Bağder Elmacı Simel, Tokatlı Mehmet, Dursun Derya, Özçelik Filiz, Şanlıbaba Pınar
Faculty of Engineering, Department of Food Engineering, Ankara University, Ankara, Turkey,
Folia Microbiol (Praha). 2015 May;60(3):241-51. doi: 10.1007/s12223-014-0363-x. Epub 2014 Nov 18.
A total of 152 lactic acid bacteria (LAB) were isolated from pickles produced in the Ankara-Çubuk region. These isolates were clustered into eight groups on the basis of their phenotypic characteristics including cell morphology, CO2 production from glucose, growth at 10 and 45 °C, growth in 6.5 % NaCl, and growth at pH 9.6. API 50 CH carbohydrate fermentation test, 16S ribosomal RNA (rRNA) sequence analysis, and sodium dodecyl sulfate-acrylamide gel electrophoresis (SDS-PAGE) whole-cell protein profile analysis were also performed for precise identification of the isolates at the species level. Molecular identification revealed that the most prevalent LAB species involved in pickle fermentation were Pediococcus ethanolidurans (46 isolates, 30.3 %), Lactobacillus brevis (37 isolates, 24.3 %), Lactobacillus plantarum (37 isolates, 24.3 %), and Lactobacillus buchneri (15 isolates, 9.9 %). Other LAB were found in minor frequencies such as Pediococcus parvulus (8 isolates, 5.3 %), Lactobacillus namurensis (6 isolates, 3.9 %), Lactobacillus diolivorans (1 isolate, 0.7 %), Lactobacillus parabrevis (1 isolate, 0.7 %), and Enterococcus casseliflavus (1 isolate, 0.7 %). When results of phenotypic and genotypic identification methods were compared, differences in the species distribution of LAB associated with pickles were defined between the API and the 16S rRNA sequencing. The API 50 CHL test coincided with the 16S rRNA results in 71 out of the 152 tested isolates, indicating that API gave unreliable identification results. A clear correlation could not be found between the results of whole-cell SDS profiles and 16S rRNA sequencing. Therefore, molecular characterization by 16S rRNA sequencing was considered to be the most reliable method for identifying isolates. The results presented in this work provide insight in to the LAB population associated with traditional Çubuk pickles and constitute a LAB strain resource for further studies involving the development of starter cultures.
从安卡拉-楚布克地区生产的泡菜中总共分离出152株乳酸菌(LAB)。这些分离株根据其表型特征分为八组,包括细胞形态、葡萄糖产二氧化碳、在10和45℃下生长、在6.5%氯化钠中生长以及在pH 9.6下生长。还进行了API 50 CH碳水化合物发酵试验、16S核糖体RNA(rRNA)序列分析和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)全细胞蛋白质谱分析,以便在种水平上精确鉴定这些分离株。分子鉴定表明,参与泡菜发酵的最常见乳酸菌种类为嗜乙醇片球菌(46株,30.3%)、短乳杆菌(37株,24.3%)、植物乳杆菌(37株,24.3%)和布氏乳杆菌(15株,9.9%)。其他乳酸菌的出现频率较低,如微小片球菌(8株,5.3%)、那慕尔乳杆菌(6株,3.9%)、二醇解乳杆菌(1株,0.7%)、副短乳杆菌(1株,0.7%)和格氏肠球菌(1株,0.7%)。当比较表型和基因型鉴定方法的结果时,发现泡菜相关乳酸菌的种分布在API和16S rRNA测序之间存在差异。在152株测试分离株中,有71株的API 50 CHL试验结果与16S rRNA结果一致,这表明API给出的鉴定结果不可靠。全细胞SDS图谱结果与16S rRNA测序之间未发现明显相关性。因此,16S rRNA测序的分子表征被认为是鉴定分离株最可靠的方法。本研究结果为了解与传统楚布克泡菜相关的乳酸菌群体提供了见解,并构成了用于进一步研究开发发酵剂培养物的乳酸菌菌株资源。