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. Author manuscript; available in PMC: 2023 Jul 22.
Published in final edited form as: ACS Synth Biol. 2021 Sep 1;10(9):2359–2370. doi: 10.1021/acssynbio.1c00258

Figure 1.

Figure 1.

Redox balance-based selection platform for NADH-dependent oxygenases. (A) Deletion of mixed acid fermentation pathways (ldhA, adhE, and frdBC), respiration associated genes (nuoF, ndh, and ubiC), and transhydrogenase component (pntB) greatly diminished native NADH oxidation. The aerobic accumulation of NADH severely inhibited growth without an alternative means of cofactor consumption. NADH-consuming oxidase Lb Nox was expressed on an arabinose inducible plasmid to alleviate NADH stress during nonselection conditions. (B, C) Growth of the NADH selection strain (MX304) is rescued by an NADH oxidase, but not by an NADPH oxidase in liquid media and on agar plates. Values are an average of at least three replicates, and the error bars represent one standard deviation.