共查询到20条相似文献,搜索用时 31 毫秒
1.
López RL García MT Abriouel H Ben Omar N Grande MJ Martínez-Cañamero M Gálvez A 《Journal of industrial microbiology & biotechnology》2007,34(12):779-785
The influence of substrate composition on the production of enterocin EJ97 and the conditions for semi-preparative bacteriocin
recovery have been studied. Final bacteriocin concentrations of 12.5 or 15.6 mg/l were obtained in the commercial media brain
heart infusion broth (BHI) and tryptic soya broth, respectively. The bacteriocin was also produced in the complex medium CM
(8.75 mg/l), in which the vitamin supplement was essential for production. Some combinations of meat peptone and yeast extract
plus either soy peptone or BHI also supported bacteriocin production, at concentrations of 6.25–7.5 mg/l. In cow milk (whole,
half-skimmed, and skimmed), the final bacteriocin concentrations obtained ranged from 7.5 to 11.25 mg/l. Highest bacteriocin
activity was obtained by using pasteurised milk whey as growth substrate (up to 25 mg/l), suggesting that this bacteriocin
can be obtained on a large scale by using this cheap food-grade industrial by-product. Highest bacteriocin titres were always
obtained after 8 h of incubation at 37 °C. Semi-preparative concentration and purification of enterocin EJ97 produced in a
complex medium was achieved by bulk cation exchange chromatography without previous cell separation, followed by reversed-phase
chromatography. This two-step procedure allowed preparation of milligram quantities of purified bacteriocin, which is an improvement
compared to purification procedures established for most other bacteriocins (35). The availability of purified enterocin EJ97
will facilitate other studies such as the elucidation of its molecular structure and its interaction with target bacteria. 相似文献
2.
Purification and amino acid sequence of a bacteriocin produced by Pediococcus acidilactici. 总被引:9,自引:0,他引:9
J C Nieto Lozano J N Meyer K Sletten C Peláz I F Nes 《Journal of general microbiology》1992,138(9):1985-1990
A bacteriocin produced by Pediococcus acidilactici has been purified to homogeneity by a rapid and simple four-step purification procedure which includes ammonium sulphate precipitation, chromatography with a cation-exchanger and Octyl Sepharose, and reverse-phase chromatography. The purification resulted in an approximately 80,000-fold increase in the specific activity and about a 6-fold increase in the total activity. The amino acid composition and sequencing data indicated that the bacteriocin contained 43-44 amino acid residues. The predicted M(r) and isolectric point of the bacteriocin are about 4600 and 8.6, respectively. Comparing the amino acid sequence of this bacteriocin with the sequences of leucocin A-UAL 187, sakacin P and curvacin A (bacteriocins produced by Leuconostoc gelidum, Lactobacillus sake and Lactobacillus curvatus, respectively) revealed that all four bacteriocins had in their N-terminal region the sequence Tyr-Gly-Asn-Gly-Val-Xaa-Cys, indicating that this concensus sequence is of fundamental importance for this group of bacteriocins. The bacteriocin from P. acidilactici and sakacin P were very similar, having at least 25 common amino acid residues. The sequence similarity was greatest in the N-terminal half of the molecules--17 of the first 19 residues were common--indicating the fundamental importance of this region. Leucocin A-UAL 187 and curvacin A had, respectively, at least 16 and 13 amino acid residues in common with the bacteriocin from P. acidilactici. 相似文献
3.
Liquid cultures of Staphylococcus epidermidis 1580 contained rather small amounts of a bacteriocin, staphylococcin 1580, which was found both in the supernatant fluid and in the cell pellet. It could be extracted from the cells with 5% NaCl solution. The staphylococcin production could not be induced by ultraviolet irradiation or treatment with mitomycin C. Bacteria grown on semisolid medium produced a much larger amount of the compound with a high specific activity. The staphylococcin was purified by ammonium sulfate precipitation, ultracentrifugation, and chromatography on Sephadex columns. The purified material was homogeneous on polyacrylamide gel electrophoresis. The molecular weight was between 150,000 and 400,000. The bacteriocin was composed of protein, carbohydrate, and lipid and consisted of subunits exhibiting a molecular weight of about 20,000. 相似文献
4.
