Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT

Hirotoshi Sushida, Naoki Ishibashi, Takeshi Zendo, Pongtep Wilaipun, Vichien Leelawatcharamas, Jiro Nakayama, Kenji Sonomoto

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

EnkT is a novel ATP-binding cassette (ABC) transporter responsible for secretion of four bacteriocins, enterocins NKR-5-3A, C, D, and Z (Ent53A, C, D, and Z), produced by Enterococcus faecium NKR-5-3. It is generally recognized that the secretion of a bacteriocin requires a dedicated ABC transporter, although molecular mechanisms of this secretion are yet to be revealed. In order to characterize the unique ability of EnkT to secrete multiple bacteriocins, the role of N-terminal leader peptides of bacteriocin precursors was evaluated using Ent53C precursor as a model. The 18-amino acid leader peptide of Ent53C (Lc) was modified by site-directed mutagenesis to generate various point mutations, truncations, or extensions, and substitutions with other leader peptides. The impact of these Lc mutations on Ent53C secretion was evaluated using a quantitative antimicrobial activity assay. We observed that Ent53C production increased with Ala substitution of the highly conserved C-terminal double glycine residues that are recognized as the cleavage site. In contrast, Ent53C antimicrobial activity decreased, with decrease in the length of the putative α-helix-forming region of Lc. Furthermore, EnkT recognized and transported Ent53C of the transformants possessing heterologous leader peptides of enterocin A, pediocin PA-1, brochocins A and B, and lactococcins Qα and Qβ. These results indicated that EnkT shows significant tolerance towards the sequence and length of leader peptides, to secrete multiple bacteriocins. This further demonstrates the functional diversity of bacteriocin ABC transporters and the importance of leader peptides as their recognition motif.

元の言語英語
ページ(範囲)23-29
ページ数7
ジャーナルJournal of Bioscience and Bioengineering
126
発行部数1
DOI
出版物ステータス出版済み - 7 1 2018

Fingerprint

Bacteriocins
Protein Sorting Signals
ATP-Binding Cassette Transporters
Adenosinetriphosphate
Amino acids
Substitution reactions
Enterococcus faecium
Mutagenesis
Site-Directed Mutagenesis
Point Mutation
Glycine
Assays
Amino Acids
Mutation

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

これを引用

Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT. / Sushida, Hirotoshi; Ishibashi, Naoki; Zendo, Takeshi; Wilaipun, Pongtep; Leelawatcharamas, Vichien; Nakayama, Jiro; Sonomoto, Kenji.

:: Journal of Bioscience and Bioengineering, 巻 126, 番号 1, 01.07.2018, p. 23-29.

研究成果: ジャーナルへの寄稿記事

Sushida, Hirotoshi ; Ishibashi, Naoki ; Zendo, Takeshi ; Wilaipun, Pongtep ; Leelawatcharamas, Vichien ; Nakayama, Jiro ; Sonomoto, Kenji. / Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT. :: Journal of Bioscience and Bioengineering. 2018 ; 巻 126, 番号 1. pp. 23-29.
@article{65762f3595ec4306abb462450a212dcb,
title = "Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT",
abstract = "EnkT is a novel ATP-binding cassette (ABC) transporter responsible for secretion of four bacteriocins, enterocins NKR-5-3A, C, D, and Z (Ent53A, C, D, and Z), produced by Enterococcus faecium NKR-5-3. It is generally recognized that the secretion of a bacteriocin requires a dedicated ABC transporter, although molecular mechanisms of this secretion are yet to be revealed. In order to characterize the unique ability of EnkT to secrete multiple bacteriocins, the role of N-terminal leader peptides of bacteriocin precursors was evaluated using Ent53C precursor as a model. The 18-amino acid leader peptide of Ent53C (Lc) was modified by site-directed mutagenesis to generate various point mutations, truncations, or extensions, and substitutions with other leader peptides. The impact of these Lc mutations on Ent53C secretion was evaluated using a quantitative antimicrobial activity assay. We observed that Ent53C production increased with Ala substitution of the highly conserved C-terminal double glycine residues that are recognized as the cleavage site. In contrast, Ent53C antimicrobial activity decreased, with decrease in the length of the putative α-helix-forming region of Lc. Furthermore, EnkT recognized and transported Ent53C of the transformants possessing heterologous leader peptides of enterocin A, pediocin PA-1, brochocins A and B, and lactococcins Qα and Qβ. These results indicated that EnkT shows significant tolerance towards the sequence and length of leader peptides, to secrete multiple bacteriocins. This further demonstrates the functional diversity of bacteriocin ABC transporters and the importance of leader peptides as their recognition motif.",
author = "Hirotoshi Sushida and Naoki Ishibashi and Takeshi Zendo and Pongtep Wilaipun and Vichien Leelawatcharamas and Jiro Nakayama and Kenji Sonomoto",
year = "2018",
month = "7",
day = "1",
doi = "10.1016/j.jbiosc.2018.01.015",
language = "English",
volume = "126",
pages = "23--29",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "The Society for Biotechnology, Japan",
number = "1",

