RRC ID 55438
著者 Sushida H, Ishibashi N, Zendo T, Wilaipun P, Leelawatcharamas V, Nakayama J, Sonomoto K.
タイトル Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter, EnkT.
ジャーナル J Biosci Bioeng
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.
巻・号 126(1)
ページ 23-29
公開日 2018-7-1
DOI 10.1016/j.jbiosc.2018.01.015
PII S1389-1723(17)30929-5
PMID 29452934
MeSH ATP-Binding Cassette Transporters* / chemistry ATP-Binding Cassette Transporters* / genetics ATP-Binding Cassette Transporters* / metabolism Amino Acid Sequence Bacteriocins / metabolism* Biological Transport / genetics Enterococcus faecium / genetics Enterococcus faecium / metabolism Mutagenesis, Site-Directed Point Mutation / physiology Protein Precursors / chemistry Protein Precursors / genetics Protein Precursors / metabolism Protein Sorting Signals / genetics Protein Sorting Signals / physiology* Secretory Pathway / genetics
IF 2.032
引用数 6
リソース情報
一般微生物 JCM 5803 JCM 5804