Appl Microbiol Biotechnol. 2012 Aug;95(4):1001-10. Physiological characterization of recombinant Saccharomyces cerevisiae expressing the Aspergillus nidulans phosphoketolase pathway: validation of activity through 13C-based metabolic flux analysis. Papini M, Nookaew I, Siewers V, 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 | N/A | R02736+R01528+R00267+R00214 |
| NADPH production rate Minium | N/A | 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 | N/A | R02736+R01528+R00267+R00214 |
| NADPH production rate Minium | N/A | R02736+R01528 |
| Growth Condition | |
| Strains | wild-type Saccharomyces cerevisiae CEN.PK 113-7D |
| Genotype | MATa ura3-52 MAL2-8cSUC2 |
| Culture medium | For batch cultivations, a previously described (Verduyn et al.1992)mineral salts mediumwasused,commonly called CBS,having the following composition (per liter): (NH4)2SO4, 5 g;KH2PO4, 3 g; MgSO4-7H2O, 0.50 g; Antifoam 289, 0.050 mL; trace metals,1 mL; and vitamins, 1 mL. The trace metal solution consisted of the following (per liter): EDTA (sodium salt), 15.0 g;ZnSO4-7H2O, 0.45 g; MnCl2-2H2O, 1 g; CoCl2-6H2O,0.3 g; CuSO4-5H2O, 0.3 g; Na2MoO4-2H2O, 0.4 g;CaCl2-2H2O, 0.45 g; FeSO4-7H2O, 0.3 g; H3BO3, 0.1 g; and KI, 0.1 g. The pH of the trace metal solution was adjusted to4.0 with 2 M NaOH prior to heat sterilization. The vitamin solution contained (per liter): biotin, 0.05 g; p-amino benzoic acid, 0.2 g; nicotinic acid, 1 g; Ca pantothenate, 1 g; pyridoxine HCl, 1 g; thiamine HCl, 1 g; and myo-inositol, 25 g. The pH of the vitamin solution was adjusted to 6.5 with 2 M NaOH. The vitamin solution was filter sterilized and stored at4 C.This medium was supplemented with 20g/Lglucose. |
| Carbon source | D-Glucose |
| Growth rate | 0.11/h |
| Case-specific description | Flux distribution of the wild-type CEN.PK 113-7D based 13C labeling. |
| Growth Condition | |
| Strains | recombinant strain MP003 |
| Genotype | Transforming S.cerevisiae CEN.PK 113-5D(MATa MAL2-8c SUC2 ura3-52) with the vector pMPa generated the strain MP003. |
| Culture medium | For batch cultivations, a previously described (Verduyn et al.1992)mineral salts mediumwasused,commonly called CBS,having the following composition (per liter): (NH4)2SO4, 5 g;KH2PO4, 3 g; MgSO4-7H2O, 0.50 g; Antifoam 289, 0.050 mL; trace metals,1 mL; and vitamins, 1 mL. The trace metal solution consisted of the following (per liter): EDTA (sodium salt), 15.0 g;ZnSO4-7H2O, 0.45 g; MnCl2-2H2O, 1 g; CoCl2-6H2O,0.3 g; CuSO4-5H2O, 0.3 g; Na2MoO4-2H2O, 0.4 g;CaCl2-2H2O, 0.45 g; FeSO4-7H2O, 0.3 g; H3BO3, 0.1 g; and KI, 0.1 g. The pH of the trace metal solution was adjusted to4.0 with 2 M NaOH prior to heat sterilization. The vitamin solution contained (per liter): biotin, 0.05 g; p-amino benzoic acid, 0.2 g; nicotinic acid, 1 g; Ca pantothenate, 1 g; pyridoxine HCl, 1 g; thiamine HCl, 1 g; and myo-inositol, 25 g. The pH of the vitamin solution was adjusted to 6.5 with 2 M NaOH. The vitamin solution was filter sterilized and stored at4 C.This medium was supplemented with 20g/Lglucose. |
| Carbon source | D-Glucose |
| Growth rate | 0.1/h |
| Case-specific description | Flux distribution of the recombinant strain MP003 based 13C labeling. |
| Specific Rate | (mmol g-1 h-1) |
| Glucose | 15.1 |
| Specific Rate | (mmol g-1 h-1) |
| Glucose | 14.1 |