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1.
A bacteriocinogenic strain identified as Enterococcus faecium SH01 was isolated from mukeunji, a Korean traditional over-ripened kimchi with antimicrobial activities against Listeria monocytogenes KCTC 3569 and Lactobacillus curvatus KFRI 166. The maximum bacteriocin titer (1,280 AU/mL) was detected at the early stationary phase and was maintained for 28 h with no activity loss. The bacteriocin activity, which disappeared after treatment with the proteolytic enzymes α-chymotrypsin, pronase E, proteinase K, and trypsin, was partially inactivated using α-amylase. The activity of bacteriocin SH01 remained after heat treatment (121°C, 15 min) and exposure to pH values from 2–12. The molecular weight of crude bacteriocin SH01 was 3 kDa. The bacteriocin production phenotype (Bac+) was linked to a 6 kb plasmid. Bacteriocin SH1 production was not induced by the co-presence of viable cells, heat treatment, or a cell-free indicator supernatant. The mode of action of bacteriocin SH1 was bactericidal.  相似文献   

2.
The aim of this work was to isolate and characterize bacteriocins produced by 2 Lactobacillus fermentum strains isolated from artisanal Mexican Cocido cheese. Fractions (F ≤3 kDa) obtained from cell-free supernatants of Lb. fermentum strains J23 and J32 were further fractionated by reversed-phase HPLC on a C18 column. Antimicrobial activities of F ≤3 kDa and bacteriocin-containing fractions (BCF), obtained from fractionation of F ≤3 kDa against 4 indicator microorganisms, were determined by the disk diffusion method and growth inhibition in milk. Subsequently, isolated BCF were analyzed by reversed-phase HPLC tandem mass spectrometry. Results showed that BCF presented antimicrobial activity against the 4 indicator microorganisms tested. For J23, one of the fractions (F3) presented the highest activity against Escherichia coli and was also inhibitory against Staphylococcus aureus, Listeria innocua, Salmonella Typhimurium, and Salmonella Choleraesuis. Similarly, fractions F3 and F4 produced by J32 presented antimicrobial activity against all indicator microorganisms. Furthermore, generation time and growth rate showed that F3 from J23 presented significantly higher antimicrobial activity against the 4 indicator microorganisms (2 gram-positive and 2 gram-negative) when inoculated in milk compared with F3 from J32. Interestingly, this fraction presented a broader antimicrobial spectrum in milk than nisin (control). Reversed-phase HPLC tandem mass spectrometry analysis revealed the presence of several peptides in BCF; however, F3 from J23 that was the most active fraction of all presented only 1 bacteriocin. The chemical characterization of this bacteriocin suggested that it was a novel peptide with 10 hydrophobic AA residues in its sequence and a molecular weight of 2,056 Da. This bacteriocin and its producing strain, J23, may find application as a biopreservative against these indicator microorganisms in dairy products.  相似文献   

3.
Bacteriocins are antimicrobial peptides acting on certain pathogens and spoilage microorganisms. They have recently been considered as natural alternative to food preservatives. The objective of this work was to study the inhibition spectrum of a bacteriocin produced by Lactobacillus buchneri, isolated from a Mexican sausage, against Listeria monocytogenes and several lactic acid bacteria, and the influence of several extrinsic parameters on bacteriocin activity. Culture media (MRS or APT) had no significant effect on bacteriocin production (p < 0.060) although higher activity was observed in MRS (220 AU/mL). Conversely, temperature significant affected production (p > 0.001). Bacteriocin activity was also increased by 100% nitrogen atmosphere (180 AU/mL) as compared to N2/CO2 (50:50) (150 AU/mL) and 100% CO2 (60 AU/mL). Maximum activity occurred at the end of the exponential growth phase. This bacteriocin inhibited some Listeria monocytogens strains and several lactic acid bacteria, mainly Lactobacillus sp.  相似文献   

4.
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.  相似文献   

5.
Lactococcus lactis 19.3 produces a bacteriocin with a wide inhibitory spectrum, including the food pathogen Listeria monocytogenes. The producing strain was able to grow and produce similar amounts of bacteriocin in MRS medium, cow's milk and soya milk, respectively. The mode of action of this bacteriocin was further investigated using two indicator strains, Lactobacillus delbrueckii subsp. bulgaricus LMG 6901T and Listeria monocytogenes ATCC 1911‐1. The bacteriocin displayed a bactericidal effect, causing a rapid decrease of the cell viability. Bacteriocin treatment of the sensitive strains resulted in major morphological changes, including pore formation, as shown by scanning electron microscopy. Finally, the bacteriocin‐encoding gene was identified by sequencing. The presence of nisin gene was confirmed. Based on all our data gathered so far, L. lactis 19.3 is a good candidate for a starter or protective culture in the manufacturing of both fermented dairy and vegetarian food products.  相似文献   

