Cytosolic NADPH balancing in Penicillium chrysogenum cultivated on mixtures of glucose and ethanol.
The in vivo flux through the oxidative branch of the pentose phosphate pathway (oxPPP) in Penicillium chrysogenum was determined during growth in glucose/ethanol carbon-limited chemostat cultures, at the same growth rate. Non-stationary (13)C flux analysis was used to measure the oxPPP flux. A nearly constant oxPPP flux was found for all glucose/ethanol ratios studied. This indicates that the cytosolic NADPH supply is independent of the amount of assimilated ethanol. The cofactor assignment in the model of van Gulik et al. (Biotechnol Bioeng 68(6):602-618, 2000) was supported using the published genome annotation of P. chrysogenum. Metabolic flux analysis showed that NADPH requirements in the cytosol remain nearly the same in these experiments due to constant biomass growth. Based on the cytosolic NADPH balance, it is known that the cytosolic aldehyde dehydrogenase in P. chrysogenum is NAD(+) dependent. Metabolic modeling shows that changing the NAD(+)-aldehyde dehydrogenase to NADP(+)-aldehyde dehydrogenase can increase the penicillin yield on substrate.
Zhao
Zheng
Z
Department of Biotechnology, Kluyver Centre for Genomics of Industrial Fermentation, Delft University of Technology, Julianalaan 67, 2628BC, Delft, The Netherlands. z.zhao@tudelft.nl
Kuijvenhoven
Karel
K
van Gulik
Walter M
WM
Heijnen
Joseph J
JJ
van Winden
Wouter A
WA
Verheijen
Peter J T
PJ
D003600:Cytosol / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D010408:Penicillium chrysogenum / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010427:Pentose Phosphate Pathway
D005656:Fungal Proteins; D000431:Ethanol; D009249:NADP; D005947:Glucose
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