Modulation of Gut Microbiota by Protein-Oligosaccharide Maillard-Reaction-Derived Probiotic Microcapsules

Abstract

As a new type of prebiotic, the Maillard reaction products of protein - oligosaccharide conjugates have emerged as a promising material for probiotic encapsulation and improve host health. However, the regulatory mechanisms of these conjugates on gut microbiota remain insufficiently explored. This study developed a probiotic microcapsule wall material through wet-heating conjugation of soy protein isolate (SPI) and galactooligosaccharide (GOS). Lactobacillus acidophilus was successfully encapsulated using emulsification technology. Physicochemical characterization (pH, intermediate products, browning intensity, and glycation degree) validated the formation of SPI-GOS conjugates. Subsequently, this study further investigated the impact of the Maillard-reaction-based microcapsule on human intestinal flora. The SGM microcapsule can significantly regulate the composition of intestinal flora. The in vitro digestion model demonstrated that the SGM microcapsules could increase the relative abundance of beneficial bacteria (Bifidobacterium in Actinobacteriota and Prevotella in the Firmicutes) and reduce the relative abundance of harmful bacteria (Desulfovibrio and Escherichia-Shigella in the Proteobacteria, Eggerthella, and Actinomyces in the Actinobacteriota). Functional prediction analysis indicated that SGM microcapsules could improve the metabolic and biosynthetic functions of the intestinal microbiota. This Maillard-derived SPI-GOS microencapsulation system establishes a novel synbiotic paradigm synergistically combining prebiotic conjugates and probiotics. The findings provide guidance for developing functional foods targeting gut microbiota modulation, with significant implications for dietary intervention strategies in metabolic health management.

Keywords

gut microbiota / in vitro fermentation / Maillard reaction / microcapsule / synbiotic

References

[1] Ao, S., W. Wei, P. O. S., et al. 2022. “Maillard Reaction of Food-Derived Peptides as a Potential Route to Generate Meat Flavor Compounds: A Review.” Food Research International 151: 110823. https://doi.org/10.1016/j.foodres.2021.110823.

[2] Arslan-Tontul, S. 2020. “The Combined Usage of β-Cyclodextrin and Milk Proteins in Microencapsulation of Bifidobacterium bifidum BB-12.” Probiotics and Antimicrobial Proteins 12, no. 2: 747–755. https://doi.org/10.1007/s12602-019-09621-x.

[3] Ashbala, S., C. Zhang, J. Xie, et al. 2022. “Maillard Reaction Chemistry in Formation of Critical Intermediates and Flavour Compounds and Their Antioxidant Properties.” Food Chemistry 393: 133416. https://doi.org/10.1016/j.foodchem.2022.133416.

[4] Chen, H., Y. Su, S. Li, et al. 2024. “Advances in Oligosaccharides and Polysaccharides With Different Structures as Wall Materials for Probiotics Delivery: A Review.” International Journal of Biological Macromolecules 277, no. 4: 134468. https://doi.org/10.1016/j.ijbiomac.2024.134468.

[5] Chen, H. W., Y. W. Chang, and W. P. Fang. 2020. “A New Approach for the Microencapsulation of Clitoria ternatea Petal Extracts by a High-Pressure Processing Method.” Pharmaceutics 13, no. 1: 23. https://doi.org/10.3390/pharmaceutics13010023.

[6] Corona-Hernandez, R. I., E. Álvarez-Parrilla, J. Lizardi-Mendoza, A. R. Islas-Rubio, L. A. de la Rosa, and A. Wall-Medrano. 2013. “Structural Stability and Viability of Microencapsulated Probiotic Bacteria: A Review.” Comprehensive Reviews in Food Science and Food Safety 12, no. 6: 614–628. https://doi.org/10.1111/1541-4337.12030.

[7] de Oliveira, F. C., J. S. R. Coimbra, E. B. de Oliveira, A. D. G. Zuñiga, and E. E. G. Rojas. 2016. “Food Protein-Polysaccharide Conjugates Obtained via the Maillard Reaction: A Review.” Critical Reviews in Food Science and Nutrition 56, no. 7: 1108–1125. https://doi.org/10.1080/10408398.2012.755669.

