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In Vivo Rate of Formaldehyde Condensation with Tetrahydrofolate

1
Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany
2
Institute of Molecular Systems Biology, ETH Zürich, Otto-Stern-Weg 3, 8093 Zürich, Switzerland
3
Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, México
4
Tecnologico de Monterrey, Centro de Biotecnología-FEMSA, Ave. Eugenio Garza Sada 2501, Monterrey N.L. 64849, México
5
Horticultural Sciences Department, University of Florida, Gainesville, FL 32611, USA
*
Author to whom correspondence should be addressed.
Metabolites 2020, 10(2), 65; https://doi.org/10.3390/metabo10020065
Received: 29 December 2019 / Revised: 10 February 2020 / Accepted: 11 February 2020 / Published: 12 February 2020
(This article belongs to the Special Issue Metabolic Engineering and Synthetic Biology Volume 2)
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.
Keywords: one-carbon metabolism; spontaneous reaction; auxotrophy; serine cycle; phenotypic phase plane one-carbon metabolism; spontaneous reaction; auxotrophy; serine cycle; phenotypic phase plane
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He, H.; Noor, E.; Ramos-Parra, P.A.; García-Valencia, L.E.; Patterson, J.A.; Díaz de la Garza, R.I.; Hanson, A.D.; Bar-Even, A. In Vivo Rate of Formaldehyde Condensation with Tetrahydrofolate. Metabolites 2020, 10, 65.

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