{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T07:58:14Z","timestamp":1784275094188,"version":"3.55.0"},"reference-count":53,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T00:00:00Z","timestamp":1747785600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T00:00:00Z","timestamp":1747785600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100008982","name":"National Science Foundation","doi-asserted-by":"publisher","award":["IOS-1449525"],"award-info":[{"award-number":["IOS-1449525"]}],"id":[{"id":"10.13039\/501100008982","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008982","name":"National Science Foundation","doi-asserted-by":"publisher","award":["IOS-1938948"],"award-info":[{"award-number":["IOS-1938948"]}],"id":[{"id":"10.13039\/501100008982","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2025,5,21]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Evidence suggests that multienzyme complexes are involved in biological methane production (methanogenesis), although the composition of the Wolfe Cycle methanogenesis complexes may vary between diverse methanoarchaeal taxa. Methylenetetrahydromethanopterin reductase (Mer) is the first committed step in C1 oxidation to CO\n              <jats:sub>2<\/jats:sub>\n              during methylotrophic methanogenesis. However, Mer is downregulated when cells use acetate as a substrate. We hypothesized that Mer overexpression during methylotrophic methanogenesis would be beneficial, while overexpression during acetoclastic methanogenesis would be detrimental for energy conservation. To test this hypothesis, we overexpressed Mer and characterized strain physiology on methanol, acetate, and when switching substrates. We found that Mer overexpression results in faster growth on methanol, with less C fixation into biomass, and no effect on methanogenesis. Growth on acetate was not affected by Mer overexpression, but switching between substrates was affected. The native Mer overexpressing strain was slower to adjust from methanol to acetate and vice-versa. These data suggest that tight regulation of Mer expression is necessary to regulate C flux through methylotrophic versus acetoclastic methanogenesis pathways in\n              <jats:italic toggle=\"yes\">Methanosarcina<\/jats:italic>\n              .\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>Methanoarchaea thrive near the \u201cthermodynamic limit of life\u201d and have likely evolved efficient mechanisms to control flux of substrates to conserve energy. Methylenetetrahydromethanopterin reductase (Mer) is a highly conserved, key enzyme in the Wood-Ljungdahl and Wolfe Cycle methanogenesis pathways. Our study sheds light on how Mer enzyme stoichiometry affects methanogenesis and suggests avenues for engineering the organism to promote renewable fuel or bioproduct synthesis.<\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/aem.00675-25","type":"journal-article","created":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T13:00:25Z","timestamp":1745586025000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Mer overexpression in\n            <i>Methanosarcina acetivorans<\/i>\n            affects growth and methanogenesis during substrate adaptation"],"prefix":"10.1128","volume":"91","author":[{"given":"Darla","family":"Brennan","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dillon","family":"Lieber","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary","family":"Walter","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan","family":"Price","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7560-973X","authenticated-orcid":true,"given":"Nicole R.","family":"Buan","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0065-2911(08)60068-5"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-144-9-2377"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.030508.152103"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2018.05.021"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.rser.2015.04.129"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1931"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf8821"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep14295"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.energy.2023.127819"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13568-015-0117-4"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1042\/ETLS20180031"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0079-6603(02)71045-3"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1023\/b:jobb.0000019598.64642.97"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00117-08"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2009.06990.x"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1213193109"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)68417-0"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1994.tb18608.x"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1003653107"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0107563"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00881-12"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.163.3.1000-1006.1985"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4838(91)90072-8"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.14.8352"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0407486101"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.00226.x"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2011.00069"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00895-13"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00189-12"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01424-07"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01162-15"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00203-007-0266-1"},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.6.2626"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.5.1449-1454.2002"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2016.02.003"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1155\/2008\/534081"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.4150102"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.19.5797-5802.1996"},{"key":"e_1_3_5_40_2","first-page":"23","volume-title":"Methods in enzymology","author":"Buan N","year":"2011","unstructured":"Buan N, Kulkarni G, Metcalf W. 2011. Genetic methods for Methanosarcina species, p 23\u201342. In Methods in enzymology. Vol. 494. Elsevier."},{"key":"e_1_3_5_41_2","unstructured":"SNAP i.d. 2.0 protein detection system for Western blotting. Available from: https:\/\/www.sigmaaldrich.com\/deepweb\/assets\/sigmaaldrich\/product\/documents\/310\/895\/snap-id-western-00000825w-ms.pdf"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02417-20"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.21769\/bioprotoc.1779"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1007783107"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1100025"},{"key":"e_1_3_5_46_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature09015"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.01322"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr049832c"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr049831k"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.2001.02358.x"},{"key":"e_1_3_5_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.1.184-188.1987"},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.15252\/msb.202110504"},{"key":"e_1_3_5_53_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1990.tb13963.x"},{"key":"e_1_3_5_54_2","doi-asserted-by":"publisher","DOI":"10.1093\/gbe\/evw114"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.00675-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.00675-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T13:04:01Z","timestamp":1747832641000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aem.00675-25"}},"subtitle":[],"editor":[{"given":"Gemma","family":"Reguera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,5,21]]},"references-count":53,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2025,5,21]]}},"alternative-id":["10.1128\/aem.00675-25"],"URL":"https:\/\/doi.org\/10.1128\/aem.00675-25","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,21]]},"assertion":[{"value":"2025-03-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00675-25"}}