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Bacteriocins: Academic Advances and Immediate Applications

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A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Antimicrobial Agents and Resistance".

Deadline for manuscript submissions: closed (31 December 2022) | Viewed by 28856

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Special Issue Editors

Prof. Dr. Djamel Drider

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Guest Editor

Unit of Research BIOECOAGRO INRA 1158, Lille University, 59655 Villeneuve d’Ascq, France
Interests: cheese; PCR; molecular biology; genetics; microbiology; antimicrobials; food safety; food microbiology; antibacterial activity; probiotics; mycology; salmonella; lactic acid bacteria; bacteriocins; listeria monocytogenes; antimicrobial peptides; foodborne pathogens; listeria
Special Issues, Collections and Topics in MDPI journals

Dr. Yanath Belguesmia

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Guest Editor

UMR Transfrontalière BioEcoAgro 1158, Université de Lille, Lille, France
Interests: bacteriocins; lactobacillales; biopreservatives

Special Issue Information

Dear Colleagues,

Bacteriocins are antimicrobial peptides ribosomally synthesized by Gram-negative bacteria, Gram-positive bacteria, and Archaea. Bacteriocins are molecules of major interest because of their potential applications in different fields such as food protection, veterinary medicine, human health, and plant protection. The application of bacteriocins is currently limited to nisin in the form of food additive E234. Nevertheless, the application of bacteriocins is foreseen to diversify and grow mainly in the medical area. Related to this point, bacteriocins offer many incentives as they can be used alternatives to antibiotics, or at least as potentiating agents to enhance the activity of aging antibiotics. This therapeutic application must take place immediately and will help to impede the antibiotic crisis from worsening. In the medical area, bacteriocins also have uses as anticancer or antiviral agents.

This Special Issue of Microorganisms will welcome original contributions, commentaries, and relevant reviews on bacteriocins. A balance between academic and applied papers will be privileged. Therefore, papers showing insightful data on mode of actions, immunity, structure, and the transport of bacteriocins will be welcomed.

On the other hand, papers with original novel applications in the aforementioned fields will be welcomed, as well.

The Guest Editors invite authors or cluster of authors with growth experience in bacteriocins to suggest a common classification scheme of bacteriocins.

Prof. Dr. Djamel Drider
Dr. Yanath Belguesmia
Guest Editors

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Keywords

bacteriocins
transport
gene regulation
immunity
functions and structure
mode of actions
applications foods
veterinary medicine
human health

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Published Papers (10 papers)

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21 pages, 1187 KiB

Open AccessArticle

In Vitro Anti-Candida albicans Mode of Action of Enterococcus mundtii and Enterococcus faecium

by Svetoslav Dimitrov Todorov, Richard Weeks, Igor Popov, Bernadette Dora Gombossy de Melo Franco and Michael Leonidas Chikindas

Microorganisms 2023, 11(3), 602; https://doi.org/10.3390/microorganisms11030602 - 27 Feb 2023

Cited by 1 | Viewed by 1968

Abstract

Candida albicans is an important vaginosis causative agent, affecting several women worldwide each year. This study reports on two strains of lactic acid bacteria (Enterococcus mundtii CRL35 and Enterococcus faecium ST88Ch) expressing bacteriocin-like inhibitor substances (BLIS) active against C. albicans 1281. Both strains were γ-hemolytic and not affected by numerous antibiotics, contraceptives, and commercial drugs, suggesting safety for human use. The recorded antimicrobial activity of semi-purified BLIS was 25,600 AU/mL for E. mundtii CRL35 and 800 AU/mL for E. faecium ST88Ch. Treatment of BLIS with 1 mg/mL proteinase K resulted in complete loss of antimicrobial activity against Listeria monocytogenes ATCC 15313 and partial loss of activity against C. albicans 1281. The killing effect of the semi-purified BLIS on cell suspensions of C. albicans 1281 after 9 h of contact was dose-dependent: for E. mundtii CRL35, 400 AU/mL to 25,600 AU/mL caused 63.61% to 79.35% lysis, while for E. faecium ST88Ch, 200 AU/mL to 800 AU/mL caused 29.32% to 31.25% cell lysis. The effects of temperature, pH, and presence of the contraceptive Nordette-28 on the adsorption levels of the BLIS to C. albicans 1281 were also evaluated. Nordette-28 (10% or 20%) promoted increased adsorption of both studied BLIS to the cells of C. albicans 1281 at pH 5.0, while a minor effect was observed at pH 3.0. Different levels of aggregation between C. albicans 1281 and E. mundtii CRL35 or E. faecium ST88Ch were recorded, and optimal adsorption levels were recorded at 37 °C. Appropriate BLIS-producing strains can effectively contribute to the equilibrium of vaginal microbial status quo and reduce negative consequences from the development of C. albicans infections. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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13 pages, 1475 KiB

