Table S1. the Simplified Metabolic Reactions Used for MFA in This Study

1 Table S1. The simplified metabolic reactions used for MFA in this study

Pathway Reaction

fSorb-S6P Sorbitol +ATP → Sorbitol-6-phosphate + ADP

fS6P-FDP Sorbitol-6-phosphate + ATP + NAD → Fructose-1,6-phosphate + ADP + NADH

fFDP-GAP Fructose-1,6-phosphate → 2 Glyceraldehyde-3-phosphate

fMeth-Form Methanol + (1/2)  O2→ Formaldehyde + H2O

fForm-CO2 Formaldehyde + 2 NAD → CO2 + 2 NADH

fForm-GAP Formaldehyde + ATP → (1/3)  Glyceraldehyde-3-phosphate + ADP

fGAP-CO2 Glyceraldehyde-3-phosphate + 5 NAD + FAD + 2 ADP + GDP →

3 CO2 + 5 NADH + FADH2 + 2 ATP + GTP

fNADH-ATP NADH + (1/2)  O2 + (P/O)  ADP → NAD + (P/O)  ATP + H2O

fFADH2-NADH FADH2 + GTP + NAD → NADH + GDP + FAD

fGAP-X (1/3)  Glyceraldehyde-3-phosphate + (1/2)(4.14/31-4.06)  NADH + YATP/X  ATP →

* X(CHNO) + (1/2)(4.0614/3-4.06)  NAD + YATP/X  ADP

2*: biomass in chemical formula CHNO ; 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26

 27 Table S2. Measured ( fexta, i ) and reconciled ( f extra,i ) specific rates (mmol/(g DCWh)) of metabolites

1 1 1 during the first transient continuous culture with the change of methanol fraction (C-mol/C-mol). 2 Methanol fraction 0 0.25 0.50 0.75

    fexta, i f extra,i fexta, i f extra,i fexta, i f extra,i fexta, i f extra,i

Methanol 0 0 -0.87 -0.80 -1.64 -1.70 -2.86 -2.71 Sorbitol -0.55 -0.49 -0.38 -0.40 -0.25 -0.26 -0.11 -0.11 X 0.92 0.95 0.92 0.91 0.92 0.91 0.92 0.94

O2 -2.88 -2.20 -3.57 -2.85 -3.83 -3.27 -4.03 -3.81

CO2 1.57 1.98 2.19 2.27 2.26 2.32 2.51 2.43

h 6.88 1.69 1.16 1.25 2 5.99 5.99 5.99 5.99 Pass/fail Fail Pass Pass Pass 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

1 2  1 Table S3. Measured ( fexta, i ) and reconciled ( f extra,i ) specific rates (mmol/(g DCWh)) of metabolites

2 during the second transient continuous culture with the change of methanol fraction (C-mol/C-mol). 3 Methanol fraction 0.84 0.65 0.50 0.27

    fexta, i f extra,i fexta, i f extra,i fexta, i f extra,i fexta, i f extra,i

Methanol -2.94 -3.09 -2.18 -2.35 -1.59 -1.71 -0.78 -0.84 Sorbitol -0.09 -0.09 -0.19 -0.20 -0.26 -0.28 -0.29 -0.33 X 0.92 0.91 0.92 0.89 0.92 0.89 0.92 0.86

O2 -4.92 -4.29 -4.49 -3.89 -4.18 -3.45 -3.13 -2.49

CO2 2.71 2.72 2.96 2.64 2.84 2.53 2.03 1.93

h 1.06 2.15 3.15 6.82 2 5.99 5.99 5.99 5.99 Pass/fail Pass Pass Pass Fail 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31