副干酪乳杆菌Q-1-4的筛选鉴定及抗菌物质特性分析

为获得应用潜力较高的抗菌菌株,对内蒙古地区传统乳制品中分离的8株乳酸杆菌进行抗菌菌株筛选,获得了一株具有广谱抗菌活性的菌株(编号为Q-1-4)。经常规生理生化及分子生物学实验,鉴定该菌为副干酪乳杆菌(Lactobacillus paracasei)。通过实验排除有机酸和过氧化氢干扰后,L.paracasei Q-1-4发酵离心后的无细胞发酵上清液(cell-free fermentation supernatant,CFS)对鼠伤寒沙门氏菌Salmonella typhimurium CMCC50115仍具有抑制作用,且经蛋白酶处理后抗菌活性下降,说明L.paracasei Q-1-4所产抗菌物质中含有蛋白类物质。其在pH 2.0~5.5范围内对鼠伤寒沙门氏菌有抑制作用,且在此pH值范围内该抗菌物质对热稳定,对紫外线不敏感;与十二烷基硫酸钠、乙二胺四乙酸共同作用效果好于单一作用,而其他的表面活性剂、有机溶剂、金属离子对其无明显影响;该抗菌物质具有一定的贮藏稳定性且抑菌谱较广。     

腊肉拮抗菌分离及抗真菌脂肽特性分析

从传统腊肉中筛选拮抗菌株,纯化鉴定新型抗菌物质并对其抗菌特性研究,为抗菌脂肽在食品防腐中的应用提供理论依据。采用抑菌圈法筛选拮抗菌株;通过酸沉淀、硅胶柱层析和半制备反相高效液相色谱分离纯化抗菌脂肽;通过傅里叶红外光谱、基质辅助激光解吸电离飞行时间质谱对脂肽鉴定。结果表明,筛选得到12?株拮抗细菌,其中菌株XLP27对革兰氏阳性菌、革兰氏阴性菌以及对1?株酵母菌和4?株霉菌有明显抑菌作用。拮抗菌株XLP27鉴定为短小芽孢杆菌,其抗菌物质主要在稳定期合成,并在发酵60?h时达到最大产量。分离纯化得到单一组分抗菌物质,并鉴定为环状脂肽Iturin?A,其由m/z?1?046.182、1?060.213、1?074.198、1?088.201和1?102.175的5?个同系物组成,对应脂肪酸链长度C14～C18。脂肽具有极高的热稳定性,在40～100?℃下保温30?min活性基本保持不变,在较宽的pH值范围（4～10）内稳定性好,且对所有蛋白水解酶不敏感。菌株XLP27所产抗菌物质主要是脂肽Iturin?A,具有广谱抑菌活性,稳定性良好,有潜力在食品保鲜和农业生物防治领域替代传统抗菌剂使用。     

Antagonistic intestinal microflora produces antimicrobial substance inhibitory to Pseudomonas species and other spoilage organisms

Chicken intestine harbors a vast number of bacterial strains. In the present study, antimicrobial substance produced by lactic acid bacteria (LAB) isolated from the gastrointestinal tract of healthy chicken was detected, characterized, and purified. Based on 16S rRNA sequencing, the bacteria were identified as Lactobacillus plantarum vN. The antimicrobial substance produced by this bacterium was designated vN-1 and exhibited a broad-spectrum of activity against many important pathogenic and spoilage microorganisms, including Pseudomonas aeruginosa, Staphylococcus aureus, Micrococcus luteus, Salmonella Typhimurium, and Erwinia amylovova. vN-1 was determined to be thermostable, insensitive to pH values ranging from 2.0 to 8.0, resistant to various organic solvents and to enzymatic inactivation. The inhibition kinetics displayed a bactericidal mode of action. This study revealed an antimicrobial substance with low molecular mass of less than 1 kDa as determined by ultrafiltration and having features not previously reported for LAB isolated from chicken intestines. The detection of this antimicrobial substance addresses an important aspect of biotechnological control agents of spoilage caused by Pseudomonas spp. and promises the possibility for preservation of refrigerated poultry meat. Practical Application: The newly characterized antimicrobial substance and designated as vN-1 may have the potential to be used in food preservation.     