Open AccessArticle

Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing

by Susana Escobedo, Ana B. Campelo, Özgün C. O. Umu, María Jesús López-González, Ana Rodríguez, Dzung B. Diep and Beatriz Martínez

Microorganisms 2023, 11(2), 501; https://doi.org/10.3390/microorganisms11020501 - 16 Feb 2023

Cited by 3 | Viewed by 1701

Abstract

In view of the current threat of antibiotic resistance, new antimicrobials with low risk of resistance development are demanded. Lcn972 is a lactococcal bacteriocin that inhibits septum formation by binding to the cell wall precursor lipid II in Lactococcus. It has a species-specific spectrum of activity, making Lcn972 an attractive template to develop or improve existing antibiotics. The aim of this work was to identify mutations present in the Lcn972-resistant clone Lactococcus cremoris D1-20, previously evolved from the sensitive strain L. cremoris MG1614. Whole-genome sequencing and comparison over the reference genome L. cremoris MG1363 identified several unexpected mutations in the parental strain MG1614, likely selected during in-house propagation. In the Lcn972R clone, two previously identified mutations were mapped and confirmed. Additionally, another transposition event deregulating cellobiose uptake was identified along with three point mutations of unknown consequences for Lcn972 resistance. Two new independent evolution experiments exposing L. cremoris MG1614 to Lcn972 revealed transposition of IS981 into the LLMG_RS12285 locus as the predominant mutation selected by Lcn972. This event occurs early during evolution and was found in 100% of the evolved clones, while other mutations were not selected. Therefore, activation of LLMG_RS12285 coding for a putative anti-ECF (extra-cytoplasmic function) sigma factor is regarded as the main Lcn972 resistance factor in L. cremoris MG1614. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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19 pages, 3673 KiB

Open AccessArticle

Nisin E Is a Novel Nisin Variant Produced by Multiple Streptococcus equinus Strains

by Ivan Sugrue, Daragh Hill, Paula M. O’Connor, Li Day, Catherine Stanton, Colin Hill and R. Paul Ross

Microorganisms 2023, 11(2), 427; https://doi.org/10.3390/microorganisms11020427 - 8 Feb 2023

Cited by 7 | Viewed by 2973

Abstract

Nisin A, the prototypical lantibiotic, is an antimicrobial peptide currently utilised as a food preservative, with potential for therapeutic applications. Here, we describe nisin E, a novel nisin variant produced by two Streptococcus equinus strains, APC4007 and APC4008, isolated from sheep milk. Shotgun whole genome sequencing and analysis revealed biosynthetic gene clusters similar to nisin U, with a unique rearrangement of the core peptide encoding gene within the cluster. The 3100.8 Da peptide by MALDI-TOF mass spectrometry, is 75% identical to nisin A, with 10 differences, including 2 deletions: Ser29 and Ile30, and 8 substitutions: Ile4Lys, Gly18Thr, Asn20Pro, Met21Ile, His27Gly, Val32Phe, Ser33Gly, and Lys34Asn. Nisin E producing strains inhibited species of Lactobacillus, Bacillus, and Clostridiodes and were immune to nisin U. Sequence alignment identified putative promoter sequences across the nisin producer genera, allowing for the prediction of genes in Streptococcus to be potentially regulated by nisin. S. equinus pangenome BLAST analyses detected 6 nisin E operons across 44 publicly available genomes. An additional 20 genomes contained a subset of nisin E transport/immunity and regulatory genes (nseFEGRK), without adjacent peptide production genes. These genes suggest that nisin E response mechanisms, distinct from the canonical nisin immunity and resistance operons, are widespread across the S. equinus species. The discovery of this new nisin variant and its immunity determinants in S. equinus suggests a central role for nisin in the competitive nature of the species. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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10 pages, 708 KiB

Open AccessArticle

The Discovery of Oropharyngeal Microbiota with Inhibitory Activity against Pathogenic Neisseria gonorrhoeae and Neisseria meningitidis: An In Vitro Study of Clinical Isolates

by Elvis Achondou Akomoneh, Jolein Gyonne Elise Laumen, Saïd Abdellati, Christophe Van Dijck, Thibau Vanbaelen, Xavier Basil Britto, Sheeba S. Manoharan-Basil and Chris Kenyon

