Characterization of Purified Bacteriocin (Plantaricin and Acidocin) Produced from Lactobacillus Isolates and Study its Effects Against Growth Pathogenic Bacteria (original) (raw)
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The study of bacteriocins is one of the most relevant areas of research. This is due to the increase in the number of pathogenic microorganisms resistant to antibiotics. The bacteriocins of lactic acid bacteria have the most common application because of their safety in use. To assess the antimicrobial potential of bacteriocins of lactic acid bacteria and the possibilities for their further use, this study deals with the intensity of bacteriocin production by strains of lactic acid bacteria, the minimal inhibitory concentration of bacteriocins produced by these strains with respect to E. coli В-6954, as well as the antimicrobial activity of the produced bacteriocins with respect to some strains of pathogenic microorganisms. This work was carried out on the basis of the Kemerovo Technological Institute of Food Industry (Russia). The study objects were strains of microorganisms Lactobacillus delbrueckii B2455, Lactobacillus paracasei B2430, Lactobacillus plantarum B884. Biomass concentration was determined by spectrophotometric method. Study of antimicrobial activity in relation to a number of pathogenic strains was carried out in a culture fluid by optical density, and also by a disk-diffusion test. As a result, the maximum productivity value, characterizing the intensity of bacteriocin production, was observed for strain B884. However, the quality of bacteriocin (antimicrobial activity relative to the E. coli strain) with a lower productivity is higher for strain B2430. The minimum inhibitory concentration for the strains studied was: B2430 – 1.0 10 6 CFU/ml, B884 – 1.0 10 8 CFU/ml, B2455 – 1.0 10 7 CFU/ml. The best indicator of the minimum inhibitory concentration was observed for strain B2430. Thus the strain B2430 had the greatest antimicrobial activity, it had an inhibitory effect on 12 test cultures: Escherichia
2020
Two broad spectrum bacteriocins producing lactobacilli isolated from dairy samples were identified as L. plantarum DP2 and L. casai DD1. Bacteriocin production from L. plantarum DP2 was recorded maximum in the presence of maltose as sole source of carbon, whereas glucose was found to be best carbon source in case of L. casai DD1. Both bacteriocin showed maximum production when three nitrogen sources tryptone, yeast extract and meat extract were present together in the medium at pH 6 and 37 o C. Bacteriocins from L. plantarum DP2 and L. casai DD1 were purified up to 4 fold and 6.3 fold with a recovery of 146.8% and 130.2% respectively. Molecular weights of purified bacteriocins were 4.8 kDa (L. plantarum DP2) and 9.2 kDa (L. casai DD1). Bacteriocin from L. plantarum DP2 was thermal stable, active at pH 4 to 8 and also showed stability at high salt concentrations (2-10%). Increased activities of this bacteriocin were also recorded with 2% NaCl, 1% EDTA and 1% tween 80. Whereas bacteri...
Plantaricin LD1: A Bacteriocin Produced by Food Isolate of Lactobacillus plantarum LD1
Applied Biochemistry and Biotechnology, 2014
Plantaricin LD1, a bacteriocin produced by Lactobacillus plantarum LD1, was characterized for biochemical and antimicrobial properties. Bacteriocin showed stability at high temperatures (100°C for 20 min and 121°C for 15 min under 15 psi pressure), in a pH range of 2.0-8.0 and also in the presence of organic solvents, surfactants and detergents. The crude preparation was not affected by catalase, amylase and lipase but activity was reduced in the presence of pepsin, trypsin and proteinase K showing proteinaceous nature of the compound. The molecular weight of bacteriocin was found to be ∼6.5 kDa, and antimicrobial activity was confirmed by bioassay. It inhibited not only related strains but also other Grampositive and Gram-negative bacteria such as Lactobacillus curvatus NRRL B-4562, Lactococcus lactis subsp. lactis NRRL B-1821, Enterococcus faecium NRRL B-2354, Enterobacter cloacae NRRL B-14298, Micrococcus luteus, Staphylococcus aureus, urogenic Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Shigella flexneri and Vibrio sp. These properties of plantaricin LD1 suggest its applications not only in food safety but in therapeutics as well.
