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1.
Lactobacillus brevis SB27, isolated from sausages, produced an antimicrobial substance active against numerous strains of heterofermentative lactobacilli and against some strains of pediococci andBacillus. The antibacterial agent was shown to be heat stable, resistant over a wide pH range, and sensitive to proteolytic enzymes. It was identified as a bacteriocin and termed brevicin 27. Dialysis and ultrafiltration suggested an apparent molecular weight between 10 and 30 kDa for the crude inhibitory molecule. Brevicin 27 exhibited a hydrophobic character. A partially purified preparation, resulting from ammonium sulfate precipitation and cation exchange chromatography, permitted confirmation of some characteristics of the bacteriocin, previously established with the crude extract. After treatment of the original brevicin 27-producing strain with novobiocin, a nonproducing mutant was obtained. This mutant was sensitive to brevicin 27, and its plasmid profile revealed the loss of a plasmid of about 3 MDa. 相似文献
2.
Interaction of the bacteriocin pediocin SJ-1 with the cytoplasmic membrane of sensitive bacterial cells as detected by ANS fluorescence 总被引:1,自引:0,他引:1
The l-anilino-8-naphthalenesulphonic acid (ANS) fluorescent probe was used to monitor alterations in the cytoplasmic membrane of sensitive Lactobacillus plantarurm cells, caused by pediocin SJ-1, a bacteriocin produced by Pediococcus acidilactici. The addition of pediocin SJ-1 to the sensitive cells caused an increase in fluorescence intensity of ANS and a blue shift in its emission maximum from 520 to 475 nm. None of these spectral changes could be detected when pediocin SJ-1 was added to cells of a Lact. plantarum variant resistant to pediocin SJ-1. Upon the addition of pediocin SJ-1, dose-dependent energy transfer took place between tryptophanyl residues in the cytoplasmic membrane of sensitive cells and ANS. Similar ANS-fluorescence changes were observed with the bacteriocin nisin. The concentrations of pediocin SJ-1 needed to effect changes in the fluorescence spectrum of ANS were of the same magnitude as those required for a bactericidal effect and the release of u.v.-absorbing material. A hypothesis on the mode of action of pediocin SJ-1 is proposed. 相似文献
3.
G. Green L.M.T. Dicks G. Bruggeman E.J. Vandamme & M. L. Chikindas 《Journal of applied microbiology》1997,83(1):127-132
Pediocin PD-1, produced by Pediococcus damnosus NCFB 1832, is inhibitory to several food spoilage bacteria and food-borne pathogens. However, pediocin PD-1 is not active against other Pediococcus spp. and differs in this respect to other pediocins produced by Pediococcus acidilactici and Pediococcus pentosaceus. Production of pediocin PD-1 starts during early growth and reaches a plateau at the end of exponential growth. Pediocin PD-1 was partially purified and its size was determined by tricine-SDS-PAGE as ≈ 3·5 kDa. The isoelectric point (pI) of pediocin PD-1 is ≈ 3·5, as determined with the Rotofor electrofocusing cell (BioRad). Pediocin PD-1 is heat-resistant (10 min at 121°C) and remains active after 30 min of incubation at pH 2–10. Pediocin PD-1 is resistant to treatment with pepsin, papain, α-chemotrypsin and trypsin, but not Proteinase K. Pediocin PD-1 is bactericidal against sensitive cells of Oenococcus oeni (previously Leuconostoc oenos ). 相似文献
4.
5.
Isolation, purification and partial characterization of plantaricin 423, a bacteriocin produced by Lactobacillus plantarum 总被引:5,自引:1,他引:4
Lactobacillus plantarum 423, isolated from sorghum beer, produces a bacteriocin (plantaricin 423) which is inhibitory to several food spoilage bacteria and food-borne pathogens, including Bacillus cereus , Clostridium sporogenes , Enterococcus faecalis , Listeria spp. and Staphylococcus spp. Plantaricin 423 is resistant to treatment at 80 °C, but loses 50% of its activity after 60 min at 100 °C and 75% of its activity after autoclaving (121 °C, 15 min). Plantaricin 423 remains active after incubation at pH 1–10 and is inactivated when treated with pepsin, papain, α-chymotrypsin, trypsin and Proteinase K. Plantaricin 423 was partially purified and its size estimated at 3·5 kDa, as determined by tricine-SDS-PAGE. The mechanism of activity of plantaricin 423 is weakly bactericidal, as determined against Oenococcus oeni (previously Leuconostoc oenos ). High DNA homology was obtained between the plasmid DNA of strain 423 and the pediocin PA-1 operon of Pediococcus acidilactici PAC 1·0, suggesting that plantaricin 423 is plasmid-encoded and related to the pediocin gene cluster. 相似文献
6.
