Non‐canonical metabolic pathways in the malaria parasite detected by isotope‐tracing metabolomics.

The malaria parasite, Plasmodiumfalciparum, proliferates rapidly in human erythrocytes by actively scavenging multiple carbon sources and essential nutrients from its host cell. However, a global overview of the metabolic capacity of intraerythrocytic stages is missing. Using multiplex 13C‐labelling coupled with untargeted mass spectrometry and unsupervised isotopologue grouping, we have generated a draft metabolome of P.falciparum and its host erythrocyte consisting of 911 and 577 metabolites, respectively, corresponding to 41% of metabolites and over 70% of the metabolic reaction predicted from the parasite genome. An additional 89 metabolites and 92 reactions were identified that were not predicted from genomic reconstructions, with the largest group being associated with metabolite damage‐repair systems. Validation of the draft metabolome revealed four previously uncharacterised enzymes which impact isoprenoid biosynthesis, lipid homeostasis and mitochondrial metabolism and are necessary for parasite development and proliferation. This study defines the metabolic fate of multiple carbon sources in P.falciparum, and highlights the activity of metabolite repair pathways in these rapidly growing parasite stages, opening new avenues for drug discovery. Synopsis: Multiplex stable‐isotope labelling defines the observable metabolome of red blood cell stages of the malaria parasite Plasmodiumfalciparum and indicates potential roles for metabolic enzymes of unknown function. A draft metabolome of P.falciparum and its host erythrocyte is presented, consisting of 911 and 577 metabolites respectively.The metabolome covers 70% of predicted metabolic reactions.92 unpredicted reactions are identified, the largest group associated with metabolite damage repair.Mediators of isoprenoid biosynthesis, lipid homeostasis, and mitochondrial metabolism are identified. [ABSTRACT FROM AUTHOR]