Purification and characterization of pedioxanthin (carotenoid pigment) produced by Pediococcus pentosaceus N33 strain isolated from pickles

Forty-two lactic acid bacteria were isolated from 28 pickles and yoghurt samples, and screened for their antimicrobial efficacy against certain multidrug resistant (MDR) bacteria using well diffusion method. A strain Pediococcus pentosaceus N33 (MF062087) showed the best inhibitory effects against both Brevibacillus brevis H18 (MG637284) and Lysinibacillus fusiformis R29 (MG637285). Pediococcus pentosaceus N33 strain produced high thermostable antimicrobial compound as indicated by heat treatment between 60 and 100°C for 20 min as well as at 121°C for 15 min. It is a proteinaceous compound highly active at acidic pH (2–6). The antimicrobial compound was extracted and precipitated by 70% ammonium sulfate and finally fractionated using Sephadex G₅₀ column. The molecular weight of pure compound was 10 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The spectroscopic analysis based on UV, IR, NMR and MS analyses were used to characterize and identify the pure antimicrobial compound. From the characterization and identification of this inhibitory compound, it was observed that the target compound contains carotenoid pigment in addition to protein. This carotenoid pigment was identified as pedioxanthin which is the first observed from this strain. Our results evaluated the potential activity of Pediococcus pentosaceus N33 strain as a probiotic agent and its potential contribution for food industry and pharmaceutical applications.     

Antimicrobial Activity and Hydrophobicity of Edible Whey Protein Isolate Films Formulated with Nisin and/or Glucose Oxidase

The use of edible antimicrobial films has been reported as a means to improve food shelf life through gradual releasing of antimicrobial compounds on the food surface. This work reports the study on the incorporation of 2 antimicrobial agents, nisin (N), and/or glucose oxidase (GO), into the matrix of Whey protein isolate (WPI) films at pH 5.5 and 8.5. The antimicrobial activity of the edible films was evaluated against Listeria innocua (ATCC 33090), Brochothrix thermosphacta (NCIB10018), Escherichia coli (JMP101), and Enterococcus faecalis (MXVK22). In addition, the antimicrobial activity was related to the hydrophobicity and water solubility of the WPI films. The greatest antibacterial activity was observed in WPI films containing only GO. The combined addition of N and GO resulted in films with lower antimicrobial activity than films with N or GO alone. In most cases, a pH effect was observed as greater antimicrobial response at pH 5.5 as well as higher film matrix hydrophobicity. WPI films supplemented with GO can be used in coating systems suitable for food preservation.     

Survival to different acid challenges and outer membrane protein profiles of pathogenic Escherichia coli strains isolated from pozol, a Mexican typical maize fermented food

In this study, the acid resistance and the changes in outer membrane protein (Omps) profiles of Escherichia coli strains isolated from pozol, an acid-fermented maize beverage consumed in Southeastern Mexico, were determined. Results showed that adaptation to acid by these E. coli strains significantly enhances their survival in acid conditions. Changes in Omp profiles were found in non-adapted acid challenged cells compared with non-challenged cells that had not been adapted to acid. Challenged adapted cells showed no significant changes in these profiles when compared with the acid adapted non-challenged strains. N-terminal sequences of some of the Omps were determined. The intensity of the main porins OmpC and OmpA was lower in the acid challenged strains, than in the non-challenged ones. The OmpF porin was identified in non-challenged K12 strain, but did not appear in adapted or non-adapted pozol strains nor in E. coli O157:H7. A protein band with an approximate molecular mass of 22 kDa corresponds to OmpW and its expression decreased in pozol strains challenged with HCl and lactic acid. OmpX was one of the main proteins expressed when strains were acid challenged with organic acids. Seventy out of seventy-three E. coli strains isolated from pozol in a previous work [Sainz, T., Wacher, C., Espinoza, J., Centurion, D., Navarro, A., Molina, J., Cravioto, A., Eslava, C., 2001. Survival and characterization of Escherichia coli strains in a typical Mexican acid-fermented food. International Journal of Food Microbiology 71, 169-176] carry this gene and belong to a reported pathogenic class of E. coli strains, or have virulence factors or survived at pH values less than 4.8. We suggest this protein could be involved in survival to stress conditions.     

