Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast
The use of natural polysaccharides as biomaterials is gaining importance in tissue engineering due to their inherent biocompatibility. In this direction, the present study aims to explore the structure and biocompatibility of the EPS produced by <i>Virgibacillus dokdonensis</i> VITP14. T...
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MDPI AG
2022-09-01
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| Online Access: | https://www.mdpi.com/2073-4360/14/19/3986 |
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| author | Monic Andrew Gurunathan Jayaraman |
| author_facet | Monic Andrew Gurunathan Jayaraman |
| author_sort | Monic Andrew |
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| description | The use of natural polysaccharides as biomaterials is gaining importance in tissue engineering due to their inherent biocompatibility. In this direction, the present study aims to explore the structure and biocompatibility of the EPS produced by <i>Virgibacillus dokdonensis</i> VITP14. This marine bacterium produces 17.3 g/L of EPS at 96 h of fermentation. The EPS was purified using ion exchange and gel permeation chromatographic methods. The porous web-like structure and elemental composition (C, O, Na, Mg, P, S) of the EPS were inferred from SEM and EDX analysis. AFM analysis revealed spike-like lumps with a surface roughness of 84.85 nm. The zeta potential value of −10 mV indicates the anionic nature of the EPS. Initial molecular characterization showed that the EPS is a heteropolysaccharide composed of glucose (25.8%), ribose (18.6%), fructose (31.5%), and xylose (24%), which are the monosaccharide units in the HPLC analysis. The FTIR spectrum indicates the presence of functional groups/bonds typical of EPSs (O-H, C-H, C-O-H, C-O, S=O, and P=O). The polymer has an average molecular weight of 555 kDa. Further, NMR analysis revealed the monomer composition, the existence of two α- and six β-glycosidic linkages, and the branched repeating unit as → 1)[α-D-Xyl<i>p</i>-(1 → 2)-α-D-Glc<i>p</i>-(1 → 6)-β-D-Glc<i>p</i>-(1 → 5)]-β-D-Fru<i>p</i>-(2 → 2)[β-D-Xyl<i>p</i>-(1 → 4)]-β-D-Xyl<i>p</i>-(1 → 6)-β-D-Fru<i>f</i>-(2 → 4)-β-D-Rib<i>p</i>-(1 →. The EPS is thermally stable till 251.4 °C. X-ray diffraction analysis confirmed the semicrystalline (54.2%) nature of the EPS. Further, the EPS exhibits significant water solubility (76.5%), water-holding capacity (266.8%), emulsifying index (66.8%), hemocompatibility (erythrocyte protection > 87%), and cytocompatibility (cell viability > 80% on RAW264.7 and keratinocyte HaCaT cells) at higher concentrations and prolongs coagulation time in APTT and PT tests. Our research unveils the significant biocompatibility of VITP14 EPS for synthesizing a variety of biomaterials. |
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| spelling | doaj.art-bf413b9d9ec84f44b39d917c73f63b492023-11-23T21:32:28ZengMDPI AGPolymers2073-43602022-09-011419398610.3390/polym14193986Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta CoastMonic Andrew0Gurunathan Jayaraman1School of Biosciences and Technology, Vellore Institute of Technology, Vellore 632014, Tamil Nadu, IndiaSchool of Biosciences and Technology, Vellore Institute of Technology, Vellore 632014, Tamil Nadu, IndiaThe use of natural polysaccharides as biomaterials is gaining importance in tissue engineering due to their inherent biocompatibility. In this direction, the present study aims to explore the structure and biocompatibility of the EPS produced by <i>Virgibacillus dokdonensis</i> VITP14. This marine bacterium produces 17.3 g/L of EPS at 96 h of fermentation. The EPS was purified using ion exchange and gel permeation chromatographic methods. The porous web-like structure and elemental composition (C, O, Na, Mg, P, S) of the EPS were inferred from SEM and EDX analysis. AFM analysis revealed spike-like lumps with a surface roughness of 84.85 nm. The zeta potential value of −10 mV indicates the anionic nature of the EPS. Initial molecular characterization showed that the EPS is a heteropolysaccharide composed of glucose (25.8%), ribose (18.6%), fructose (31.5%), and xylose (24%), which are the monosaccharide units in the HPLC analysis. The FTIR spectrum indicates the presence of functional groups/bonds typical of EPSs (O-H, C-H, C-O-H, C-O, S=O, and P=O). The polymer has an average molecular weight of 555 kDa. Further, NMR analysis revealed the monomer composition, the existence of two α- and six β-glycosidic linkages, and the branched repeating unit as → 1)[α-D-Xyl<i>p</i>-(1 → 2)-α-D-Glc<i>p</i>-(1 → 6)-β-D-Glc<i>p</i>-(1 → 5)]-β-D-Fru<i>p</i>-(2 → 2)[β-D-Xyl<i>p</i>-(1 → 4)]-β-D-Xyl<i>p</i>-(1 → 6)-β-D-Fru<i>f</i>-(2 → 4)-β-D-Rib<i>p</i>-(1 →. The EPS is thermally stable till 251.4 °C. X-ray diffraction analysis confirmed the semicrystalline (54.2%) nature of the EPS. Further, the EPS exhibits significant water solubility (76.5%), water-holding capacity (266.8%), emulsifying index (66.8%), hemocompatibility (erythrocyte protection > 87%), and cytocompatibility (cell viability > 80% on RAW264.7 and keratinocyte HaCaT cells) at higher concentrations and prolongs coagulation time in APTT and PT tests. Our research unveils the significant biocompatibility of VITP14 EPS for synthesizing a variety of biomaterials.https://www.mdpi.com/2073-4360/14/19/3986halophilesmarine bacteriaexopolysaccharidesfermentationstructural characterizationbiomaterial |
| spellingShingle | Monic Andrew Gurunathan Jayaraman Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast Polymers halophiles marine bacteria exopolysaccharides fermentation structural characterization biomaterial |
| title | Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast |
| title_full | Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast |
| title_fullStr | Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast |
| title_full_unstemmed | Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast |
| title_short | Molecular Characterization and Biocompatibility of Exopolysaccharide Produced by Moderately Halophilic Bacterium <i>Virgibacillus dokdonensis</i> from the Saltern of Kumta Coast |
| title_sort | molecular characterization and biocompatibility of exopolysaccharide produced by moderately halophilic bacterium i virgibacillus dokdonensis i from the saltern of kumta coast |
| topic | halophiles marine bacteria exopolysaccharides fermentation structural characterization biomaterial |
| url | https://www.mdpi.com/2073-4360/14/19/3986 |
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