Mireille ?ngela Bernardes Sousa Luiz de Macêdo Farias Patrícia Luciana de Oliveira Jaqueline Silvana Moreira Ana Carolina Morais Apol?nio Jamil Silvano Oliveira Marcelo Matos Santoro Edilberto Nogueira Mendes Paula Prazeres Magalh?es 《Memórias do Instituto Oswaldo Cruz》2013,108(6):724-729
Bacteriocins are antibacterial, proteinaceous substances that mediate microbial
dynamics. Bacteriocin production is a highly disseminated property among all
major lineages of bacteria, including Shigella. In this paper,
we addressed the purification and characterisation of a bacteriocin produced by
a Shigella sonnei strain (SS9) isolated from a child with acute
diarrhoea. The substance was purified through ammonium-sulphate precipitation
and sequential steps of chromatography. The intracellular fraction obtained at
75% ammonium sulphate maintained activity following exposure to pH values from
1-11 and storage at -80ºC for more than two years and was inactivated by high
temperatures and proteases. The molecular mass of the purified bacteriocin was
determined by mass spectrometry to be 18.56 kDa. The N-terminal sequence of the
bacteriocin did not match any other antibacterial proteins described. A putative
new bacteriocin produced by S. sonnei has been detected. This
bacteriocin may represent a newly described protein or a previously described
protein with a newly detected function. Considering that SS9 expresses
antagonism against other diarrhoeagenic bacteria, the bacteriocin may contribute
to S. sonnei virulence and is potentially applicable to either
preventing or controlling diarrhoeal disease. 相似文献
5.
Detection and characterization of a novel antibacterial substance produced by a Lactobacillus delbrueckii strain 1043 总被引:2,自引:0,他引:2
V. Miteva I. Ivanova I. Budakov A. Pantev T. Stefanova S. Danova P. Moncheva V. Mitev X. Dousset & P. Boyaval 《Journal of applied microbiology》1998,85(3):603-614
A novel antibacterial substance produced by a strain isolated from Bulgarian yellow cheese was characterized. The producer strain was identified by molecular typing to belong to the species Lactobacillus delbrueckii , which is a rare producer of bacteriocins. The inhibitory agent was heat stable and active against lactic acid bacteria species and several food-borne pathogens : Listeria monocytogenes , Staphylococcus aureus , Enterococcus faecalis , Escherichia coli , Yersinia enterocolitica and Y. pseudotuberculosis . Its sensitivity to amylolitic enzymes and lipase suggested that a lipid and carbohydrate moiety could be important for the activity. The amino acid content of the purified bacteriocin was estimated to 29 amino acids. The bacteriocin was shown to be small (3·6–6 kDa) by three different methods : HPLC gel-filtration, SDS-PAGE and amino acid contents. 相似文献
6.
The developmentally regulated carbohydrate binding protein discoidin (from Dictyostelium discoideum) has been purified in a nonagglutinating form. While substantial agglutination activity is present in cell lysates, this activity is consistently lost upon affinity purification of discoidin. The lack of agglutination activity is not due to a mutational event or a nutritional deficiency. The carbohydrate binding site of the protein is functional, and dissociation of the oligomeric protein into subunits has not occurred. The addition of aqueous dispersions of a CHCl3/CH3OH extract of a slime-mold particulate fraction to the purified discoidin reconstitutes agglutination activity in a concentration-dependent manner. The reconstituted agglutination activity has the specificity of discoidin's carbohydrate binding sites. The reconstitutive ability of the CHCl3/CH3OH extract is due to a lipid component. Treatments of the lipid extract and fractionation of the active species suggest that it may be unsaturated fatty acid. Of many purified lipids tested, only high concentrations of cisvaccenic acid (C18:1 delta11) or oleic acid (C18:1 delta9) significantly reconstituted agglutination activity. 相似文献
7.
