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Biochemical and genetic evidence of enterocin P production by two Enterococcus faecium-like strains isolated from fermented sausages

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CURRENT MICROBIOLOGY 1999年第5期39卷 282-290页

作者： Herranz, C Mukhopadhyay, S Casaus, P Martínez, JM Rodríguez, JM Nes, IF Cintas, LM Hernández, PE Univ Complutense Dept Nutr & Bromatol 3 Fac Vet E-28040 Madrid Spain Agr Univ Norway Lab Microbial Gene Technol Dept Biotechnol Sci N-1432 As Norway

Two bacteriocin-producing Enterococcus faecium-like strains were independently isolated from fermented sausages. Bacteriocins were purified to homogeneity by ammonium sulfate precipitation, gel filtration, cationic exchange, hydrophobic interaction, and reverse-phase liquid chromatography. Two peptide inhibitory fractions were purified from each strain, denominated A and B for E. faecium AA13, and C and D for E. faecium G16. Fraction B was blocked for amino acid sequencing by Edman degradation, while the amino acid sequences obtained from peptides A, C, and D contained the YGNGV consensus motif in positions 5 to 9, and the ATRS sequence in positions 1 to 4. By use of PCR techniques and nucleotide sequencing, the structural gene of enterocin P was found both in E. faecium AA13 and E. faecium G16. Metabolic and genetic features of the two strains suggest that they are slightly different, they may produce more than one bacteriocin, and both produce enterocin P.

关键词：氨基酸序列细菌素类/生物合成细菌素类/化学细菌素类/遗传学细菌素类/分离和提纯肠球菌屎/遗传学肠球菌屎/分离和提纯肠球菌屎/代谢发酵肉制品/微生物学分子序列数据聚合酶链反应序列分析 DNA

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Characterization, production, and purification of leucocin H, a two-peptide bacteriocin from Leuconostoc MF215B

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CURRENT MICROBIOLOGY 1999年第1期39卷 43-48页

作者： Blom, H Katla, T Holck, A Sletten, K Axelsson, L Holo, H Norwegian Food Res Inst MATFORSK N-1430 As Norway Norwegian Dairies R&D Ctr TINE Oslo Norway Agr Univ Norway Lab Microbial Gene Technol N-1432 As Norway Univ Oslo Dept Biochem N-0367 Oslo Norway

Leuconostoc MF215B was found to produce a two-peptide bacteriocin referred to as leucocin H. The two peptides were termed leucocin H alpha and leucocin H beta. When acting together, they inhibit, among others, Listeria monocytogenes, Bacillus cereus, and Clostridium perfringens. Production of leucocin H in growth medium takes place at temperatures down to 6 degrees C and at pH below 7. The highest activity of leucocin H in growth medium was demonstrated in the late exponential growth phase. The bacteriocin was purified by precipitation with ammonium sulfate, ion-exchange (SP Sepharose) and reverse phase chromatography. Upon purification, specific activity increased 10(5)-fold, and the final specific activity was 2 X 10(7) BU/OD280. Amino acid composition analyses of leucocin H alpha and leucocin H beta indicated that both peptides consisted of around 40 amino acid residues. Their N-termini were blocked for Edman degradation, and the methionin residues of leucocin H beta did not respond to Cyanogen Bromide (CNBr) cleavage. Absorbance at 280 nm indicated the presence of tryptophan residues and tryptophan-fracturing opened for partial sequencing by Edman degradation. From leucocin H alpha, the sequence of 20 amino acids was obtained;from leucocin H beta the sequence of 28 amino acid residues was obtained. No sequence homology to other known bacteriocins could be demonstrated. It also appeared that the two peptides themselves shared little or no sequence homology. The presence of soy oil did not affect the activity of leucocin H in agar.