6.
Characterization of some bacteriocins with regard to possible applications in food industry. Part 1. Selection of strains. The ability to produce bacteriocins is common in all genera of lactic acid bacteria. Thirty-four out of 223 strains of lactic acid bacteria from the culture collection of the University Potsdam produced bacteriocins active against one or more indicator strains. Seventeen Gram-positive and Gram-negative strains, frequent occur as spoilage bacteria in food industry, were chosen as indicator strains, and 11 of them were inhibited by one or several bacteriocin producers. Most of the bacteriocin producers belong to the genera Enterococcus or Lactobacillus. Bacteriocins derived from Enterococcus faecium and Pediococcus acidilactici exhibited the widest inhibitory spectrum. The properties of Enterococcus faecium 10.051 and Enterococcus faecium 10.211 bacteriocins were further investigated to evaluate their potential use as natural food preservatives. The bacteriocin produced by Enterococcus faecium 10.211 exhibited a very wide inhibitory spectrum against food-spoiling strains. It was heat stable (100 °C for 60 min) and stable in a wide range of pH (1.2–10.0). The bacteriocin produced by Enterococcus faecium 10.051 showed a smaller inhibitory spectrum, and it was less stable against temperature and pH as the bacteriocin from strain 10.211. Both bacteriocins were inactivated by proteinase K. pronase E, and α-chymotrypsin. Bacteriocin from strain 10.211 will be the object of further investigations for application of bacteriocins in food industry.  相似文献   

7.
Bacteriocins are natural antimicrobial agents mainly act against closely related bacteria and at times unrelated organisms including various food spoilage and pathogenic organisms. Bacteriocins from Lactic Acid Bacteria (LAB) have been widely used for food preservation because of their safety nature. In this study, bacteriocin from Pediococcus pentosaceus KJBC11 was purified with the recovery of 15%, using cell adsorption‐desorption technique, gel permeation chromatography, hydrophobic interaction chromatography, and reversed phase HPLC successively. Molecular weight of bacteriocin KJBC11 was determined to be 4.5 kDa using Tricine SDS‐PAGE. The bacteriocin KJBC11 showed strong inhibitory activity against various Gram‐positive and Gram‐negative pathogens. Bacteriocin KJBC11 activity was not altered when treated with amylase, lipase, and trypsin but inactivated by protease and proteinase enzyme treatment. It was heat stable (100°C, 1 hr) and exhibited strong antimicrobial activity within the pH 3–7. Its mode of action was bactericidal in nature as revealed against Listeria monocytogenes showing leakage of internal contents out of the cell. Bacteriocin KJBC11 could belong to Class IIa‐pediocin like bacteriocins based on its characteristics as well as the presence of pediocin gene in the genome of the isolate KJBC11.

Practical applications

Studies on Lactic Acid Bacteria and bacteriocin have attracted increasing attention for a long time, due to their established role in food fermentation and preservation. The antimicrobial properties of bacteriocin became a trademark approach to achieve food safety and to counter the menace of antibiotic resistant pathogens. This study revealed a potent bacteriocin with wide range of antibacterial activity against various foodborne and clinically important pathogens to have potential application in food preservation and biomedicine.  相似文献   

8.
This study was conducted to optimize bacteriocin producing enterococcal strains isolated from indigenous fermented dairy products of Pakistan. Isolates IJ-06, IJ-21, and IJ-31 were identified as Enterococcus faecium and IJ-11 was identified as Enterococcus faecalis. All of these enterococcal isolates were catalase, gelatinase negative, and non-hemolytic on sterile sheep blood. Bacteriocin production trials confirmed E. faecium IJ-06, IJ- 21, and IJ- 31 as the potential producer of bacteriocin showing antimicrobial activity in cell-free supernatant (CFS). E. faecalis IJ-11 displayed activity after partial purification. Optimization of culture conditions for the production of bacteriocin by the selected strains showed that maximum production was achieved at 35–37°C with 1% inoculum after 16 h of incubation and at pH 7–8. The molecular mass of the partially purified enterocins falls in the range of 4.5–4.7 kDa. These enterocins were close to class IIa of bacteriocins and were highly active against Listeria monocytogenes, Bacillus subtilis, Bacillus cereus and other closely related strains.  相似文献   

