Altering glycopeptide antibiotic biosynthesis through mutasynthesis allows incorporation of fluorinated phenylglycine residues.

Autor:	Voitsekhovskaia I; Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, University of Tübingen Tübingen Germany evi.stegmann@uni-tuebingen.de., Ho YTC; Department of Biochemistry and Molecular Biology, The Monash Biomedicine Discovery Institute, Monash University Clayton VIC 3800 Australia max.cryle@monash.edu.; EMBL Australia, Monash University Clayton VIC 3800 Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia., Klatt C; Institute of Organic Chemistry, University of Tübingen Tübingen Germany., Müller A; Institute for Pharmaceutical Microbiology, University Hospital Bonn, University of Bonn Bonn Germany., Machell DL; Department of Biochemistry and Molecular Biology, The Monash Biomedicine Discovery Institute, Monash University Clayton VIC 3800 Australia max.cryle@monash.edu.; EMBL Australia, Monash University Clayton VIC 3800 Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia., Tan YJ; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia.; Research School of Chemistry, The Australian National University Acton ACT 2601 Australia., Triesman M; Department of Biochemistry and Molecular Biology, The Monash Biomedicine Discovery Institute, Monash University Clayton VIC 3800 Australia max.cryle@monash.edu.; EMBL Australia, Monash University Clayton VIC 3800 Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia., Bingel M; Institute for Pharmaceutical Microbiology, University Hospital Bonn, University of Bonn Bonn Germany., Schittenhelm RB; Monash Proteomics and Metabolomics Platform, Monash University Clayton VIC 3800 Australia., Tailhades J; Department of Biochemistry and Molecular Biology, The Monash Biomedicine Discovery Institute, Monash University Clayton VIC 3800 Australia max.cryle@monash.edu.; EMBL Australia, Monash University Clayton VIC 3800 Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia., Kulik A; Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, University of Tübingen Tübingen Germany evi.stegmann@uni-tuebingen.de., Maier ME; Institute of Organic Chemistry, University of Tübingen Tübingen Germany., Otting G; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia.; Research School of Chemistry, The Australian National University Acton ACT 2601 Australia., Wohlleben W; Microbiology/Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, University of Tübingen Tübingen Germany., Schneider T; Institute of Organic Chemistry, University of Tübingen Tübingen Germany.; Institute for Pharmaceutical Microbiology, University Hospital Bonn, University of Bonn Bonn Germany., Cryle M; Department of Biochemistry and Molecular Biology, The Monash Biomedicine Discovery Institute, Monash University Clayton VIC 3800 Australia max.cryle@monash.edu.; EMBL Australia, Monash University Clayton VIC 3800 Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia., Stegmann E; Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, University of Tübingen Tübingen Germany evi.stegmann@uni-tuebingen.de.; ARC Centre of Excellence for Innovations in Peptide and Protein Science Australia.; German Centre for Infection Research (DZIF), Partner Site Tübingen Tübingen Germany.; Cluster of Excellence 'Controlling Microbes to Fight Infections' (CMFI), University of Tübingen Tübingen Germany.
Jazyk:	angličtina
Zdroj:	RSC chemical biology [RSC Chem Biol] 2024 Aug 12. Date of Electronic Publication: 2024 Aug 12.
DOI:	10.1039/d4cb00140k
Abstrakt:	Glycopeptide antibiotics (GPAs) are peptide natural products used as last resort treatments for antibiotic resistant bacterial infections. They are produced by the sequential activities of a linear nonribosomal peptide synthetase (NRPS), which assembles the heptapeptide core of GPAs, and cytochrome P450 (Oxy) enzymes, which perform a cascade of cyclisation reactions. The GPAs contain proteinogenic and nonproteinogenic amino acids, including phenylglycine residues such as 4-hydroxyphenylglycine (Hpg). The ability to incorporate non-proteinogenic amino acids in such peptides is a distinctive feature of the modular architecture of NRPSs, with each module selecting and incorporating a desired amino acid. Here, we have exploited this ability to produce and characterise GPA derivatives containing fluorinated phenylglycine (F-Phg) residues through a combination of mutasynthesis, biochemical, structural and bioactivity assays. Our data indicate that the incorporation of F-Phg residues is limited by poor acceptance by the NRPS machinery, and that the phenol moiety normally present on Hpg residues is essential to ensure both acceptance by the NRPS and the sequential cyclisation activity of Oxy enzymes. The principles learnt here may prove useful for the future production of GPA derivatives with more favourable properties through mixed feeding mutasynthesis approaches. Competing Interests: There are no conflicts to declare. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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