关键词：氨基酸序列芽孢杆菌蜡状/药物作用细菌素类/遗传学细菌素类/分离和提纯细菌素类/代谢细菌素类/药理学色谱法离子交换梭菌产气荚膜/药物作用明串珠菌属/代谢利斯特菌单核细胞增生/药物作用分子序列数据

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Solvent extraction of bacteriocins from liquid cultures

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LETTERS IN APPLIED MICROBIOLOGY 2000年第3期31卷 193-197页

作者： Burianek, LL Yousef, AE Ohio State Univ Dept Food Sci & Technol Columbus OH 43210 USA

A solvent extraction method was developed to concentrate lacidin from the culture of Lactobacillus acidophilus OSU133. The new method concentrates the bacteriocin at the interface between chloroform and the aqueous culture of the producing bacterium. Compared with other extraction procedures, the new method effectively recovers higher bacteriocin yield and results in relatively clean preparations. Recovery of lacidin by the chloroform extraction procedure, compared with ammonium sulphate precipitation and cell acidification methods, was > 10-fold and about 100-fold greater, respectively. The new extraction procedure saves time and is easy to perform. This method is also effective in recovering subtilin, bacillicin, pediocin and nisin from cultures of Bacillus subtilis ATCC 6633, B. subtilis OSY1115/C, Pediococcus acidilactici PO2 and Lactococcus lactis ATCC 11454, respectively.

关键词：硫酸铵细菌素类/化学细菌素类/分离和提纯细菌素类/药理学化学沉淀氯仿培养基/化学革兰氏阳性菌嗜酸乳杆菌/生长和发育溶剂

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A method for bacteriocin quantification

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JOURNAL OF APPLIED MICROBIOLOGY 1999年第6期87卷 907-914页

作者： Cabo, ML Murado, MA González, MP Pastoriza, L CSIC Inst Invest Marinas Vigo 36208 Pontevedra Spain

Different aspects of the most commonly used assay methods in the study of bacteriocins were examined. The conditions under which extraction and incubation (including exposure time) take place were analysed, and several different formal models that are usually employed to calculate ID50 were compared. As an alternative designed to overcome the problems which characterize the response of micro-organisms that are sensitive to bacteriocins, an operative procedure in a liquid medium and a modified re-parameterized logistic equation is proposed. When applied to the inhibition of Leuconostoc mesenteroides by nisin, the model allows an optimal experimental procedure to be defined.

关键词：细菌素类/分离和提纯细菌素类/药理学培养基食品微生物学氢离子浓度半数抑制浓度乳乳球菌/化学明串珠菌属/药物作用明串珠菌属/生长和发育微生物敏感性试验模型生物学鲑/微生物学动物

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Characterization of the mesB gene and expression of bacteriocins by Leuconostoc mesenteroides Y105

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CURRENT MICROBIOLOGY 1999年第5期39卷 265-269页

作者： Héchard, Y Berjeaud, JM Cenatiempo, Y CNRS Mol Biol Lab ESA 6031 IBMIG F-86022 Poitiers France

Leuconostoc mesenteroides Y105, previously described for production of mesentericin Y105, an anti-listeria bacteriocin, was shown to secrete a second bacteriocin. The latter was purified, and its molecular mass of 3446 Da, obtained by mass spectrometric analysis, indicates that this bacteriocin should be identical to mesenterocin 52B [Revol-Junelles et al;, Lett Appl Microbiol 23:120, 1996]. This second bacteriocin produced by L. mesenteroides Y105 was named mesentericin B105. Its structural gene, mesB, was then localized by a reverse genetic approach, cloned. and sequenced. MesB was found on the pHY30 plasmid, next to mesY gene clusters. Curing experiments led to isolation of two L. mesenteroides Y105 derivatives, named L. mesenteroides Y29 and Y30. The latter had lost pHY30 plasmid, encoding bacteriocin determinants, therefore explaining its phenotype (MesY-, MesB-). On the contrary, Y29 derivative still harbors the pHY30 but did not produce any bacteriocin. Thus, its phenotype could Likely result from a point mutation within a gene, probably encoding a protein involved in production of both mesentericin Y105 and mesentericin B105.

