DpgC Is a Metal- and Cofactor-Free 3,5-Dihydroxyphenylacetyl-CoA 1,2-Dioxygenase in the Vancomycin Biosynthetic Pathway Original Research Article

3,5-Dihydroxyphenylglycine is a crucial amino acid monomer in the nonribosomal glycopeptide antibiotic vancomycin. This nonproteinogenic amino acid is constructed from malonyl-CoA by a set of four enzymes, DpgA–D, in the biosynthetic cluster. DpgC is an unusual metal-free, cofactor-free enzyme that consumes O2 during the conversion of 3,5-dihydroxyphenylacetyl-CoA (DPA-CoA) to the penultimate intermediate 3,5-dihydroxyphenylglyoxylate (DPGx). We show that in anaerobic incubations, DpgC catalyzes the exchange of the C2-methylene hydrogens of DPA-CoA at unequal rates, consistent with enzyme-mediated formation of the substrate-derived C2-carbanion as an early intermediate. Incubations with 18O2 reveal that DpgC transfers both atoms of an O2 molecule to DPGx product. This establishes DpgC as a 1,2-dioxygenase that mediates thioester cleavage by the oxygen transfer process. These results are consistent with a DPA-CoA C2-peroxy intermediate, followed by enzyme-directed α-peroxylactone formation and collapse by O-O bond cleavage.