Nisin expression production from Lactococcus lactis in milk whey medium

BACKGROUND: Nisin is a commercially available bacteriocin produced by Lactococcus lactis ATCC 11454 and used as a natural agent in the biopreservation of food. In the current investigation, milk whey, a byproduct from dairy industries was used as a fermentation substrate for the production of nisin. Lactococcus lactis ATCC 11454 was developed in a rotary shaker (30 °C/36 h/100 rpm) using two different media with milk whey (i) without filtration, pH 6.8, adjusted with NaOH 2 mol L^?1 and without pH adjustment, both autoclaved at 121 °C for 30 min, and (ii) filtrated (1.20 µm and 0.22 µm membrane filter). These cultures were transferred five times using 5 mL aliquots of broth culture for every new volume of the respective media. RESULTS: The results showed that culture media composed of milk whey without filtration supplied L. lactis its adaptation needs better than filtrated milk whey. Nisin titers, in milk whey without filtration (pH adjusted), was 11120.13 mg L^?1 in the second transfer, and up to 1628‐fold higher than the filtrated milk whey, 6.83 mg.L^?1 obtained in the first^t transfer. CONCLUSIONS: Biological processing of milk byproducts (milk whey) can be considered a profitable alternative, generating high‐value bioproducts and contributing to decreasing river disposals by dairy industries. Copyright © 2008 Society of Chemical Industry     

产乳酸菌素菌株的筛选鉴定及其特性分析

本实验对广西巴马村天然发酵米粉中的乳酸菌进行分离鉴定,筛选出一株具有抑菌活性的植物乳杆菌M44,并研究了其所产乳酸菌素的基本性质。采用微生物形态学和16S r DNA序列分析法鉴定菌株M44为植物乳杆菌,其发酵上清液具有抑菌活性,抑菌物质确定为蛋白类。采用盐析法粗提所产乳酸菌素,确定最佳硫酸铵沉淀浓度为60%。由乳酸菌素产量与菌株生长的关系实验得出乳酸菌素的最佳收获时间为发酵培养12 h。管碟法抑菌实验表明,该乳酸菌素在p H范围2~8时有良好的抑菌活性;121℃处理15 min后仍具有抑菌活性;胃蛋白酶、胰蛋白酶处理后抑菌性消失;对单核增生李斯特细菌、大肠杆菌、沙门氏菌,金黄色葡萄球菌均有抑制作用。该乳酸菌素为天然生物防腐剂的开发与应用提供了研究基础。      

丙酸杆菌素的发酵及其分离纯化的初步探究

本文以薛氏丙酸杆菌(Propionibacterium shermanii)发酵液为原料,以抑菌活性为指标,利用分离纯化技术提取发酵液中小分子丙酸杆菌素。丙酸杆菌素初步分离纯化确定的最佳脱盐条件为:上样量5 m L,流速0.5 m L/min,收集方式为手动收集,电导率达到1.5 ms/cm时停止收集;Superdex peptide最佳凝胶过滤条件为:上样量500μL,流速0.5 m L/min,深孔板固定体积收集,每孔收集体积200μL。最终得到一种分子量为698.9556 u的丙酸杆菌素,其单位抑菌活性为初始的6.8倍,抑菌活性得率为10.9%。酶解实验表明纯化后的丙酸杆菌素仍保留抑菌活性,且对胃蛋白酶敏感。      

乳酸乳球菌KLDS4.0325产细菌素最佳培养条件的研究

以分离自新疆牧民家庭自制酸马奶中的乳酸乳球菌KLDS4.0325为研究对象,以大肠杆菌为指示菌,以抑菌圈直径为考察指标,考察了不同温度、p H、转速及培养时间等培养条件对该菌株细菌素产量的影响,经单因素实验优化得出乳酸乳球菌KLDS4.0325在培养温度33℃,培养基初始p H7.0,120 r/min摇床培养20 h的条件下,无细胞发酵上清液对大肠杆菌的抑菌圈直径最大,抑菌活性最强,细菌素产量最高。在此条件下,抑菌圈直径为25.26 mm,细菌素效价可达5383.07 IU/m L。      