Lactobacillus fermentum isolated from human colonic mucosal biopsy inhibits the growth and adhesion of enteric and foodborne pathogens

A number of Lactobacillus species are used as probiotic strains in order to benefit health. We have isolated L. fermentum from human colonic mucosal biopsy samples that possess antimicrobial activities against entroinvasive and foodborne pathogens such as Escherichia coli, Salmonella paratyphi A, Shigella sonnei, Staphylococcus aureus, Enterococcus faecalis, Proteus mirabilis, Pseudomonas aeruginosa, and Vibrio sp. In addition to lactic acid, L. fermentum secretes antimicrobial proteinacious compound(s) that was found to be active even at neutral pH (pH 7.0). The compound was sensitive to heat treatment and trypsin digestion. Lactobacillus fermentum inhibited the adhesion of enteropathogens to intestinal epithelial cells in vitro. Isolated cell surface associated proteins (SAPs) from L. fermentum were sufficient for the adhesion exclusions of enteropathogenic E. coli. Our results indicate that L. fermentum produces antimicrobial compounds and SAPs to inhibit the growth and adhesion of enteropathogens, respectively.     

LACTOBACILLUS CURVATUS PRODUCES A BACTERIOCIN-LIKE AGENT ACTIVE AGAINST GRAM-NEGATIVE PATHOGENIC BACTERIA

Isolates from fermented foods were screened for antimicrobial activity against gram‐negative bacteria. The most active isolate was identified as a Lactobacillus curvatus by biochemical analysis and ribotyping, and the isolate was designated as OSY‐HJC6. Lactobacillus curvatus OSY‐HJC6 was further tested for intracellular and extracellular production of antimicrobial agents. A reduction of > 8 log₁₀ cfu/mL was observed when cell suspensions of Salmonella enterica serovar Enteritidis and Escherichia coli O157:H7 were treated with equal volumes of Lb. curvatus culture supernatant. Gram‐positive bacteria were not sensitive to the culture supernatant, but antimicrobial activity was detected when the cell extract was tested against several gram‐positive and gram‐negative bacteria. Culture supernatant and cell extract retained the antimicrobial activity after heating at 60–100C for 10 min but not after protease treatment. The cell extract of Lb. curvatus retarded the growth of E. coli p220 in broth medium and food extracts (i.e., bacteriostatic action) but showed bactericidal activity against the bacterium in phosphate buffer.     

Characterization of a Bacteriocin from Lactobacillus plantarum Strain LR/14

A rhizospheric isolate of a lactic acid bacterium, identified as Lactobacillus plantarum strain LR/14, was characterized to produce a bacteriocin. A supernatant from 20h culture growth was used as the source of bacteriocin. The antimicrobial compound showed remarkable stability at high temperatures (100°C for 30 min and 121°C for 15 min under 15 psi pressure) and to the presence of organic solvents, detergents and surfactants. It was also active in the pH regime of pH 2.0–6.0. Moreover, the compound was stable under different storage temperatures as tested up to 24 months. While antimicrobial function was not lost by catalase or β-glycerophosphate treatment, the same was sensitive to a number of proteolytic enzymes. The crude preparation inhibited not only related strains but also other gram-positive and gram-negative bacteria, such as Staphylococcus aureus, Listeria monocytogenes and urogenic E. coli. Bacteriocinogenic activity co-migrated as a single protein band on tricine-SDS-PAGE with molecular mass of ～3.6 kDa.     