[8] De Vos, W. M., M. Nguyen Trung, M. Davids, et al. 2024. “Phytate Metabolism Is Mediated by Microbial Cross-Feeding in the Gut Microbiota.” Nature Microbiology 9, no. 7: 1812–1827. https://doi.org/10.1038/s41564-024-01698-7.

[9] Dubois, P. C. A., G. Trynka, L. Franke, et al. 2010. “Multiple Common Variants for Celiac Disease Influencing Immune Gene Expression.” Nature Genetics 42, no. 4: 295–302. https://doi.org/10.1038/ng.543.

[10] Finot, P. A. 2005. “The Absorption and Metabolism of Modified Amino Acids in Processed Foods.” Journal of AOAC International 88, no. 3: 894–903. https://doi.org/10.1093/jaoac/88.3.894.

[11] Gu, F., J. M. Kim, K. Hayat, S. Xia, B. Feng, and X. Zhang. 2009. “Characteristics and Antioxidant Activity of Ultrafiltrated Maillard Reaction Products From a Casein–Glucose Model System.” Food Chemistry 117, no. 1: 48–54. https://doi.org/10.1016/j.foodchem.2009.03.074.

[12] Guo, Y., Y. H. Bao, and Y. Y. Chai. 2019. “Preparation of Microcapsule Antioxidative Wall Materials of Pine Nut Oil by the Maillard Reaction.” Journal of the Science of Food and Agriculture 99, no. 6: 2793–2801. https://doi.org/10.1002/jsfa.9488.

[13] Han, K., Y. Yao, S. Dong, et al. 2017. “Chemical Characterization of the Glycated Myofibrillar Proteins From Grass Carp (Ctenopharyngodon idella) and Their Impacts on the Human Gut Microbiota In Vitro Fermentation.” Food & Function 8, no. 3: 1184–1194. https://doi.org/10.1039/c6fo01632d.

[14] Hernandez-Hernandez, O., F. J. Moreno, S. Kolida, R. A. Rastall, and M. L. Sanz. 2011. “Effect of Glycation of Bovine β-Lactoglobulin With Galactooligosaccharides on the Growth of Human Faecal Bacteria.” International Dairy Journal 21, no. 12: 949–952. https://doi.org/10.1016/j.idairyj.2011.06.002.

[15] How, Y., and L. Pui. 2021. “Effect of Prebiotics Encapsulated With Probiotics on Encapsulation Efficiency, Microbead Size, and Survivability: A Review.” Journal of Food Measurement and Characterization 15, no. 6: 4899–4916. https://doi.org/10.1007/s11694-021-01059-6.

[16] Huang, L. S., C. Kong, R. Y. Gao, et al. 2018. “Analysis of Fecal Microbiota in Patients With Functional Constipation Undergoing Treatment With Synbiotics.” European Journal of Clinical Microbiology & Infectious Diseases 37, no. 3: 555–563. https://doi.org/10.1007/s10096-017-3149-7.

[17] Jandhyala, S. M. 2015. “Role of the Normal Gut Microbiota.” World Journal of Gastroenterology 21, no. 29: 8787–8803. https://doi.org/10.3748/wjg.v21.i29.8787.

[18] Kan, X., G. Chen, W. Zhou, et al. 2021. “Application of Protein-Polysaccharide Maillard Conjugates as Emulsifiers: Source, Preparation and Functional Properties.” Food Research International 150, no. A: 110740. https://doi.org/10.1016/j.foodres.2021.110740.

[19] Kan, X., Y. Hu, Y. Hang, et al. 2022. “Characterization of Whey Protein Isolate-Gum Arabic Maillard Conjugate and Evaluation of the Effects of Conjugate-Stabilized Emulsion on Microbiota of Human Fecal Cultures.” Food Hydrocolloids 134, no. 1: 108060. https://doi.org/10.1016/j.foodhyd.2022.108060.

[20] Lau, S. K., P. C. Woo, G. K. Woo, et al. 2004. “Eggerthella hongkongensis sp. nov. and Eggerthella sinensis sp. nov., Two Novel Eggerthella Species, Account for Half of the Cases of Eggerthella bacteremia.” Diagnostic Microbiology and Infectious Disease 49, no. 4: 255–263. https://doi.org/10.1016/j.diagmicrobio.2004.04.012.

