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Download filePrecursor Manipulation in Glycopeptide Antibiotic Biosynthesis: Are β‑Amino Acids Compatible with the Oxidative Cyclization Cascade?
journal contribution
posted on 2018-04-30, 18:20 authored by Melanie Schoppet, Julien Tailhades, Ketav Kulkarni, Max J. CryleNatural
products such as the glycopeptide antibiotics (GPAs, including
vancomycin and teicoplanin) are of great pharmaceutical importance
due to their use against Gram-positive bacteria such as methicillin-resistant Staphylococcus aureus. GPAs are assembled in a complex
process based on nonribosomal peptide synthesis and late-stage, multistep
cross-linking of the linear heptapeptide performed by cytochrome P450
monooxygenases. These P450 enzymes demonstrate varying degrees of
substrate selectivity toward the linear peptide precursor, with limited
information available about their tolerance regarding modifications
to amino acid residues within the essential antibiotic core of the
GPA. In order to test the acceptance of altered residues by the P450-catalyzed
cyclization cascade, we have explored the use of β-amino acids
in both variable and highly conserved positions within GPA peptides.
Our results indicate that the incorporation of β-amino acids
at the C-terminus of the peptide leads to a dramatic reduction in
the efficiency of peptide cyclization by the P450s during GPA biosynthesis,
whereas replacement of residue 3 is well tolerated by the same enzymes.
These results show that maintaining the C-terminal 3,5-dihydroxyphenylglycine
residue is of key importance to maintain the efficiency of this complex
and essential enzymatic cross-linking process.
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methicillin-resistant Staphylococcus aureusGlycopeptide Antibiotic Biosynthesiscytochrome P 450 monooxygenasesresults showGram-positive bacteriaNatural productsP 450 enzymesP 450-catalyzed cyclization cascadepeptide cyclizationmultistep cross-linkingPrecursor Manipulationglycopeptide antibioticssubstrate selectivitypeptide precursorβ-importanceefficiencyGPA biosynthesisnonribosomal peptide synthesisacid residuesGPA peptidesP 450sresidue 3antibiotic coreOxidative Cyclization Cascadecross-linking process