乳酸菌中抗李斯特细菌素的筛选及理化性质分析

从鱼肉丸中分离筛选到1株具有抑菌活性的乳酸菌菌株Y153,对单核细胞增生李斯特菌ATCC54003的生长具有良好的抑制作用。16SrRNA序列同源性分析鉴定乳酸菌Y153为乳酸链球菌属（Streptococcus）。在排除有机酸、过氧化氢的干扰后,确定该抑菌物质为蛋白类物质,即细菌素,命名为细菌素Y153。理化性质分析表明,细菌素Y153具有较好的热稳定性、酸碱稳定性可被人体内蛋白酶降解的特性。由此对乳酸链球菌所产细菌素Y153有了初步的研究,为开发天然的食品防腐剂奠定理论基础。     

植物乳杆菌KLDS1.0391在酸奶体系中的细菌素产生特点

植物乳杆菌KLDS1.0391能够合成细菌素,也是益生菌,在食品中既可作为益生菌使用,也可作为辅助发酵剂用于生物防控,该研究主要考察了KLDS1.0391菌株在酸奶体系中细菌素的产生特点。研究结果表明,在发酵的6 h期间,抑菌活性随发酵时间延长而增强,发酵结束时,添加植物乳杆菌KLDS1.0391酸奶组的抑菌活性显著高于仅使用酸奶发酵剂的对照组(P<0.01)。与对照组相比,加入辅助发酵剂的实验组的感官品质未发生明显的变化。植物乳杆菌KLDS1.0391具备开发益生酸奶的潜力。     

1 株水开菲尔粒中植物乳杆菌的鉴定及产细菌素基因分析

目的：对从水开菲尔粒中分离得到1 株能产生抑菌物质的菌株QF01进行菌种鉴定,并对其产细菌素进行实验、鉴定和基因序列分析。方法：通过牛津杯法测定抑菌能力,采用聚合酶链式反应（polymerase chain reaction,PCR）扩增16S rDNA和细菌素相关基因并测序,利用ProParam tool、TMHMM 2.0、InterProScan、SOPM和SWISS-MODEL在线软件对基因编码产物进行分析。结果：该菌株产的细菌素对大肠杆菌（Escherichia coli JM109）有明显的抑制作用,通过16S rDNA序列分析初步确定该菌株属于植物乳杆菌（Lactobacillus plantarum）。该菌株含有plnD、plnEF、plnV、plnR四种基因,其中plnV基因编码产物有跨膜螺旋结构,其结构存在信号肽特征。此外,从三级结构的模型预测得到4?种基因的编码产物均存在α-螺旋、β-转角、伸展链和无规则卷曲。结论：从水开菲尔粒分离得到的L. plantarum QF01菌株,含有plnD、plnEF、plnV、plnR细菌素基因,对大肠杆菌有很好的抑制作用。     

1株产细菌素植物乳杆菌的筛选及所产细菌素的理化性质分析

从浙江省生产的一种火腿制品中分离筛选到1株具有抑菌活性的乳酸菌菌株LH-09,对单核细胞增生李斯特菌ATCC54003的生长具有良好的抑制作用.16SrRNA序列同源性分析鉴定乳酸菌LH-09为植物乳杆菌.在排除有机酸、过氧化氢的干扰后,确定该抑菌物质为蛋白类物质,即细菌素.理化性质分析表明细菌素LH-09具有较好的热稳定性、酸碱稳定性,可被人体内蛋白酶降解.N-羟甲基甲基甘氨酸-SDS-PAGE电泳确定细菌素LH-09的分子质量在2～6 kDa之间.     

Novel bacteriocinogenic Enterococcus hirae and Pediococcus pentosaceus strains with antilisterial activity isolated from Brazilian artisanal cheese

We isolated and characterized bacteriocin producers Enterococcus hirae ST57ACC and Pediococcus pentosaceus ST65ACC from raw milk artisanal cheeses. Their bacteriocins were tolerant to temperatures from 4°C to 100°C and under sterilization conditions (121°C for 15 min). Additionally, the tested bacteriocins remained active after being exposed to pH 2.0 to 10.0 for 2 h. The activity of the bacteriocins was affected by proteolytic enzymes but remained stable after treatment with EDTA, sodium dodecyl sulfate, NaCl, skim milk, and Tween 80. Cell-free supernatants were capable of inhibiting Listeria innocua and several strains of Listeria monocytogenes obtained from different sources and belonging to different serotypes. When L. monocytogenes 211 and L. monocytogenes 422 were treated with bacteriocins, growth was completely inhibited over 12 h. Cocultures of bacteriocinogenic strains and L. monocytogenes 422 in skim milk showed that E. hirae ST57ACC could control the growth of the pathogen in the matrix after 48 h. None of the selected isolates presented positive results on a screening panel for 25 bacteriocin-related genes, however, indicating that both strains might express novel bacteriocins.