9.
Leuconostoc mesenteroides E131, isolated from Greek traditional fermented sausage, prepared without the addition of starters, produces a bacteriocin which is active against the pathogen Listeria monocytogenes. The bacteriocin was purified by 50% ammonium sulphate precipitation, cation exchange, and reverse-phase chromatography. Bacteriocin is active at pH values between 4.0 and 9.0 and retains activity after incubation for 1 h at 100 °C. Proteolytic enzymes inactivated the bacteriocin after 1 h of incubation, while renin resulted in full inactivation only after 24 h. Lipase resulted in full inactivation after 4 h. Applying molecular methods, it was determined that the bacteriocin produced, named as mesenterocin E131, was identical to mesenterocin Y105 and was expressed during the exponential growth phase.  相似文献   

10.
Bacteriocin KU24 produced by Lactococcus lactis KU24 exhibited an inhibitory effect against methicillin‐resistant Staphylococcus aureus (MRSA). Bacteriocin KU24 was inactivated by protease XIV, showing that it has a proteinaceous nature on S. aureus ATCC 33591. Also, bacteriocin KU24 exhibited a strong heat stability (121 °C for 15 min) and pH stability (pH 3 to 9). The mode of inhibition was determined for S. aureus ATCC 33591 by treatment of 0, 250, and 500 AU/mL of bacteriocin KU24. S. aureus ATCC 33591 was inhibited by added bacteriocin KU24, while control was increased. The cell membranes of S. aureus ATCC 33591 were damaged with treatment of 500 AU/mL of bacteriocin KU24. Also, bacteriocin KU24 inhibited the occurrence of mecA gene, the methicillin resistance gene in S. aureus ATCC 33591. Bacteriocin KU24 was purified by C18 Sep‐Pack column, cation exchange chromatography, and gel filtration chromatography, and molecular mass is approximately 6.5 kDa by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. These results demonstrate that bacteriocin KU24 can be used as an alternative antimicrobial agent for the treatment of infection of MRSA in the food industry.  相似文献   

11.
The dynamics of cell growth and bacteriocin production by Lactobacillus curvatus CWBI‐B28 in modified De Man/Rogosa/Sharp (mMRS) broth with various concentrations of glucose and complex nitrogen source (CNS; peptone, yeast extract and meat extract) was investigated in flask fermentations and in a laboratory fermentor using batch and fed‐batch cultivations. In fed‐batch fermentation the rate of feeding of the reactor with the substrates was either maintained constant (0.12 L h?1) or varied exponentially as a function of time. The results showed that both cell growth and bacteriocin activity were influenced by changes in the concentrations of glucose and CNS. Optimal growth and bacteriocin activity were obtained in mMRS broth containing 40 g L?1 glucose and 40 g L?1 CNS (mMRS40/40). A bacteriocin titre of 4266 AU mL?1 and a cell count of 8.7 log colony‐forming units (cfu) mL?1 were recorded when this medium was used for cultivation. In batch fermentation using the same medium, a higher cell count (9.5 log cfu mL?1) and twice as much bacteriocin as in flask fermentation were produced. The highest bacteriocin titre (8533 AU mL?1) was obtained with fed‐batch fermentation at an exponentially varying rate of feeding. Bacteriocin activity and cell dry mass did not always correlate. Copyright © 2007 Society of Chemical Industry  相似文献   

12.
Pediococcus species were isolated from ogi, fermented cow and sheep milk. Functional properties such as hydrogen peroxide production, tolerance to simulated gastric transit with pepsin and bile salts, bile salt hydrolytic (BSH) activity, in vitro adherence assay and antimicrobial characteristics were carried out. The strains tolerated bile salts and BSH activity was positive. Pediococcus acidilactici OB4 survived gastric transit after 180min comparable to a probiotic strain, L. acidophilus CNRZ1923. The strains exhibited good adhesion to the three extracellular matrices. Two isolates (P. acidilactici OB4 and P. pentosaceus SM3) produced bacteriocin. Bacteriocins were stable at pH 4-9 and on treatment with lipase, catalase, α-amylase and lysozyme, while their activity was lost on treatment with proteinase K, pronase E, pepsin and trypsin. The bacteriocins produced by P. pentosaceus SM3 was heat stable at 100°C for 10 min while P. acidilactici OB4 was stable at 100°C for 30 min. The bacteriocin produced by P. acidilactici OB4 was identified as PedA while P. pentosaceus SM3 was PedB. The bacteriocins had relative heat stability, high anti-listerial activity and a good spectrum of activity against some pathogenic microorganisms. The results demonstrated possible inclusion in a starter culture fermentation process of food and dairy products and safety characteristics.  相似文献   

