Discovery of a new type of methylenetetrahydrofolate reductase family that couples with tetrahydrofolate-dependent demethylases

Discovery of a new type of methylenetetrahydrofolate reductase family that couples with tetrahydrofolate-dependent demethylases | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Discovery of a new type of methylenetetrahydrofolate reductase family that couples with tetrahydrofolate-dependent demethylases Toshiya Senda, HongYang Yu, Naofumi Kamimura, Ryo Kato, Miki Senda, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4402079/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in one-carbon (1C) metabolism, catalyzing the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate. Interestingly, Sphingobium lignivorans SYK-6, a model bacterium for the catabolism of lignin-derived aromatic compounds, has a unique MTHFR (S6MTHFR) that has been proposed to catalyze the reverse reaction of typical MTHFRs—namely, the oxidation of 5-methyltetrahydrofolate. However, no direct evidence for this has been obtained. Here, we demonstrated that S6MTHFR catalyzes the oxidation of 5-methyltetrahydrofolate and elucidated the molecular mechanism underlying the unique enzymatic properties of S6MTHFR based on its crystal structure. Furthermore, a database search revealed that a group of bacteria, including S. lignivorans SYK-6, utilize tetrahydrofolate-dependent demethylases to produce 5-methyltetrahydrofolate, which in turn is oxidized by an S6MTHFR-type enzyme. We propose that the combination of a demethylase with an S6MTHFR-type enzyme forms a new type of 1C metabolism that may regulate methionine biosynthesis. Biological sciences/Structural biology/X-ray crystallography Biological sciences/Biochemistry/Enzymes/Oxidoreductases Full Text Additional Declarations There is NO Competing Interest. Supplementary Files 2SupplementaryTablesv2.0.xlsx Supplementary Tables Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4402079","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":302001683,"identity":"73f6fba3-1017-41f1-b448-936895fed1fe","order_by":0,"name":"Toshiya 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Interestingly, Sphingobium lignivorans SYK-6, a model bacterium for the catabolism of lignin-derived aromatic compounds, has a unique MTHFR (S6MTHFR) that has been proposed to catalyze the reverse reaction of typical MTHFRs—namely, the oxidation of 5-methyltetrahydrofolate. However, no direct evidence for this has been obtained. Here, we demonstrated that S6MTHFR catalyzes the oxidation of 5-methyltetrahydrofolate and elucidated the molecular mechanism underlying the unique enzymatic properties of S6MTHFR based on its crystal structure. Furthermore, a database search revealed that a group of bacteria, including S. lignivorans SYK-6, utilize tetrahydrofolate-dependent demethylases to produce 5-methyltetrahydrofolate, which in turn is oxidized by an S6MTHFR-type enzyme. 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