Three hundred and thirty-five lactic acid bacteria were isolated from sour doughs and screened for antagonistic activity. Of these 145 showed activity against one or several of the indicator strains used in the screening. The antimicrobial activity of 18 isolates were due to a proteinaceous compound. These 18 isolates belonged to three different Lactobacillus species: Lactobacillus bavaricus, Lactobacillus curvatus and Lactobacillus plantarum. The spectrum of antimicrobial activity for the three species suggested that the inhibitory components were different. The inhibitory compound from Lact. bavaricus MI401 was chosen for further study. The proteinaceous nature, antimicrobial activity against closely-related species, heat resistance and sensitivity to alkaline treatment strongly indicated that this substance was a bacteriocin, which we designated bavaricin A. The bacteriocin was purified to homogeneity by ammonium sulphate precipitation, ion exchange, hydrophobic interaction and reverse-phase chromatography. The purification resulted in 193000-fold increase in specific activity. SDS-PAGE of bavaricin A showed a molecular weight of 3500—4000 Da. By amino acid sequencing 41 amino acids were determined. Bavaricin A had a bactericidal mode of action and inhibited nine out of 10 Listeria monocytogenes. Lactobacillus bavaricus MI401 produced bavaricin A at temperatures from 4°C to 30°C. The production of active bavaracin A was inhibited at increasing sodium chloride concentration. In the presence of 3% sodium chloride at 4°C no active bavaricin A could be detected. Nitrite (100 ppm) did not affect the production of active bavaricin A. 相似文献
8.
Lipsy Chopra Gurdeep Singh Vikas Choudhary Debendra K. Sahoo 《Applied and environmental microbiology》2014,80(10):2981-2990
Marine environments are the greatest fronts of biodiversity, representing a resource of unexploited or unknown microorganisms and new substances having potential applications. Among microbial products, antimicrobial peptides (AMPs) have received great attention recently due to their applications as food preservatives and therapeutic agents. A new marine soil isolate producing an AMP was identified as Bacillus sonorensis based on 16S rRNA gene sequence analysis. It produced an AMP that showed a broad spectrum of activity against both Gram-positive and Gram-negative bacteria. The peptide, named sonorensin, was purified to homogeneity using a combination of chromatographic techniques. The intact molecular mass of the purified peptide, 6,274 Da, as revealed by matrix-assisted laser desorption ionization–time of flight (MALDI-TOF), was in agreement with Tricine-SDS-PAGE analysis. A PCR array of primers was used to identify AMP structural genes, which allowed the successful amplification of the related genes from strain MT93. The putative open reading frame of sonorensin was amplified, cloned into the pET-32a(+) vector, expressed as a thioredoxin (Trx) fusion protein in Escherichia coli, and then purified. Sequence alignment analysis revealed that the bacteriocin being reported could belong to new subfamily of bacteriocins, heterocycloanthracin. The peptide indicated its potential as a biocontrol agent or food antimicrobial agent, due to its antimicrobial activity against bacteria such as Listeria monocytogenes and Staphylococcus aureus. This is the first report of the production, purification, and characterization of wild-type and recombinant bacteriocin by B. sonorensis and the first bacteriocin of the heterocycloanthracin subfamily to be characterized. 相似文献
9.
Cell of Enterobacter cloacae (Clo DF13) produce a bacteriocin which is characterized by its very effective killing activity against sensitive bacteria. Purification and characterization of the excreted bacteriocin has revealed that this bacteriocin consists of an equimolar complex of two plasmid-specific gene products: the cloacin and its inhibitor the immunity protein. Dissociation of the complex by treatment with sodium dodecylsulfate induces the endonucleolytic activity of the cloacin but strongly reduces the killing activity. The purified complex possesses no activity in vitro. Both cloacin and immunity protein isolated from the complex were functionally identical to cloacin and immunity protein purified from the bacteriocinogenic cells by other methods. Reconstitution of the complex results in a partial restoration of killing activity. 相似文献
10.
