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. 2023 Oct 30;89(11):e01106-23. doi: 10.1128/aem.01106-23

FIG 8.

FIG 8

The effect of introducing an ATP regeneration system on NMN yield. (A) The scheme of the synthesis route for NMN from ribose-5-phosphate (R5P) via coupling BaPRS and HdNAMPT. The AMP was a byproduct of R5P phosphorylation. The ATP regeneration system was applied for the recycling of AMP back into ATP. The NMN generation (B), ATP consumption (C), and AMP generation (D) in the absence and in the presence of an ATP regeneration system containing wild-type ChPPK or the variant ChPPK/A79G/S106C/I108F/L285P at indicated time points. In B, C, and D, the reactions without an ATP regeneration system (the black and blue lines) consisted of 0.2 or 20.0 mM ATP, 30 mM R5P, 50 mM nicotinamide (NAM), 8 mg/mL BL21-BaPRS cell lysate supernatant, and 12 mg/mL BL21-HdNampt cell lysate supernatant. The reactions with an ATP regeneration system (the red and green lines) consisted of 0.2 mM ATP, 30 mM R5P, 50 mM NAM, 4.8 g/L PPA, 8 mg/mL BL21-BaPRS cell lysate supernatant, 12 mg/mL BL21-HdNampt cell lysate supernatant, and 8 mg/mL cell lysate supernatant of wild-type BL21-ChPPK or BL21-ChPPK/A79G/S106C/I108F/L285P.