产细菌素苏云金芽孢杆菌的鉴定及其所产抗菌物质性质

从腌制蔬菜表面分离到一株产细菌素的菌株K2,其中和后的无细胞发酵液主要抑制革兰氏阳性细菌,特别是对芽孢杆菌有强烈的抑制作用。发酵上清液经硫酸铵盐析和透析后,仍然有很强的抗菌活性,并对多种蛋白酶敏感,表明抗菌活性物质为蛋白类物质。通过形态培养特征、生理生化特征、16S rDNA 序列比对及系统发育分析,鉴定菌株K2 是苏云金芽孢杆菌(Bacillus thuringiensis)。菌株K2 产生的抗菌物质在pH6～9 条件下80℃处理30min 仍保持稳定的抗菌活性,其对敏感菌的作用主要是杀菌,而且对芽孢萌发有很好的抑制作用。该抗菌物质在菌体生长对数中期产生,在稳定期中期抗菌活性达到最大。     

一种抗猪链球菌细菌素的生物学特性研究

为了开发新型天然抗菌物质,本研究以猪粪便中分离出的一株产抗猪链球菌细菌素产生菌屎肠球菌LPL420P06为试材,对其产生的细菌素进行了理化特性研究。结果表明,该细菌素具有良好的热稳定性,在90℃、30min热处理的条件下仍能保持90%以上的活性;易被胰蛋白酶、蛋白酶K、木瓜蛋白酶、中性蛋白酶失活,表明该细菌素是一类蛋白类物质;在pH5～9范围内37℃、4h处理,活性保持稳定;抑菌谱实验结果表明,该细菌素不仅对单增李斯特氏菌、金黄色葡萄球菌、猪链球菌等多种革兰氏阳性病原菌有明显的抑制作用,而且对沙门氏菌、大肠杆菌等部分革兰氏阴性菌有不同程度的抑制作用,显示出该细菌素具有很好的开发应用前景。     

Garvicin KS,a Broad-Spectrum Bacteriocin Protects Zebrafish Larvae against Lactococcus garvieae Infection

Bacteriocins are emerging as a viable alternative to antibiotics due to their ability to inhibit growth or kill antibiotic resistant pathogens. Herein, we evaluated the ability of the bacteriocin Garvicin KS (GarKS) produced by Lactococcus garvieae KS1546 isolated from cow milk to inhibit the growth of fish and foodborne bacterial pathogens. We found that GarKS inhibited the growth of five fish L. garvieae strains isolated from infected trout and eels. Among fish pathogens, GarKS inhibited the growth of Streptococcus agalactiae serotypes Ia and Ib, and Aeromonas hydrophila but did not inhibit the growth of Edwardsiella tarda. In addition, it inhibited the growth of A. salmonicida strain 6421 but not A. salmonicida strain 6422 and Yersinia ruckeri. There was no inhibition of three foodborne bacterial species, namely Salmonella enterica, Klebsiella pneumoniae, and Escherichia coli. In vitro cytotoxicity tests using different GarKS concentrations showed that the highest concentration of 33 µg/mL exhibited low cytotoxicity, while concentrations ≤3.3 µg/mL had no cytotoxicity on CHSE-214 and RTG-2 cells. In vivo tests showed that zebrafish larvae treated with 33 µg/mL and 3.3 µg/mL GarKS prior to challenge had 53% and 48% survival, respectively, while concentrations ≤0.33 µg/mL were nonprotective. Altogether, these data show that GarKS has a broad inhibitory spectrum against Gram positive and negative bacteria and that it has potential applications as a therapeutic agent for a wide range of bacterial pathogens. Thus, future studies should include clinical trials to test the efficacy of GarKS against various bacterial pathogens in farmed fish.     

Immunity gene pedB enhances production of pediocin PA-1 in naturally resistant Lactococcus lactis strains

Heterologous production of the antilisterial bacteriocin pediocin PA-1 in lactococci is an attractive objective to increase the safety of dairy products. In a previous paper, we developed a system for the heterologous production of the bacteriocin pediocin PA-1 in pediocin-resistant lactococcal hosts through a leader exchange strategy. The system was based on 3 genes, 1 encoding the fusion between the lactococcin A leader and propediocin PA-1, and the other 2 encoding the lactococcin A secretion machinery. In this study, we investigated whether the addition of the pediocin PA-1 immunity gene (pedB) to this system has any effect on pediocin production. Introduction of the plasmid(s) carrying the genes described above into nisinproducing and non-nisinproducing lactococcal hosts led to a significant increase in the production of pediocin compared with the equivalent pedB-devoid systems. In addition, we obtained a nisin-producing strain with the ability to secrete pediocin PA-1 at a level equivalent to that of the parental strain Pediococcus acidilactici 347, which represents a notable improvement over our previous systems.     

