Processing and secretion of non-cognate bacteriocins by EnkT, an ABC transporter from a multiple-bacteriocin producer, Enterococcus faecium NKR-5-3
Section snippets
Bacterial strains, plasmids, and culture conditions
The strains used in this study are listed in Table 1. The heterologous expression host strain, Lactococcus lactis NZ9000 was cultured in M17 medium (Merck, Darmstadt, Germany) supplemented with 0.5 % glucose (Nacalai Tesque, Kyoto, Japan) (GM17) and propagated at 30°C. The indicator strains, E. faecalis JCM 5803T and L. lactis subsp. lactis IL1403 were cultured in MRS broth (Oxoid, Basingstoke, UK) at 37°C, and L. lactis subsp. lactis ATCC 19435T and Lactobacillus sakei subsp. sakei JCM 1157T
Production of non-cognate bacteriocins by EnkT
EnkT is an ABC transporter protein, which is responsible for the processing and secretion of multiple bacteriocins belonging to different subclasses. This allowed us to evaluate the capacity of EnkT to process and secrete non-cognate bacteriocins. Sequence analysis of the non-cognate precursor peptides of EntA, PedA, LcnA, and LaqQ did not reveal the presence of significantly conserved motifs in their leader peptide moieties relative to their Ent53 bacteriocin counterparts except the presence
Discussion
In this present study, we demonstrated the wide substrate tolerance of EnkT for processing different non-cognate bacteriocin precursor peptides to yield their respective mature and active forms. The heterologous non-cognate bacteriocin precursor peptides despite significantly varying in both, the length and the sequence, were still successfully processed and secreted by EnkT (13). Although, previous studies have already described about some ABC transporters that can process heterologous
Acknowledgments
This study was supported in part by the Grants-in-Aid for Scientific Research provided by the Japan Society for the Promotion of Science (JSPS KAKENHI, Grant Number JP17H03797) and the “Innovation inspired by Nature” Research Support Program, Sekisui Chemical Co., Ltd.
References (36)
- et al.
Further observations on an inhibitory substance (nisin) from lactic streptococci
Lancet
(1947) - et al.
Monitoring of the multiple bacteriocin production by Enterococcus faecium NKR-5-3 through a developed liquid chromatography and mass spectrometry-based quantification system
J. Biosci. Bioeng.
(2012) - et al.
Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter
EnkT. J. Biosci. Bioeng.
(2018) - et al.
Purification and primary structure of pediocin PA-1 produced by Pediococcus acidilactici PAC-1.0
Arch. Biochem. Biophys.
(1992) - et al.
Characterisation of the action mechanism of a Lactococcus-specific bacteriocin
lactococcin Z. J. Biosci. Bioeng.
(2018) - et al.
Utilization of the leucocin A export system in Leuconostoc gelidum for production of a Lactobacillus bacteriocin
FEMS Microbiol. Lett.
(1995) - et al.
NisT, the transporter of the lantibiotic nisin, can transport fully modified, dehydrated, and unmodified prenisin and fusions of the leader peptide with non-lantibiotic peptides
J. Biol. Chem.
(2004) - et al.
Processing and export of peptide pheromones and bacteriocins in Gram-negative bacteria
Trends Microbiol.
(2001) - et al.
Novel bacteriocins from lactic acid bacteria (LAB): various structures and applications
Microb. Cell Fact.
(2014) - et al.
Bacteriocins: developing innate immunity for food
Nat. Rev. Microbiol.
(2005)
Circular and leaderless bacteriocins: biosynthesis, mode of action, applications, and prospects
Front. Microbiol.
The expanding structure variety among bacteriocins from Gram-positive bacteria
FEMS Microbiol. Rev.
Purification and characterization of multiple bacteriocins and an inducing peptide produced by Enterococcus faecium NKR-5-3 from Thai fermented fish
Biosci. Biotechnol. Biochem.
Identification of enterocin NKR-5-3C, a novel class IIa bacteriocin produced by a multiple bacteriocin producer, Enterococcus faecium NKR-5-3
Biosci. Biotechnol. Biochem.
Identification, characterization, and the three-dimensional structure of the novel circular bacteriocin, enterocin NKR-5-3B, from Enterococcus faecium
Biochemistry
Gene cluster responsible for secretion of and immunity to multiple bacteriocins, the NKR-5-3 enterocins
Appl. Environ. Microbiol.
Functional analysis of the genes involved in the biosynthesis of enterocin NKR-5-3B, a novel circular bacteriocin
J. Bacteriol.
Mutations near the cleavage site of enterocin NKR-5-3B prepeptide reveal new insights into its biosynthesis
Microbiology
Cited by (1)
Multiple bacteriocin production in lactic acid bacteria
2022, Journal of Bioscience and BioengineeringCitation Excerpt :The capacity of EnkT to process the heterogenous module 1 bacteriocins of strain NKR-5-3 contradicts the common understanding that each bacteriocin requires a dedicated ABC-transporter for its processing and secretion. The unique capacity of EnkT in processing heterologous bacteriocin precursor peptides as its substrates was extensively investigated by Sushida et al. (51,58). It was demonstrated that the bioactivity of EnkT requires a specific structural confirmation of the leader peptide rather than specific residues in recognizing and processing of its substrate (bacteriocin precursor peptide).