Beaulieu L Groleau D Miguez CB Jetté JF Aomari H Subirade M 《Protein expression and purification》2005,43(2):3314-125
Expression of the pedA gene from Pediococcus acidilactici, coding for mature bacteriocin Pediocin PA-1, was investigated using the yeast Pichia pastoris to obtain larger quantities of pediocin to support additional studies, including structure-function research. Following various cloning strategies, a KM71H (Mut(s)) strain was selected. A significant concentration (74 microg/ml) of extracellular recombinant pediocin was obtained but the pediocin showed no biological activity. Supernatant fluids from P. pastoris cultures, harboring or not pedA, inhibited the biological activity of natural pediocin PA-1. The recombinant pediocin appeared as a mixture of three main fractions (7-8, 11, 20 kDa vs. 4.6 kDa for natural pediocin PA-1). The recombinant pediocin was also less hydrophobic and behaved differently when subjected to isoelectric focusing. Strong evidence indicated that some "collagen-like" material was tightly associated, most probably via covalent binding, to the recombinant pediocin. The "collagen-like" material was most probably responsible for the lack of biological activity of the recombinant pediocin and for the differences observed regarding some of the physico-chemical properties. Both the recombinant pediocin and natural pediocin were sensitive to collagenase, suggesting that pediocin PA-1 may possess a somewhat "collagen-like" nature. Interestingly, recombinant pediocin preparations showed the ability to assemble into fibrils. 相似文献
7.
H D Paik S S Bae S H Park J G Pan 《Journal of industrial microbiology & biotechnology》1997,19(4):294-298
Bacillus thuringiensis subsp tochigiensis HD868 was identified as a bacteriocin producer which exhibited a bactericidal effect against closely related species. This
bacteriocin designated as tochicin, was partially purified by 75% ammonium sulfate precipitation followed by subsequent
dialysis. This partially purified tochicin showed a narrow antibacterial spectrum of activity against most of 20 typical
B. thuringiensis strains and a strain of B. cereus, but not against other bacteria and yeasts tested. The antibacterial activity of tochicin on sensitive indicator cells disappeared
completely by proteinase K treatment (1 mg ml−1), which indicates its proteinaceous nature. Tochicin was very stable throughout the range of pH 3.0–9.0 and was relatively
heat-stable at 90°C, but bacteriocin activity was not detected after boiling for 30 min. The relationship between cell growth
and bacteriocin production was studied in a semi-defined medium. Tochicin activity was detected at the mid-log growth phase,
reached the maximum at the early stationary phase, but decreased after the stationary phase. Direct detection of tochicin
activity on sodium dodecyl sulfate-polyacrylamide gel suggested it has an apparent molecular mass of about 10.5 kDa. Tochicin
exhibited a bactericidal activity against B. thuringiensis subsp thompsoni HD522 in phosphate buffer (pH 7.0).
Received 02 December 1996/ Accepted in revised form 25 August 1997 相似文献
8.
Prakash M. Halami A. Ramesh A. Chandrashekar 《World journal of microbiology & biotechnology》2005,21(8-9):1351-1358
Summary A strain of Pediococcus acidilactici CFR K7 isolated from cucumber, produced an antimicrobial peptide which acted against Leuconostoc mesenteroides, selected strains of Lactobacillus spp., Pediococcus spp. and Enterococcus spp. The partially purified bacteriocin had molecular weight of ~4.6 kDa, heat stability in a range of 40–121 °C and was
active over a wide range of pH (2.0–9.0). This bacteriocin possessed strong antilisterial activity and was susceptible to
proteolytic enzymes. Southern hybridization using the PCR-generated pedA probe established that the gene for the bacteriocin was plasmid-borne as in the case of pediocin PA-1. Nucleotide sequence
of the pedAB gene indicated 100% homology to a pediocin AcH/PA-1. Certain bacteriocinogenic strains isolated from naturally fermented
cucumber were tested by colony hybridization using the pedA gene probe. Nine out of twenty colonies reacted with the probe indicating their ability to produce the pediocin-like bacteriocin.
These nine colonies were further tested for their antimicrobial spectrum, proteolytic inactivation and plasmid profile. It
was found that a few of them were active against Bacillus cereus, Micrococcus luteus and Listeria monocytogenes. Their proteolytic inactivation showed that the antimicrobial compound was susceptible to proteinase K. Colony hybridization
could thus enable rapid detection of pediocin and pediocin-like bacteriocin producers among a population of bacteriocinogenic
strains. 相似文献
9.
