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 N-acetylglucosamine β-O-linked to Ser37 and Thr46. In this study, the biosynthetic gene cluster of enterocin F4-9 was cloned and expressed in Enter...

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Autores principales:	Mohamed Abdelfattah Maky, Naoki Ishibashi, Jiro Nakayama, Takeshi Zendo
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	enterocin F4-9 glycocin bacteriocin biosynthesis glycosylation Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/462444ca16cc4e758c49ce9ed72e0f70
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spelling	oai:doaj.org-article:462444ca16cc4e758c49ce9ed72e0f702021-11-25T18:24:47ZCharacterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin10.3390/microorganisms91122762076-2607https://doaj.org/article/462444ca16cc4e758c49ce9ed72e0f702021-11-01T00:00:00Zhttps://www.mdpi.com/2076-2607/9/11/2276https://doaj.org/toc/2076-2607Enterocin 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.Mohamed Abdelfattah MakyNaoki IshibashiJiro NakayamaTakeshi ZendoMDPI AGarticleenterocin F4-9glycocinbacteriocinbiosynthesisglycosylationBiology (General)QH301-705.5ENMicroorganisms, Vol 9, Iss 2276, p 2276 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	enterocin F4-9 glycocin bacteriocin biosynthesis glycosylation Biology (General) QH301-705.5
spellingShingle	enterocin F4-9 glycocin bacteriocin biosynthesis glycosylation Biology (General) QH301-705.5 Mohamed Abdelfattah Maky Naoki Ishibashi Jiro Nakayama Takeshi Zendo Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
description	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.
format	article
author	Mohamed Abdelfattah Maky Naoki Ishibashi Jiro Nakayama Takeshi Zendo
author_facet	Mohamed Abdelfattah Maky Naoki Ishibashi Jiro Nakayama Takeshi Zendo
author_sort	Mohamed Abdelfattah Maky
title	Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
title_short	Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
title_full	Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
title_fullStr	Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
title_full_unstemmed	Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin
title_sort	characterization of the biosynthetic gene cluster of enterocin f4-9, a glycosylated bacteriocin
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/462444ca16cc4e758c49ce9ed72e0f70
work_keys_str_mv	AT mohamedabdelfattahmaky characterizationofthebiosyntheticgeneclusterofenterocinf49aglycosylatedbacteriocin AT naokiishibashi characterizationofthebiosyntheticgeneclusterofenterocinf49aglycosylatedbacteriocin AT jironakayama characterizationofthebiosyntheticgeneclusterofenterocinf49aglycosylatedbacteriocin AT takeshizendo characterizationofthebiosyntheticgeneclusterofenterocinf49aglycosylatedbacteriocin
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Characterization of the Biosynthetic Gene Cluster of Enterocin F4-9, a Glycosylated Bacteriocin

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