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ORIGINAL RESEARCH article

Front. Microbiol.

Sec. Extreme Microbiology

Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1622325

This article is part of the Research TopicExtremophiles in Biotechnology: Challenges and Advancements in Sustainable ApplicationsView all 4 articles

Genomic Basis and Functional Characterization of the Exopolysaccharide Production by a Thermotolerant Bacillus isolated from Tolhuaca hot spring

Provisionally accepted
Cynthia MezaCynthia Meza1Benjamin SepulvedaBenjamin Sepulveda2Nicolás Flores-CastañónNicolás Flores-Castañón3Francisca ValenzuelaFrancisca Valenzuela3Catherine OrmeñoCatherine Ormeño4Alexis Castillo BrunaAlexis Castillo Bruna5Alex Echeverría-VegaAlex Echeverría-Vega5Sura JasemSura Jasem6Saja Mohsen AlardhiSaja Mohsen Alardhi7Alex R GonzalezAlex R Gonzalez8Bárbara Mora-LagosBárbara Mora-Lagos9Aparna BanerjeeAparna Banerjee4*
  • 1Doctorado en Biotecnología Traslacional, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Talca, Chile, Talca, Chile
  • 2Ingeniería en Biotecnología, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Talca, Chile, Talca, Chile
  • 3Doctorado en Ciencias Biomédicas, Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Talca, Chile, Talca, Chile
  • 4Functional Polysaccharides Research Group, Instituto de Ciencias Aplicadas, Facultad de Ingeniería, Universidad Autónoma de Chile, Sede Talca, Talca, Chile, Talca, Chile
  • 5Centro de Investigación de Estudios Avanzados del Maule (CIEAM), Vicerrectoría de Investigación y Postgrado, Universidad Católica del Maule, Talca, Chile, Talca, Chile
  • 6Biology Department, Al-Rasheed University College, Baghdad, Iraq, Baghdad, Iraq
  • 7Nanotechnology and Advanced Materials Research Center, University of Technology, Iraq, Baghdad, Iraq
  • 8Laboratorio de Microbiología Ambiental y Extremófilos, Departamento de Ciencias Biológicas y Biodiversidad, Universidad de Los Lagos, Osorno 5290000, Chile, Osorno, Chile
  • 9Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, 4810101, Temuco, Chile

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Bacillus licheniformis Tol1, a thermotolerant bacterial strain isolated from the Tolhuaca hot spring in Chile, was investigated for its genomic features and the functional properties of its exopolysaccharide (EPS). The whole-genome sequencing revealed ∼4.25 Mbp genome with a GC content of 45.9% and a rich repertoire of genes associated with environmental stress adaptation, antibiotic resistance, sporulation, biofilm formation, and EPS biosynthesis, including the presence of epsD and epsC. The strain also harbored intact prophage elements and a Type I-A CRISPR-Cas system, indicating potential horizontal gene transfer and genome plasticity. Confocal microscopy revealed robust biofilm formation at 45–55 °C under neutral to slightly alkaline pH, with strong EPS matrix development. EPS production was optimized using OFAT and Response Surface Methodology (RSM), achieving a yield of 2.11 g L⁻¹ under optimized conditions, which was further validated using an Artificial Neural Network (ANN) model (R² = 0.9909). The EPS exhibited promising antioxidant activity and significant emulsification potential across various vegetable oils, which were comparable or superior to commercial bacterial EPS xanthan gum. Notably, the EPS also showed cytotoxic effects against AGS gastric adenocarcinoma cells, reducing viability by 38.38% and 37% at 50–100 μg μL⁻¹ concentrations, respectively, suggesting potential anticancer activity. Altogether, the study highlights B. licheniformis Tol1 as a multifunctional thermophile with valuable biotechnological potential, particularly for applications in food, pharmaceutical, and biomedical industries

Keywords: Bacillus, Exopolysaccharide, Anticancer activity, Emulsifying activity, Genomics

Received: 03 May 2025; Accepted: 17 Jul 2025.

Copyright: © 2025 Meza, Sepulveda, Flores-Castañón, Valenzuela, Ormeño, Bruna, Echeverría-Vega, Jasem, Alardhi, Gonzalez, Mora-Lagos and Banerjee. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Aparna Banerjee, Functional Polysaccharides Research Group, Instituto de Ciencias Aplicadas, Facultad de Ingeniería, Universidad Autónoma de Chile, Sede Talca, Talca, Chile, Talca, Chile

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Supplementary Material