}

TY - JOUR

T1 - Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT

AU - Sushida, Hirotoshi

AU - Ishibashi, Naoki

AU - Zendo, Takeshi

AU - Wilaipun, Pongtep

AU - Leelawatcharamas, Vichien

AU - Nakayama, Jiro

AU - Sonomoto, Kenji

PY - 2018/7/1

Y1 - 2018/7/1

N2 - EnkT is a novel ATP-binding cassette (ABC) transporter responsible for secretion of four bacteriocins, enterocins NKR-5-3A, C, D, and Z (Ent53A, C, D, and Z), produced by Enterococcus faecium NKR-5-3. It is generally recognized that the secretion of a bacteriocin requires a dedicated ABC transporter, although molecular mechanisms of this secretion are yet to be revealed. In order to characterize the unique ability of EnkT to secrete multiple bacteriocins, the role of N-terminal leader peptides of bacteriocin precursors was evaluated using Ent53C precursor as a model. The 18-amino acid leader peptide of Ent53C (Lc) was modified by site-directed mutagenesis to generate various point mutations, truncations, or extensions, and substitutions with other leader peptides. The impact of these Lc mutations on Ent53C secretion was evaluated using a quantitative antimicrobial activity assay. We observed that Ent53C production increased with Ala substitution of the highly conserved C-terminal double glycine residues that are recognized as the cleavage site. In contrast, Ent53C antimicrobial activity decreased, with decrease in the length of the putative α-helix-forming region of Lc. Furthermore, EnkT recognized and transported Ent53C of the transformants possessing heterologous leader peptides of enterocin A, pediocin PA-1, brochocins A and B, and lactococcins Qα and Qβ. These results indicated that EnkT shows significant tolerance towards the sequence and length of leader peptides, to secrete multiple bacteriocins. This further demonstrates the functional diversity of bacteriocin ABC transporters and the importance of leader peptides as their recognition motif.

AB - EnkT is a novel ATP-binding cassette (ABC) transporter responsible for secretion of four bacteriocins, enterocins NKR-5-3A, C, D, and Z (Ent53A, C, D, and Z), produced by Enterococcus faecium NKR-5-3. It is generally recognized that the secretion of a bacteriocin requires a dedicated ABC transporter, although molecular mechanisms of this secretion are yet to be revealed. In order to characterize the unique ability of EnkT to secrete multiple bacteriocins, the role of N-terminal leader peptides of bacteriocin precursors was evaluated using Ent53C precursor as a model. The 18-amino acid leader peptide of Ent53C (Lc) was modified by site-directed mutagenesis to generate various point mutations, truncations, or extensions, and substitutions with other leader peptides. The impact of these Lc mutations on Ent53C secretion was evaluated using a quantitative antimicrobial activity assay. We observed that Ent53C production increased with Ala substitution of the highly conserved C-terminal double glycine residues that are recognized as the cleavage site. In contrast, Ent53C antimicrobial activity decreased, with decrease in the length of the putative α-helix-forming region of Lc. Furthermore, EnkT recognized and transported Ent53C of the transformants possessing heterologous leader peptides of enterocin A, pediocin PA-1, brochocins A and B, and lactococcins Qα and Qβ. These results indicated that EnkT shows significant tolerance towards the sequence and length of leader peptides, to secrete multiple bacteriocins. This further demonstrates the functional diversity of bacteriocin ABC transporters and the importance of leader peptides as their recognition motif.

UR - http://www.scopus.com/inward/record.url?scp=85041961405&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041961405&partnerID=8YFLogxK

U2 - 10.1016/j.jbiosc.2018.01.015

DO - 10.1016/j.jbiosc.2018.01.015

M3 - Article

VL - 126

SP - 23

EP - 29

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 1

ER -