Microorganisms 2022, 10(12), 2497; https://doi.org/10.3390/microorganisms10122497 - 16 Dec 2022

Cited by 2 | Viewed by 2217

Abstract

With increasing incidence of pathogenic Neisseria infections coupled with emerging resistance to antimicrobials, alternative approaches to limit the spread are sought. We investigated the inhibitory effect of oropharyngeal microbiota on the growth of N. gonorrhoeae and N. meningitidis and the impact of the essential oil-based mouthwash Listerine Cool Mint^® (Listerine). Oropharyngeal swabs from 64 men who have sex with men (n = 118) from a previous study (PReGo study) were analysed (ClinicalTrials.gov, NCT03881007). These included 64 baseline and 54 samples following three months of daily use of Listerine. Inhibition was confirmed by agar overlay assay, and inhibitory bacteria isolated using replica plating and identified using MALDI-TOF. The number of inhibitory isolates were compared before and after Listerine use. Thirty-one pharyngeal samples (26%) showed inhibitory activity against N. gonorrhoeae and/or N. meningitidis, and 62 inhibitory isolates were characterised. Fourteen species belonging to the genera Streptococci and Rothia were identified. More inhibitory isolates were observed following Listerine use compared to baseline, although this effect was not statistically significant (p = 0.073). This study isolated and identified inhibitory bacteria against pathogenic Neisseria spp. and established that daily Listerine use did not decrease their prevalence. These findings could provide a new approach for the prevention and treatment of pharyngeal Neisseria infections. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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15 pages, 1539 KiB

Open AccessArticle

Streptococcus salivarius 24SMBc Genome Analysis Reveals New Biosynthetic Gene Clusters Involved in Antimicrobial Effects on Streptococcus pneumoniae and Streptococcus pyogenes

by Gaia Vertillo Aluisio, Ambra Spitale, Luca Bonifacio, Grete Francesca Privitera, Aldo Stivala, Stefania Stefani and Maria Santagati

Microorganisms 2022, 10(10), 2042; https://doi.org/10.3390/microorganisms10102042 - 16 Oct 2022

Cited by 4 | Viewed by 2890

Abstract

Streptococcus salivarius 24SMBc is an oral probiotic with antimicrobial activity against the otopathogens Streptococcus pyogenes and Streptococcus pneumoniae. Clinical studies have reinforced its role in reducing the recurrence of upper respiratory tract infections (URTIs) and rebalancing the nasal microbiota. In this study, for the first time, we characterized 24SMBc by whole genome sequencing and annotation; likewise, its antagonistic activity vs. Streptococcus pneumoniae and Streptococcus pyogenes was evaluated by in vitro co-aggregation and competitive adherence tests. The genome of 24SMBc comprises 2,131,204 bps with 1933 coding sequences (CDS), 44 tRNA, and six rRNA genes and it is categorized in 319 metabolic subsystems. Genome mining by BAGEL and antiSMASH tools predicted three novel biosynthetic gene clusters (BGCs): (i) a Blp class-IIc bacteriocin biosynthetic cluster, identifying two bacteriocins blpU and blpK; (ii) an ABC-type bacteriocin transporter; and (iii) a Type 3PKS (Polyketide synthase) involved in the mevalonate pathway for the isoprenoid biosynthetic process. Further analyses detected two additional genes for class-IIb bacteriocins and 24 putative adhesins and aggregation factors. Finally, in vitro assays of 24SMBc showed significant anti-adhesion and co-aggregation effects against Streptococcus pneumoniae strains, whereas it did not act as strongly against Streptococcus pyogenes. In conclusion, we identified a novel blpU-K bacteriocin-encoding BGC and two class-IIb bacteriocins involved in the activity against Streptococcus pneumoniae and Streptococcus pyogenes; likewise the type 3PKS pathway could have beneficial effects for the host including antimicrobial activity. Furthermore, the presence of adhesins and aggregation factors might be involved in the marked in vitro activity of co-aggregation with pathogens and competitive adherence, showing an additional antibacterial activity not solely related to metabolite production. These findings corroborate the antimicrobial activity of 24SMBc, especially against Streptococcus pneumoniae belonging to different serotypes, and further consolidate the use of this strain in URTIs in clinical settings. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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18 pages, 3220 KiB

