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Microorganisms
Special Issues
Benefical Properties and Safety of Lactic Acid Bacteria
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Benefical Properties and Safety of Lactic Acid Bacteria

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Food Microbiology".

Deadline for manuscript submissions: closed (31 July 2022) | Viewed by 42188

Special Issue Editors

Prof. Dr. Wilhelm Holzapfel

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Guest Editor
AGEE, Handong Global University, Pohang, Korea
Interests: beneficial lactic acid bacteria; probiotics; bioactive peptides; gut microbiota; taxonomy and safety of the lactic acid bacteria
Dr. Svetoslav Todorov

E-Mail Website
Guest Editor
ProBacLab, Laboratório de Microbiologia de Alimentos, Departamento de Alimentos e Nutrição Experimental, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo 05508-000, SP, Brazil
Interests: fermented food products; lactic acid bacteria; probiotics; bacteriocins; traditional food products
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear colleagues,

Application of LAB in different sectors of the biotechnical and food industry, human and veterinary practice, health promoting practices and cosmetics is currently highly explored. Different traditional and innovative aspects of application of LAB were subject of intensive research all around the World. Rediscovery of old and establishing new processes based on production and application of different metabolites produced by LAB, and new approaches in fermentation processes by LAB have become hot topics. Different antimicrobial peptides, including bacteriocins, were proposed as alternatives to antibiotics or suggested to be their synergetic partners Applications of probiotics are being widened to new innovative areas targeted towards personalized applications for improvement of human health. Deeper understanding of bioactive peptides has opened the way for their application as alternative or complimentary to Western medicine. Biopreservation approaches require less chemical preservatives and is highly explored in food science research. Enrichment and fortification of food products with biologically active metabolites, including vitamins, antimicrobials, and immunomodulators are only some of the topics explored as options for application of different LAB in the food industry.

Prof. Dr. Wilhelm Holzapfel
Prof. Dr. Svetoslav Todorov
Guest Editors

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Keywords

  • Lactic acid bacteria
  • bacteriocins
  • antimicrobial peptides
  • bioactive peptides
  • beneficial properties
  • probiotics
  • virulence factors
  • antibiotic resistance
  • functional models
  • starter cultures
  • biopreservation

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Published Papers (11 papers)

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Editorial

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5 pages, 216 KiB  
Editorial
Special Issue: Beneficial Properties and Safety of Lactic Acid Bacteria
by Wilhelm Heinrich Holzapfel and Svetoslav Dimitrov Todorov
Microorganisms 2023, 11(4), 871; https://doi.org/10.3390/microorganisms11040871 - 28 Mar 2023
Cited by 1 | Viewed by 1646
Abstract
The application of LAB in various sectors, including in the biotechnical and food industry, in human and veterinary practice, and in health-promoting practices and cosmetics, has been the subject of intensive research across the globe, with a range of traditional and innovative methods [...] Read more.
The application of LAB in various sectors, including in the biotechnical and food industry, in human and veterinary practice, and in health-promoting practices and cosmetics, has been the subject of intensive research across the globe, with a range of traditional and innovative methods currently being explored [...] Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)

