Metabolic perturbation of an essential pathway: evaluation of a glycine precursor of coenzyme A

Pantetheine, and its corresponding disulfide pantethine, play a key role in metabolism as a building block of coenzyme A (CoA), an essential cofactor utilized in ~4% of primary metabolism and central to fatty acid, polyketide, and non-ribosomal peptide synthases. Using a combination of recombinant engineering and chemical synthesis, we demonstrate that the disulfide of N-pantoylglycyl-2-aminoethanethiol (GlyPan), with one carbon deletion from pantetheine, can rescue a mutant E. coli strain MG1655CΔpanC lacking a functional pantothenate synthetase. Using mass spectral methods, we demonstrate that the GlyPan variant is accepted by the downstream CoA biosynthetic machinery, ultimately being incorporated into essential acyl carrier proteins. These findings point to further flexibility in CoA-dependent pathways and offer the opportunity to incorporate orthogonal analogs.