A family of diiron monooxygenases catalyzing amino acid beta-hydroxylation in antibiotic biosynthesis

Thomas M. Makris, Mrinmoy Chakrabarti, Eckard Münck, John D. Lipscomb

Research output: Contribution to journalArticlepeer-review

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The biosynthesis of chloramphenicol requires a β-hydroxylation tailoring reaction of the precursor L-p-aminophenylalanine (L-PAPA). Here, it is shown that this reaction is catalyzed by the enzyme CmlA from an operon containing the genes for biosynthesis of L-PAPAand the nonribosomal peptide synthetase CmlP. EPR, Mössbauer, and optical spectroscopies reveal that CmlA contains an oxo-bridged dinuclear iron cluster, a metal center not previously associated with nonribosomal peptide synthetase chemistry. Single-turnover kinetic studies indicate that CmlA is functional in the diferrous state and that its substrate is L-PAPA covalently bound to CmlP. Analytical studies show that the product is hydroxylated L-PAPA and that O 2 is the oxygen source, demonstrating a monooxygenase reaction. The gene sequence of CmlA shows that it utilizes a lactamase fold, suggesting that the diiron cluster is in a protein environment not previously known to effect monooxygenase reactions. Notably, CmlA homologs are widely distributed in natural product biosynthetic pathways, including a variety of pharmaceutically important beta-hydroxylated antibiotics and cytostatics.

Original languageEnglish (US)
Pages (from-to)15391-15396
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number35
StatePublished - Aug 31 2010


  • Beta-hydroxylation
  • Dinuclear iron cluster
  • Nonheme oxygenase
  • Nonribosomal peptide synthetase
  • Spectroscopy

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