Research

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19 pages, 8720 KiB  
Article
Effect of Lactobacteria on Bioactive Peptides and Their Sequence Identification in Mature Cheese
by Marina Kurbanova, Roman Voroshilin, Oksana Kozlova and Victor Atuchin
Microorganisms 2022, 10(10), 2068; https://doi.org/10.3390/microorganisms10102068 - 19 Oct 2022
Cited by 13 | Viewed by 2995
Abstract
An in silico study that featured the effect of starter cultures on the bioactivity and other health benefits of peptides in semi-hard cheese is presented in this contribution. Model Caciotta-type cheese samples were obtained in laboratory conditions in two variations. Sample A included [...] Read more.
An in silico study that featured the effect of starter cultures on the bioactivity and other health benefits of peptides in semi-hard cheese is presented in this contribution. Model Caciotta-type cheese samples were obtained in laboratory conditions in two variations. Sample A included starter cultures of Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris. Sample B included starter cultures of Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and a culture of lactobacilli Lacticaseibacillus casei. The in silico method showed that the peptides inhibited angiotensin-converting enzymes (ACE) and ipeptidyl peptidase IV (DPP-4), as well as possessed antioxidant properties. Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris had a greater effect on the formation of bioactive peptides. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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16 pages, 312 KiB  
Article
Safety Assessment of Lactiplantibacillus plantarum TWK10 Based on Whole-Genome Sequencing, Phenotypic, and Oral Toxicity Analysis
by Han-Yin Hsu, Yi-Chu Liao, Shih-Hsuan Lin, Jin-Seng Lin, Chia-Chia Lee and Koichi Watanabe
Microorganisms 2022, 10(4), 784; https://doi.org/10.3390/microorganisms10040784 - 7 Apr 2022
Cited by 18 | Viewed by 3561
Abstract
Lactiplantibacillus plantarum TWK10 (TWK10), isolated from Taiwanese pickled cabbage, has been demonstrated to exert beneficial probiotic effects in both mice and humans. Here, we comprehensively assessed the safety of TWK10 using both in vivo and in vitro approaches, including whole-genome sequence analysis, an [...] Read more.
Lactiplantibacillus plantarum TWK10 (TWK10), isolated from Taiwanese pickled cabbage, has been demonstrated to exert beneficial probiotic effects in both mice and humans. Here, we comprehensively assessed the safety of TWK10 using both in vivo and in vitro approaches, including whole-genome sequence analysis, an assessment of hemolytic activity, and performing an antimicrobial susceptibility test, the Ames bacterial reverse mutation assay, the chromosomal aberration test, a rodent peripheral blood micronucleus test, and the 28-day subacute oral toxicity assay. The results showed that there was no significant increase in the incidence of reverse mutations or chromosomal aberrations following exposure to TWK10. Moreover, no significant changes were detected either in the number of reticulocytes or the incidence of micronuclei in ICR mice, and no subacute toxicity was recorded in SD rats at the oral TWK10 dosage of 2000 mg/kg body weight/day repeated for 28 days. Additionally, TWK10 exhibited no hemolytic activity and was susceptible to all the antibiotics tested, except kanamycin. However, no antimicrobial resistance genes, virulence factors, or genes involved in biogenic amine synthesis were found in the genome of TWK10. Our findings demonstrated that TWK10 has high potential of being safe for human consumption as a probiotic. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
13 pages, 2404 KiB  
Article
Safety Evaluation of Weissella cibaria JW15 by Phenotypic and Genotypic Property Analysis
by Ye-Ji Jang, Hee-Min Gwon, Woo-Soo Jeong, Soo-Hwan Yeo and So-Young Kim
Microorganisms 2021, 9(12), 2450; https://doi.org/10.3390/microorganisms9122450 - 27 Nov 2021
Cited by 19 | Viewed by 3171
Abstract
Weissella cibaria is one of the bacteria in charge of the initial fermentation of kimchi and has beneficial effects such as immune-modulating, antagonistic, and antioxidant activities. In our study, we aimed to estimate the safety of W. cibaria JW15 for the use of [...] Read more.