13.
从植物性材料中筛选到1株对大肠杆菌有明显抑制作用的乳酸菌,在排除有机酸和过氧化氢的干扰后,该菌株的发酵上清液仍有较强的抑菌性;胃蛋白酶处理后,抑菌活性明显降低,说明其抑菌物质为蛋白质类物质,是一种细菌素。通过形态学和生理生化分析,初步鉴定该菌株为植物乳杆菌。生物学特性研究表明,该菌株发酵上清液经高温、吐温-80、SDS、EDTA处理后,仍保持较好的抑菌活性;在酸性条件下稳定;对蛋白酶K和胰蛋白酶较胃蛋白酶敏感。抑菌谱显示,该菌株的发酵上清液具有广谱抑菌性。  相似文献   

14.
The aim of this work was the characterization of new strains of lactic acid bacteria (LAB) from farmed fish and with potential application as biopreservatives against both Listeria monocytogenes and Staphylococcus aureus. Twenty-five strains of LAB isolated from the muscle of farmed turbot were investigated. Genetic identification of the bacteriocin-producing LAB strains was performed by means of a PCR method using novel BAL1/BAL2 16S ribosomal-RNA-targeted primers. Maximum bacteriocin production by Lactococcus lactis ssp. lactis USC-39, Enterococcus faecium USC-46 and Enterococcus mundtii USC-51 was detected in the stationary phase of growth. Both acidification and the production of hydrogen peroxide by LAB were ruled out as the source of the inhibition. In contrast, the antimicrobial activity of all three LAB strains was inactivated by the addition of proteinase K, thus confirming the proteinaceous nature of the inhibition. The activity against L. monocytogenes was maintained in the 3.5–5.5 or 3.5–6.5 pH range, depending on the LAB strain. Likewise, inhibition of S. aureus strains was observed in the 3.5–4.5 and in the 3.5–5.5 pH ranges, depending on the LAB strain and on the S. aureus strain tested. Bacteriocin activity was stable in all three strains after heating the cell-free extract for 60 min at 100 °C, or even for 15 min at 121 °C, in all the three LAB strains. The acidic and heat-resistant bacteriocins produced by the three LAB strains isolated from turbot, able to inhibit the growth of both L. monocytogenes and S. aureus may find application as biopreservatives in fermented and/or heated food products.  相似文献   

15.
In this study, the growth of chloramphenicol‐resistant bacteriocin‐sensitive indicator strain Lactobacillus casei DPC 2048CM was evaluated in Cheddar cheese made with bacteriocin‐producing Lactobacillus paracasei DPC 4715. No suppression of growth of the indicator strain was observed in the cheese during ripening, and no bacteriocin production by L. paracasei DPC 4715 was detected by the well diffusion method in cheese and cheese extracts. The bacteriocin produced by L. paracasei DPC 4715 was sensitive to chymosin and cathepsin D, and it may have been hydrolysed by the rennet used for cheese manufacture or by indigenous milk proteases.  相似文献   

16.
Bacteriocin‐producing (Bac+) Lactococcus lactis ssp. lactis CCMM/IAV/BK1 isolated from traditional lben was used in the preparation of lben from pasteurized milk to assess its potential inhibitory activity against Listeria monocytogenes ATCC 7644. Production of bacteriocin (arbitrary units, AU) in MRS broth fortified with yeast extract (MRSY) in a fermentor under controlled and uncontrolled pH conditions was also investigated. This Bac+ strain yielded about 35 times more bacteriocin when the pH was maintained constant at 6.5 than under varying pH conditions. To test the effect of in situ bacteriocin production against L. monocytogenes, lben was made from cow's milk artificially contaminated with approximately 107 cfu/mL and fermented with a mixed mesophilic starter culture consisting of the lactococcal Bac+ organism and Lc. lactis ssp. lactis biovar diacetylactis 66, a diacetyl‐producing strain, in a ratio of 1 : 1. Numbers of L. monocytogenes were monitored during fermentation and storage of lben at refrigeration temperature (c. 7°C) for up to 6 days. Performances of the Bac+ starter were compared to those of an isogenic Bac? derivative strain obtained from the Bac+ starter by curing with ethidium bromide. The results showed that the amount of L. monocytogenes decreased to below the detectable level in a 1‐mL sample within 24 h of storage at 7°C in lben fermented with the Bac+ starter culture. On the contrary, L. monocytogenes survived for 6 days of storage at 7°C in lben made with the Bac? starter. The Bac+ wild strain of the starter studied could be adequately used to produce lben or similar indigenous fermented milks of improved hygienic quality on an industrial scale. Alternatively, it could be used as an adjunct in minimally processed products or in products obtained from raw milk to add a safety factor.  相似文献   