[21] Li, S., W. Jiang, C. Zheng, et al. 2020. “Oral Delivery of Bacteria: Basic Principles and Biomedical Applications.” Journal of Controlled Release 327: 801–833. https://doi.org/10.1016/j.jconrel.2020.09.011.

[22] Liu, L., P. Chen, W. Zhao, X. Li, H. Wang, and X. Qu. 2017. “Effect of Microencapsulation With the Maillard Reaction Products of Whey Proteins and Isomaltooligosaccharide on the Survival Rate of Lactobacillus rhamnosus in White Brined Cheese.” Food Control 79: 44–49. https://doi.org/10.1016/j.foodcont.2017.03.016.

[23] Liu, L., X. Li, Y. Zhu, et al. 2016. “Effect of Microencapsulation With Maillard Reaction Products of Whey Proteins and Isomaltooligosaccharide on the Survival of Lactobacillus rhamnosus.” LWT 73: 37–43. https://doi.org/10.1016/j.lwt.2016.05.030.

[24] Liu, S. C., D. J. Yang, S. Y. Jin, C. H. Hsu, and S. L. Chen. 2008. “Kinetics of Color Development, pH Decreasing, and Anti-Oxidative Activity Reduction of Maillard Reaction in Galactose/Glycine Model Systems.” Food Chemistry 108, no. 2: 533–541. https://doi.org/10.1016/j.foodchem.2007.11.006.

[25] Lorelei, M., D. Ghislain, and A. Remi. 2022. “Chitosan Chemistry Review for Living Organisms Encapsulation.” Carbohydrate Polymers 29, no. 5: 119877. https://doi.org/10.1016/j.carbpol.2022.119877.

[26] Lu, G., Y. Zhang, Y. Ren, J. S. Shi, Z. H. Xu, and Y. Geng. 2023. “Diversity and Comparison of Intestinal Desulfovibrio in Patients With Liver Cirrhosis and Healthy People.” Microorganisms 11, no. 2: 276. https://doi.org/10.3390/microorganisms11020276.

[27] Lund, M. N., and C. A. Ray. 2017. “Control of Maillard Reactions in Foods: Strategies and Chemical Mechanisms.” Journal of Agricultural and Food Chemistry 65, no. 23: 4537–4552. https://doi.org/10.1021/acs.jafc.7b00882.

[28] Martinez-Alvarenga, M. S., E. Y. Martinez-Rodriguez, L. E. Garcia-Amezquita, et al. 2014. “Effect of Maillard Reaction Conditions on the Degree of Glycation and Functional Properties of Whey Protein Isolate – Maltodextrin Conjugates.” Food Hydrocolloids 38: 110–118. https://doi.org/10.1016/j.foodhyd.2013.11.006.

[29] Minekus, M., M. Alminger, P. Alvito, et al. 2014. “A Standardised Static In Vitro Digestion Method Suitable for Food - An International Consensus.” Food & Function 5, no. 6: 1113–1124. https://doi.org/10.1039/c3fo60702j.

[30] Muthaiyan, A., O. Hernandez-Hernandez, F. J. Moreno, M. L. Sanz, and S. C. Ricke. 2012. “Hydrolyzed Caseinomacropeptide Conjugated Galactooligosaccharides Support the Growth and Enhance the Bile Tolerance in Lactobacillus Strains.” Journal of Agricultural and Food Chemistry 60, no. 27: 6839–6845. https://doi.org/10.1021/jf301392y.

[31] Nooshkam, M., and A. Madadlou. 2016. “Maillard Conjugation of Lactulose With Potentially Bioactive Peptides.” Food Chemistry 192: 831–836. https://doi.org/10.1016/j.foodchem.2015.07.094.

[32] Nooshkam, M., and M. Varidi. 2020. “Maillard Conjugate-Based Delivery Systems for the Encapsulation, Protection, and Controlled Release of Nutraceuticals and Food Bioactive Ingredients: A Review.” Food Hydrocolloids 100: 105389. https://doi.org/10.1016/j.foodhyd.2019.105389.