Weissella cibaria is one of the bacteria in charge of the initial fermentation of kimchi and has beneficial effects such as immune-modulating, antagonistic, and antioxidant activities. In our study, we aimed to estimate the safety of W. cibaria JW15 for the use of probiotics according to international standards based on phenotypic (antibiotic resistance, hemolysis, and toxic metabolite production) and genotypic analysis (virulence genes including antibiotic resistance genes). The results of the safety assessment on W. cibaria JW15 were as follows; (1) antibiotic resistance genes (ARGs) (kanamycin and vancomycin etc.) were intrinsic characteristics; (2) There were no acquired virulence genes including Cytolysin (cylA), aggregation substance (asa1), Hyaluronidase (hyl), and Gelatinase (gelE); (3) this strain also lacked β-hemolysis and the production of toxic metabolites (D-lactate and bile salt deconjugation). Consequently, W. cibaria JW15 is expected to be applied as a functional food ingredient in the food market. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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13 pages, 2899 KiB  
Article
Lactic Acid Bacteria Exert a Hepatoprotective Effect against Ethanol-Induced Liver Injury in HepG2 Cells
by Ji Yeon Lee, Hyemin Kim, Yulah Jeong and Chang-Ho Kang
Microorganisms 2021, 9(9), 1844; https://doi.org/10.3390/microorganisms9091844 - 31 Aug 2021
Cited by 32 | Viewed by 4721
Abstract
Alcoholic liver fatty disease (ALFD) is caused by excessive and chronic alcohol consumption. Alcohol consumption causes an imbalance in the intestinal microflora, leading to liver disease induced by the excessive release of endotoxins into the hepatic portal vein. Therefore, research on the intestinal [...] Read more.
Alcoholic liver fatty disease (ALFD) is caused by excessive and chronic alcohol consumption. Alcohol consumption causes an imbalance in the intestinal microflora, leading to liver disease induced by the excessive release of endotoxins into the hepatic portal vein. Therefore, research on the intestinal microflora to identify treatments for ALFD is increasing. In this study, the protective effects of lactic acid bacteria (LAB) strains, including Levilactobacillus brevis, Limosilactobacillus reuteri, and Limosilactobacillus fermentum, were evaluated in ethanol-induced HepG2 cells. Among the evaluated LAB, nine strains increased aldehyde dehydrogenase (ALDH) levels and downregulated lipid peroxidation and liver transferase in the ethanol-induced HepG2 cells. Moreover, L. brevis MG5280 and MG5311, L. reuteri MG5458, and L. fermentum MG4237 and MG4294 protected against ethanol-induced HepG2 cell damage by regulating CYP2E1, antioxidant enzymes (SOD, CAT, and GPX), lipid synthesis factors (SREBP1C and FAS), and lipid oxidation factors (PPARα, ACO, and CPT-1). Moreover, five LAB were confirmed to be safe probiotics based on antibiotic susceptibility and hemolysis assays; their stability and adhesion ability in the gastrointestinal tract were also established. In conclusion, L. brevis MG5280 and MG5311, L. reuteri MG5458, and L. fermentum MG4237 and MG4294 may be useful as new probiotic candidates for ALFD prevention. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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12 pages, 3608 KiB  
Article
In Vivo Confirmation of the Antimicrobial Effect of Probiotic Candidates against Gardnerella vaginalis
by Hyemin Kim, YongGyeong Kim and Chang-Ho Kang
Microorganisms 2021, 9(8), 1690; https://doi.org/10.3390/microorganisms9081690 - 9 Aug 2021
Cited by 15 | Viewed by 3650
Abstract
Bacterial vaginosis (BV) is caused by a microbial imbalance of the vaginal ecosystem, causing genital discomfort and potentially even various complications in women. Moreover, research on the treatment or prevention of BV is increasing. In this study, we evaluated the antimicrobial and anti-inflammation [...] Read more.
