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Flexizyme-Enabled Benchtop Biosynthesis of Thiopeptides.

ABSTRACT: Thiopeptides are natural antibiotics that are fashioned from short peptides by multiple layers of post-translational modification. Their biosynthesis, in particular the pyridine synthases that form the macrocyclic antibiotic core, has attracted intensive research but is complicated by the challenges of reconstituting multiple-pathway enzymes. By combining select RiPP enzymes with cell free expression and flexizyme-based codon reprogramming, we have developed a benchtop biosynthesis of thiopeptide scaffolds. This strategy side-steps several challenges related to the investigation of thiopeptide enzymes and allows access to analytical quantities of new thiopeptide analogs. We further demonstrate that this strategy can be used to validate the activity of new pyridine synthases without the need to reconstitute the cognate prior pathway enzymes.

SUBMITTER: Fleming SR

PROVIDER: S-EPMC6642631 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Flexizyme-Enabled Benchtop Biosynthesis of Thiopeptides.

Fleming Steven R SR Bartges Tessa E TE Vinogradov Alexander A AA Kirkpatrick Christine L CL Goto Yuki Y Suga Hiroaki H Hicks Leslie M LM Bowers Albert A AA

Journal of the American Chemical Society 20190108 2

Thiopeptides are natural antibiotics that are fashioned from short peptides by multiple layers of post-translational modification. Their biosynthesis, in particular the pyridine synthases that form the macrocyclic antibiotic core, has attracted intensive research but is complicated by the challenges of reconstituting multiple-pathway enzymes. By combining select RiPP enzymes with cell free expression and flexizyme-based codon reprogramming, we have developed a benchtop biosynthesis of thiopeptid ...[more]

PMID: 30602112

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