Isolation of lipid A by acid hydrolysis of Shigella flexneri lipopolysaccharide resulted in a product that consisted of a heterogeneous mixture of bands when visualized by thin layer chromatography. Differential extraction with ethyl acetate and chloroform, or extraction with EDTA, followed by chloroform-methanol-water (Bligh-Dyer extraction), or a combination of both extraction schemes, resulted in partial purification of immunologically active lipid A. Eight fractions were purified further by preparative thin layer chromatography, and each of the fractions had phosphate, carbohydrate, and esterified fatty acids. Upon incorporation into liposomes, five of the eight purified fractions reacted with anti-lipid A serum, but the three fractions with the most number of esterified fatty acids failed to react with anti-lipid A serum. At least one fraction that originally was unreactive with anti-lipid A serum became reactive as a hapten inhibitor upon removal of esterified fatty acids by alkaline hydrolysis. Alkali-treated fractions from "unreactive" and "reactive" lipid A had similar activities as hapten inhibitors. Our data suggest that lipid A can exist in multiple forms that differ by the number and placement, and possibly by the type, of fatty acids linked to the carbohydrate of lipid A. Highly acylated forms of lipid A do not react with antiserum against the unpurified lipid A mixture, but removal of fatty acids does expose immunoreactive groups. 相似文献
11.
Ramakrishnan Srinivasan Dinesh K. Kumawat Sunil Kumar Anil Kumar Saxena 《Annals of microbiology》2013,63(1):387-392
Bacteriocins are ribosomally synthesized peptides having considerable potential as a food preservative because of their strong antagonistic activity against many food spoilage and pathogenic organisms. A bacteriocin from Lactobacillus rhamnosus isolates was purified using ammonium sulphate precipitation and molecular exclusion chromatography techniques. Ammonium sulphate precipitation resulted in higher yield of bacteriocin, but the specific activity and fold purification were higher for molecular exclusion chromatography. The bacteriocin exhibited inhibition against food-borne pathogens and spoilage microorganisms, including both Gram-positive and -negative bacteria. Amylase, lipase and catalase did not alter the antimicrobial activity but proteolytic enzymes inactivated the bacteriocin. It was heat stable and exhibited activity in a pH range of 2–8 with maximum activity at pH 5.0. Molecular weight of bacteriocin was found to be ~5.6 kDa using SDS-PAGE. HPLC profile showed a single peak further attesting the purity of the bacteriocin. 相似文献
12.
CTP:phosphorylcholine cytidylyltransferase from rat liver. Isolation and characterization of the catalytic subunit 总被引:3,自引:0,他引:3
We reported previously the purification of CTP:phosphorylcholine cytidylyltransferase from rat liver (Weinhold, P. A., Rounsifer, M. E., and Feldman, D. A. (1986) J. Biol. Chem. 261, 5104-5110). The purified enzyme appeared to contain equal amounts of two nonidentical proteins, with Mr of about 38,000 and 45,000. We have now separated and purified these proteins. Polyacrylamide electrophoresis in the presence of sodium dodecyl sulfate indicated that each protein was homogeneous. The 45,000 protein contained the catalytic activity. Analysis by gel filtration chromatography and glycerol gradient centrifugation indicated that the 38,000 and 45,000 proteins in the purified cytidylyltransferase were independently associated with Triton X-100 micelles. The apparent Mr of the complexes suggested that a tetramer of each protein was bound to one Triton X-100 micelle. The isolated 45,000 catalytic protein had the same lipid requirement and kinetic properties as the purified cytidylyltransferase containing both proteins. Enzyme activity was stimulated to maximal values by phosphatidylcholine vesicles containing 9 mol % of either oleic acid, phosphatidylinositol, or phosphatidylglycerol. The amino acid compositions of the isolated 38,000 and 45,000 proteins were distinctly different. Overall, the results suggested that a tetramer of the 45,000 protein possessed nearly optimal catalytic activity. A functional role of the 38,000 protein as part of a cytidylyltransferase enzyme complex could not be documented. However, the need for stabilizing concentrations of Triton X-100 in the purified enzyme preparation may have prevented the association of the two proteins. 相似文献
13.
