{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T15:29:40Z","timestamp":1780586980339,"version":"3.54.1"},"reference-count":40,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,2,12]],"date-time":"2020-02-12T00:00:00Z","timestamp":1581465600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004189","name":"Max-Planck-Gesellschaft","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100004189","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Formaldehyde is a highly reactive compound that participates in multiple spontaneous reactions, but these are mostly deleterious and damage cellular components. In contrast, the spontaneous condensation of formaldehyde with tetrahydrofolate (THF) has been proposed to contribute to the assimilation of this intermediate during growth on C1 carbon sources such as methanol. However, the in vivo rate of this condensation reaction is unknown and its possible contribution to growth remains elusive. Here, we used microbial platforms to assess the rate of this condensation in the cellular environment. We constructed Escherichia coli strains lacking the enzymes that naturally produce 5,10-methylene-THF. These strains were able to grow on minimal medium only when equipped with a sarcosine (N-methyl-glycine) oxidation pathway that sustained a high cellular concentration of formaldehyde, which spontaneously reacts with THF to produce 5,10-methylene-THF. We used flux balance analysis to derive the rate of the spontaneous condensation from the observed growth rate. According to this, we calculated that a microorganism obtaining its entire biomass via the spontaneous condensation of formaldehyde with THF would have a doubling time of more than three weeks. Hence, this spontaneous reaction is unlikely to serve as an effective route for formaldehyde assimilation.<\/jats:p>","DOI":"10.3390\/metabo10020065","type":"journal-article","created":{"date-parts":[[2020,2,25]],"date-time":"2020-02-25T08:12:22Z","timestamp":1582618342000},"page":"65","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["In Vivo Rate of Formaldehyde Condensation with Tetrahydrofolate"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1223-2813","authenticated-orcid":false,"given":"Hai","family":"He","sequence":"first","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology, Am M\u00fchlenberg 1, 14476 Potsdam-Golm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elad","family":"Noor","sequence":"additional","affiliation":[{"name":"Institute of Molecular Systems Biology, ETH Z\u00fcrich, Otto-Stern-Weg 3, 8093 Z\u00fcrich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Perla A.","family":"Ramos-Parra","sequence":"additional","affiliation":[{"name":"Tecnologico de Monterrey, Escuela de Ingenier\u00eda y Ciencias, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"},{"name":"Tecnologico de Monterrey, Centro de Biotecnolog\u00eda-FEMSA, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liliana E.","family":"Garc\u00eda-Valencia","sequence":"additional","affiliation":[{"name":"Tecnologico de Monterrey, Escuela de Ingenier\u00eda y Ciencias, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"},{"name":"Tecnologico de Monterrey, Centro de Biotecnolog\u00eda-FEMSA, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenelle A.","family":"Patterson","sequence":"additional","affiliation":[{"name":"Horticultural Sciences Department, University of Florida, Gainesville, FL 32611, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0006-5694","authenticated-orcid":false,"given":"Roc\u00edo I.","family":"D\u00edaz de la Garza","sequence":"additional","affiliation":[{"name":"Tecnologico de Monterrey, Escuela de Ingenier\u00eda y Ciencias, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"},{"name":"Tecnologico de Monterrey, Centro de Biotecnolog\u00eda-FEMSA, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew D.","family":"Hanson","sequence":"additional","affiliation":[{"name":"Horticultural Sciences Department, University of Florida, Gainesville, FL 32611, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arren","family":"Bar-Even","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology, Am M\u00fchlenberg 1, 14476 Potsdam-Golm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1146\/annurev-biochem-030409-143718","article-title":"Enzyme promiscuity: A mechanistic and evolutionary perspective","volume":"79","author":"Khersonsky","year":"2010","journal-title":"Annu. 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