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Eukaryot Cell. 2003 Jun;2(3):599-608. Identification of in vivo enzyme activities in the cometabolism of glucose and acetate by Saccharomyces cerevisiae by using 13C-labeled substrates. dos Santos MM, Gombert AK, Christensen B, et al.



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ATP production rate Maximum N/A -R01786-R04779+2.5*R01061+R01512+R00200+2.5*R00209
+2.5*R00267+2.5*R08549-R00405+1.5*R00402+2.5*R00342
+2.5*R00214
ATP production rate Minium N/A -R01786-R04779+2.5*R01061+R01512+R00200+2.5*R00209
+2.5*R08549-R00405+1.5*R00402+2.5*R00342
NADPH production rate Maximum 143.1 R02736+R01528+R00267+R00214
NADPH production rate Minium 84.2 R02736+R01528
ATP production rate Maximum N/A -R01786-R04779+2.5*R01061+R01512+R00200+2.5*R00209
+2.5*R00267+2.5*R08549-R00405+1.5*R00402+2.5*R00342
+2.5*R00214
ATP production rate Minium N/A -R01786-R04779+2.5*R01061+R01512+R00200+2.5*R00209
+2.5*R08549-R00405+1.5*R00402+2.5*R00342
NADPH production rate Maximum 155 R02736+R01528+R00267+R00214
NADPH production rate Minium 88.4 R02736+R01528
Growth Condition
Strains Saccharomyces cerevisiae maeI delta strain
Genotype isogenic malic enzyme deletion
Culture medium Both strains were maintained at 4 Celsius on YPD agar plates (containing, per liter, 10 g of Difco yeast extract, 20 g of Difco peptone, 20 g of glucose, and 20 g of agar), which were prepared monthly. Chemostat cultivations. Aerobic carbon-limited chemostats were operated at 30 Celsius with a dilution rate (which equals the specific growth rate in steady-state cultures) of 0.1/h, using homemade fermentors with a working volume of approximately 170 ml. The volume was kept constant by withdrawing liquid with a continuously operating peristaltic pump. The pH was controlled to 5 with NaOH, except for cultivations with an acetic acid fraction in the feed of 0.95 mol/C of substrate, for which a pH of 6 was kept constant by addition of HCl (pH 6 was used because acetic acid inhibition is lower at this pH than at pH 5).
Carbon source Glucose
Growth rate 0.1/h
Case-specific description Distribution of metabolic fluxes for a malic enzyme deletion strain during chemostat cultivation with [1-13C]glucose,sum was 61.6 for the deletion mutant.
Growth Condition
Strains Saccharomyces cerevisiae CEN.PK113-7D
Genotype (MATa MAL2-8c SUC2)
Culture medium Both strains were maintained at 4 Celsius on YPD agar plates (containing, per liter, 10 g of Difco yeast extract, 20 g of Difco peptone, 20 g of glucose, and 20 g of agar), which were prepared monthly. Chemostat cultivations. Aerobic carbon-limited chemostats were operated at 30 Celsius with a dilution rate (which equals the specific growth rate in steady-state cultures) of 0.1/h, using homemade fermentors with a working volume of approximately 170 ml. The volume was kept constant by withdrawing liquid with a continuously operating peristaltic pump. The pH was controlled to 5 with NaOH, except for cultivations with an acetic acid fraction in the feed of 0.95 mol/C of substrate, for which a pH of 6 was kept constant by addition of HCl (pH 6 was used because acetic acid inhibition is lower at this pH than at pH 5).
Carbon source Glucose
Growth rate 0.1/h
Case-specific description Distribution of metabolic fluxes for the reference strain during chemostat cultivation with [1-13C]glucose, sum was 62.7 .
Specific Rate (mmol g-1 h-1)
Specific Rate (mmol g-1 h-1)
Source