Pediocin SA-1, an antimicrobial peptide from Pediococcus acidilactici NRRL B5627: production conditions, purification and characterization 总被引:1,自引:0,他引:1
Anastasiadou S Papagianni M Filiousis G Ambrosiadis I Koidis P 《Bioresource technology》2008,99(13):5384-5390
Fermentation broths of Pediococcus acidilactici NRRL B5627 exhibited a certain antimicrobial activity due to a bacteriocin produced during early growth and until the stationary phase of growth was reached (at optimum of 60% dissolved oxygen saturation). Its size was determined by electrospray ionization mass spectrometric analysis as 3.660 kDa. N-terminal sequencing showed that the bacteriocin had 19 amino acid residues in the order KYYGXNGVXTXGKHSXVDX. The purified bacteriocin is similar to pediocins isolated by various Pediococci and therefore, it belongs to the class IIa of bacteriocins and is thus designated pediocin SA-1. Sensitivity of the purified pediocin to various storage temperatures and enzyme treatments was examined. Purified pediocin SA-1 is heat stable for up to 60 min at 121 °C. Pediocin SA-1 is inhibitory to several food-borne pathogens and food spoilage bacteria. It appears to be significantly more effective against Listeria spp. compared to pediocin PD-1 produced by P. damnosus. The mode of action of the purified bacteriocin appears to be bactericidal. 相似文献
10.
A. Aktypis G. Kalantzopoulos J.H.J. Huis in't Veld & B. ten Brink 《Journal of applied microbiology》1998,84(4):568-576
ACA-DC 0040 produced an antimicrobial agent, which was named thermophilin T, active against several lactic acid bacteria strains of different species and food spoilage bacteria, such as Clostridium sporogenes C22/10 and Cl. tyrobutyricum NCDO-1754. The crude antimicrobial compound is sensitive to proteolytic enzymes and α-amylase, heat-stable (100 °C for 30 min), resistant to pH exposure at pH 1–12 and demonstrates a bactericidal mode of action against the sensitive strain Lactococcus cremoris CNRZ-117. The production of bacteriocin was optimized approximately 10-fold in an aerobic fermenter held at constant pH 5·8 and 6·2. Ultrafiltration experiments with culture supernatant fluids containing the bacteriocin, and further estimation of molecular weight with gel filtration chromatography, revealed that bacteriocin in the native form has a molecular weight in excess of 300 kDa. SDS-gel electrophoresis of partially purified thermophilin T showed that bacteriocin activity was associated with a protein band of approximately 2·5 kDa molecular mass. 相似文献
11.
A.M. Strasser de Saad S.E. Pasteris María C. Manca de Nadra 《Journal of applied microbiology》1995,78(5):473-476
The production and stability of pediocin N5p from Pediococcus pentosaceus , isolated from wine, were examined in grape juice medium. Maximum growth and higher titre (4000 U ml-1 ) were observed at a initial pH of 7·5 and 30°C. The activity of the inhibitory substance was stable between pH values from 2·0 to 5·0 at 4° and 30°C. At pH 10·0 it was completely inactivated. When submitted to 30 min at 80°, 100° and 115°C, maximal stability was observed at pH 2·0. Ethanol up to 10% did not affect pediocin activity at acid pH, nor did 40–80 mg 1-1 SO2 , independently or combined with different ethanol concentrations, affect inhibitory activity. 相似文献
12.
AIMS: To characterize polyfermenticin SCD, a newly identified bacteriocin of Bacillus polyfermenticus SCD. METHODS AND RESULTS: Bacillus polyfermenticus SCD was identified as a bacteriocin producer with a bactericidal activity against Bacillus subtilis IFO 12113. Polyfermenticin SCD, named tentatively as the bacteriocin produced by B. polyfermenticus SCD, showed a narrow spectrum of activity against Gram-positive and Gram-negative bacteria, a yeast and moulds. Production of polyfermenticin SCD in a 5 l jar fermenter followed typical kinetics of primary metabolite synthesis. The antibacterial activity of polyfermenticin SCD on sensitive indicator cells disappeared completely by treatment with proteinase K, which indicates its proteinaceous nature. Polyfermenticin SCD seemed to be very stable throughout the pH range of 2.0 to 9.0, and it was relatively heat labile compared with other bacteriocins. Direct detection of polyfermenticin SCD activity on SDS-PAGE suggested that it had an apparent molecular mass of about 14.3 kDa. CONCLUSIONS: Bacillus polyfermenticus SCD produced relatively heat-labile polyfermenticin SCD with a narrow spectrum of activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus polyfermenticus SCD is a commercial probiotic which has been used for the treatment of long-term intestinal disorders. New findings on polyfermenticin SCD will be valuable in the evaluation of commercial probiotics. Polyfermenticin SCD can be used to control Bacillus spoilage organisms as a biological control agent. 相似文献
13.