关键词：氨基酸序列细菌素类/生物合成细菌素类/遗传学细菌素类/分离和提纯细菌素类/药理学染色体图克隆分子基因细菌明串珠菌属/药物作用明串珠菌属/遗传学明串珠菌属/代谢利斯特菌属/药物作用质谱分析法微生物敏感性试验分子序列数据序列分析 DNA

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PURIFICATION AND EFFECTS OF FULVOCIN-C, A BACTERIOCIN FROM MYXOCOCCUS-FULVUS MX-F16

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ARCHIVES OF MICROBIOLOGY 1978年第3期119卷 279-286页

作者： HIRSCH, HJ TSAI, H GEFFERS, I Gesellschaft für Biotechnologische Forschung mbH

Fulvocin C is a bacteriocin from M. fulvus Mx f16. It has a MW of 4672 and is one of the smallest bacteriocins known. Four disulfide bonds give the molecule a tight structure, so that its native form was not attacked by chymotrypsin or Pronase. Fulvocin C was stable in various organic solvents and could tolerate ***. C in aqueous solution without loss of activity. The killing effect of fulvocin C was observed only at concentrations higher than 0.25 .***/l. Macromolecular synthesis (DNA, RNA, protein) was affected very gradually. Viability in growing cultures decreased slowly from 100-25% during 1 generation (8 h). Cell division was affected early. After 1 generation v-shaped cell pairs had accumulated in the culture. Electron micrographs revealed extended membrane systems connected with the inner membrane. The most striking effect was that often the outer membranes of neighboring cells seemed to have fused laterally. With further incubation many cells lost their rod shape and empty bags became predominant.

关键词：细菌蛋白质类/生物合成细菌素类/生物合成细菌素类/分离和提纯细菌素类/药理学细胞分裂/药物作用 DNA 细菌/生物合成黏球菌目/药物作用黏球菌目/代谢黏球菌目/超微结构 RNA 细菌/生物合成

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Inhibition of Bacillus cereus ATCC 14579 by plantaricin UG1 in vitro and in food

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NAHRUNG-FOOD 2000年第5期44卷 364-367页

作者： Enan, G Zagazig Univ Fac Sci Dept Bot & Microbiol Zagazig Egypt

The inhibition of Bacillus cereus ATCC 14579 viable growth by Lactobacillus plantarum UG1 bacteriocin (plantaricin UG1) in vitro and in food (pasteurized milk and minced meat) was studied. The inhibitory effect against B. cereus food-borne pathogen noticed in this study was due to plantaricin UG1, but not due to lactic acid produced by the L. plantarum UG1 culture. Plantaricin UG1 negative clone did not affect viable growth of B. cereus in both broth and meat or pasteurized milk. The inhibitory effect of L. plantarum UG1 and its bacteriocion was apparently more in liquid systems (BHI broth & pasteurized milk) than in minced meat. The inhibitory effect of plantaricin UG1 against B. cereus was dependent on its concentration. The 22880 AU/ml concentration appeared to be an ideal preservative against B. cereus ATCC 14579 in liquid systems.

关键词：芽孢杆菌蜡状/药物作用细菌素类/生物合成细菌素类/化学细菌素类/分离和提纯细菌素类/药理学培养基乳杆菌属/代谢肉/微生物学乳/微生物学动物

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Production, recovery and purification of bacteriocins from lactic acid bacteria

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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 1999年第5期52卷 628-638页

作者： Parente, E Ricciardi, A Univ Basilicata Dipartimento Biol Difesa & Biotecnol Agroforestal I-85100 Potenza Italy

