Facile Synthetic Access to Glycopeptide Antibiotic Precursor Peptides for the Investigation of Cytochrome P450 Action in Glycopeptide Antibiotic Biosynthesis

Clara Brieke, Veronika Kratzig, Madeleine Peschke, Max J Cryle

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

6 Citations (Scopus)


The glycopeptide antibiotics are an important class of complex, medically relevant peptide natural products. Given that the production of such compounds all stems from in vivo biosynthesis, understanding the mechanisms of the natural assembly system-consisting of a nonribosomal-peptide synthetase machinery (NRPS) and further modifying enzymes-is vital. In order to address the later steps of peptide biosynthesis, which are catalyzed by Cytochrome P450s that interact with the peptide-producing nonribosomal peptide synthetase, peptide substrates are required: these peptides must also be in a form that can be conjugated to carrier protein domains of the nonribosomal peptide synthetase machinery. Here, we describe a practical and effective route for the solid phase synthesis of glycopeptide antibiotic precursor peptides as their Coenzyme A (CoA) conjugates to allow enzymatic conjugation to carrier protein domains. This route utilizes Fmoc-chemistry suppressing epimerization of racemization-prone aryl glycine derivatives and affords high yields and excellent purities, requiring only a single step of simple solid phase extraction for chromatographic purification. With this, comprehensive investigations of interactions between various NRPS-bound substrates and Cytochrome P450s are enabled.
Original languageEnglish
Title of host publicationNonribosomal Peptide and Polyketide Biosynthesis
Subtitle of host publicationMethods and Protocols
EditorsBradley S Evans
Place of PublicationNew York NY USA
PublisherHumana Press
Number of pages18
ISBN (Electronic)9781493933754
ISBN (Print)9781493933730
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology: Springer Protocols
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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