Potential of lactic acid bacteria isolated from specific natural niches in food production and preservation

Autochthonous strains of lactic acid bacteria (LAB) have been isolated from traditionally homemade cheeses collected from specific ecological localities across Serbia and Montenegro. Genetic and biochemical analysis of this LAB revealed that they produce bacteriocins, proteinases and exopolysaccharides. LAB produces a variety of antimicrobial substances with potential importance for food fermentation and preservation. Apart from the metabolic end products, some strains also secrete antimicrobial substances known as bacteriocins. Among the natural isolates of LAB from homemade cheeses, bacteriocin producers were found in both lactococci and lactobacilli. Lactococcus lactis subsp. lactis BGMN1-5 was found to produce three narrow spectrum class II heat-stable bacteriocins. In addition to bacteriocin production, BGMN1-5 synthesized a cell envelope-associated proteinase (CEP) and shows an aggregation phenotype. Another isolate, L. lactis subsp. lactis BGSM1-19 produces low molecular mass (7 kDa) bacteriocin SM19 that showed antimicrobial activity against Staphylococcus aureus, Micrococcus flavus and partially against Salmonella paratyphi. Production of bacteriocin reaches a plateau after 8 h of BGSM1-19 growth. Bacteriocin SM19 retained activity within the wide pH range from 1 to 12 and after the treatment at 100 degrees C for 15 min. Among collection of lactobacilli, the isolate Lactobacillus paracasei subsp. paracasei BGSJ2-8 produces heat-stable bacteriocin SJ (approx. 5 kDa) polypeptide. It retained activity after treatment for 1 h at 100 degrees C, and in the pH range from 2 to 11. In addition to isolates from cheeses, bacteriocin-producing human oral lactobacilli were detected. Most of them showed antimicrobial activity against streptococci, staphylococci and micrococci, but not against Candida. Isolate BGHO1 that showed the highest antimicrobial activity was determined as L. paracasei. Interestingly, Lactobacillus helveticus BGRA43, which was isolated from the human intestine showed strong activity against Clostridium sporogenes, but it was not possible to detect any bacteriocin production in this isolate by using standard procedures. Further analysis of antimicrobial activity revealed that BGRA43 has a relatively broad spectrum. Lactobacilli resistant to nisin were also detected among natural isolates. They produce bacteriocins, which have no activity against nisin producing lactococci.     

Aflatoxins in pozol, a nixtamalized, maize-based food

To determine whether pozol, a nixtamalized maize-based food was contaminated with aflatoxins, samples of non-fermented pozol were collected during the period November 2002 to April 2003 from local markets at Comitan in Chiapas, Mexico. The samples were analyzed for the presence of aflatoxins. Nineteen out of one hundred and eleven samples were contaminated with aflatoxin B2 (AFB2) and traces of aflatoxin B1 (AFB1). The percentage of samples contaminated with AFB2 in pozol prepared with white maize was 5.4%. Pozol mixed with toasted cacao paste had a contamination rate of 41.5%. No aflatoxins were detected in pozol prepared with yellow maize. It was found that only 1 of 19 contaminated samples had aflatoxin concentrations above 20 ppb.     

花生壳乙醇提取物(EEPH)抗菌性的研究

测定了花生壳乙醇提取物(EEPH)对食品中常见污染菌的抗菌活性，探讨了热处理和pH值对其抗菌活性的影响，阐明了花生壳乙醇提取物的抗菌特点以及作为食品天然防腐剂的开发价值。结果发明：花生壳乙醇提取物的抑菌作用显著，其抗菌活性随着其浓度增加而增强，对细菌和真菌的最低抑制浓度(MIC)分别为0．28‰和0．50‰；花生壳乙醇提取物的抗菌活性具有热稳定性，能忍受高温短时的热处理，并且在pH5～9的范围内均具有抗菌活性。     

广谱拮抗菌株的筛选诱变及抗菌物质分离鉴定

分离和鉴定产生广谱抗菌活性物质的菌株,并对抗菌物质分离鉴定.通过形态学、生理生化特征和16S rDNA序列分析鉴定菌株.采用紫外线.硫酸二乙酯化学复合诱变的方法,获得抗菌能力强、遗传稳定的高产菌株.分别用甲醇提取、凝胶过滤色谱(Sephadex G-10)和半制备高效液相色谱分离纯化抗菌物质,基质辅助激光解吸电离飞行时...     