Aim
This study aimed to develop a simplified chemically defined medium that could sustain the growth and bacteriocin (enterolysin A) production by Enterococcus faecalis B9510.Methods and Results
The nutritional requirements of E. faecalis B9510 in a chemically defined medium were determined by single omission experiments. It was observed that eight amino acids (arginine, glycine, histidine, isoleucine, leucine, methionine, tryptophan and valine), three B vitamins (nicotinic acid, Ca‐pantothenic acid and pyridoxal) and magnesium sulphate were essential for growth. Based on this information, a Simplified Defined Medium (SDM) was formed consisting of 26 components. Comparison of SDM with M‐17 showed that growth and bacteriocin production in SDM was similar to that in M‐17. The bacteriocin from SDM was then purified by ultrafiltration. The retentate of ultrafiltration step was analysed by SDS‐PAGE and the results showed a single active band in the gel, which was excised and analysed by mass spectrometry, which indicated that the active band was enterolysin A, a cell wall degrading bacteriocin.Conclusions
A simplified defined medium can be formulated for the growth and bacteriocin production by Enterococcus faecalis, whose efficiency is comparable with that of a complex commercial medium.Significance and Impact of the Study
The development of such a medium can be useful for bacteriocin production and subsequent purification in a simplified manner and, therefore, helpful in the identification of novel bacteriocins. 相似文献14.
Simple method of purification and sequencing of a bacteriocin produced by Pediococcus acidilactici UL5 总被引:5,自引:2,他引:3
H. Daba C. Lacroix J. Huang R.E. Simard L. Lemieux 《Journal of applied microbiology》1994,77(6):682-688
H. DABA, C. LACROIX, J. HUANG, R.E. SIMARD AND L. LEMIEUX. 1994. A bacteriocin produced by a strain of Pediococcus acidilactici was successfully purified sequentially by acid extraction (at pH 2) and reverse-phase high-performance liquid chromatography (HPLC). Cell extracts of derivative strains deficient in bacteriocin production exhibited a similar HPLC elution profile to the active extracts except for the two peaks containing bacteriocin activity. The sequence of the antibacterial peptide consisted of 44 amino acid residues of which 42 were identified, and its molecular weight was 4624 Da, as determined by mass spectrometry. Moreover, according to the molecular weight of the peptide, the unidentified residues in the bacteriocin sequence must correspond to two tryptophan residues, confirming that the peptide isolated from Ped. acidilactici UL5 is pediocin PA-1. However, oxidized forms of the bacteriocin produced during storage also showed bacteriocin activity and resulted in a second peak with activity in the chromatograms. HPLC chromatograms of cell surface preparations from the active and from the deficient strains were confirmed by capillary electrophoresis. The purification method used is simple and effective in comparison with traditional methods, permitting a selective recovery of cell-associated bacteriocin at low pH, and its isolation in pure form for sequencing. 相似文献
15.