Open AccessArticle

Effect of Polydextrose on the Growth of Pediococcus pentosaceus as Well as Lactic Acid and Bacteriocin-like Inhibitory Substances (BLIS) Production

by Maria Carolina Wanderley Porto, Pamela Oliveira de Souza de Azevedo, Felipe Rebello Lourenço, Attilio Converti, Michele Vitolo and Ricardo Pinheiro de Souza Oliveira

Microorganisms 2022, 10(10), 1898; https://doi.org/10.3390/microorganisms10101898 - 24 Sep 2022

Cited by 1 | Viewed by 1932

Abstract

Pediococcus pentosaceus was cultivated in MRS medium supplemented or not with polydextrose under different conditions in order to evaluate its effect on cell growth, lactic acid and bacteriocin-like inhibitory substance (BLIS) production. Independent variables were pH (4.0, 5.0, 6.0), rotational speed (50, 100, 150 rpm), polydextrose concentration (0.5, 1.0, 1.5%) and temperature (25, 30, 35 °C), while cell concentration and productivity after 24 h, maximum specific growth rate, specific rate of substrate (glucose) consumption, volumetric and specific lactic acid productivities, yields of biomass and lactic acid on consumed substrate were the dependent. The maximum cell concentration (10.24 ± 0.16 g_X L⁻¹) and productivity (0.42 ± 0.01 g_X L⁻¹ h⁻¹) were achieved at pH 6.0, 35 °C, 150 rpm using 1.5% polydextrose, while the maximum specific growth rate (0.99 ± 0.01 h⁻¹) and yield of biomass (2.96 ± 0.34 g_X g_S⁻¹) were achieved at the same pH and polydextrose concentration, but at 25 °C and 50 rpm. The specific substrate consumption rate (0.09 ± 0.02 g_S g_X⁻¹ h⁻¹) and the volumetric lactic acid productivity (0.44 ± 0.02 g_P L⁻¹ h⁻¹) were maximized at pH 6.0, 35 °C, 50 rpm and 0.5% polydextrose. BLIS produced in this last run displayed the highest antibacterial activity against Escherichia coli, while the same activity was displayed against Enterococcus faecium using 1.5% polydextrose. These results appear to be quite promising in view of possible production of this BLIS as an antibacterial agent in the food industry. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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12 pages, 591 KiB

Open AccessCommunication

Bacteriocin-Producing Escherichia coli Isolated from the Gastrointestinal Tract of Farm Animals: Prevalence, Molecular Characterization and Potential for Application

by Marina V. Kuznetsova, Veronika S. Mihailovskaya, Natalia B. Remezovskaya and Marjanca Starčič Erjavec

Microorganisms 2022, 10(8), 1558; https://doi.org/10.3390/microorganisms10081558 - 2 Aug 2022

Cited by 7 | Viewed by 2325

Abstract

Due to the spread of antibiotic-resistant bacteria, new alternatives to antibiotics and ways to prevent infections are being sought. Bacteriocin-producing bacteria are therefore attracting attention due to their probiotic potential as a safe alternative to antimicrobial drugs. The aim of this work was to determine the prevalence of bacteriocin-encoded genes among Escherichia coli strains from healthy farm animals and to characterize the presence of virulence-associated genes, the possibility of prophage induction, and hemolytic and bacterial antagonistic activity of the bacteriocin-producing E. coli in order to reveal their potential for application. It was found that 17 of 72 E. coli strains (23.6%) produced bacteriocins. Among them, 18 out of 30 bacteriocin genes were detected: the most prevalent genes were those for microcin M (58.8%), colicin E1 (52.9%), and colicin M (35.3%). Colicin Ia (29.4%), colicin E9, colicin Ib, colicin B (23.5%), and colicin E9 (17.7%) genes were also frequent, while the prevalence of genes encoding microcins V, B17, and H47 and colicins E3, K, N, U, Y, 5, and 10 did not exceed 11.8%. At least two different bacteriocin genes were detected in all 17 bacteriocinogenic strains; the highest number of different bacteriocin genes detected in one strain was seven genes. E. coli strains with combinations of colicin E1 and E or microcin M and colicin E1 genes were more prevalent than others (17.7%). Among the 17 bacteriocin-producing E. coli strains, 5.9% were hemolytic, 47.1% contained prophages, and 58.8% carried genes encoding toxins. Cell-free supernatants of bacteriocin-producing strains were shown to inhibit the growth of pathogenic E. coli strains belonging to the APEC, STEC, and ETEC pathotypes. Thus, among the studied bacteriocin-producing E. coli isolated from the gastrointestinal tract of farm animals, three strains with high antagonistic bacterial activity and the absence of pathogenicity genes, prophages, and hemolytic activity were identified and therefore have potential for application. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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20 pages, 7742 KiB

