Fig. 4

DEGs and intracellular formaldehyde accumulation analyses of the evolved strain HTX-33 at 33 °C compared to the starting strain X-33 at 30 °C in 0.5% methanol minimal medium. a, Cluster analysis related to DEGs from methanol metabolism, PPP, EMP, TCA and cell wall synthesis; b, Changes in intracellular formaldehyde accumulation (µM per OD); c, Validation of selected DEGs by RT-qPCR; d, Correlation of the RNA-Seq data (X-axis) and the RT-qPCR data (Y-axis). Data are average values and standard deviations of triplicate experiments. An asterisk indicates a significant difference with ^***P < 0.001; e, Sketch of DEGs in methanol metabolism. Genes in green showed significant down-regulation in expression. Bold border represented enhanced cell wall. Enzymes: Aox, alcohol oxidase; Fld1, formaldehyde dehydrogenase; Fgh1, S-formylglutathione hydrolase; Fdh1, formate dehydrogenase; Das, dihydroxyacetone synthase; Dhak, dihydroxyacetone kinase; Fba, fructosebisphosphate aldolase/sedoheptulose-bisphosphate aldolase; Fbp, fructose bisphosphatase; Sbp, sedoheptulose bisphosphatase; Rpe, ribulose-phosphate 3-epimerase; RpiA, ribose-5-phosphate isomerase; Metabolites: Xu5P, xylulose-5-phosphate; Ru5P, ribulose-5-phosphate; R5P, ribose-5-phosphate; DHA, dihydroxyacetone; DHAP, dihydroxyacetone phosphate; F16dP, fructose-1,6-bisphosphate; F6P, fructose-6-phosphate; G3P, glyceraldehyde-3-phosphate; E4P, erythrose-4-phosphate; S7P, sedoheptulose-7-phosphate; S17dP, sedoheptulose-1,7-bisphosphate