Survival and characterization of Escherichia coli strains in a typical Mexican acid-fermented food.

In this study, the presence and pathogenic characteristics of Escherichia coli strains in pozol, an acid-fermented maize beverage consumed in South-eastern Mexico, were determined. Seventy-three E. coli strains were isolated at early and late times (6 and 48 h) during the pozol fermentation process, when pH values of the doughs were 6.7-4.7 (6 h) and 4.7-3.7 (48 h). Serotypes that belong to diarrheagenic E. coli serogroups O18, O88, O8, O11, O20, O173 were identified. HEp-2 cell adherence in vitro assays showed localized, diffuse and aggregative adherence patterns among some of these strains. A DNA colony hybridization analysis with different probes showed the presence of virulence genes related to diarrheal pathogenesis. Thirty-three percent of the E. coli strains were tetracycline-resistant and 95% had a 20 kb plasmid. The presence and survival of potentially pathogenic E. coli in acid-fermented pozol suggest that such foods may be a potential source of foodborne outbreaks.     

玫瑰红景天挥发性成分分析及其抗氧化和抗菌活性

目的:提取玫瑰红景天根茎中的挥发性物质,测试抗氧化和抗菌活性,为玫瑰红景天的综合利用提供科学依据。方法:采用水蒸气蒸馏法提取玫瑰红景天根茎中的挥发性物质,经GC-MS分离和鉴定,借助GC用系列正构烷烃对各组分进行定性,面积归一化法确定各组分的相对含量;通过H₂O₂/Fe2+体系考察对羟基自由基的清除作用;用微量肉汤稀释法测定最小抑菌浓度(MIC)和平板转种法测定最小杀菌浓度(MBC)确定抗菌活性。结果:从玫瑰红景天挥发性物质中鉴定出53种化合物,主要组分有1-辛醇(42.72%)、香叶醇(29.85%)、苯乙醇(9.67%)、桃金娘烯醇(2.69%)、芳樟醇(1.89%)等,玫瑰红景天挥发性物质对羟基自由基有清除作用,其半抑制浓度IC₅₀为0.0073 mg/mL,对抗菌试验所选的7个菌株均有抑制作用,对金黄色葡萄球菌ATTCC25925的MIC值是0.12 mg/mL、MBC值是0.36 mg/mL,对大肠杆菌ATCC25922的MIC值是0.58 mg/mL、MBC值是1.90 mg/mL。结论:通过水蒸气蒸馏法提取得到的玫瑰红景天根茎挥发性物质的组分丰富多样,是一种天然的抗氧化、抗菌活性物质。     

海洋链霉菌Streptomyces sp.MCCC 1A09830抗菌成分的分离与鉴定

目的:研究海洋链霉菌Streptomyces sp. MCCC 1A09830与其它菌种的亲缘关系,并分离纯化Streptomyces sp. MCCC 1A09830产生的抗菌成分,鉴定抗菌成分的结构。方法:通过构建系统发育树研究Streptomyces sp. MCCC 1A09830与其它菌种的亲缘关系;采用甲醇提取、硅胶柱层析、高效液相色谱法等分离技术纯化抗菌成分,鉴定抗菌成分的结构。结果:Streptomyces sp. MCCC 1A09830与Streptomyces sp. NEAE-126存在很近的亲缘关系;Streptomyces sp. MCCC 1A09830产生的抗菌成分得到了纯化,获得了化合物1。化合物1经高分辨质谱(High Resolution Mass Spectrometry,HRMS)、一维及二维核磁共振(Nuclear Magnetic Resonance,NMR)分析鉴定为Borrelidin A(疏螺旋体素)。结论:从Streptomyces sp. MCCC 1A09830的发酵液中分离得到1种抗菌成分,丰富了Borrelidin的生产菌库,为海洋菌株活性产物的分离提供了支持。     