Open AccessArticle

Combined Action of Antibiotics and Bacteriocins against Vancomycin-Resistant Enterococci

by Jorge Enrique Vazquez Bucheli, Joanna Ivy Irorita Fugaban, Wilhelm Heinrich Holzapfel and Svetoslav Dimitrov Todorov

Microorganisms 2022, 10(7), 1423; https://doi.org/10.3390/microorganisms10071423 - 14 Jul 2022

Cited by 11 | Viewed by 2223

Abstract

Antibiotics have been one of the most important discoveries in the area of applied medical microbiology; however, as a result of various factors, we are currently facing a dramatic and relatively dangerous increase in the number of cases of antibiotic resistance, and the need for new types of antimicrobials continues to grow. New approaches are needed to combat antibiotic-resistant pathogens. Bacteriocins, as part of the group of antimicrobial peptides, can be considered as alternatives and/or complements to known antibiotics. Their narrow spectra of activity can be explored for the control of various pathogens, such as vancomycin-resistant enterococci (VRE), as single therapies or in combination with known antibiotics. In the present study, we isolated bacteriocins from different lactic acid bacteria (LAB) strains, including Enterococcus and Pediococcus, and explored the possible synergistic inhibition of growth by bacteriocins and vancomycin. It was observed in the growth dynamics with previously selected VRE strains that the bacteriocins had a high specificity and a promising inhibitory effect against the VRE strains, and these results were validated by a propidium iodide viability test using flow cytometry. The data obtained indicate that the selected bacteriocins can be used to control VRE in the food industry or even as an alternative treatment to combat infections with antibiotic-resistant bacteria. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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9 pages, 1753 KiB

Open AccessCommunication

Genomic Analyses of Weissella cibaria W25, a Potential Bacteriocin-Producing Strain Isolated from Pasture in Campos das Vertentes, Minas Gerais, Brazil

by Camila Gonçalves Teixeira, Rafaela da Silva Rodrigues, Ricardo Seiti Yamatogi, Anca Lucau-Danila, Djamel Drider, Luís Augusto Nero and Antônio Fernandes de Carvalho

Microorganisms 2022, 10(2), 314; https://doi.org/10.3390/microorganisms10020314 - 28 Jan 2022

Cited by 6 | Viewed by 2815

Abstract

Weissella is a genus containing Gram-positive, heterofermentative bacteria belonging to the lactic acid bacteria (LAB) group. These bacteria are endowed with promising technological and antimicrobial attributes. Weissella cibaria W25 was isolated from a dairy environment where raw milk cheeses are produced. Therefore, we sequenced and assembled the W25 draft genome sequence, which consists of 41 contigs totaling ~2.4 Mbp, with a G + C content of 45.04%. Then we carried out a comprehensive comparative genomic analysis with W. cibaria 110, known to produce the weissellicin 110 bacteriocin, and four other non-bacteriocin-producing W. cibaria strains. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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Review

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24 pages, 4250 KiB

Open AccessReview

Current Knowledge of the Mode of Action and Immunity Mechanisms of LAB-Bacteriocins

by Adrián Pérez-Ramos, Désiré Madi-Moussa, Françoise Coucheney and Djamel Drider

Microorganisms 2021, 9(10), 2107; https://doi.org/10.3390/microorganisms9102107 - 7 Oct 2021

Cited by 71 | Viewed by 6354

Abstract

Bacteriocins produced by lactic acid bacteria (LAB-bacteriocins) may serve as alternatives for aging antibiotics. LAB-bacteriocins can be used alone, or in some cases as potentiating agents to treat bacterial infections. This approach could meet the different calls and politics, which aim to reduce the use of traditional antibiotics and develop novel therapeutic options. Considering the clinical applications of LAB-bacteriocins as a reasonable and desirable therapeutic approach, it is therefore important to assess the advances achieved in understanding their modes of action, and the resistance mechanisms developed by the producing bacteria to their own bacteriocins. Most LAB-bacteriocins act by disturbing the cytoplasmic membrane through forming pores, or by cell wall degradation. Nevertheless, some of these peptides still have unknown modes of action, especially those that are active against Gram-negative bacteria. Regarding immunity, most bacteriocin-producing strains have an immunity mechanism involving an immunity protein and a dedicated ABC transporter system. However, these immunity mechanisms vary from one bacteriocin to another. Full article

(This article belongs to the Special Issue Bacteriocins: Academic Advances and Immediate Applications)

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