Bacterial vaginosis (BV) is caused by a microbial imbalance of the vaginal ecosystem, causing genital discomfort and potentially even various complications in women. Moreover, research on the treatment or prevention of BV is increasing. In this study, we evaluated the antimicrobial and anti-inflammation effects of the lactic acid bacteria (LAB) Ligilactobacillus salivarius MG242, Limosilactobacillus fermentum MG901, and Lactiplantibacillus plantarum MG989 in a BV-induced mice model. The oral administration of the LAB significantly inhibited the growth of Gardnerella vaginalis up to 43% (p < 0.05). The LAB downregulated the expression of pro-inflammatory cytokines (IL-1β and TNF-α) and myeloperoxidase (p < 0.05). Upon histological examination, the exfoliation of epithelial cells in the vaginal tissues was found to be reduced in the probiotic administration group compared to the infected group. In addition, the LAB tolerated the gastric and/or intestinal simulated conditions and proliferated, showing potential in promoting health based on hemolysis activity, antibiotic susceptibility, enzyme activity, and lactic acid production. Altogether, our results showed that the investigated LAB may be a good food ingredient candidate for ameliorating BV in women. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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23 pages, 3730 KiB  
Article
Characterization of Partially Purified Bacteriocins Produced by Enterococcus faecium Strains Isolated from Soybean Paste Active Against Listeria spp. and Vancomycin-Resistant Enterococci
by Joanna Ivy Irorita Fugaban, Jorge Enrique Vazquez Bucheli, Wilhelm Heinrich Holzapfel and Svetoslav Dimitrov Todorov
Microorganisms 2021, 9(5), 1085; https://doi.org/10.3390/microorganisms9051085 - 18 May 2021
Cited by 24 | Viewed by 4125
Abstract
Three out of one hundred eighty putative LAB isolates from Korean traditional fermented soybean paste were identified to be unique and bacteriocinogenic strains. Based on phenotypic and 16S rRNA sequencing analysis, selected strains were identified as Enterococcus faecium ST651ea, E. faecium ST7119ea and [...] Read more.
Three out of one hundred eighty putative LAB isolates from Korean traditional fermented soybean paste were identified to be unique and bacteriocinogenic strains. Based on phenotypic and 16S rRNA sequencing analysis, selected strains were identified as Enterococcus faecium ST651ea, E. faecium ST7119ea and E. faecium ST7319ea. The bacteriocinogenic properties of the studied strains were evaluated against Listeria monocytogenes ATCC15313, Listeria innocua ATCC33090 and vancomycin-resistant E. faecium VRE19 of clinical origin. The strains E. faecium ST651ea, ST7119ea and ST7319ea expressed bacteriocins with an activity of 12,800 AU/mL, 25,600 AU/mL and 25,600 AU/mL, respectively, recorded against L. monocytogenes ATCC15131. According to the PCR-based screening of bacteriocin-related genes, which was further confirmed through amplicon sequencing, showed that strain E. faecium ST651ea carries entB and entP genes, whereas both E. faecium ST7119ea and ST7319ea strains harbor entA and entB genes. The molecular size of expressed bacteriocins was estimated by tricine-SDS-PAGE showing an approximative protein size of 4.5 kDa. The assessment of the spectrum of activity of bacteriocins ST651ea, ST7119ea and ST7319ea showed strong activity against most of clinical VRE isolates, majority of other Enterococcus spp. and Listeria spp. Bacteriocins ST651ea, ST7119ea and ST7319ea were partially purified by combination of 60% ammonium sulfate precipitation and hydrophobic chromatography on the SepPakC18 column. Challenge test with semi-purified (60% 2-propanol fraction) bacteriocins resulted in a significant reduction of viable cells for all test organisms. Thus, indicating that all the bacteriocins evaluated can be used as potential biocontrol in food and feed industries as well as an alternative treatment for VRE-related infections in both veterinary and clinical settings. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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21 pages, 1618 KiB  
Article
Rehydration before Application Improves Functional Properties of Lyophilized Lactiplantibacillus plantarum HAC03
by Karina Arellano-Ayala, Juhwan Lim, Subin Yeo, Jorge Enrique Vazquez Bucheli, Svetoslav Dimitrov Todorov, Yosep Ji and Wilhelm Heinrich Holzapfel
Microorganisms 2021, 9(5), 1013; https://doi.org/10.3390/microorganisms9051013 - 8 May 2021
Cited by 15 | Viewed by 4106
Abstract
