Bacterial Cytochrome P450-catalyzed Post-translational Macrocyclization

He, Bei-Bei; Cheng, Zhuo; Liu, Fei; Zhong, Zheng; Gao, Ying; Liu, Hongyan; Li, Yong-Xin

doi:10.1101/2023.05.08.539676

Cited by 3 publications

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

He,

Liu,

Cheng

et al. 2023

Angewandte Chemie

Self Cite

View full text Add to dashboard Cite

Ribosomally synthesized and post‐translationally modified peptides (RiPPs) are a fascinating group of natural products that exhibit diverse structural features and bioactivities. P450‐catalyzed RiPPs stand out as a unique but underexplored family. Here, we introduce a rule‐based genome mining strategy that harnesses the intrinsic biosynthetic principles of RiPPs, including the co‐occurrence, co‐conservation, and interactions between precursors and P450s, successfully facilitating the identification of diverse P450 catalyzed RiPPs. Intensive BGC characterization revealed four new P450s, KstB, ScnB, MciB, and SgrB, that can respectively catalyze Trp‐Trp‐Tyr (one C‐C and two C‐N bonds), Tyr‐Trp (C‐C bond), Trp‐Trp (C‐N bond), and His‐His (ether bond) crosslinks within three or four residues. KstB, ScnB, and MciB could accept non‐native precursors, suggesting they could be promising starting templates for bioengineering to construct macrocycles. Our study highlights the potential of P450s in expanding the chemical diversity of strained macrocyclic peptides and enriching biocatalytic tools for peptide macrocyclization.

show abstract

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

He,

Liu,

Cheng

et al. 2023

Angewandte Chemie

Self Cite

View full text Add to dashboard Cite

show abstract

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

He,

Liu,

Cheng

et al. 2023

Angew Chem Int Ed

Self Cite

View full text Add to dashboard Cite

Ribosomally synthesized and post‐translationally modified peptides (RiPPs) are a fascinating group of natural products that exhibit diverse structural features and bioactivities. P450‐catalyzed RiPPs stand out as a unique but underexplored family. Herein, we introduce a rule‐based genome mining strategy that harnesses the intrinsic biosynthetic principles of RiPPs, including the co‐occurrence and co‐conservation of precursors and P450s and interactions between them, successfully facilitating the identification of diverse P450‐catalyzed RiPPs. Intensive BGC characterization revealed four new P450s, KstB, ScnB, MciB, and SgrB, that can catalyze the formation of Trp‐Trp‐Tyr (one C−C and two C−N bonds), Tyr‐Trp (C−C bond), Trp‐Trp (C−N bond), and His‐His (ether bond) crosslinks, respectively, within three or four residues. KstB, ScnB, and MciB could accept non‐native precursors, suggesting they could be promising starting templates for bioengineering to construct macrocycles. Our study highlights the potential of P450s to expand the chemical diversity of strained macrocyclic peptides and the range of biocatalytic tools available for peptide macrocyclization.

show abstract

Biosynthesis of macrocyclic peptides with C-terminal β-amino-α-keto acid groups by three different metalloenzymes

Nguyen,

Zhu,

Gray

et al. 2023

Preprint

View full text Add to dashboard Cite

Advances in genome sequencing and bioinformatics methods have identified myriad biosynthetic gene clusters (BGCs) encoding uncharacterized molecules. By examining genomic databases for BGCs containing a prevalent peptide-binding domain used for the biosynthesis of approximately half of ribosomally synthesized and post-translationally modified peptides (RiPPs), we uncovered a new class involving modifications installed by a cytochrome P450, a multi-nuclear iron-dependent non-heme oxidative enzyme (MNIO, formerly DUF692), a cobalamin- and radicalS-adenosyl-L-methioninedependent enzyme (B12-rSAM), and a methyltransferase. The activity of each enzyme was reconstitutedin vivousingBurkholderiasp. FERM BP-3421. Structural characterization demonstrated that the P450 catalyzed a biaryl C-C crosslink formation between two Tyr residues. The B12-rSAM generated β-methyltyrosine, while the MNIO transformed a C-terminal Asp residue into a C-terminal aminopyruvic acid. The methyltransferase acted on the β-carbon of the α-keto acid. Marfey’s method and exciton-coupled circular dichroism spectroscopy were used to elucidate the stereochemical configurations for the chiral carbons and the atropisomer that formed upon biaryl crosslinking. To the best of our knowledge, the MNIO featured in this biosynthetic pathway is the first to act on a non-Cys residue. Our study underscores that the pace of discovery of new macrocyclic peptides deriving from ribosomal peptides continues to accelerate and that RiPP BGCs remain a significant source of previously undiscovered enzyme chemistry.

show abstract

Bacterial Cytochrome P450-catalyzed Post-translational Macrocyclization

Cited by 3 publications

References 35 publications

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

Bacterial Cytochrome P450 Catalyzed Post‐translational Macrocyclization of Ribosomal Peptides**

Biosynthesis of macrocyclic peptides with C-terminal β-amino-α-keto acid groups by three different metalloenzymes

Contact Info

Product

Resources

About