Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring

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...

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Main Authors: Cynthia Meza, Benjamin Sepulveda, Nicolás Flores-Castañón, Francisca Valenzuela, Catherine Ormeño, Alexis Castillo, Alex Echeverría-Vega, Sura Jasem Mohammed Breig, Saja Mohsen Alardhi, Alex Gonzalez, Bárbara Mora-Lagos, Aparna Banerjee
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1622325/full
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author Cynthia Meza
Benjamin Sepulveda
Nicolás Flores-Castañón
Francisca Valenzuela
Catherine Ormeño
Alexis Castillo
Alex Echeverría-Vega
Sura Jasem Mohammed Breig
Saja Mohsen Alardhi
Alex Gonzalez
Alex Gonzalez
Bárbara Mora-Lagos
Aparna Banerjee
author_facet Cynthia Meza
Benjamin Sepulveda
Nicolás Flores-Castañón
Francisca Valenzuela
Catherine Ormeño
Alexis Castillo
Alex Echeverría-Vega
Sura Jasem Mohammed Breig
Saja Mohsen Alardhi
Alex Gonzalez
Alex Gonzalez
Bárbara Mora-Lagos
Aparna Banerjee
author_sort Cynthia Meza
collection DOAJ
description 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.
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spelling doaj-art-0ee5da254c0f4f9bbf3c7330c67591542025-08-20T03:00:54ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-08-011610.3389/fmicb.2025.16223251622325Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot springCynthia Meza0Benjamin Sepulveda1Nicolás Flores-Castañón2Francisca Valenzuela3Catherine Ormeño4Alexis Castillo5Alex Echeverría-Vega6Sura Jasem Mohammed Breig7Saja Mohsen Alardhi8Alex Gonzalez9Alex Gonzalez10Bárbara Mora-Lagos11Aparna Banerjee12Doctorado en Biotecnología Traslacional, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Talca, ChileIngeniería en Biotecnología, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Talca, ChileDoctorado en Ciencias Biomédicas, Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Talca, ChileDoctorado en Ciencias Biomédicas, Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Talca, ChileFunctional Polysaccharides Research Group, Instituto de Ciencias Aplicadas, Facultad de Ingeniería, Universidad Autónoma de Chile, Sede Talca, Talca, ChileCentro de Investigación de Estudios Avanzados del Maule (CIEAM), Vicerrectoría de Investigación y Postgrado, Universidad Católica del Maule, Talca, ChileCentro de Investigación de Estudios Avanzados del Maule (CIEAM), Vicerrectoría de Investigación y Postgrado, Universidad Católica del Maule, Talca, ChileBiology Department, Al-Rasheed University College, Baghdad, IraqNanotechnology and Advanced Materials Research Center, University of Technology, Baghdad, IraqLaboratorio de Microbiología Ambiental y Extremófilos, Departamento de Ciencias Biológicas y Biodiversidad, Universidad de Los Lagos, Osorno, ChilePrograma de Investigación en Biomedicina y Biotecnología en Salud, Departamento de Salud, Universidad de los Lagos, Osorno, Chile0Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Temuco, ChileFunctional Polysaccharides Research Group, Instituto de Ciencias Aplicadas, Facultad de Ingeniería, Universidad Autónoma de Chile, Sede Talca, Talca, ChileBacillus 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.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1622325/fullBacillusexopolysaccharideanticancer activityemulsifying activitygenomics
spellingShingle Cynthia Meza
Benjamin Sepulveda
Nicolás Flores-Castañón
Francisca Valenzuela
Catherine Ormeño
Alexis Castillo
Alex Echeverría-Vega
Sura Jasem Mohammed Breig
Saja Mohsen Alardhi
Alex Gonzalez
Alex Gonzalez
Bárbara Mora-Lagos
Aparna Banerjee
Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
Frontiers in Microbiology
Bacillus
exopolysaccharide
anticancer activity
emulsifying activity
genomics
title Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
title_full Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
title_fullStr Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
title_full_unstemmed Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
title_short Genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant Bacillus isolated from Tolhuaca hot spring
title_sort genomic basis and functional characterization of the exopolysaccharide production by a thermotolerant bacillus isolated from tolhuaca hot spring
topic Bacillus
exopolysaccharide
anticancer activity
emulsifying activity
genomics
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1622325/full
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