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Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia

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Abstract

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.

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All the data we have, we deposited on the NCBI database and put in the supplementary material.

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Funding

This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia [Grant No. 451–03-47/2023–01/200042].

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042, Belgrade, Serbia

    Milka Malesevic, Nemanja Stanisavljevic, Danka Matijasevic, Jovana Curcic & Milan Kojic

  2. Faculty of Informatics and Computing, Singidunum University, Belgrade, Serbia

    Vukasin Tasic

  3. The Academy of Applied Technical and Preschool Studies, Nis, Serbia

    Srdjan Tasic

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  1. Milka Malesevic
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  2. Nemanja Stanisavljevic
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Contributions

Conceptualization, M.M., and M.K.; Methodology, S.T., V.T., and M.K.; Investigation, M.K., N.S., M.M. and D.M; Data Curation, M.M.; Writing—Original Draft Preparation, M.M. and M.K.; Writing—Review and Editing, D.M., N.S., and J.C.; Visualization, M.K., and M.M.; Supervision, M.K. and S.T. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Milka Malesevic.

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Malesevic, M., Stanisavljevic, N., Matijasevic, D. et al. Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. Microb Ecol 86, 2344–2356 (2023). https://doi.org/10.1007/s00248-023-02242-6

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