Preservation of probiotics by lyophilization is considered a method of choice for developing stable products. However, both direct consumption and reconstitution of dehydrated probiotic preparations before application “compromise” the survival and functional characteristics of the microorganisms under the stress of the upper gastro-intestinal [...] Read more.
Preservation of probiotics by lyophilization is considered a method of choice for developing stable products. However, both direct consumption and reconstitution of dehydrated probiotic preparations before application “compromise” the survival and functional characteristics of the microorganisms under the stress of the upper gastro-intestinal tract. We evaluated the impact of different food additives on the viability, mucin adhesion, and zeta potential of a freeze-dried putative probiotic, Lactiplantibacillus (Lp.) plantarum HAC03. HAC03-compatible ingredients for the formulation of ten rehydration mixtures could be selected. Elevated efficacy was achieved by the B-active formulation, a mixture of non-protein nitrogen compounds, sugars, and salts. The survival of Lp. plantarum HAC03 increased by 36.36% compared rehydration with distilled water (4.92%) after passing simulated gastro-intestinal stress conditions. Cell viability determined by plate counting was confirmed by flow cytometry. B-active formulation also influenced Lp. plantarum HAC03 functionality by increasing its adherence to a Caco-2 cell-line and by changing the bacterial surface charge, measured as zeta potential.Hydrophobicity, mucin adhesion and immunomodulatory properties of Lp. plantarum HAC03 were not affected by the B-active formulation. The rehydration medium also effectively protected Lp. plantarum ATCC14917, Lp. plantarum 299v, Latilactobacillus sakei (Lt.) HAC11, Lacticaseibacillus (Lc.) paracasei 532, Enterococcus faecium 200, and Lc. rhamnosus BFE5263. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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16 pages, 343 KiB  
Article
Safety Evaluation, Biogenic Amine Formation, and Enzymatic Activity Profiles of Autochthonous Enterocin-Producing Greek Cheese Isolates of the Enterococcus faecium/durans Group
by Charikleia Tsanasidou, Stamatia Asimakoula, Nikoletta Sameli, Christos Fanitsios, Elpiniki Vandera, Loulouda Bosnea, Anna-Irini Koukkou and John Samelis
Microorganisms 2021, 9(4), 777; https://doi.org/10.3390/microorganisms9040777 - 8 Apr 2021
Cited by 29 | Viewed by 4360
Abstract
Autochthonous single (Ent+) or multiple (m-Ent+) enterocin-producing strains of dairy enterococci show promise for use as bioprotective adjunct cultures in traditional cheese technologies, provided they possess no pathogenic traits. This study evaluated safety, decarboxylase activity, and enzymatic (API ZYM) activity profiles of nine [...] Read more.
Autochthonous single (Ent+) or multiple (m-Ent+) enterocin-producing strains of dairy enterococci show promise for use as bioprotective adjunct cultures in traditional cheese technologies, provided they possess no pathogenic traits. This study evaluated safety, decarboxylase activity, and enzymatic (API ZYM) activity profiles of nine Ent+ or m-Ent+ Greek cheese isolates previously assigned to four distinct E. faecium (represented by the isolates KE64 (entA), GL31 (entA), KE82 (entA-entB-entP) and KE77 (entA-entB-entP-bac31)) and two E. durans (represented by the isolates KE100 (entP) and KE108 (entP-bac31-cyl)) strain genotypes. No strain was β-hemolytic or harbored vanA and vanB or the virulence genes agg, ace, espA, IS16, hyl, or gelE. All strains were of moderate to high sensitivity to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, penicillin, tetracycline, and vancomycin, except for the E. faecium KE64 and KE82 strains, which were resistant to erythromycin and penicillin. All cheese strains showed moderate to strong esterase-lipase and aminopeptidase activities and formed tyramine, but none formed histamine in vitro. In conclusion, all Ent+ or m-Ent+ strain genotypes of the E. faecium/durans group, except for the cyl-positive E. durans KE108, were safe for use as adjunct cultures in traditional Greek cheeses. Further in situ biotechnological evaluations of the strains in real cheese-making trials are required. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
10 pages, 248 KiB  