Bacteriocins produced by lactic acid bacteria are a heterogeneous group of peptide inhibitors which include lantibiotics (class I, e.g. nisin), small heat-stable peptides (class II, e.g. pediocin AcH/PA1) and large heat-labile proteins (class III, e.g. helveticin J). Many bacteriocins belonging to the first two groups can be successfully used to inhibit undesirable microorganisms in foods, but only nisin is produced industrially and is licensed for use as a food preservative in a partially purified form. This review focuses on the production and purification of class I and class II bacteriocins from lactic acid bacteria. Bacteriocin production is growth associated but the yield of bacteriocin per unit biomass is affected by several Factors, including the producing strain, media (carbohydrate and nitrogen sources, cations, etc.) and fermentation conditions (pH, temperature, agitation, aeration and dilution rate in continuous fermentations). Continuous fermentation processes with cell recycle or immobilized cells can result in a dramatic improvement in productivity over batch fermentations. Several simple recovery processes, based on adsorbing bacteriocin on resins or silica compounds, have been developed and can be used to build integrated production processes.

关键词：吸附氨基酸序列细菌素类/遗传学细菌素类/分离和提纯细菌素类/代谢培养基/化学氢离子浓度乳杆菌属/代谢分子序列数据树脂类植物/代谢二氧化硅/代谢温度

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PHYSICOCHEMICAL PROPERTIES OF PYOCIN-F1

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JOURNAL OF BIOCHEMISTRY 1979年第1期85卷 21-28页

作者： KURODA, K KAGEYAMA, M MAEDA, T FUJIME, S Mitsubishi-Kasei Institute of Life SciencesMinamiooya Machida Tokyo 194

The physicochemical properties of pyocin F1 from Pseudomonas aeruginosa were studied. Pyocin F1 consists of flexuous rod-like particles homogeneous in size. Each particle was composed of rod and fiber parts. The rod part was 105.5 .+-. 9.5 nm long and 10.0 .+-. 1.4 nm wide, and showed regular striations amounting to 23 layers. The fiber part was composed of several filaments;the length of the longest filament was 43.0 .+-. 12.0 nm. The amino acid composition, the partial specific volume (0.720 ml/g), the sedimentation coefficient (SO20,W = 35.1S) and the translational diffusion constant (0.94 .+-. 0.01 .times. 10-7 cm2/s) were determined. The particle weight was 3.23 .times. 106 daltons.

关键词：氨基酸类/分析细菌素类/分离和提纯显微镜检查电子分子量蛋白质构象铜绿假单胞菌/分析脓菌素类/分离和提纯

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ISOLATION, HOMOGENEITY, AND PROPERTIES OF CORE PARTICLE FROM PYOCIN-R1

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JOURNAL OF BIOCHEMISTRY 1979年第2期85卷 403-411页

作者： HASEGAWA, T ISHII, S HOKKAIDO UNIV FAC PHARMACEUT SCI DEPT BIOCHEM KITA KU SAPPORO HOKKAIDO 060 JAPAN

By a mild alkaline treatment of pyocin R1, the core particle was released from the contracted sheath. After sucrose density-gradient centrifugation, core-rich fractions were treated with anti-sheath serum and by a second density-gradient centrifugation, purified core particles were isolated. Homogeneity of the preparation was confirmed by observation under the electron microscope, immuno-precipitation reaction, and sucrose density-gradient centrifugation. The core particle exhibited a sedimentation coefficient of 37S. The quaternary structure of the core consists of a single kind of subunit protein with a molecular weight of 18,000. No contamination by other proteins was detected by SDS-disc electrophoreses. Amino acid analysis revealed that the core is rich in glycine, alanine, valine, leucine, aspartic acid (or asparagine), glutamic acid (or glutamine), and serine. This amino acid composition bears some resemblance to that of T-even bacteriophage tail-core.

关键词：氨基酸类/分析细菌素类/分离和提纯细菌噬菌体大肠杆菌免疫扩散大分子物质显微镜检查电子分子量铜绿假单胞菌/分析脓菌素类/分离和提纯分光光度法紫外线病毒蛋白质类

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