Bacteriocinogenic Lactobacillus plantarum ST16Pa isolated from papaya (Carica papaya) — From isolation to application: Characterization of a bacteriocin

Strain ST16PA, isolated from papaya was identified as Lactobacillus plantarum based on biochemical tests, PCR with species-specific primers and 16S rDNA sequencing. L. plantarum ST16PA produces a 6.5 kDa bacteriocin, active against different species from genera Enterobacter, Enterococcus, Lactobacillus, Pseudomonas, Streptococcus and Staphylococcus and different serotypes of Listeria spp. The peptide is inactivated by proteolytic enzymes, but not when treated with ??-amylase, catalase, lipase, Triton X-100, SDS, Tween 20, Tween 80, urea, NaCl and EDTA. However, presence of 1% Triton X-114 deactivates the bacteriocin. No change in activity was recorded after 2 h at pH values between 2.0 and 12.0, and after treatment at 100 °C for 120 min or 121 °C for 20 min. The mode of activity against Lactobacillus sakei ATCC 15521, Enterococcus faecalis ATCC 19443 and Listeria innocua 2030C was bactericidal, resulting in cell lysis and enzyme-leakage. No significant differences in cell growth and bacteriocin production were observed when strain ST16Pa was cultured in MRS broth at 26 °C and 30 °C for 24 h (25 600 AU/ml). However, even though strain ST16PA grows well in MRS broth at 15 °C and 37 °C, a reduction of bacteriocin production was observed (400 AU/ml and 1600 AU/ml, respectively). In addition, effect of MRS medium components, different initial pH and additions of glycerol or vitamins to the media on bacteriocin ST16Pa production was studied.Peptide ST16PA adsorbs (400 AU/ml) to producer cells. However, bacteriocin ST16Pa was adsorbed at 50% to cells of L. innocua 2030C and at 75% to L. sakei ATCC 15521 and E. faecalis ATCC 19433 when experiments were conducted at 30 °C and pH 6.5. Adsorption of bacteriocin ST16Pa to target cells at different temperatures, pH and in presence of potassium sorbate, sodium nitrate, sodium chloride, ascorbic acid, Tween 80 and Tween 20 were also studied. To the best of our knowledge, this is the first report on detection of L. plantarum in papaya.     

Effect of inulin on growth and bacteriocin production by Lactobacillus plantarum in stationary and shaken cultures

The prebiotic effect of inulin added to MRS medium on growth and bacteriocin production by L. plantarum ST16 Pa was investigated in stationary cultures in anaerobic jars with medium containing 0.025% sodium thioglycolate or in flasks shaken at 100 rpm. In the presence of 1% inulin in anaerobic stationary cultures, this strain produced lactic acid at a level that was 36.5% higher than in the absence of the polysaccharide. In shaken cultures without inulin, cell count was 54% higher than in the stationary ones. Under stationary conditions in anaerobic jars, the addition of inulin increased the maximum specific growth rate from 0.37 to 0.49 h^?1 and reduced the generation time from 1.85 h to 1.40 h. Consequently, the exponential phase was shortened from 12 to 9 h when the cells were grown in stationary cultures with the oxygen scavenger. Despite this effect of inulin on growth rate, stationary cultures without inulin displayed higher antimicrobial activity against Listeria monocytogenes L104 (3200 AU/mL) than cultures with inulin (1600 AU/mL); therefore, inulin behaved as a compound able to accelerate growth rather than to stimulate bacteriocin production. The results presented in this study are very promising, as L. monocytogenes is a well‐known foodborne pathogenic microorganism. Moreover, L. plantarum ST16 Pa has proven to be a potential producer of a natural food preservative at an industrial level.     

