cs0c01719_si_001.pdf (29.92 MB)
Mapping and Exploiting the Promiscuity of OxyB toward the Biocatalytic Production of Vancomycin Aglycone Variants
journal contribution
posted on 2020-08-05, 17:06 authored by Clarissa
C. Forneris, Andy K. L. Nguy, Mohammad R. SeyedsayamdostVancomycin is one
of the most important clinical antibiotics in
the fight against infectious disease. Its biological activity relies
on three aromatic cross-links, which create a cup-shaped topology
and allow tight binding to nascent peptidoglycan chains. The cytochrome
P450 enzymes OxyB, OxyA, and OxyC have been shown to introduce these
synthetically challenging aromatic linkages. The ability to utilize
the P450 enzymes in a chemo-enzymatic scheme to generate vancomycin
derivatives is appealing but requires a thorough understanding of
their reactivities and mechanisms. Herein, we systematically explore
the scope of OxyB biocatalysis and report installation of diverse
diaryl ether and biaryl cross-links with varying macrocycle sizes
and compositions, when the enzyme is presented with modified vancomycin
precursor peptides. The structures of the resulting products were
determined using one-dimensional/two-dimensional nuclear magnetic
resonance spectroscopy, high-resolution mass spectrometry (HR-MS),
tandem HR-MS, and isotopic labeling, as well as ultraviolet–visible
light absorption and fluorescence emission spectroscopies. An exploration
of the biological activities of these alternative OxyB products surprisingly
revealed antifungal properties. Taking advantage of the promiscuity
of OxyB, we chemo-enzymatically generated a vancomycin aglycone variant
containing an expanded macrocycle. Mechanistic implications for OxyB
and future directions for creating vancomycin analogue libraries are
discussed.
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Keywords
antifungal propertiesdiaryl etherBiocatalytic Productionvancomycin analogue librariesHR-MSmacrocycle sizesfuture directionsOxyB biocatalysismass spectrometryVancomycin Aglycone Variants Vancomycinbiaryl cross-linkschemo-enzymatic schemecytochrome P 450 enzymes OxyBvancomycin aglycone variantresonance spectroscopypeptidoglycan chainsreport installationMechanistic implicationsalternative OxyB productsvancomycin derivativesfluorescence emission spectroscopiescup-shaped topologyvancomycin precursor peptidesP 450 enzymes
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