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Methanogens are of biotechnological importance due to the ability to produce methane (CH\n                    <jats:sub>4<\/jats:sub>\n                    ) from molecular hydrogen (H\n                    <jats:sub>2<\/jats:sub>\n                    ) and carbon dioxide (CO\n                    <jats:sub>2<\/jats:sub>\n                    ) and to excrete proteinogenic amino acids. This study focuses on analyzing the link between biological methanogenesis and amino acid excretion under N\n                    <jats:sub>2<\/jats:sub>\n                    -fixing conditions. Among five hydrogenotrophic, autotrophic methanogens,\n                    <jats:italic>Methanothermobacter marburgensis<\/jats:italic>\n                    was prioritized and further cultivated in closed batch cultivation mode under N\n                    <jats:sub>2<\/jats:sub>\n                    -fixing conditions.\n                    <jats:italic>M. marburgensis<\/jats:italic>\n                    was grown on chemically defined minimal medium with different concentrations of ammonium in a H\n                    <jats:sub>2<\/jats:sub>\n                    \/CO\n                    <jats:sub>2<\/jats:sub>\n                    \/N\n                    <jats:sub>2<\/jats:sub>\n                    atmosphere. This enabled the quantification of ammonia uptake, N\n                    <jats:sub>2<\/jats:sub>\n                    -fixation, amino acid excretion and the conversion of H\n                    <jats:sub>2<\/jats:sub>\n                    \/CO\n                    <jats:sub>2<\/jats:sub>\n                    to CH\n                    <jats:sub>4<\/jats:sub>\n                    . To quantify N\n                    <jats:sub>2<\/jats:sub>\n                    -fixation rates in a mass balance setting a novel method has been established. The method utilizes the pressure drop below a certain threshold pressure in closed batch cultivation mode \u2013 the\n                    <jats:italic>th<\/jats:italic>\n                    reshold\n                    <jats:italic>p<\/jats:italic>\n                    ressure for\n                    <jats:italic>N<\/jats:italic>\n                    <jats:sub>\n                      <jats:italic>2<\/jats:italic>\n                    <\/jats:sub>\n                    <jats:italic>-fix<\/jats:italic>\n                    ation (THp\n                    <jats:sub>N2fix<\/jats:sub>\n                    ). Using the THp\n                    <jats:sub>N2fix<\/jats:sub>\n                    method, volumetric N\n                    <jats:sub>2<\/jats:sub>\n                    -fixation rates of\n                    <jats:italic>M. marburgensis<\/jats:italic>\n                    as high as 0.91 mmol L\n                    <jats:sup>\u22121<\/jats:sup>\n                    h\n                    <jats:sup>\u22121<\/jats:sup>\n                    were determined. Excretion of amino acids was found with highest detected values of glutamic acid, alanine, glycine and asparagine. The highest total amino acid excretion of 7.5 \u00b5mol L\n                    <jats:sup>\u22121<\/jats:sup>\n                    h\n                    <jats:sup>\u22121<\/jats:sup>\n                    was detected with H\n                    <jats:sub>2<\/jats:sub>\n                    \/CO\n                    <jats:sub>2<\/jats:sub>\n                    \/N\n                    <jats:sub>2<\/jats:sub>\n                    at an ammonium concentration of 40 mmol L\n                    <jats:sup>\u22121<\/jats:sup>\n                    . This study sheds light on the link between methanogenesis, biological N\n                    <jats:sub>2<\/jats:sub>\n                    -fixation, and proteinogenic amino acid excretion. The concomitant production of amino acids and CH\n                    <jats:sub>4<\/jats:sub>\n                    could become of biotechnological relevance in an integrated approach coupling biomethanation and N\n                    <jats:sub>2<\/jats:sub>\n                    -fixation in a biorefinery concept.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-87686-1","type":"journal-article","created":{"date-parts":[[2025,1,30]],"date-time":"2025-01-30T06:08:54Z","timestamp":1738217334000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Quantitative analysis of amino acid excretion by Methanothermobacter marburgensis under N2-fixing conditions"],"prefix":"10.1038","volume":"15","author":[{"given":"Barbara","family":"Reischl","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Schupp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hayk","family":"Palabikyan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Steger-M\u00e4hnert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Fink","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon K.-M. 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