Engineering of acyl ligase domain in non-ribosomal peptide synthetases to change fatty acid moieties of lipopeptides

• Published in: Communications chemistry Vol. 8; no. 1; pp. 17 - 14
• Main Authors: Aoki, Rina, Kumagawa, Eri, Kamata, Kazuaki, Ago, Hideo, Sakai, Naoki, Hasunuma, Tomohisa, Taoka, Naoaki, Ohta, Yukari, Kobayashi, Shingo
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21.01.2025; Nature Publishing Group; Nature Portfolio
• Subjects: 101/58; 631/45/611; 631/92/60/1167; 639/638/77/603; 639/638/92/349; 82/47; Amino acids; Antimicrobial agents; Binding; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Fatty acids; In vivo methods and tests; Mutation; Peptides
• DOI: 10.1038/s42004-024-01379-w
• ISSN: 2399-3669

Cyclic lipopeptides (CLPs) produced by the genus Bacillus are amphiphiles composed of hydrophilic amino acid and hydrophobic fatty acid moieties and are biosynthesised by non-ribosomal peptide synthetases (NRPSs). CLPs are produced as a mixture of homologues with different fatty acid moieties, whose length affects CLP activity. Iturin family lipopeptides are a family of CLPs comprising cyclic heptapeptides and β-amino fatty acids and have antimicrobial activity. There is little research on how the length of the fatty acid moiety of iturin family lipopeptides is determined. Here, we demonstrated that the acyl ligase (AL) domain determines the length of the fatty acid moiety in vivo. In addition, enzyme assays revealed how mutations in the substrate-binding pocket of the AL domain affected substrate specificity in vitro. Our findings have implications for the design of fatty acyl moieties for CLP synthesis using NRPS. Iturin antimicrobial lipopeptides are a family of cyclic lipopeptides (CLPs) biosynthesized by non-ribosomal peptide synthetases (NRPSs) and comprising cyclic heptapeptides and β-amino fatty acids, however, determination of the length of the fatty acid moiety is underexplored. Here, the authors demonstrate in vivo that the acyl ligase (AL) domain in NRPS determines the length of the fatty acid moiety, and show in vitro that the mutations in the substrate-binding pocket of the AL domain affect substrate specificity.