Physicochemical factors differentially affect the biomass and bacteriocin production by bovine Enterococcus mundtii CRL1656

Bovine Enterococcus mundtii CRL1656 (Centro de Referencia para Lactobacilos Culture Collection) produces an anti-Listeria and anti-Streptococcus dysgalactiae bacteriocin identified as mundticin CRL1656. The strain and its bacteriocin are candidates to be included in a beneficial product to prevent bovine mastitis as an alternative to antimicrobial agents. To optimize the production of biomass and mundticin CRL1656 by E. mundtii CRL1656, a complete 3 × 2⁴ factorial design was applied. The effect of culture medium, initial pH, inoculum size, incubation temperature, and agitation conditions on biomass and bacteriocin production was evaluated simultaneously. Growth parameters were determined using the modified Gompertz model. A nonlinear model was used to estimate the effects of the variables on growth parameters. Bacteriocin production was analyzed using a linear mixed model. Optimal biomass and mundticin CRL1656 production by E. mundtii CRL1656 were obtained in different conditions. Maximal growth was recorded in autolyzed yeast, peptone, tryptone, Tween 80, and glucose or M17 broths, pH 6.5, 5.0% inoculum, 30°C, with agitation. However, bacteriocin titers were higher in autolyzed yeast, peptone, tryptone, Tween 80, and glucose or de Man-Rogosa-Sharpe (MRS) broths, pH 6.5, 30°C, both with or without agitation. Knowledge of the optimum conditions for growth and bacteriocin production of E. mundtii CRL1656 will allow the obtainment of high levels of biomass and mundticin CRL1656 as bioingredients of potential products to prevent bovine mastitis.     

Biochemical,antimicrobial and molecular characterization of a noncytotoxic bacteriocin produced by Lactobacillus plantarum ST71KS

Lactobacillus (Lb.) plantarum ST71KS was isolated from homemade goat feta cheese and identified using biochemical and molecular biology techniques. As shown by Tricine-SDS-PAGE, this lactic acid bacterium produces a bacteriocin (ST71KS) with an estimated molecular weight of 5.0 kDa. Bacteriocin ST71KS was not affected by the presence of α-amylase, catalase and remained stable in a wide range of pH and after treatment with Triton X-100, Triton X-114, Tween 20, Tween 80, NaCl, SDS, urea and EDTA. This bacteriocin also remained active after being heated at 100 °C for 2 h and even after 20 min at 121 °C; however, it was inactivated by proteolitic enzymes. Production of bacteriocin ST71KS reached 6400 AU/mL during stationary growth phase of Lb. plantarum cultivated in MRS at 30 °C and 37 °C. Bacteriocin ST71KS displayed a bactericidal effect against Listeria monocytogenes strains 603 and 607 and did not adsorb to the producer cells. Lb. plantarum ST71KS harbors two bacteriocin genes with homology to plantaricin S and pediocin PA-1. These characteristics indicate that bacteriocin ST71KS is a class IIa bacteriocin. The peptide presented no toxic effect when tested in vitro with kidney Vero cells, indicating safe technological application to control L. monocytogenes in foods.     

Induction of bacteriocin production by coculture is widespread among plantaricin-producing Lactobacillus plantarum strains with different regulatory operons

We describe the bacteriocin-production phenotype in a group of eight singular bacteriocinogenic Lactobacillus plantarum strains with three distinct genotypes regarding the plantaricin locus. Genotyping of these strains revealed the existence of two different plantaricin-production regulatory operons, plNC8-plNC8HK-plnD or plnABCD, involving three-component systems controlled each of them by a specific autoinducer peptide (AIP), i.e. PLNC8IF or PlnA. While all of the strains produced antimicrobial activity when growing on solid medium, most of them halted this production when cultured in broth, thus reflecting the functionality of regulatory mechanisms. Antimicrobial activity in broth cultures was re-established or enhanced when the specific AIP was added to the culture or by coculturing with specific bacterial strains. The latter trait appeared to be widespread in bacteriocinogenic L. plantarum strains independently of the regulatory system used to regulate bacteriocin production or the specific bacteriocins produced. The induction spectrum through coculture, i.e. the pattern of bacterial strains able to induce bacteriocin production, was characteristic of each individual L. plantarum strain. Also, the ability of some bacteria to induce bacteriocin production in L. plantarum by coculture appeared to be strain specific. The fact that induction of bacteriocin production by coculturing appeared to be a common feature in L. plantarum can be exploited accordingly to enhance the viability of this species in food and feed fermentations, as well as to contribute to probiotic functionality when colonising the gastrointestinal tract.     