[33] Orive, G., S. K. Tam, J. L. Pedraz, and J. P. Hallé. 2006. “Biocompatibility of Alginate-Poly-L-Lysine Microcapsules for Cell Therapy.” Biomaterials 27, no. 20: 3691–3700. https://doi.org/10.1016/j.biomaterials.2006.02.048.

[34] Olabisi, R. M. 2015. “Cell Microencapsulation With Synthetic Polymers.” Journal of Biomedical Materials Research. Part A 103, no. 2: 846–859. https://doi.org/10.1002/jbm.a.35205.

[35] Rodrigues, F. J., M. F. Cedran, J. L. Bicas, and H. H. Sato. 2020. “Encapsulated Probiotic Cells: Relevant Techniques, Natural Sources as Encapsulating Materials and Food Applications -A Narrative Review.” Food Research International 137: 109682. https://doi.org/10.1016/j.foodres.2020.109682.

[36] Shin, N. R., T. W. Whon, and J. W. Bae. 2015. “Proteobacteria: Microbial Signature of Dysbiosis in Gut Microbiota.” Trends in Biotechnology 33, no. 9: 496–503. https://doi.org/10.1016/j.tibtech.2015.06.011.

[37] Vhangani, L. N., and J. Van Wyk. 2013. “Antioxidant Activity of Maillard Reaction Products (MRPs) Derived From Fructose–Lysine and Ribose–Lysine Model Systems.” Food Chemistry 137, no. 1–4: 92–98. https://doi.org/10.1016/j.foodchem.2012.09.030.

[38] Wang, G., J. Zhu, L. Liu, et al. 2020. “Optimization for Galactooligosaccharides Synthesis: A Potential Alternative for Gut Health and Immunity.” Life Sciences 245, no. C: 117353. https://doi.org/10.1016/j.lfs.2020.117353.

[39] Wang, Y., H. F. Sheng, Y. He, et al. 2012. “Comparison of the Levels of Bacterial Diversity in Freshwater, Intertidal Wetland, and Marine Sediments by Using Millions of Illumina Tags.” Applied and Environmental Microbiology 78, no. 23: 8264–8271. https://doi.org/10.1128/aem.01821-12.

[40] Wen, C., J. Zhang, W. Qin, et al. 2020. “Structure and Functional Properties of Soy Protein Isolate-Lentinan Conjugates Obtained in Maillard Reaction by Slit Divergent Ultrasonic Assisted Wet Heating and the Stability of Oil-in-Water Emulsions.” Food Chemistry 331: 127374. https://doi.org/10.1016/j.foodchem.2020.127374.

[41] Xu, X., H. Fu, H. Quan, et al. 2025. “Effects of Fructooligosaccharides and Lactobacillus reuteri on the Composition and Metabolism of Gut Microbiota in Students.” Food & Function 16, no. 4: 1562–1575. https://doi.org/10.1039/d4fo03763d.

[42] Zhang, C., A. Yin, H. Li, et al. 2015. “Dietary Modulation of Gut Microbiota Contributes to Alleviation of Both Genetic and Simple Obesity in Children.” EBioMedicine 2, no. 8: 968–984. https://doi.org/10.1016/j.ebiom.2015.07.007.

[43] Zhong, L., N. Ma, Y. Wu, et al. 2019. “Characterization and Functional Evaluation of Oat Protein Isolate-Pleurotus Ostreatus β-Glucan Conjugates Formed via Maillard Reaction.” Food Hydrocolloids 87: 459–469. https://doi.org/10.1016/j.foodhyd.2018.08.034.

[44] Zhong, S. R., M. F. Li, Z. H. Zhang, M. H. Zong, X. L. Wu, and W. Y. Lou. 2021. “Novel Antioxidative Wall Materials for Lactobacillus Casei Microencapsulation via the Maillard Reaction Between the Soy Protein Isolate and Prebiotic Oligosaccharides.” Journal of Agricultural and Food Chemistry 69, no. 46: 13744–13753. https://doi.org/10.1021/acs.jafc.1c02907.

RIGHTS & PERMISSIONS

2025 The Author(s). Food Bioengineering published by John Wiley & Sons Australia, Ltd. on behalf of State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology.