Fig. 3. PRS, as a metabolic valve, reallocates carbon flows through nucleotide anabolism and methylotrophic assimilation by up-regulating the PKT pathway.

a Comparison of the intermediates involved in the central methylotrophic metabolism and nucleotide synthesis of the AM1^PTR and AM1^WT strains grown on 10 mM methanol (n = 3 biologically independent samples.). b Transcriptional comparison of gene expression between the AM1^PTR strain and AM1^WT strain grown on 10 mM methanol (n = 3 biologically independent samples with five technical repeats). c Specific activities of FDH in the strains of AM1^PTR and AM1^WT grown on 10 mM methanol (n = 3 biologically independent samples with three technical repeats). d Knocking out the gene xfp2 (AM1^PTR 04) decreased the growth advantage of the AM1^PTR strain on low methanol concentrations of 5 mM, 10 mM, and 15 mM, while it did not affect the growth of the AM1^WT strain (n = 5 biologically independent samples with three technical repeats.). e Schematic central methylotrophic network in the AM1^PTR strain on low methanol concentration. Partially turning off the PRS valve decreased the allocation of limited methanol for nucleotide synthesis and up-regulated the PKT pathway to channel more methanol through the assimilation pathways. Reversible reactions are shown as double arrows. Fold changes of gene transcriptional levels in the AM1^PTR strain versus the AM1^WT are indicated by colored arrows. Fold changes of intermediate pools in the AM1^PTR strain versus the AM1^WT are indicated by colored metabolites. “NA” is abbreviated for “not analyzed”. F6P fructose-6-phosphate, G6P glucose-6-phosphate, G1P glucose-1-phosphate, Pyr pyruvate, FBP fructose-1,6-biphosphate, 3-HB-CoA 3-hydroxybutyryl-CoA, AcP acetyl-phosphate, PHB poly-β-hydroxybutyrate. Data are presented as mean ± SD. Significance was analyzed using a two-tailed t-test analysis. Source data are provided as a Source Data file.