Detection and partial characterization of a broad-range bacteriocin produced by Lactobacillus plantarum (ATCC 8014)

Lactobacillus plantarum (ATCC 8014) was tested for production of antimicrobial compounds. This study demonstrates that the cell-free supernatant of L. plantarum is effective in inhibiting the growth of a wide range of Gram-positive and Gram-negative bacteria including, Staphylococcus aureus, Escherichia coli, Listeria innocua, and Pseudomonas aeruginosa. The inhibitory compound lost activity when heated to temperatures greater than 30°C and when subjected to pH changes that lowered the pH below 4 or above 5. In addition, the inhibitory protein was susceptible to digestion by various proteases. The apparent molecular weight based on SDS-PAGE analysis is 122 KDa. These findings support the idea that the inhibitory compound is a protein and can be considered a bacteriocin secreted from the Gram-positive bacterium L. plantarum.     

The role of plNC8HK-plnD genes in bacteriocin production in Lactobacillus plantarum KLDS1.0391

The function of plNC8HK gene and plnD gene on the bacteriocin production of Lactobacillus plantarum KLDS1.0391 was studied. The results of bioinformatics analysis showed that PlNC8HK protein was a histidine protein kinase and a cytoplasmic membrane protein with six transmembrane helices. PlnD protein was a response regulator of the LytR/AlgR family. Quantitative real-time PCR revealed that plNC8HK and plnD genes of L. plantarum KLDS1.0391 were up-regulated significantly (P < 0.01) when L. plantarum KLDS1.0391 co-cultured with Lactobacillus helveticus KLDS1.9207. The plNC8HK-plnD mutant was also constructed. The change of antibacterial activity, AI-2 activity and cell number of plNC8HK-plnD mutant in co-culture and mono-culture demonstrated that plNC8HK and plnD genes were essential for bacteriocin production in L. plantarum KLDS1.0391, and the increase of bacteriocin production correlated closely with plNC8HK and plnD genes in co-culture. These results are beneficial to understand the role of two components system in the bacteriocin production of L. plantarum.     

Physical properties and antilisterial activity of bioactive edible films containing Lactobacillus plantarum

Bioactive polymeric films were developed from the incorporation of Lactobacillus plantarum into polysaccharide or protein edible films. Lactic acid bacteria (LAB) were added directly to the film forming solution and films were obtained by casting. Four different biopolymers were tested: sodium caseinate, pea protein, methylcellulose and hydroxypropylmethylcellulose. In order to study the impact of the incorporation of the protective culture into the biopolymer matrix, the water vapour permeability (WVP), optical and mechanical properties of the dry films were evaluated. Furthermore, survival of L. plantarum and the antimicrobial potential of bioactive films against Listeria innocua were studied. Whereas optical and mechanical properties of the films were not notably altered by the LAB incorporation, WVP values increased for all biopolymers. Viability of the strain was lower in cellulose derivative matrices than in protein films, although bacteriocin production in the newly prepared films was greater in these matrices. As a result, a significant antimicrobial activity against L. innocua was observed for polysaccharide matrices, which was not observed for protein films, where the bacteriocin production is delayed. In pea protein films the maximum bacteriocin production occurs at 15 storage days and afterwards decreases. So, the nature of the biopolymer and the knowledge of time when an adequate bacteriocin concentration is produced are key to use bioactive films in an effective way.     