Postępy Higieny i Medycyny Doświadczalnej, 2012
In this paper, an attempt was made to evaluate the antibacterial potential of standard strains of lactic acid bacteria (LAB) producing bacteriocins of various classes, thus demonstrating various mechanisms of cell membrane damages against the Streptococcus agalactiae strains (Group B Streptococcus, GBS), depending on surface polysaccharides and surface alpha-like protein genes. Materials/Methods: Antimicrobial property of the strains of L. plantarum C 11, L. sakei DSMZ 6333, and L. lactis ATCC 11454 producing bacteriocins: JK and EF plantaricins, sakacin and nisin, respectively, against the GBS strains was evaluated. The chosen to the study GBS strains were represented by serotypes Ia, Ib, II, III, V and they had bca, epsilon, rib, alp2 or alp3 alpha-like protein genes. The experiment was conducted by means of suspension culture and the bacteria count was determined using the serial dilution method. Results: A great ability of L. plantarum C 11 strain was proven to inhibit the GBS growth. The strain of L. sakei DSMZ 6333 did not demonstrate any ability to inhibit the growth of GBS, whereas L. lactis ATCC 11454 inhibited the growth of S. agalactiae indicator strains to a minor extent. Statistically significant differences were demonstrated between the GBS strains representing various serotypes against the antimicrobial activity of model LAB strains. The least sensitive to the activity of bacteriocins were the strains representing serotypes Ib and III, whereas the strains representing serotype II were the most sensitive. The sensitivity of the GBS strains to the antimicrobial activity of LAB was not dependent on alpha-like protein genes. Discussion: Among the LAB standard strains producing bacteriocins, the strongest antimicrobial property was observed in the strain of L. plantarum C 11. Because of the generally known and verified strong antagonistic property of the strains of L. plantarum species against indicator bacteria, it is necessary to further pursue the research presented in this paper.
Detection and characterization of a bacteriocin produced by Lactobacillus plantarum C19
Canadian Journal of Microbiology, 1993
Strain C19, isolated from fermented cucumbers and identified as Lactobacillus plantarum, produced a bacteriocin. This bacteriocin, named plantaricin C19, was stable at acidic pH, was relatively thermostable, and had a molecular mass of about 3.5 kDa. It inhibited some pathogenic (i.e., Listeria spp.) and spoilage Gram-positive bacteria but had weak or no action against lactic acid bacteria. Its adsorption on sensitive and resistant bacteria was pH dependent and was reduced by pretreatment of cells with lipase or lysozyme. Curing treatments with acriflavine or novobiocin yielded nonproducing mutants sensitive to plantaricin C19.Key words: lactic acid bacteria, Lactobacillus plantarum, bacteriocin, protein.
OPTIMIZATION OF BACTERIOCIN PRODUCTION BY LACTOBACILLUS PLANTARUM AA135
ROMANIAN ARCHIVES, 1971
The presence of the specific antibodies for some enterobacteria -Yersinia, Salmonella and Shigella was investigated in patients hospitalized in the period 2000 -2007 with reactive arthritis and other rheumatoid diseases. The antibacterial antibodies in the diagnosis titres were found in 172 out of 1102 (15.6 %) patients. Increased antibodies levels to Yersinia were detected in the sera from 113 (65.7 %) of the 172 patients, for Shigella in 47 (27.3 %) and for Salmonella in 12 (7.0 %) cases. From all serologically positive patients 126 (73.2 %) had clinical diagnosis of reactive arthritis and 46 (26.7%) other rheumatoid diseases (ankylosing spondilytis, Reiter's syndrome, sacroilitis). The most serologically positive cases (63.9 %) were of middle-age (30-50 years). There were no significant differences between sexes among serologically positive cases. ABSTRACT Growth of Lactobacillus plantarum AA135 in MRS broth at 30 o C yielded maximal plantaricin AA13 activity of 10000 AU/ml at pH 3.8 after 16 hours. Maximal activity (10000 AU/ml) of plantaricin AA13 was recorded in MRS broth at initial pH of 5 or 5.5. Optimal production (10000 AU/ml) was recorded in the presence of tryptone (20 g/l), a combination of tryptone plus meat extract (1:0.6) or tryptone with yeast extract (1:0.6) as sole nitrogen source. Growth of Lactobacillus plantarum AA135 in the presence of 20 g/l glucose yielded bacteriocin activity of 10000 AU/ml. Moreover, when 40 g/l glucose was added, the plantaricin AA13 levels doubled to 20000 AU/ml. Concentrations of 5, 10 and 20 g/l K 2 HPO 4 lowered bacteriocin activity by 60 %. Supplementing MRS with 1 g/l or more glycerol repressed bacteriocin production. Maximal activity of 20000 AU/ml was recorded in MRS supplemented with Vitamin DL-6,8-thioctic acid. ABSTRACT Introduction. The emergence of the bacterial antibiotic multi-resistance made more and more stringent the developing of new anti-microbial strategies. The purpose of the present study was to investigate the antimicrobial potential of six (6) newly synthesized chemical compounds (derivating from phenantroline and dimethylguanin-copper complex combinations) versus 97 enterobacterial strains isolated from the hospital environment. The qualitative screening of the antimicrobial activity of the chemical compounds was performed by an adapted diffusion method. The minimal inhibitory concentrations (MIC) of the active chemical compounds were established by Mueller Hinton broth microdillution method. The tested chemical compounds were also tested for their ability to inhibit microbial adherence and biofilm development on inert substrata by a simple microtiter method. All six chemical compounds proved to have antimicrobial activity versus the most of the tested strains, the phenantroline derivatives exhibiting higher antimicrobial activity than the dimethylguanidine-copper complex combinations. The subinhibitory concentrations of the tested chemical products slightly inhibited the adherence ability of the bacterial strains to the inert substratum. Our results demonstrated that phenantroline derivatives may represent a new strategy of antimicrobial treatment, simultaneously with the bactericidal effect, the subinhibitory concentrations of these newly synthesized chemical compounds decreasing the adherence ability of bacteria to the inert substratum.
