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Elucidation of a Self-Sustaining Cycle in Escherichia colil-Serine Biosynthesis That Results in the Conservation of the Coenzyme, NAD+

Grant, Gregory A.
Biochemistry 2018 v.57 no.11 pp. 1798-1806
Escherichia coli, Mycobacterium tuberculosis, NAD (coenzyme), alpha-ketoglutaric acid, biochemical pathways, biosynthesis, humans, liver, phosphoglycerate dehydrogenase, serine, transferases
The equilibrium of the reaction catalyzed by d-3-phosphoglycerate dehydrogenase (PGDH), the first enzyme in the l-serine biosynthetic pathway, is far in the direction away from serine synthesis. As such, the enzyme is usually assayed in this direction. To easily assay it in the direction of l-serine synthesis, it can be coupled to the next enzyme in the pathway, phosphoserine aminotransferase (PSAT), with the activity monitored by the conversion of NAD⁺ to NADH by PGDH. However, when PGDHs from several different species were coupled to PSAT, it was found that one of them, ecPGDH, conserves the coenzyme in the production of l-serine by utilizing an intrinsic cycle of NAD⁺/NADH interconversion coupled with the conversion of α-ketoglutarate (αKG) to α-hydroxyglutarate. Furthermore, the cycle can be maintained by production of αKG by the second enzyme in the pathway, PSAT, and does not require any additional enzymes. This is not the case for PGDH from another bacterial source, Mycobacterium tuberculosis, and a mammalian source, human liver, where net consumption of NAD⁺ occurs. Both NAD⁺ and NADH appear to remain tightly bound to ecPGDH during the cycle, effectively removing a requirement for the presence of an exogenous coenzyme pool to maintain the pathway and significantly reducing the energy requirement needed to maintain this major metabolic pathway.