Purification and Characterization of Multiple Bacteriocins and an Inducing Peptide Produced by<i>Enterococcus faecium</i>NKR-5-3 from Thai Fermented Fish

Ishibashi, Naoki; Himeno, Kunio; Fujita, Koji; Masuda, Yoshito; Pérez, Rodney; Zendo, Takeshi; Wilaipun, Pongtep; Leelawatcharamas, Vichien; Nakayama, Jiro; Sonomoto, Kenji

doi:10.1271/bbb.110972

Cited by 64 publications

(57 citation statements)

References 33 publications

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“…To determine the minimum inhibitory concentrations (MICs) of the Ent53B Leu1 derivatives, the bacteriocins were purified from their respective phenotypes following our previously reported purification method of NKR-5-3 enterocins [22,23] and were subsequently quantified using the BCA Protein Assay kit (Thermo Scientific). The MICs were determined using the spot-on-lawn assay, as described previously [21,22], using serial dilutions of the purified bacteriocins of known concentrations against a set of indicator bacteria.…”

Section: Rna Isolation and Semi-quantitative Reverse Transcription Pcmentioning

confidence: 99%

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Mutations near the cleavage site of enterocin NKR-5-3B prepeptide reveal new insights into its biosynthesis

et al. 2017

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Enterocin NKR-5-3B (Ent53B) is a 64-residue novel circular bacteriocin synthesized from an 87-residue prepeptide. Albeit through a still unknown mechanism, the EnkB1234 biosynthetic enzyme complex processes the prepeptide to yield its mature active, circular form. To gain insights into the key region/residue that plays a role in Ent53 maturation, several mutations near the cleavage site on the precursor peptide were generated. The interaction of the precursor peptide and EnkB1234 appeared to be hydrophobic in nature. At the Leu1 position, only mutations with helix structure-promoting hydrophobic residues (Ala, Ile, Val or Phe) were able to yield the mature Ent53B derivative. In this study, we also highlight the possible conformation-stabilizing role of the Ent53B leader peptide on the precursor peptide for its interaction with its biosynthetic enzyme complex. Any truncations of the leader peptide moiety interfered in the processing of the prepeptide. However, when propeptides of other circular bacteriocins (circularin A, leucocyclicin Q or lactocyclicin Q) were cloned at the C-terminus of the leader peptide, EnkB1234 could not process them to yield a mature bacteriocin. Taken together, these findings offer new perspectives in our understanding of the possible molecular mechanism of the biosynthesis of this circular bacteriocin. These new perspectives will help advance our current understanding to eventually elucidate circular bacteriocin biosynthesis. Understanding the biosynthetic mechanism of circular bacteriocins will materialize their application potential.

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Section: Rna Isolation and Semi-quantitative Reverse Transcription Pcmentioning

confidence: 99%

“…Enterocin NKR-5-3B (Ent53B) is a 64-amino acid novel circular bacteriocin produced by Enterococcus faecium NKR-5-3 [21][22][23][24]. The Ent53B structural gene, enkB, encodes an 87-amino acid prepeptide, which consists of a 23-amino acid leader peptide and a 64-amino acid propeptide domain.…”

Section: Introductionmentioning

confidence: 99%

Mutations near the cleavage site of enterocin NKR-5-3B prepeptide reveal new insights into its biosynthesis

et al. 2017

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“…The genome of Enterococcus faecium NKR-5-3, isolated from pla-ra Thai fermented fish, encodes 5 enterocins, NKR-5-3 -A, B, C, D and Z and produces at least four of them, that is NKR-5-3 -A, B, C and D. Enterocin NKR-5-3C was confirmed to be a class IIa bacteriocin which exhibits potent antilisterial activity. The other bacteriocins are proposed to represent different classes but further investigations are required to establish this definitively [16,17].…”

Section: The Continuing Search For Novel Bacteriocinsmentioning

confidence: 99%

Antimicrobial antagonists against food pathogens: a bacteriocin perspective

O’Connor

Ross

Hill

et al. 2015

Current Opinion in Food Science

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Efforts are continuing to find novel bacteriocins with enhanced specificity and potency. Traditional plating techniques are still being used for bacteriocin screening studies, however, the availability of ever more bacterial genome sequences and the use of in silico gene mining tools have revealed novel bacteriocin gene clusters that would otherwise have been overlooked. Furthermore, synthetic biology and bioengineering-based approaches are allowing scientists to harness existing and novel bacteriocin gene clusters through expression in different hosts and by enhancing functionalities. The same principles apply to bacteriocin producing probiotic cultures and their application to control pathogens in the gut. We can expect that the recent developments on bacteriocins from Lactic Acid Bacteria (LAB) described here will contribute greatly to increased commercialisation of bacteriocins in food systems.

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“…10) Purified enterocin y To whom correspondence should be addressed. Fax: +81-92-642-3019; E-mail: sonomoto@agr.kyushu-u.ac.jp NKR-5-3C was analyzed by electrospray ionization time-of-flight mass spectrometry and the molecular weight was determined to be 4,512.8 Da.…”

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confidence: 99%

“…and other Gram-positive species with MICs in the nanomolar range, like other class IIa bacteriocins. 10) In this study, enterocin NKR-5-3C was further investigated for structure. Purified enterocin NKR-5-3C obtained by several runs of the four-step purification procedure was analyzed for N-terminal amino acid sequence by Edman degradation with protein sequencer model PPSQ-21 (Shimadzu, Kyoto, Japan).…”

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Identification of Enterocin NKR-5-3C, a Novel Class IIa Bacteriocin Produced by a Multiple Bacteriocin Producer, <i>Enterococcus faecium</i> NKR-5-3

Himeno

Fujita

Zendo

et al. 2012

Bioscience, Biotechnology, and Biochemistry

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Purification and Characterization of Multiple Bacteriocins and an Inducing Peptide Produced byEnterococcus faeciumNKR-5-3 from Thai Fermented Fish

Cited by 64 publications

References 33 publications

Mutations near the cleavage site of enterocin NKR-5-3B prepeptide reveal new insights into its biosynthesis

Mutations near the cleavage site of enterocin NKR-5-3B prepeptide reveal new insights into its biosynthesis

Antimicrobial antagonists against food pathogens: a bacteriocin perspective

Identification of Enterocin NKR-5-3C, a Novel Class IIa Bacteriocin Produced by a Multiple Bacteriocin Producer, <i>Enterococcus faecium</i> NKR-5-3

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