Isomelezitose Overproduction by Alginate-Entrapped Recombinant E. coli Cells and In Vitro Evaluation of Its Potential Prebiotic Effect
Abstract
:1. Introduction
2. Results and Discussion
2.1. Production and Characterisation of a Biocatalyst Based on Alginate-Entrapped E. coli Cells
2.2. Isomelezitose Overproduction Employing the Immobilised E. coli Cells in Alginate Beads
2.3. Potential Prebiotic Effect of the Hetero-GlcOS Generated by the Alginate-Entrapped E. coli Cells Expressing Mr-αGlu
2.4. Analysis of the Potential Prebiotic Activity of Isomelezitose
3. Materials and Methods
3.1. Materials and Reagents
3.2. Microorganisms and Culture Conditions
3.3. Optimal Conditions for Mr-αGlu Expression in E. coli
3.4. Standard Hydrolytic Activity Assay
3.5. Entrapment of E. coli Cells Harbouring the Mr-αGlu Activity into Alginate Beads
3.6. Thermal and pH Stability of the Alginate-Entrapped E. coli Cells
3.7. Production of Honey Hetero-GlcOS and Operational Stability of Immobilised Biocatalysts in Transglucosylation Reactions
3.8. Partial Purification of Hetero-GlcOS Mixture by Alginate-Entrapped K. phaffii and Isomelezitose Purification
3.9. Effect of Isomelezitose and Commercial Prebiotic Sugars on the Grow and Metabolic Activity of Probiotic Bacteria
3.10. Analytical Analysis
3.11. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Glc + Fru | Suc | Tre | Isom | Rest of Hetero-GlcOS | Total Hetero-GlcOS | |
---|---|---|---|---|---|---|
Entrapped E. coli cells | 217 ± 8.9 | 65 ± 1.6 | 95 ± 3.8 | 75 ± 1.1 | 48 ± 1 | 218 ± 6 |
Soluble Mr-αGlu | 97 ± 2 | 146 ± 16 | 61 ± 2 | 173 ± 14 | 23 ± 1.2 | 256 ± 17 |
Bacteria | Grow Parameters on MRS Media | |||||
---|---|---|---|---|---|---|
ΔmaxOD600a | g (h) b | |||||
NCS | FOS | Isom | NCS | FOS | Isom | |
L. casei | 0.63 ± 0.03 | 0.71 ± 0.03 | 0.76 ± 0.01 * | 1.75 ± 0.01 | 1.34 ± 0.09 | 1.41 ± 0.08 * |
L. rhamnosus | 0.94 ± 0.01 | 1.09 ± 0.02 | 1.13 ± 0.02 * | 1.30 ± 0.07 | 0.87 ± 0.04 | 0.80 ± 0.02 * |
E. faecium | 0.58 ± 0.03 | 0.79 ± 0.03 | 0.89 ± 0.05 *,# | 1.70 ± 0.20 | 0.71 ± 0.14 | 0.87 ± 0.06 * |
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Garcia-Gonzalez, M.; Cervantes, F.V.; Ipiales, R.P.; de la Rubia, A.; Plou, F.J.; Fernández-Lobato, M. Isomelezitose Overproduction by Alginate-Entrapped Recombinant E. coli Cells and In Vitro Evaluation of Its Potential Prebiotic Effect. Int. J. Mol. Sci. 2022, 23, 12682. https://doi.org/10.3390/ijms232012682
Garcia-Gonzalez M, Cervantes FV, Ipiales RP, de la Rubia A, Plou FJ, Fernández-Lobato M. Isomelezitose Overproduction by Alginate-Entrapped Recombinant E. coli Cells and In Vitro Evaluation of Its Potential Prebiotic Effect. International Journal of Molecular Sciences. 2022; 23(20):12682. https://doi.org/10.3390/ijms232012682
Chicago/Turabian StyleGarcia-Gonzalez, Martin, Fadia V. Cervantes, Ricardo P. Ipiales, Angeles de la Rubia, Francisco J. Plou, and María Fernández-Lobato. 2022. "Isomelezitose Overproduction by Alginate-Entrapped Recombinant E. coli Cells and In Vitro Evaluation of Its Potential Prebiotic Effect" International Journal of Molecular Sciences 23, no. 20: 12682. https://doi.org/10.3390/ijms232012682