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bioRxiv · 10.1101/2024.01.28.577108

Rubisco supplies pyruvate for the 2-C-methyl-D-erythritol-4-phosphate pathway in Arabidopsis

Evans, S. E.·Xu, Y.·Bergman, M. E.·Ford, S.·Liu, Y.·Sharkey, T.·Phillips, M. A.
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Abstract

Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) produces pyruvate in the chloroplast through beta-elimination of the aci-carbanion intermediate. Here we show that this side reaction supplies pyruvate for isoprenoid, fatty acid, and branched chain amino acid biosynthesis in photosynthetically active tissue. 13C labeling studies of whole Arabidopsis plants demonstrate that the total carbon commitment to pyruvate is too large for phosphoenolpyruvate (PEP) to serve as precursor. Low oxygen stimulates rubisco carboxylase activity and increased pyruvate production and flux through the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway, which supplies the precursors for plastidic isoprenoid biosynthesis. Metabolome analysis of mutants defective in PEP or pyruvate import further supported rubisco as the main source of pyruvate in chloroplasts. Rubisco beta-elimination leading to pyruvate constituted 0.7% of the product profile in in vitro assays, which translates to 2% of the total carbon leaving the Calvin-Benson-Bassham cycle (CBC). These insights solve the so-called pyruvate paradox, improve the fit of metabolic models for central metabolism, and connect the MEP pathway directly to carbon assimilation.

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Evans, S. E., Xu, Y., Bergman, M. E., Ford, S., Liu, Y., Sharkey, T., Phillips, M. A.. 2024-01-31. Rubisco supplies pyruvate for the 2-C-methyl-D-erythritol-4-phosphate pathway in Arabidopsis. https://doi.org/10.1101/2024.01.28.577108

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