Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
Enterocin F4-9 belongs to the glycocin family having post-translational modifications by two molecules of <i>N</i>-acetylglucosamine β-<i>O</i>-linked to Ser37 and Thr46. In this study, the biosynthetic gene cluster of enterocin F4-9 was cloned and expressed in <i>Enter...
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| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/462444ca16cc4e758c49ce9ed72e0f70 |
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| Sumario: | Enterocin F4-9 belongs to the glycocin family having post-translational modifications by two molecules of <i>N</i>-acetylglucosamine β-<i>O</i>-linked to Ser37 and Thr46. In this study, the biosynthetic gene cluster of enterocin F4-9 was cloned and expressed in <i>Enterococcus faecalis</i> JH2-2. Production of glycocin by the JH2-2 expression strain was confirmed by expression of the five genes. The molecular weight was greater than glycocin secreted by the wild strain, <i>E. faecalis</i> F4-9, because eight amino acids from the N-terminal leader sequence remained attached. This N-terminal extension was eliminated after treatment with the culture supernatant of strain F4-9, implying an extracellular protease from <i>E</i>. <i>faecalis</i> F4-9 cleaves the N-terminal sequence. Thus, leader sequences cleavage requires two steps: the first via the EnfT protease domain and the second via extracellular proteases. Interestingly, the long peptide, with N-terminal extension, demonstrated advanced antimicrobial activity against Gram-positive and Gram-negative bacteria. Furthermore, <i>enfC</i> was responsible for glycosylation, a necessary step prior to secretion and cleavage of the leader peptide. In addition, <i>enfI</i> was found to grant self-immunity to producer cells against enterocin F4-9. This report demonstrates specifications of the minimal gene set responsible for production of enterocin F4-9, as well as a new biosynthetic mechanism of glycocins. |
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