Carrier Protein‐Free Enzymatic Biaryl Coupling in Arylomycin A2 Assembly and Structure of the Cytochrome P450 AryC

Abstract

What is the most significant result of this study?Many potent glycopeptide antibiotics, such as vancomycin, contain biaryl structural features. These are biosynthetically introduced by dedicated cytochrome P450 enzymes, which require their substrate to be bound to a carrier protein as a precondition for substrate recognition. AryC, by contrast, accepts free, untethered substrates to install the biaryl bond in arylomycin antibiotics. It is thus a unique enzyme for biaryl peptide construction with huge potential for the streamlined chemo-enzymatic synthesis of peptide antibiotics.

Structural diversity, biosynthesis, and biological functions of lipopeptides fromStreptomyces

“…A cytochrome P450 protein (AryC) was recently conrmed to generate the biaryl bond, in a process that occurs aer cleavage of the peptide from the NRPS. 142 2.1.15 K97-0239s and enamidonin. K97-0239A and K97-0239B are cyclic lipopeptides isolated from Streptomyces sp.…”

“…A cytochrome P450 protein (AryC) was recently confirmed to generate the biaryl bond, in a process that occurs after cleavage of the peptide from the NRPS. 142…”

Non-Native Site-Selective Enzyme Catalysis

“…It was recently discovered that the enzyme AryC accomplishes a similar intramolecular C−C coupling reaction using only a lipophilic chain instead of a PCP tether to anchor the substrate within the enzyme active site. 368 Recently, a P450 from Streptomyces sp. MG-AR, originally identified for its ability to hydroxylate testosterone, was engineered to enable gram-scale oxidative coupling of the arylomycin core, demonstrating scalable production of this medicinally relevant motif (Scheme 55A).…”

Development of a P450 Fusion Enzyme for Biaryl Coupling in Yeast

“…16 Alternatively, P450s exert control over the bond formation step following the initial oxidation, providing a platform for site-and atroposelective biaryl coupling reactions. 17 Although a number of these P450s are carrier protein-dependent 18 or limited to intramolecular biaryl bond forming events (such as in the formation of 3), 19 a growing number of P450s have been identified to perform intermolecular oxidative biaryl coupling reactions. 13,20 Despite the prevalence of biaryl natural products in all kingdoms of life, to date, the only P450s that have been discovered to catalyze intermolecular biaryl coupling reactions are from microbial sources.…”

“…Alternatively, P450s exert control over the bond formation step following the initial oxidation, providing a platform for site- and atroposelective biaryl coupling reactions . Although a number of these P450s are carrier protein-dependent or limited to intramolecular biaryl bond forming events (such as in the formation of 3 ), a growing number of P450s have been identified to perform intermolecular oxidative biaryl coupling reactions. , Despite the prevalence of biaryl natural products in all kingdoms of life, to date, the only P450s that have been discovered to catalyze intermolecular biaryl coupling reactions are from microbial sources . Among these, fungal P450s have demonstrated particularly impressive control over the site- and atroposelective formation of biaryl metabolites. , However, the membrane-bound nature and catalytic-dependence on a separate reductase enzyme has limited the use of fungal P450s as biocatalysts in organic synthesis …”