Determination of antimicrobial activity of bacteriocin of lactic acid producing bacteria
A study was conducted to explore the potential of bacteriocins, produced by lactic acid producing bacteria, against Staphylococcus aureus, Eneterococcus fecalis and Escherichia coli. Different lactic acid producing bacteria viz. Lactobacillus fermentum, Lactobacillus acidophilus and Lactobacillus rhamnosus were isolated from milk, milk products of dairy animals and intestinal contents of healthy broiler chicken. The isolated bacterial species were identified morphologically and biochemically by conventional methods. From these bacteria, bacteriocins were isolated by precipitation method and antibacterial activity was determined by disk diffusion method. Isolated bacteriocin showed the inhibitory activity against Staphylococcus aureus and Enterococcus feacalis. Whereas bacteriocins showed very poor inhibitory activity (2mm) against Eschrechia coli. The study showed that bacteriocins are quite useful against commonly known food pathogens and thus can be used for the preservation of fo...
The Iraqi Journal of Veterinary Medicine, 2012
Two isolates of Lactobacillus acidophilus were isolated from locally homemade soft cheese and identified in accordance with their main features as described by bergeys manual of determinative bacteriology .Agar well diffusion bioassay was used for the evaluation of antimicrobial activity of cell free supernatant (culture filtrates) of Lactobacillus acidophilus isolates against the sensitive strain Lactobacillus acidophilus R0052 and the diameter of the inhibition zone was proportional to the bacteriocin concentration. The antimicrobial activity of the bacteriocin was proportional to the reciprocal of the highest dilution factor producing a detectable zone of inhibition . The storage temperature of the crude bacteriocin had a significant(P˂ 0.01) effect on its potency against the sensitive strain .The potential of the antimicrobial activities of the crude bacteriocin that was stored for 10 days at refrigeration temperature (4°C) was significantly (P˂ 0.01) higher than that stored at ...
Antimicrobial Activity Studies of Bactoriocin Produced by Lactobacilli Isolates from Carrot Kanji
Online Journal of Biological Sciences, 2012
In the present study, Staphylococcus aureus a causative agent of food poisoning is selected as a test organism to study the antimicrobial effect of bacteriocin. S. aureus produces number of exotoxins and enterotoxins which enters the body via contaminated food causing illness. Approach: In this case the use of antibiotics is one of the ways of treatment, but in addition to this if we advise such patients to consume the carrot kanji then it will cause better effect because carrot kanji is the naturally fermented food beverage consisting of microflora mainly the Lactobacilli. Results: The Lactobacilli have ability to produce antimicrobial compounds called bacteriocin. Isolation of bacteriocin was carried out from the naturally fermented carrot kanji. The bacteriocin produced by Lactobacilli was dialysed and used for the further studies. The well diffusion method is used to study the antimicrobial activity, effect of temperature, pH, enzymes on bacteriocin. From the diameter of zone of inhibition the activity of bacteriocin was determined. The sensitivity of bacteriocin at different pH range showed that at neutral pH the diameter of inhibition zone was greater than that at alkaline as well as acidic pH. Upto 100°C the bacteriocin activity was 80% but as temperature range increased upto 121°C it reduced sharply to 28%. Conclusion/Recommendations: In addition to this the effect of alpha amylase, trypsin, catalase enzyme on bacteriocin activity was also studied which shows positive results with alpha amylase, reduced activity with trypsin and catalase remained unaffected.
Determination of Antibacterial Activity of Bacteriocins of Lactic Acid Producing Bacteria
2013
Received: 2013-01-30 Revised: 2013-02-27 Accepted: 2013-02-28 This study was conducted to explore the potential of bacteriocins, produced by lactic acid producing bacteria, against Staphylococcus aureus, Eneterococcus fecalis and Escherichia coli. Different lactic acid producing bacteria viz. Lactobacillus fermentum, Lactobacillus acidophilus and Lactobacillus rhamnosus were isolated from milk, milk products of dairy animals and intestinal contents of healthy broiler chicken. These isolates were identified morphologically and biochemically by conventional methods. Bacteriocins were obtained from these bacteria, by precipitation method. Antibacterial activity of bacteriocins was determined by disk diffusion method. Isolated bacteriocin showed the inhibitory activity against Staphylococcus aureus and Enterococcus feacalis but showed very poor inhibitory activity (2mm) against Eschrechia coli. The result of this study showed that bacteriocins are quite useful against commonly known foo...