Cultural conditions and nutritional components affecting the growth and bacteriocin production of Lactobacillus plantarum KC21

This study was conducted to investigate the effect of cultural conditions and nutritional components on the cell growth and bacteriocin production of Lactobacillus plantarum KC21. In cultures without pH control, the bacteriocin activity of L. plantarum KC21 was higher at 30oC, however, the cell growth rate was higher at 37°C. In MRS broth with an initial pH 6.0, the cell growth was lower, but high bacteriocin levels were recorded than at pH 7.0. The bacteriocin activity was maximal in medium containing 1.0 or 1.5% glucose or 1.0% lactose. Yeast extract (0.25 or 0.5%) added to MRS broth increased the bacteriocin activity, moreover, the bacteriocin in the presence of 1.0 or 3.0% NaCl, 0.5% NH₄PO₄, or 0.25 or 0.5% KH₂PO₄ resulted in the activity of 12,800 BU/mL, but excessively high salts concentration hindered the cell growth and bacteriocin production significantly. Besides 1.0 or 2.0 mM MgSO₄ highly increased the bacteriocin activity without affecting the growth of L. plantarum KC21, and ascorbic acid (1.0 or 3.0 ppm) enhanced the bacteriocin activity up to 2 fold.     

High-salt brines compromise autoinducer-mediated bacteriocinogenic Lactobacillus plantarum survival in Spanish-style green olive fermentations

The effect of NaCl on plantaricin production by five Lactobacillus plantarum strains was investigated. Plantaricin production in these strains was found to be regulated by three-component regulatory systems ruled by two different autoinducer peptides (AIPs), either PLNC8IF or Plantaricin A. Bacteriocin activity exhibited by these strains came to a halt in liquid medium containing NaCl concentrations of or above 2%. In contrast, bacteriocin activity was still observed when the producing strains were growing on solid medium containing up to 4% NaCl. Bacteriocin activity in liquid medium containing up to 2% NaCl could be restored by coculturing the producing strains with a selected plantaricin-production inducing strain of Lactococcus lactis. Growth of these bacteriocinogenic L. plantarum strains was monitored in traditional Spanish-style green olive fermentations. Survival of these strains could not be enhanced when provided with a range of plantaricin-production inducing mechanisms previously described, such as constitutive AIP production or coinoculation with a specific bacteriocin-production inducing strain of L. lactis. Our results suggest that it is advisable the use of constitutive bacteriocin producers, or at least non-AIP-dependant ones, as starters for olive fermentations due to the intrinsic physical characteristics of this food fermentation, especially the high salt concentration of the brines currently used.     

Use of Byproducts of Food Industry for Production of Antimicrobial Activity by Bacillus sp. P11

Production of antimicrobial activity by Bacillus sp. P11 was tested on different byproducts of food industry, like fish meal, grape waste, an industrial fibrous soybean residue, soybean meal, and cheese whey. Bacillus sp. P11 produced the higher amount of antimicrobial activity on soybean meal, followed by fish meal and fibrous soybean residue. Soybean meal was the selected medium to determine the effect of three variables (temperature, initial pH, and substrate concentration) on bacteriocin activity by response surface methodology, using a 2³ factorial design. Statistical analysis showed good adequacy to the model (R ² of 0.8268). In the range studied, temperature and initial pH of the medium have a significant effect on bacteriocin production, and substrate concentration has no significant effect. Response surface data showed maximum bacteriocin production at initial pH between 7.0 and 8.5 and temperature between 39 and 42 °C. In the optimum conditions (initial pH 7.0 and 42 °C), production of bacteriocin activity by Bacillus sp. P11 was compared using a commercial medium (BHI broth) and soybean meal. Maximum activity achieved with the soybean meal-based medium was similar to that obtained with BHI, indicating that soybean meal may be a cost-effective substrate for production of antimicrobial activity by Bacillus sp. P11.     

