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bioRxiv · 10.64898/2026.03.24.713513

The Role of Phosphoenolpyruvate Carboxylase-Protein Kinase in C4 Photosynthesis: Insights from Zea mays Mutant Analysis

Enyew, M.·Studer, A. J.·Woodford, R.·Ermakova, M.·von Caemmerer, S.·Cousins, A. B.
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Abstract

Understanding the regulation of enzyme activity involved in photosynthesis is essential for engineering enhanced carbon fixation in crops. In C4 plants, the enzyme phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is one of the most abundant leaf enzymes and plays an essential role in photosynthetic carbon dioxide (CO2) fixation. The enzyme also plays a key role in central metabolism (e.g., providing intermediates to the citric acid cycle) and therefore must be highly regulated to coordinate its activity. The regulation of PEPC activity can occur allosterically by glucose 6-phosphate activation and malate inhibition, which is in part influenced by reversible phosphorylation. A specific light-dependent phosphorylation of PEPC at an N-terminal serine residue by the PEPC-protein kinase (PEPC-PK) can regulate its sensitivity to this allosteric regulation. However, the impact of this PEPC phosphorylation has not been tested in a C4 crop. Therefore, we created PEPC-PK mutant lines in Zea mays to assess the impact of PEPC phosphorylation on its allosteric regulation, photosynthesis, and growth. While the maximum PEPC activity was unchanged, PEPC in the PEPC-PK mutant plants was not phosphorylated under light and was more sensitive to malate inhibition. However, gas exchange, electron transport, and field biomass analyses showed no differences in the PEPC-PK mutant plants. These results demonstrate that in Z. mays PEPC phosphorylation affects enzyme sensitivity to malate in vitro but does not substantially alert photosynthetic performance or growth under field conditions suggesting additional regulation of PEPC activity in planta.

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Enyew, M., Studer, A. J., Woodford, R., Ermakova, M., von Caemmerer, S., Cousins, A. B.. 2026-03-27. The Role of Phosphoenolpyruvate Carboxylase-Protein Kinase in C4 Photosynthesis: Insights from Zea mays Mutant Analysis. https://doi.org/10.64898/2026.03.24.713513

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