Halil Dündar Mehmet Atakay Ömür Çelikbıçak Bekir Salih Faruk Bozoğlu 《Preparative biochemistry & biotechnology》2013,43(8):796-809
This study aimed to compare two different approaches for the purification of enterocin B from Enterococcus faecium strain W3 based on the observation that the bacteriocin was found both in cell associated form and in culture supernatant. The first approach employed ammonium sulfate precipitation, cation-exchange chromatography, and sequential reverse-phase high-performance liquid chromatography. The latter approach exploited a pH-mediated cell adsorption–desorption method to extract cell-bound bacteriocin, and one run of reverse-phase chromatography. The first method resulted in purification of enterocin B with a recovery of 4% of the initial bacteriocin activity found in culture supernatant. MALDI-TOF MS analysis and de novo peptide sequencing of the purified bacteriocin confirmed that the active peptide was enterocin B. The second method achieved the purification of enterocin B with a higher recovery (16%) and enabled us to achieve pure bacteriocin within a shorter period of time by avoiding time consuming purification protocols. The purity and identity of the active peptide were confirmed again by matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry (MS) analysis. Although both approaches were satisfactory to obtain a sufficient amount of enterocin B for use in MS and amino acid sequence analysis, the latter was proved to be applicable in large-scale and rapid purification of enterocin B. 相似文献
16.
Lactobacillus fermentum strain L23 produced a small bacteriocin, designated bacteriocin L23, with an estimated molecular mass of < 7000 Da. Isolation,
purification, and partial characterization of bacteriocin L23 are described. It displayed a wide inhibitory spectrum including
both Gram-negative and Gram-positive pathogenic strains and two species of Candida. The antibacterial activity of cell-free culture supernatant fluid was not affected by catalase or urease but was abolished
by the proteolytic enzymes trypsin and protease VI. Bacteriocin L23 was heat stable (60 min at 100°C) and showed inhibitory
activity over a wide pH range (4.0 to 7.0). The proteinaceous compound was isolated from cell-free culture supernatant fluid
and purified. Crude bacteriocin sample was prepared by a process of ammonium sulfate precipitation, gel filtration, thin-layer
chromatography, bioautography, and reversed-phase HPLC. 相似文献
17.
H Okazaki C Tani M Ando K Ishii S Ishibashi Y Nishimura K Kato 《Journal of biochemistry》1992,112(3):409-413
A previously found proteinase possibly involved in the modification of hexokinase to eliminate the mitochondria-binding ability without appreciable change in the catalytic activity (called hexokinase-processing enzyme hereafter), was purified by sequential chromatographies from rat liver and its properties were examined. The hexokinase-processing enzyme had carbohydrate moieties as evidenced by adsorption on immobilized concanavalin A, and had a molecular weight of about 23,000 as estimated by SDS-PAGE and gel filtration chromatography. Benzyloxycarbonyl-phenylalanyl-L-arginine-4-methylcoumaryl-7-amide (Z-Phe-Arg-MCA)-hydrolyzing activity was co-purified with this processing activity throughout the purification, while the hydrolyzing activity for benzyloxycarbonyl-L-arginyl-L-arginine-4-methylcoumaryl-7-amide (Z-Arg-Arg-MCA) was not. The processing activity, as well as Z-Phe-Arg-MCA hydrolyzing activity, was highly sensitive to cysteine proteinase inhibition, for example, by leupeptin and N-[N-3-(trans-carboxirane-2-carbonyl)-L-leucyl]agmatine (E-64). Furthermore, the enzyme preparation reacted with an antibody against cathepsin L purified from rat kidney. These results indicated that cathepsin L may be involved in the above-mentioned processing of hexokinase. 相似文献
18.
The goal of this study was to demonstrate the ability of carbohydrate nanoparticle-stabilized emulsion to prolong the efficacy of bacteriocin against food pathogens. An amphiphilic, negatively charged carbohydrate nanoparticle, phytoglycogen octenyl succinate (PG-OS), was used to form oil-in-water emulsion for delivering bacteriocin nisin against the food pathogen Listeria monocytogenes. Dynamic light scattering test showed that in emulsion all PG-OS nanoparticles were adsorbed at the surface of oil droplets. Zeta-potential analysis indicated an effective adsorption of positively charged nisin molecules at the surface of PG-OS interfacial layer. Nisin depletion model showed that, during 50 days of storage, the anti-listerial activity of nisin-containing PG-OS-stabilized emulsion was substantially greater than that of nisin solution. In contrast, the emulsion stabilized with a neutral, small-molecule surfactant (Tween 20) or negatively charged, hyperbranched carbohydrate polymer (modified starch) was either ineffective or less effective than the nanoparticle-stabilized emulsion to retain nisin activity during storage. 相似文献
19.