Potential of bacteriocinogenic Lactococcus lactis subsp. lactis inhabiting low pH vegetables to produce nisin variants

Antimicrobial behavior of lactic acid bacteria (LAB) has been explored since many years to assess their ability to produce bacteriocin, a natural preservative, to increase the shelf life of food. This study aims to characterize bacteriocin producing strains of lactic acid bacteria isolated from acidic to slightly acidic raw vegetables including tomato, bell pepper and green chili and to investigate their potential to inhibit food related bacteria. Among twenty nine LAB screened for antimicrobial activity, three exhibited antagonism against closely related bacterial isolates which was influenced by varying temperature and pH. They were identified up to strain level as Lactococcus lactis subsp. lactis TI-4, L. lactis subsp. lactis CE-2 and L. lactis subsp. lactis PI-2 based on 16S rRNA gene sequence. Their spectrum of inhibition was observed against food associated strains of Bacillus subtilis and Staphylococcus aureus. Moreover, L. lactis subsp. lactis PI-2 selected on the basis of higher antimicrobial activity was further evaluated for bacteriocin production which was detected as nisin A and nisin Z. These findings suggest the possible use of L. lactis strains of vegetable origin as protective cultures in slightly acidic as well as slightly alkaline food by the bio-preservative action of bacteriocins.     

Coculture with specific bacteria enhances survival of Lactobacillus plantarum NC8, an autoinducer-regulated bacteriocin producer,in olive fermentations

Bacteriocin production in Lactobacillus plantarum NC8 is activated by coculture with specific bacteriocin production-inducing bacterial strains. The system is further regulated by a three-component regulatory system involving a specific autoinducer peptide (PLNC8IF). We have used L. plantarum NC8 as a starter culture in Spanish-style green olive fermentations and examined the influence of coculturing in its survival. We found that L. plantarum NC8 greatly enhanced its growth and survival in the olive fermentations when coinoculated with two specific bacteriocin-production inducing strains, i.e. Enterococcus faecium 6T1a-20 and Pediococcus pentosaceus FBB63, when compared to singly-inoculated fermentations. In addition, a constitutive bacteriocin-producer NC8-derivative strain was used as a control in the olive fermentations and showed also better viability than the parental NC8 strain. Our results suggest the involvement of bacteriocin production in the viability enhancement found in both cases. We postulate that the presence of specific bacteria is recognized by L. plantarum NC8 as an environmental stimulus to switch a specific adaptive response on, most probably involving bacteriocin production. The design of novel bacteriocin-producing starter cultures for food fermentations should consider their constitutive versus regulated character.     

Statistical Optimization of Culture Components for Enhanced Bacteriocin Production by Lactobacillus plantarum LR/14

Statistical optimization of bacteriocin production by a natural isolate of Lactobacillus plantarum LR/14 was carried out in TGYE medium at 37°C and 200 rpm for 20h. In the first step of optimization using Plackett-Burman design, yeast extract, glucose and incubation period were identified as the most important factors for bacteriocin production. These factors were further optimized by response surface methodology (RSM) to understand their interaction and to determine their optimal levels. Results indicated that the maximum bacteriocin production was achieved in a medium containing glucose 2.0% and yeast extract 2.5%, and after an incubation period of 20h. An overall approximatley eightfold improvement in bacteriocin production was achieved as a result of optimization. These results indicated the importance of statistical tools in designing culture conditions for enhancing the production of bacteriocin from L. plantarum LR/14. Such an improved production will facilitate the application of bacteriocin, especially in food preservation.     