群体感应调控下的乳酸菌细菌素合成

细菌素是一种在新陈代谢过程中由核糖体合成的具有抑菌作用的抗菌肽,因此被作为天然、无毒抗菌剂并广泛应用到食品行业中。群体感应是细菌细胞间通过对自诱导物浓度的感知,从而对基因表达进行调控的行为,现已证明乳酸菌的群体感应是细菌素合成的关键调控机制。作者主要综述了目前乳酸菌细菌素的研究现状、细菌素的系统分类、群体感应信号的转导机制及其对乳酸菌细菌素合成的调节,以促进对细菌素的研究及应用。     

产细菌素嗜盐四联球菌的筛选及鉴定

从传统露天酿造工艺的酱油中分离出12株乳球菌,对其进行形态、生理生化特性研究及16S rDNA序列分析,鉴定为嗜盐四联球菌。在排除酸作用后,WZ-3菌株发酵液对革兰氏阳性和革兰氏阴性细菌具有较好的抑菌活性,经胃蛋白酶、胰蛋白酶处理后,发酵液抑菌活性略有下降。因此初步认为该菌株为产细菌素的嗜盐四联球菌。     

产广谱细菌素乳酸菌的筛选

从四川传统发酵食品、市售酸奶、自制泡菜、香肠中分离出267株乳酸菌,采用平板挖井法从中筛选了64株对大肠杆菌、金黄色葡萄球菌、藤黄微球菌、铜绿假单胞杆菌、枯草芽孢杆菌有抑制作用的广谱抑菌性乳酸菌菌株,再对这些乳酸菌采用牛津杯抑菌实验,排除酸、过氧化氢干扰后,部分菌株的发酵上清液仍有很强的抑菌作用;进行胰蛋白酶、木瓜蛋白酶处理后,发酵上清液抑菌活性降低,因而确定产生的抑菌物质为蛋白质类细菌素。     

乳酸片球菌细菌素的活性及特性的研究

试验采用TGE（trypticase glucose yeast extlact）肉汤培养基作为乳酸片球菌的培养基，培养条件为37℃过夜静止培养。利用改变pH的方法，即通过菌体对细菌素的吸附与解吸，对细菌素进行分离纯化，试验表明细菌素对植物乳杆菌，绿脓杆菌，单核细胞增多症李氏杆菌，沙门氏菌，枯草杆菌，金黄色葡萄球菌和大肠杆菌O157均有抑制作用。并且还具有耐热，耐盐的特性．     

一株产细菌素乳杆菌的鉴定及其细菌素编码基因的获得

应用双层平板打孔法,从自制樱桃酒里分离的乳杆菌中筛选到一株对革兰氏阳性细菌和革兰氏阴性细菌均有明显抑制作用的菌株,命名为LD 1.0008。通过API 50 CHL糖发酵产酸实验、16S rDNA基因序列分析及其特异性recA基因多重聚合酶链式反应（polymerase chain reaction,PCR）,鉴定LD 1.0008为植物乳杆菌。排除有机酸的干扰,用多种蛋白酶处理后,其抑菌活性均有明显降低,而用过氧化氢酶处理后其抑菌活性基本不变,从而确定LD 1.0008所产生的抑菌物质为细菌素。使用已报道的10 对植物乳杆菌细菌素基因片段设计的引物对菌株LD 1.0008进行PCR扩增,发现其至少含有4 个植物乳杆菌素相关编码基因,即plnD、plnO、plnV和plnW。     

筛选自泡菜的发酵乳杆菌细菌素纯化及抑菌特性分析

从中国传统发酵蔬菜中筛选鉴定对食源性致病菌有广谱抗性的细菌素（命名为BLF52）产生菌株并对细菌素分离纯化和抑菌特性分析。分子筛凝胶过滤层析和半制备高效液相色谱（RP-HPLC）纯化细菌素并用Tricine-SDS-PAGE法测定其分子量。筛选得到产细菌素的发酵乳杆菌（Lactobacilus fermentum） CH-16。MRS培养基中34℃发酵至稳定期前期即24~30 h对S.aureus抑制活性最大（875 AU/mL）。BLF52纯化至单一组分,分子量5.6 kDa,40~80℃（30 min）、pH2.0~7.0（2 h）有良好稳定性。胃蛋白酶、木瓜蛋白酶、胰蛋白酶和蛋白酶K处理后,抑菌活性完全丧失,α-淀粉酶对其无影响,表明BLF52有典型的细菌素特性。BLF52对食源性腐败菌和致病菌如单增李斯特菌、金黄色葡萄球菌、蜡状芽孢杆菌、鼠伤寒沙门氏菌、铜绿假单胞菌、大肠杆菌和副溶血性弧菌均有抑制活性,通过杀菌模式抑制培养中的S.aureus生长。细菌素BLF52有潜力开发为适合酸性或中性食品使用的食品防腐剂。