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Production and characterization of exopolysaccharide (EPS) from marine Bacillus halotolerans and its antibacterial activity against clinical pathogens

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Abstract

This study investigates the antibacterial activity of exopolysaccharides (EPS) produced by the marine bacterium Bacillus halotolerans against clinical strains of Pseudomonas aeruginosa and Serratia marcescens. EPS production was optimized by exploring different carbon sources with glucose, peptone, and calcium chloride identified as significant factors for enhanced production. The optimized medium achieved a production rate of 0.612 g/L, surpassing the unoptimized medium’s production of 0.556 g/L. The EPS exhibited noteworthy hydroxyl scavenging activity and demonstrated hydrogen-donating ability by scavenging DPPH radicals. Fourier-transform infrared spectroscopy analysis revealed characteristic peaks representing hydroxyl and carboxyl groups, glycosidic linkages, and sulfate groups within the EPS structure. Nuclear magnetic resonance analysis confirmed the complex and heterogeneous nature of the EPS, with glucose and galactose residues and the presence of polysaccharides identified. Monomer units of the EPS structure were identified as glucose and galactose, and a prominent peak at 7.942 min indicated the presence of beta-1,5-O-Dibenzoyl-ribofuranose. This study provides valuable insights into optimizing EPS production, characterizing its properties, and evaluating its antibacterial activity against Pseudomonas sp. strains. Further investigations are necessary to explore the potential applications and underlying mechanisms of these antibacterial EPS, contributing to advancements in microbiology and biotechnology.

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Abbreviations

EPS:

Exopolysaccharides

g/L:

Grams per litre

μg/mL:

Micrograms per milliliter

FTIR:

Fourier-transform infrared spectroscopy

NMR:

Nuclear magnetic resonance

DPPH:

2,2-diphenyl-1-picrylhydrazyl

min:

minutes

g/mL:

grams per millilitre

RBC:

Red blood cells

PBS:

Phosphate buffer saline

XRD:

X-ray diffraction

GCMS:

Gas chromatography-mass spectrometry

MHz:

megahertz

2θ:

2 theta

CRA:

Congo Red Agar

ZMB:

Zobel Marine Agar

RPM:

Revolutions Per Minute

NCBI:

National Center for Biotechnology Information

AST:

Antibiotic Susceptibility Testing

DNA:

Deoxyribonucleic Acid

RSM:

Response Surface Methodology

CCRD:

Central Composite Rotatable Design

BLAST:

Basic Local Alignment Search Tool

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Acknowledgments

The authors are thankful to RUSA 2.0, Alagappa University. The project was funded by Researchers Supporting Project number (RSP2023R231), King Saud University, Riyadh, Saudi Arabia.

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Correspondence to Vellaisamy Balasubramanian.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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Ravindran, A., Manivannan, A.C., Bharathi, G.S.J. et al. Production and characterization of exopolysaccharide (EPS) from marine Bacillus halotolerans and its antibacterial activity against clinical pathogens. Biologia 79, 605–619 (2024). https://doi.org/10.1007/s11756-023-01580-7

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  • DOI: https://doi.org/10.1007/s11756-023-01580-7

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