Probing the role of active site residues in NikD, an unusual amino acid oxidase that catalyzes an aromatization reaction important in nikkomycin biosynthesis.

ABSTRACT: NikD catalyzes a remarkable aromatization reaction that converts piperideine 2-carboxylate (P2C) to picolinate, a key component of the nonribosomal peptide in nikkomycin antibiotics. The enzyme exhibits a FAD-Trp355 charge-transfer band at weakly alkaline pH that is abolished upon protonation of an unknown ionizable residue that exhibits a pK(a) of 7.3. Stopped-flow studies of the reductive half-reaction with wild-type nikD and P2C show that the enzyme oxidizes the enamine tautomer of P2C but do not distinguish among several possible paths for the initial two-electron oxidation step. Replacement of Glu101 or Asp276 with a neutral residue does not eliminate the ionizable group, although the observed pK(a) is 1 or 2 pH units higher, respectively, compared with that of wild-type nikD. Importa

SUBMITTER: Kommoju PR 

PROVIDER: S-EPMC2764525 | biostudies-literature | 2009 Jul

REPOSITORIES: biostudies-literature