Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042, Belgrade, Serbia.
Faculty of Informatics and Computing, Singidunum University, Belgrade, Serbia.
Microb Ecol. 2023 Nov;86(4):2344-2356. doi: 10.1007/s00248-023-02242-6. Epub 2023 May 24.
The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63-95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.
弗尔尼亚奇卡巴尼亚温泉是巴尔干半岛上最热的温泉，其水温为 63-95°C，原位 pH 值为 7.1。根据理化分析，弗尔尼亚奇卡巴尼亚温泉属于重碳酸硫酸盐高温水。该温泉微生物群落的结构仍在很大程度上尚未被探索。为了确定和监测弗尔尼亚奇卡巴尼亚温泉的微生物区系多样性，首次同时进行了全面的非培养宏基因组分析和培养依赖方法。使用扩增子测序分析进行微生物分析揭示了存在从种到门的进化新颖分类群。基于培养的方法导致分离出 17 株属于 Anoxybacillus、Bacillus、Geobacillus 和 Hydrogenophillus 属的菌株。然后对五个代表性菌株进行全基因组测序。基因组特征和 OrthoANI 分析表明，弗尔尼亚奇卡巴尼亚温泉蕴藏着进化新颖的 Anoxybacillus 属物种，证明其独特性。此外，这些分离物包含应激反应基因，使它们能够在温泉的恶劣条件下生存。计算机分析的结果表明，大多数测序菌株具有产生耐热酶（蛋白酶、脂肪酶、淀粉酶、植酸酶、几丁质酶和葡聚糖酶）和各种抗菌分子的潜力，这些酶和分子在工业、农业和生物技术应用中具有重要意义。最后，这项研究为进一步研究和了解这些微生物的代谢潜力提供了基础。
