μ</jats:italic>M, and for sulfopyruvate, it was 196 <jats:italic>μ</jats:italic>M (for sulfopyruvate the<jats:italic>V</jats:italic><jats:sub>max</jats:sub>was 93.9 <jats:italic>μ</jats:italic>mol min<jats:sup>−1</jats:sup> mg<jats:sup>−1</jats:sup>and<jats:italic>k</jats:italic><jats:sub>cat</jats:sub>was 62.8 s<jats:sup>−1</jats:sup>). The critical role of ComC in two separate cofactor pathways makes this enzyme a potential means of developing methanogen-specific inhibitors for controlling ruminant methane emissions which are increasingly being recognized as contributing to climate change.</jats:p>","is_dataset_classified":null,"base_score":2.3978952727983707,"endowment":2.3978952727983707,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"29234237","pmcid":"PMC5695019","openalex_id":"https://openalex.org/W2767840396","authors":[],"funders":[{"funder_name":"Pastoral Greenhouse Gas Research Consortium","grant_id":"","title":null}],"total_grants":1,"fwci":0.1015,"citation_percentile":0.48349614,"influential_citations":0,"citation_trend":[{"year":2020,"count":1},{"year":2021,"count":3},{"year":2022,"count":1},{"year":2023,"count":2},{"year":2025,"count":1},{"year":2026,"count":2}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"http://downloads.hindawi.com/journals/archaea/2017/5793620.pdf","host_type":"journal"},{"url":"http://downloads.hindawi.com/journals/archaea/2017/5793620.pdf","host_type":"publisher"},{"url":"http://downloads.hindawi.com/journals/archaea/2017/5793620.xml","host_type":"publisher"},{"url":"https://doi.org/10.1155/2017/5793620","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/29234237","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/5695019","host_type":"repository"},{"url":"https://doaj.org/article/ccc9da2c0f4044c0afd1646aca6ce185","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC5695019","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC5695019?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Microbial Metabolic Engineering and Bioproduction","Microbial metabolism and enzyme function","Biofuel production and bioconversion","Biosynthetic Pathways","Cloning, Molecular","Enzyme Stability","Escherichia coli","Gene Expression","Hydrogen-Ion Concentration","Kinetics","Lactates","Mesna","Methanobrevibacter","Oxidoreductases","Pyruvates","Recombinant Proteins","Substrate Specificity"],"mesh_terms":["Cloning, Molecular","Enzyme Stability","Escherichia coli","Hydrogen-Ion Concentration","Kinetics","Lactates","Oxidoreductases","Pyruvates","Recombinant Proteins","Substrate Specificity","Mesna","Gene Expression","Methanobrevibacter","Biosynthetic Pathways"],"keywords":["Methanogen","Biology","Rumen","Carbon monoxide dehydrogenase","Archaea","Methanomicrobiales","Euryarchaeota","Molecular biology","Gene","Biochemistry","Genetics","Bacteria","Methanosarcina","Fermentation"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Climate action"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-11T22:25:31.742334Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}