Purification, amino acid sequence and characterization of Bacthuricin F4, a new bacteriocin produced by Bacillus thuringiensis 总被引:1,自引:0,他引:1
Kamoun F Mejdoub H Aouissaoui H Reinbolt J Hammami A Jaoua S 《Journal of applied microbiology》2005,98(4):881-888
AIMS: Purification and characterization of a new bacteriocin, Bacthuricin F4 of Bacillus thuringiensis. METHODS AND RESULTS: A newly isolated B. thuringiensis subsp. kurstaki strain BUPM4, was shown to produce a novel bacteriocin named Bacthuricin F4. The highest bacteriocin activity was found in the growth medium and evidenced in the late exponential growth phase. Bacthuricin F4 could be purified by a two-step procedure: ammonium sulphate precipitation of protein from culture supernatant followed by a reverse phase chromatography. Upon purification, the specific activity was increased 100-fold. This bacteriocin was heat-stable up to 70 degrees C and resisted up to pH 3.0. Bacthuricin F4 was sensitive to proteases demonstrating its proteinaceous nature. Its molecular mass, determined by mass spectrometry was 3160.05 Da. Direct N-terminal sequencing of Bacthuricin F4 revealed the following sequence: DWTXWSXL. The latter was unique in the databases. Bacthuricin F4 was active against Bacillus species while it had little or no effect on Gram-negative bacteria. CONCLUSIONS: A strain BUPM4 of B. thuringiensis subsp. kurstaki, was shown to produce a new bacteriocin named Bacthuricin F4 of both new molecular mass (3160.05 Da) and new amino acid terminal sequence. This is, to our knowledge, the first bacteriocin exhibiting such characteristics reported to be produced by B. thuringiensis. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacteriocin produced by the B. thuringiensis strain BUPM4 respond to both criteria of thermostability and stability to low pHs. Thus, it could be used for the control of the related species of Bacillus harmful for agricultural products. 相似文献
20.
Pyocin, a bacteriocin obtained from lysates of ultraviolet-induced cultures of Pseudomonas aeruginosa was characterized in vitro and in vivo after 1,000-fold purification by chemical, column, and differential centrifugation procedures. Electron micrographs of negatively stained pyocin preparations contained rod-shaped particles which resembled the contractile tail protein of the T-even phages of Escherichia coli. Although two separate and distinct pyocin fractions were eluted from diethylaminoethyl cellulose (pH 7.5) during the purification procedure, the particles appeared identical. In addition, the two fractions exhibited a close correlation between their titers and the particle numbers as observed in the electron microscope. The particles were approximately 20 by 90 mmu with a core diameter of 5 mmu and a sheath length of 50 mmu. Neither intact phage nor ghosts were seen in any of the preparations, although ringlets of two different diameters, which appeared to correspond to the diameters of the sheath and inner core, were observed. Other studies indicated that, although crude preparations were stable to freezing and thawing, purified preparations lost all of their activity under similar treatment. However, the addition of 50% glycerol to purified preparations completely protected activity. Conversely, aged normal human or rabbit sera enhanced the antibacterial activity of pyocin approximately fourfold, although serum albumin and hemoglobin had no effect. In vivo studies indicated that purified pyocin was not lethal for mice when injected intraperitoneally in concentrations of 28,000 to 1,400,000 units (5.6 to 276 mug of protein), nor was 7,200 to 36,000 units dermonecrotic for rabbits. 相似文献