Bulut Cisem, Gunes Hatice, Okuklu Burcu, Harsa Sebnem, Kilic Sevda, Coban Hatice Sevgi, Yenidunya Ali Fazil
Izmir Institute of Technology, Biotechnology Programme, 35430 Gulbahce, Urla, Izmir, Turkey.
J Dairy Res. 2005 Feb;72(1):19-24. doi: 10.1017/s0022029904000536.
Comlek peyniri is a typical artisanal cheese in Central Anatolia. This type of cheese was made by using the indigenous lactic acid bacteria (LAB) flora of cow or ewes' milk. Majority of the samples were taken from fresh cheese because the aim was to isolate homofermentative LAB. Initially 661 microbial isolates were obtained from 17 cheese samples. Only 107 were found to be homofermentative LAB. These isolates were selected and identified by using both phenotypic and molecular methods. Phenotypic identification included curd formation from skim milk, catalase test, Gram staining and light microscopy, growth at different temperatures and salt concentrations, arginine hydrolysis, gas production from glucose, and carbohydrate fermentation. Molecular identification was based on the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the 16S rRNA gene-ITS (internally transcribed spacer) region. By combining the phenotypic and molecular identification results, isolates belonging to each of the following genera were determined at species or subspecies level: 54 Lactococcus lactis subsp. lactis, 21 Enterococcus faecium, 3 Ec. faecalis, 2 Ec. durans, 10 Ec. sp., 15 Lactobacillus paracasei subsp. paracasei, and 2 Lb. casei strains. Technological characterisation was also performed by culturing each of the strains in UHT skim milk, and by monitoring pH change and lactic acid production at certain time intervals through the 24 h incubation. Results of the technological characterisation indicated that 33% of the isolates (35 strains) were capable of lowering the pH of UHT milk below 5.3 after 6 h incubation at 30 degrees C. Thirty four of these strains were Lc. lactis subsp. lactis, and only one was an Ec. faecium strain.
科姆莱克奶酪是安纳托利亚中部一种典型的手工制作奶酪。这种奶酪是利用牛或羊奶中的本土乳酸菌菌群制作而成。由于目标是分离同型发酵乳酸菌,大多数样本取自新鲜奶酪。最初从17个奶酪样本中获得了661株微生物分离株。结果发现只有107株是同型发酵乳酸菌。通过表型和分子方法对这些分离株进行了筛选和鉴定。表型鉴定包括脱脂牛奶凝乳形成试验、过氧化氢酶试验、革兰氏染色和光学显微镜观察、在不同温度和盐浓度下的生长情况、精氨酸水解试验、葡萄糖产气试验以及碳水化合物发酵试验。分子鉴定基于16S rRNA基因-ITS(内部转录间隔区)区域的聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析。综合表型和分子鉴定结果,在种或亚种水平确定了以下各属的分离株:54株乳酸乳球菌乳酸亚种、21株粪肠球菌、3株屎肠球菌、2株耐久肠球菌、10株肠球菌属、15株副干酪乳杆菌副干酪亚种以及2株干酪乳杆菌菌株。还通过在超高温灭菌脱脂牛奶中培养各菌株,并在24小时培养期间的特定时间间隔监测pH变化和乳酸产生情况,对各菌株进行了工艺特性分析。工艺特性分析结果表明,33%的分离株(35株)在30℃培养6小时后能够将超高温灭菌牛奶的pH值降至5.3以下。其中34株为乳酸乳球菌乳酸亚种,只有1株是粪肠球菌菌株。