Fig. 4: Deregulation of glycolysis module.

a Schematic illustration of the gluconeogenesis and glycolysis pathways. The gluconeogenesis pathway is marked in blue, while the linear glycolytic pathway is divided into ‘upstream glycolysis’ (green) and ‘downstream glycolysis’ (purple). b Deletion of 6 individual genes within the gluconeogenesis pathway. c Single and combinatorial overexpression of 9 genes within the glycolysis pathway. d Schematic depicting the mutated enzyme within the glycolysis pathway. Two allosterically regulated enzymes, Pfk1/2 and Pyk1, are highlighted in pink, while two phosphorylation-modified enzymes, Fba1 and Gpm1, are shown in green. The NADPH biosensor is marked yellow. e Regulation of Pfk1 and Pfk2, or their mutant versions, affects the flux through glycolysis. f Overexpression of mutated Pyk1, Fba1, and Gpm1. g Engineering of the cofactor metabolism for NADPH and NADH within the glycolysis pathway. Enzymes shown in the pathways: Mls1 malate synthase, Cit2 citrate synthase, Pyc1/2 pyruvate carboxylase, Pck1 phosphoenolpyruvate carboxykinase, Fbp1 fructose-1,6-bisphosphatase, Pgi1 phosphoglucose isomerase, Tdh3 glyceraldehyde-3-phosphate dehydrogenase, Pgk1 3-phosphoglycerate kinase, Eno2 phosphopyruvate hydratase. See Fig. 1 and its accompanying legend for details on other enzyme information. For (b, c) and (f, g), strain R30c was used as the control. All strains were cultivated in a minimal medium containing 2% xylose as the sole carbon source. All data presented in this figure represent the mean from n = 3 biologically independent samples, with error bars indicating the standard deviation. Source data are provided as a Source Data file.