Bacteriocin-producing lactobacilli in Spanish-style fermented sausages: characterization of bacteriocins

Three bacteriocins were studied. Sakacin K, a bacteriocin from Lactobacillus sake CTC494 was purified to homogeneity by a four-step system involving ammonium sulphate precipitation, binding to a cation exchanger, hydrophobic interaction and reverse phase chromatography in FPLC (fast performance liquid chromatography) system. The peptide sequence was determined by Edman degradation. The first 30 amino acid residues from the N-terminus of sakacin K were identical to those of curvacin A from Lactobacillus curvatus LTH1174 and sakacin A from Lactobacillus sakei Lb706. The structural gene of sakacin K in Lb. sake CTC494 was located on a 60 Kbp plasmid (as has been previously reported) by curvacin A in Lb. curvatus LTH1174 and sakacin A in Lb. sakei Lb706. Plantaricin D, a bacteriocin-like compound isolated from Lb. plantarum CTC305 was purified and sequenced using the same procedure as described for sakacin K. The first 15 amino acid residues of plantaricin D were identical to those obtained from the bacteriocin inducer peptide (termed plantaricin A) of the bacteriocinogenic system in Lb. plantarum C11. The structural gene of the plantaricin D peptide was located on a similar EcoRl chromosomal fragment when compared to plantaricin A in Lb. plantarum C11. These two isolated bacteriocin-like peptides (sakacin K and plantaricin D) were purified from two new different strains obtained from fermented meat, confirming the ecological importance of these substances. Lactobacillus sakei CTC372, the third bacteriocinogenic strain selected in this study from fermented sausages, produced a bacteriocin named sakacin T. The bacteriocin is not found free in the supernatant of the cells. It is active against Listeria monocytogenes and Staphylococcus aureus. The genetic determinant of this bacteriocin was localized in a 84·4 Kbp plasmid by conjugative transfer and curing assays.     

植物乳杆菌M1-NTG300产细菌素培养条件优化

以植物乳杆菌素的效价作为特征性考察指标,对植物乳杆菌M1-NTG300产细菌素培养条件进行优化。通过响应面分析结合实际值确定培养基最佳组分为:葡萄糖浓度3.5%、蛋白胨浓度3.5%、吐温80浓度0.35%、氯化镁浓度0.20%,此培养条件下植物乳杆菌M1-NTG300所产细菌素的效价为1 269.4IU/mL,细菌素效价提高了19.8%。     

Antibacterial activity and mechanism of a novel bacteriocin produced by Lactiplantibacillus plantarum against Escherichia coli and Staphylococcus aureus

The purpose of the study was to explore the antibacterial effect and mechanism of a novel bacteriocin of Lactiplantibacillus plantarum (L. plantarum) against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The antibacterial activity was determined by the Oxford cup method, and the dynamic growth curves were conducted with continuous shaking incubation. The result showed that the bacteriocin was a protein or protein-like antibacterial substance susceptible to pepsin and trypsin. And it had good antibacterial activity, pH stability, thermostability and enzyme treatment stability against E. coli and S. aureus. The SEM, flow cytometry and nucleic acid leakage showed that the bacteriocin disrupted the cell structures of the two bacteria by damaging cell walls and cell membranes. An agarose gel electrophoresis and SDS-PAGE analysis showed that the bacteriocin inhibited DNA replication and interfered with the protein formation, which resulted in the inhibition of the two bacteria's growth. Therefore, the use of the L. plantarum bacteriocin might be a promising biocontrol strategy to inhibit the pollution of E. coli and S. aureus simultaneously in foods.     

Isolation and characterisation of the bacteriocin-producing Leuconostoc citreum HW02 from malts

Herein, a lactic acid bacterium which produces bacteriocin was isolated from malted barley. The isolated strain was identified by 16 S rRNA analysis and designated Leuconostoc citreum HW02. Bacteriocin HW02 was shown to have growth inhibition against health-threatening Gram-positive microorganisms such as Staphylococcus aureus, Listeria monocytogenes, and Latilactobacillus curvatus. Bacteriocin HW02 was reported to be incapable of inhibiting the growth of health-threatening microorganisms after treatment with proteolytic enzymes; thus, it was indicated that bacteriocin is proteinaceous in nature. Bacteriocin HW02 showed stability against a broad range of temperatures (60–121 °C), pH (2–10), various detergents, or organic solvents. Its antimicrobial activity against the indicator strain, Lactiplantibacillus plantarum NCDO 955, was maximised (320 AU mL⁻¹) midway through the stationary phase at 12 h. The crude bacteriocin HW02 molecular weight was estimated to be about 7.7 kDa. After the treatment of the indicator strain with bacteriocin, the maintenance of the optical density of the culture, the morphological analysis of cell-structure abnormalities, and cell shrinkage implied that bacteriocin showed bactericidal activity.     

