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- W2902826360 abstract "Insufficient rate of NADPH regeneration often limits the activity of biosynthetic pathways. Expression of NADPH-regenerating enzymes is commonly used to address this problem and increase cofactor availability. Here, we construct an Escherichia coli NADPH-auxotroph strain, which is deleted in all reactions that produce NADPH with the exception of 6-phosphogluconate dehydrogenase. This strain grows on a minimal medium only if gluconate is added as NADPH source. When gluconate is omitted, the strain serves as a “biosensor” for the capability of enzymes to regenerate NADPH in vivo. We show that the NADPH-auxotroph strain can be used to quantitatively assess different NADPH-regenerating enzymes and provide essential information on expression levels and concentrations of reduced substrates required to support optimal NADPH production rate. The NADPH-auxotroph strain thus serves as an effective metabolic platform for evaluating NADPH regeneration within the cellular context." @default.
- W2902826360 created "2018-12-11" @default.
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- W2902826360 date "2018-11-26" @default.
- W2902826360 modified "2023-09-25" @default.
- W2902826360 title "NADPH-Auxotrophic <i>E. coli</i>: A Sensor Strain for Testing <i>in Vivo</i> Regeneration of NADPH" @default.
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- W2902826360 doi "https://doi.org/10.1021/acssynbio.8b00313" @default.
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