Carrier Protein‐Free Enzymatic Biaryl Coupling in Arylomycin A2 Assembly and Structure of the Cytochrome P450 AryC. Issue 2 (26th November 2021)

• Record Type: Journal Article
• Authors: Aldemir, Hülya; Shu, Shuangjie; Schaefers, Francoise; Hong, Hanna; Richarz, René; Harteis, Sabrina; Einsiedler, Manuel; Milzarek, Tobias M.; Schneider, Sabine; Gulder, Tobias A. M.
• Is Part Of: Chemistry. Volume 28:Issue 2(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-11-26
• Subjects: arylomycin -- biaryl coupling -- chemo-enzymatic synthesis -- crystal structure -- cytochrome P450; Chemistry -- Periodicals; 540
• DOI: 10.1002/chem.202103389
• Languages: English
• ISSNs: 0947-6539

Abstract

Abstract: The arylomycin antibiotics are potent inhibitors of bacterial type I signal peptidase. These lipohexapeptides contain a biaryl structural motif reminiscent of glycopeptide antibiotics. We herein describe the functional and structural evaluation of AryC, the cytochrome P450 performing biaryl coupling in biosynthetic arylomycin assembly. Unlike its enzymatic counterparts in glycopeptide biosynthesis, AryC converts free substrates without the requirement of any protein interaction partner, likely enabled by a strongly hydrophobic cavity at the surface of AryC pointing to the substrate tunnel. This activity enables chemo‐enzymatic assembly of arylomycin A2 that combines the advantages of liquid‐ and solid‐phase peptide synthesis with late‐stage enzymatic cross‐coupling. The reactivity of AryC is unprecedented in cytochrome P450‐mediated biaryl construction in non‐ribosomal peptides, in which peptidyl carrier protein (PCP)‐tethering so far was shown crucial both in vivo and in vitro. Abstract : The arylomycins are antibiotic natural products that inhibit bacterial signal peptidase. Within this work, the biaryl‐bond forming cytochrome P450 of arylomycin biosynthesis, AryC, is structurally and functionally characterized. Convergent preparation of the linear lipopeptide precursor followed by enzymatic installation of the biaryl structural element by AryC results in a streamlined chemo‐enzymatic access to arylomycin A2. Unlike oxidative cross‐coupling enzymes in glycopeptide biosynthesis, AryC accepts carrier protein‐free peptide substrates, a tremendous advantage for future biocatalytic applications in complex peptide synthesis.