Production of salami from beef, horse, mutton, Blesbok (Damaliscus dorcas phillipsi) and Springbok (Antidorcas marsupialis) with bacteriocinogenic strains of Lactobacillus plantarum and Lactobacillus curvatus

Lactobacillus plantarum 423, producer of bacteriocin 423, Lactobacillus curvatus DF38, producer of curvacin DF38, and a bacteriocin-negative mutant of L. plantarum 423 (423m) were evaluated as starter cultures in the production of salami from beef, horse, mutton, Blesbok (Damaliscus dorcas phillipsi) and Springbok (Antidorcas marsupialis). Growth of L. plantarum 423 and L. curvatus DF38 was best supported in Blesbok salami, as revealed by the highest growth rate during sweating, cold smoking and maturation, and final cell numbers after 70 days (1 × 10⁸ and 5 × 10⁷ cfu/g, respectively). Growth of Listeria innocua was the best suppressed in Blesbok salami fermented with L. plantarum 423 and L. curvatus DF38. Growth of L. innocua in horse salami was best suppressed when fermented with L. curvatus DF38. The final pH of salami fermented with L. plantarum 423 and L. plantarum 423m was slightly lower (4.4) compared to the pH of salami produced with L. curvatus DF38 (pH 4.7). No significant differences (P > 0.05) were recorded by a trained sensory taste panel amongst the three starter cultures regarding colour and venison like aroma. Horse, Blesbok and Springbok salami were rated significantly higher (P ? 0.05) in salami flavour than mutton salami, which was rated the lowest for this attribute. Blesbok salami was rated the highest for sour meat aroma, while beef salami was rated the lowest. Springbok salami was rated the highest in terms of oily mouth feel. Beef salami had the most compact structure and horse salami the softest structure of all meat types fermented. In general, salami produced with L. plantarum 423 yielded the best sour meat aroma, colour, texture, venison like flavour, sour meat flavour and oily mouthfeel and is considered superior to the L. plantarum mutant (strain 423m) and L. curvatus DF38.     

Characterisation of an antiviral pediocin-like bacteriocin produced by Enterococcus faecium

The bacteriocin-producing strain Enterococcus faecium ST5Ha was isolated from smoked salmon and identified by biomolecular techniques. Ent. faecium ST5Ha produces a pediocin-like bacteriocin with activity against several lactic acid bacteria, Listeria spp. and some other human and food pathogens, and remarkably against HSV-1 virus. Bacteriocin ST5Ha was produced at high levels in MRS broth at 30 °C and 37 °C, reaching a maximum production of 1.0 × 10⁹ AU/ml, checked against Listeria ivanovii ATCC19119 as target strain and surrogate of pathogenic strain Listeria monocytogenes. The molecular weight of bacteriocin ST5Ha was estimated to be 4.5 kDa according to tricine-SDS-PAGE data. Ent. faecium ST5Ha harbors a 1.044 kb chromosomal DNA fragment fitting in size to that of pediocin PA-1/AcH. In addition, the sequencing of bacteriocin ST5Ha gene indicated 99% of DNA homology to pediocin PA-1/AcH. The combined application of low levels (below MIC) of ciprofloxacin and bacteriocin ST5Ha resulted in a synergetic effect in the inhibition of target strain L. ivanovii ATCC19119. Bacteriocin ST5Ha displayed antiviral activity against HSV-1, an important human pathogen, with a selectivity index of 173. To the best of our knowledge, this is the first report on Ent. faecium as a potential producer of pediocin-like bacteriocin with antiviral activity.