Characterization of a novel killer toxin encoded by a double-stranded linear DNA plasmid of Kluyveromyces lactis 总被引:11,自引:0,他引:11
Y Sugisaki N Gunge K Sakaguchi M Yamasaki G Tamura 《European journal of biochemistry》1984,141(2):241-245
A novel killer toxin, encoded by a double-stranded linear DNA plasmid pGK l-1 (5.4 MDa) in Kluyveromyces lactis IFO 1267 was purified 320 000-fold from the culture broth of yeast. The toxin was obtained in an electrophoretically homogeneous state with a yield of 24% by hydroxyapatite column chromatography, chromatofocusing and polyacrylamide gel electrophoresis. The purified toxin was dissociated into two subunits with molecular masses of 27 kDa and above 80 kDa, as estimated by Laemmli's sodium dodecylsulfate gel electrophoresis; the exact composition ratio of the two subunits remains unestablished. The isoelectric point was between 4.4 and 4.8. As compared with the reported narrow pH range of action and instability of k1 killer toxin encoded by a double-stranded RNA plasmid of Saccharomyces cerevisiae, the K. Lactis toxin was effective with sensitive strains of S. cerevisiae in a relatively wider pH range between 4 and 8; it was stable for several months at pH 6.0 when stored below -20 degrees C. In contrast to the simple protein nature of the k1 killer toxin with a molecular mass of 11.47 kDa, the K. lactis toxin maintained a mannoprotein nature, as it was absorbed by a ConA-Sepharose column and eluted by methyl alpha-D-mannoside. The growth inhibitory activity of K. lactis toxin was enhanced 2-35-fold by the presence of 4-60% glycerol. 相似文献
14.
X.L. Liu K.H. Huang J.Z. Zhou L. Meng Y. Wang L.X. Zhang 《Letters in applied microbiology》2012,55(6):399-406
Aims: The aim of this study is to isolate and identify an endophytic fungus with antibacterial activity from the Asian medicinal and culinary plant Lilium lancifolium and to study the characteristics of its major antibacterial fractions. Methods and Results: After strict sample sterilization, an endophytic fungus BH‐3 with great antibacterial activity against Leuconostoc mesenteroides was isolated from the bulbs of L. lancifolium and was identified as Fusarium oxysporum on the basis of internal transcribed spacer (ITS) rDNA sequence and morphological traits. After partial purification including superfiltration and gel filtration, the major antibacterial fractions were found to be the substances with the molecular mass ranging from 35 to 60 kDa, mainly 55 kDa. The partially purified antibacterial fractions were stable at thermal processes, with more than 80% of activity left at 60°C for 1 h, and even 70·75% left at 121°C for 15 min. 90·33–98·97% of activity was observed in the pH range of 4·0–7·0. But the fractions were sensitive to different proteases. Conclusions: Endophytic strain F. oxysporum BH‐3 isolated from the bulbs of L. lancifolium produced protein‐like antibacterial metabolites. The antibacterial assay against Leuc. mesenteroides indicated that the fractions were stable at thermal processes and wide pH conditions, but sensitive to proteolyses. Significance and Impact of the Study: This study provides an increasing understanding of endophytic F. oxysporum in L. lancifolium and its metabolites, which have a great potential in food industry as antibacterial agents. 相似文献
15.
Fifty-five bacteriocinogenic lactic acid bacteria (LAB) isolated from seven different sources. Eight isolates were found to
produce pediocin PA-1 like bacteriocin as detected by pedB gene PCR and dot-blot hybridization. The culture filtrate (CF) activity of these isolates exhibited strong antilisterial,
antibacterial activity against tested food-borne pathogens and LAB. The identification and genetic diversity among the selected
LAB was performed by conventional morphological and molecular tools like RFLP, RAPD, and 16S rDNA gene sequencing. The isolates
were identified as, 1 each of Pediococcus acidilactici Cb1, Lactobacillus
plantarum Acr2, and Streptococcus equinus AC1, 2 were of P. pentosaceus Cb4 and R38, and other 3 were Enterococcus
faecium Acr4, BL1, V3. Partial characterization of the bacteriocins revealed that the peptide was heat-stable, active at acidic to
alkaline pH, inactivated by proteolytic enzymes, and had molecular weight around 4.6 kDa and shared the properties of class
IIa pediocin-family. The bacteriocin production at different temperatures, pH, and salt concentrations was studied to investigate
the optimal condition for application of these isolates as a starter culture or as a biopreservative in either acidic or non-acidic
foods. 相似文献
16.
