Poster Presentation 8th Modern Solid Phase Peptide Synthesis & Its Applications Symposium 2022

In vitro chemoenzymatic approach reveals the recognition and cyclisation mechanism of biarylitide P450 (#148)

Yongwei Zhao 1 , Julien Tailhades 1 , Max Cryle 1
  1. Monash University, Melbourne, VICTORIA, Australia

Cyclic peptide natural products are a valuable source of biologically active molecules for clinical and industrial use. Ribosomally synthesised and post-translationally modified peptides (RiPPs) are examples of such products (e.g., nisin, microcins and polytheonamides) and are attracting significant interest due to their leader peptide-directed biosynthesis (LDB). Such carrier protein-free peptide maturation machinery provides a much simpler system compared to non-ribosomal peptide synthesis while being capable of introducing similar modifications, for example side-chain crosslinks catalysed by cytochrome P450 enzymes(1, 2). Recently, a unique family of RiPPs named biarylitides has been revealed, which achieve a rare, crosslinked structure via P450 catalysed peptide maturation using a 2 amino acid leader peptide(3). Here, we present a chemoenzymatic approach, utilising both solid phase peptide synthesis and P450-mediated cyclisation, to access various novel biarylitide analogues in vitro. The results, quantified via high resolution mass spectrometry (HRMS), indicate the enzymatic tolerance and hence the utility of this system aswell as revealing the mechanistic basis for substrate recognition and reaction mechanism, which offers a route to pre-functionalised biaryl building blocks.

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  3. 3. Zdouc MM, Alanjary MM, Zarazua GS, Maffioli SI, Crusemann M, Medema MH, et al. A biaryl-linked tripeptide from Planomonospora reveals a widespread class of minimal RiPP gene clusters. Cell Chem Biol. 2020.