Characterisation of physical,chemical and antimicrobial properties of allicin–chitosan complexes

Formation of allicin (freshly used after prepared from fresh garlic cloves) and chitosan (78%DD with 1100 kDa, and 88%DD and 1020 kDa) complexes were conducted under the optimum conditions. After complete complexation (pH 4.5–4.8, 4 °C, and 48 h), the optimum weight ratio was resulted (1:1). The highest yield after freeze drying was 64.98%. The light yellow complexes (L* 81.0–87.3, a* 0.4–2.2 and b* 16.8–21.2) can be dissolved in wide pH range (3–11). Scanning electron micrographs revealed the round shape complexes with diameter between 5 and 15 μm. The FT‐IR analysis showed the main functional groups, including alkyl group, alkyl group‐hydroxy or possibly amino substituent, hydroxyl or amino compound, and aliphatic alcohol with carbonyl substitution. XRD results showed that the complex crystalinity was amorphous. The clear zone results and MIC (mg mL^?1) revealed that allicin–chitosan complexes exhibited the antibacterial activity against spoilage bacteria indicated the possibility to use as antimicrobial agent in foods.     

Physical,antimicrobial and antioxidant properties of starch‐based film containing ethanolic propolis extract

Propolis is a natural product that meets the requirements as functional additive for food packaging due to its antioxidant and antimicrobial activities. In this work, ethanolic propolis extract (EPE) was incorporated in cassava starch films, and characterisations with respect to their microstructure, mechanical properties, water vapour permeability (WVP), moisture sorption kinetics as well as antimicrobial and antioxidant capacities were performed. The results showed that tensile strength was not affected (P > 0.05) by the presence of EPE but Young's modulus decreased about 50% when compared to control films possibly because of EPE plasticiser effect. When 1% EPE was used, changes in moisture sorption properties were detected by a slightly hydrophobic character at films WVP. When extracted from the films, propolis retained its antioxidant activity. The films exhibited antimicrobial activity against Staphylococcus aureus and Escherichia coli even at low EPE concentrations (0.5%) mainly due to its phenolic compounds.     

Cluster analysis, richness and biodiversity indexes derived from denaturing gradient gel electrophoresis fingerprints of bacterial communities demonstrate that traditional maize fermentations are driven by the transformation process

The bacterial communities of maize fermented foods (pozol, poto-poto and ogi) from Mexico, Congo and Benin was compared using a culture-independent approach [denaturing gradient gel electrophoresis (DGGE) analysis of total DNA]. Foods produced following the same flow chart (i) grouped in distinct clusters, (ii) shared similar richness and biodiversity indexes and (iii) exhibited a high intra-specific variability. Structural biodiversity was higher in pozol samples, probably due to oxic conditions and higher initial pH. DGGE bands found in foods of different origins suggest that Lactobacillus plantarum, Lb. delbrueckii and Lb. fermentum are particularly well adapted to the fermentation of maize.     

Bacteriocinogenic activity from starter cultures used in Spanish meat industry

Thirty-nine bacterial strains, obtained from commercial starter cultures and commonly used by the meat industry in Spain, have been examined for their ability to produce bacteriocins. Fourteen (35%) of them showed antagonism against at least one of the indicator strains, by the agar spot test. The strains showing an inhibitory action against pathogenic indicator strains were identified as Pediococcus acidilactici, Lactobacillus curvatus, Lactobacillus pentosus and Lactobacillus plantarum, which showed an inhibitory action against a wide range of Gram-positive bacteria. The only strain which showed reliable inhibitory activity against pathogenic indicator strains, by the well diffusion assay, was P. acidilactici. This strain produces an inhibitory compound, which reaches its maximum activity at the beginning of the stationary phase of growth. This antimicrobial substance (bacteriocin) has a proteinaceous nature, is stable over a broad range of pH, resistant to heat and shows a bactericidal action.