Activity of plantaricin SA6, a bacteriocin produced by Lactobacillus plantarum SA6 isolated from fermented sausage 总被引:3,自引:2,他引:1
N. REKHIF, A ATRIH AND G. LEFEBVRE. 1995. Plantaricin SA6, a bacteriocin produced by Lactobacillus plantarum SA6, exhibited an inhibitory action against several mesophilic lactobacilli. It was stable at 90–100°C at pH 2–4 and it remained stable in the presence of several organic solvents, urea or β-mercaptoethanol. Plantaricin SA6 bound specifically to the cell surface of only plantaricin SA6-sensitive bacteria. The putative receptors are not destroyed by different hydrolytic enzymes added to the phosphate buffer. Plantaricin SA6 acted as a bactericidal agent lysing sensitive strains, that became more permeable to ortho-nitro-phenol-β-galactoside and lost their intracellular K+ ions and u.v.-absorbing materials. Both the adsorption and lethal action of plantaricin SA6 were maximal between pH 4 and 7, but the range of temperature tested (5–37βC) had no effect. Ions (of several salts such as MgCl2 ) inhibited the binding of plantaricin SA6 and protected cells against bacteriocin action. 相似文献
17.
Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici 总被引:4,自引:4,他引:4
An antimicrobial peptide designated pediocin AcH was isolated from Pediococcus acidilactici strain H. The pediocin AcH was purified by ion exchange chromatography. The molecular weight of pediocin AcH was determined by SDS-PAGE to be about 2700 daltons. Pediocin AcH was sensitive to proteolytic enzymes, resistant to heat and organic solvents, and active over a wide range of pH. Pediocin AcH exhibited inhibition against several food spoilage bacteria and foodborne pathogens including Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes. It was bactericidal to sensitive cells and acted very rapidly. The bactericidal effect was not produced by either cell lysis or apparent loss of membrane permeability. 相似文献
18.
Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici 总被引:25,自引:0,他引:25
An antimicrobial peptide designated pediocin AcH was isolated from Pediococcus acidilactici strain H. The pediocin AcH was purified by ion exchange chromatography. The molecular weight of pediocin AcH was determined by SDS-PAGE to be about 2700 daltons. Pediocin AcH was sensitive to proteolytic enzymes resistant to heat and organic solvents, and active over a wide range of pH. Pediocin AcH exhibited inhibition against several food spoilage bacteria and foodborne pathogens including Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes. It was bactericidal to sensitive cells and acted very rapidly. The bactericidal effect was not produced by either cell lysis or apparent loss of membrane permeability. 相似文献
19.
Location of tyrosine phenol-lyase in some Gram-negative bacteria 总被引:2,自引:0,他引:2
Abstract From various habitats (plant material, fruits, soil), yeasts belonging to the species of Pichia kluyveri and Hanseniaspora uvarum were isolated that showed killer activity. According to the activity spectrum against other yeasts these strains belonged to 11 different groups that were distinguishable from the killer strains K1 -K10 . The isoelectric points of the killer proteins were in the range of pH 3.5–3.9, the activity optimum was observed at pH 4.2–4.6. Above pH 5 and above a temperature of 25–35°C the killer proteins were inactivated. 相似文献
20.
Purification and characterization of an endo-polygalacturonase from Verticillium albo-atrum 总被引:2,自引:0,他引:2
A polygalacturonase was isolated from the culture filtrate of the fungal plant pathogen Verticillium albo-atrum and purified 22-fold to homogeneity as judged by SDS-electrophoresis. The enzyme was a basic protein with a molecular weight of 37 kDa, an isoelectric point ≥8·6 and containing 1·7% carbohydrate. The enzyme was an endo-polygalacturonase and hydrolysed a wide range of pectic substrates including polygalacturonic acid, 93% methylated pectin and pectins in tomato cell walls. The best substrate was 31% methylated pectin. Relative reaction rates on pectins with different degrees of methylation could be explained by considering both the number of susceptible bonds and non-specific enzyme-substrate interactions. The principal products of long-term hydrolysis were di- and mono-galacturonate. Maximum activity was observed at pH 4·6–5·0 and 46 °C. However, the enzyme lost activity above 30 °C in the absence of substrate. Enzyme activity was very sensitive to changes in ionic strength at low salt levels. It was stable in the pH range 3–11 at 30 °C. 相似文献