Coban Hatice Sevgi, Olgun Dicle, Temur İnci, Durak Muhammed Zeki
Department of Food Engineering, Faculty of Chemical and Metallurgical Engineering, Yıldız Technical University, 34220 İstanbul, Türkiye.
Department of Molecular Biology and Genetics, Faculty of Science and Letters, İstanbul Technical University, 34485 İstanbul, Türkiye.
Microorganisms. 2024 Nov 25;12(12):2428. doi: 10.3390/microorganisms12122428.
The aim of this study was to obtain data on Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) profiles of () isolates resulting from acquired immune memory in addition to their technological starter properties for the selection of potential starter cultures from local yogurt samples. A total of 24 isolates were collected from six local yogurt samples including Afyon/Dinar, Uşak, Konya/Karapınar, and Tokat provinces of Türkiye. Strain-specific CRISPR I-II-III and IV primers were used to determine the CRISPR profiles of the isolates. The isolates commonly had CRISPR II and IV profiles, while only one isolate had a CRISPR III profile. Polymerase chain reaction (PCR)-based and culture-based analyses were also carried out to obtain data on the technological properties of the isolates. The PCR analyses were performed for the gene for protease activity, the gene for urease enzyme, the gene for glutamate dehydrogenase, the gene for competence frequency, the gene involved in heat-shock stress resistance of the isolates with specific primers. Culture-based analyses including antimicrobial activity and acid-production ability of the isolates were completed, and proteolytic and lipolytic properties were also screened. Native spacer sequences resulting from acquired immune memory were obtained for CRISPR IV profiles of yogurt samples from the Konya-Karapınar and Tokat provinces and CRISPR III profiles of yogurt samples from the Uşak province. In conclusion, our study results suggest that it is possible to select the isolates with the desired level of technological characteristics, prioritizing the ones with the most diverse CRISPR profiles and with native spacers for potential industrial application as starter cultures. We believe that this study provides data for further biological studies on the impact of centuries of human domestication on evolutionary adaptations and how these microorganisms manage survival and symbiosis.
本研究的目的是获取因获得性免疫记忆而产生的（）分离株的成簇规律间隔短回文重复序列（CRISPR）图谱，以及它们作为技术起始菌株的特性，以便从当地酸奶样品中选择潜在的起始培养物。总共从土耳其的六个当地酸奶样品中收集了24株分离株，这些样品来自阿菲永/迪纳尔、屈塔希亚、科尼亚/卡拉皮纳尔和托卡特省。使用菌株特异性的CRISPR I-II-III和IV引物来确定分离株的CRISPR图谱。分离株通常具有CRISPR II和IV图谱，而只有一株分离株具有CRISPR III图谱。还进行了基于聚合酶链反应（PCR）和基于培养的分析，以获取有关分离株技术特性的数据。使用特异性引物对分离株的蛋白酶活性基因、脲酶基因、谷氨酸脱氢酶基因、感受态频率基因、参与热休克应激抗性的基因进行PCR分析。完成了包括分离株抗菌活性和产酸能力在内的基于培养的分析，并筛选了蛋白水解和脂肪水解特性。获得了科尼亚-卡拉皮纳尔省酸奶样品CRISPR IV图谱以及屈塔希亚省酸奶样品CRISPR III图谱因获得性免疫记忆产生的天然间隔序列。总之，我们的研究结果表明，有可能选择具有所需技术特征水平的分离株，优先选择具有最多样化CRISPR图谱和天然间隔序列的分离株，作为潜在的工业应用起始培养物。我们相信，本研究为进一步开展生物学研究提供了数据，这些研究涉及数百年人类驯化对进化适应的影响，以及这些微生物如何实现生存和共生。