17.
Forty‐two strains of Lactobacillus bulgaricus isolated from locally made yogurts were examined and compared for bacteriocin producing ability using spot on lawn assay which improved by taking photo and image processing. Lb. bulgaricus K41 exhibited the highest inhibition level against indicators. K41 Bacteriocin‐like inhibitory substance is sensitive to proteolytic enzymes (proteinase K, pepsin, and trypsin) but α‐amylase makes slight reduction in its activity and it is resistant to lipase. This antibacterial peptide is extremely heat‐stable (121 °C for 15 min) and remains active over a wide pH range (pH = 2 to 10); also nonionic detergents (Tween‐20, Tween‐80, and Triton X100) showed no effect on its activity. The inhibitory spectrum is against Gram‐positive bacteria (except Staphylococcus aureus) with extremely antilisterial activity and it is almost ineffective against Gram‐negative bacteria. The mode of its action was identified as bactericidal against Listeria monocytogenes. The properties of K41 bacteriocin‐like inhibitory substance add to its safety as a biopreservative produced by a generally recognized as safe (GRAS) bacterium suggesting it can be used in hurdle technology for ready‐to‐eat foods as one of the main sources of Listeria contaminations.  相似文献   

18.
The effect of food preservatives on the activities of acidocin CH5 and bacteriocin D10 was evaluated. Lactobacillus delbrueckii subsp. lactis LTI30 was used as an indicator strain. Methyl paraben and propyl paraben at concentrations higher than 0.075 g/l enhanced the activity of acidocin CH5 up to 45% and 50% respectively. As regards bacteriocin D10, the synergistic effect was less pronounced. The inhibitory effect of EDTA applied alone at 0.1–0.2 mmol/l against the indicator atrain increased from 4% to 10%. EDTA (0.1–0.2 mmol/l) in combination with acidocin CH5 or bacteriocin D10 increased their inhibitory effects from 50% to 70% and from 10% to 50% respectively.  相似文献   

19.
This work focused on Listeria monocytogenes growth inhibition and growth rebound in raw and cooked pork meat inoculated with Lactobacillus curvatus strains. During storage of raw meat homogenates in the presence of the bacteriocin-producing strain Lactobacillus curvatus CWBI-B28wt, the Listeria monocytogenes cfu count was initially reduced to an undetectable level, but a growth rebound occurred after two weeks, coinciding with loss of 70% of the bacteriocin activity present at the end of week 2. The Listeria growth rebound was suppressed when proteolysis of bacteriocin was countered by the absence of proteases (bacteriocin addition to cooked meat) or the presence of 1% soy flour (added to provide competing substrates). Further experiments confirmed that bacteriocin is sensitive to the action of proteolytic enzymes isolated from both Lactobacillus curvatus CWBI-B28wt and the meat matrix. Bacteriocin proteolysis thus emerges as a cause of Listeria growth rebound.  相似文献   

20.
目的:对从水开菲尔粒中分离得到1 株能产生抑菌物质的菌株QF01进行菌种鉴定,并对其产细菌素进行实验、鉴定和基因序列分析。方法:通过牛津杯法测定抑菌能力,采用聚合酶链式反应(polymerase chain reaction,PCR)扩增16S rDNA和细菌素相关基因并测序,利用ProParam tool、TMHMM 2.0、InterProScan、SOPM和SWISS-MODEL在线软件对基因编码产物进行分析。结果:该菌株产的细菌素对大肠杆菌(Escherichia coli JM109)有明显的抑制作用,通过16S rDNA序列分析初步确定该菌株属于植物乳杆菌(Lactobacillus plantarum)。该菌株含有plnD、plnEF、plnV、plnR四种基因,其中plnV基因编码产物有跨膜螺旋结构,其结构存在信号肽特征。此外,从三级结构的模型预测得到4?种基因的编码产物均存在α-螺旋、β-转角、伸展链和无规则卷曲。结论:从水开菲尔粒分离得到的L. plantarum QF01菌株,含有plnD、plnEF、plnV、plnR细菌素基因,对大肠杆菌有很好的抑制作用。  相似文献   

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