Article
Effects of a Lactobacilli Probiotic on Reducing Duration of URTI and Fever, and Use of URTI-Associated Medicine: A Re-Analysis of a Randomized, Placebo-Controlled Study
by Tatiana Altadill, Jordi Espadaler-Mazo and Min-Tze Liong
Microorganisms 2021, 9(3), 528; https://doi.org/10.3390/microorganisms9030528 - 4 Mar 2021
Cited by 8 | Viewed by 3409
Abstract
We previously reported on the effects of Lactoplantibacillus plantarum DR7 on reducing Upper Respiratory Tract Infections (URTI) symptoms’ score and frequency in 109 adults upon a 12-week consumption at 109 colony-forming units (CFU)/day, but several limitations were detected in the publication. Thus, [...] Read more.
We previously reported on the effects of Lactoplantibacillus plantarum DR7 on reducing Upper Respiratory Tract Infections (URTI) symptoms’ score and frequency in 109 adults upon a 12-week consumption at 109 colony-forming units (CFU)/day, but several limitations were detected in the publication. Thus, the present study re-analyzed some data with the aim to address some of these weaknesses, and presents new data on duration of URTI and consumption of URTI-associated medication, as compared to the placebo. Our re-analyses found probiotic administration significantly reduced the proportion of patient days of URTI and of fever (all p < 0.05). Recent history of URTI was a prevalent co-factor in affecting duration of URTI symptoms and fever, while other demographic and clinical factors had no influence. Exploratory analyses suggested probiotic had an earlier benefit in patients without a recent history of URTI compared to those with a recent history of URTI. Therefore, recent history of infections could have a modulatory effect on probiotic efficacy. Average number of months with reported use of URTI-related medication was 3.4-times lower in the probiotic group as compared to placebo (p = 0.016) during the intervention. Taken together, our present new data further support previous findings that DR7 probiotic had a beneficial effect on URTI. Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
16 pages, 6138 KiB  
Article
Assessment of Pb(II), Cd(II), and Al(III) Removal Capacity of Bacteria from Food and Gut Ecological Niches: Insights into Biodiversity to Limit Intestinal Biodisponibility of Toxic Metals
by Fanny George, Séverine Mahieux, Catherine Daniel, Marie Titécat, Nicolas Beauval, Isabelle Houcke, Christel Neut, Delphine Allorge, Frédéric Borges, Gwénaël Jan, Benoît Foligné and Anne Garat
Microorganisms 2021, 9(2), 456; https://doi.org/10.3390/microorganisms9020456 - 22 Feb 2021
Cited by 41 | Viewed by 4673
Abstract
Toxic metals (such as lead, cadmium, and, to a lesser extent, aluminum) are detrimental to health when ingested in food or water or when inhaled. By interacting with heavy metals, gut and food-derived microbes can actively and/or passively modulate (by adsorption and/or sequestration) [...] Read more.
Toxic metals (such as lead, cadmium, and, to a lesser extent, aluminum) are detrimental to health when ingested in food or water or when inhaled. By interacting with heavy metals, gut and food-derived microbes can actively and/or passively modulate (by adsorption and/or sequestration) the bioavailability of these toxins inside the gut. This “intestinal bioremediation” involves the selection of safe microbes specifically able to immobilize metals. We used inductively coupled plasma mass spectrometry to investigate the in vitro ability of 225 bacteria to remove the potentially harmful trace elements lead, cadmium, and aluminum. Interspecies and intraspecies comparisons were performed among the Firmicutes (mostly lactic acid bacteria, including Lactobacillus spp., with some Lactococcus, Pediococcus, and Carnobacterium representatives), Actinobacteria, and Proteobacteria. The removal of a mixture of lead and cadmium was also investigated. Although the objective of the study was not to elucidate the mechanisms of heavy metal removal for each strain and each metal, we nevertheless identified promising candidate bacteria as probiotics for the intestinal bioremediation of Pb(II) and Cd(II). Full article
(This article belongs to the Special Issue Benefical Properties and Safety of Lactic Acid Bacteria)
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