{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T14:45:49Z","timestamp":1782398749168,"version":"3.54.5"},"reference-count":27,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T00:00:00Z","timestamp":1782345600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Amway Research and Development"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Phytonutrient-enriched prebiotic mixtures (PEPs), composed of phytonutrients and prebiotics serving as substrates for gut microbes, are recognized for their potential to modulate gut microbial metabolic activity. However, direct evidence of enhanced effects following co-administration with probiotics remains limited. Using a three-phase design integrating ex vivo evaluation and clinical validation, we assessed how PEP components influence microbial responses and whether co-administration with probiotics enhances these effects. PEP components increased acetate, butyrate, total short-chain fatty acids (SCFAs), and lactate, with fiber-rich components showing the strongest effects (all q &lt; 0.0001 relative to negative control). Co-treatment with probiotics further enhanced butyrate and total SCFAs in a dose-dependent manner. In a randomized clinical study, all groups showed increases in fecal metabolites, with the combined group exhibiting the greatest increases in butyrate (+6.0 \u00b5mol\/g, ~1.5-fold, p &lt; 0.05) and total SCFAs (+22.9 \u00b5mol\/g, ~1.3-fold, p &lt; 0.05). Participants with constipation-type stool patterns shifted toward normal stool types across all groups. These findings support the utility of combined PEP and probiotic interventions for enhancing microbiome-derived metabolic activity.<\/jats:p>","DOI":"10.3390\/biology15131006","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T12:37:55Z","timestamp":1782391075000},"page":"1006","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Phytonutrient-Enriched Prebiotic Mixture Primes the Gut Environment to Enhance Probiotic Efficacy: Ex Vivo Screening and a Human Clinical Trial"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-9098-048X","authenticated-orcid":false,"given":"Hyo-Jin","family":"Lee","sequence":"first","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Ho","family":"Suh","sequence":"additional","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3207-2308","authenticated-orcid":false,"given":"Sunyoung","family":"Lee","sequence":"additional","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3669-1419","authenticated-orcid":false,"given":"Wilhelm H.","family":"Holzapfel","sequence":"additional","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"},{"name":"Department of Advanced Convergence, Handong Global University, Pohang 37554, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yosep","family":"Ji","sequence":"additional","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew K.","family":"Runyon","sequence":"additional","affiliation":[{"name":"Amway Research and Development, Ada, MI 49355, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hae","family":"Jo","sequence":"additional","affiliation":[{"name":"Amway Research and Development, Ada, MI 49355, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung-Yoon","family":"Hur","sequence":"additional","affiliation":[{"name":"Amway Research and Development, Ada, MI 49355, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ri","family":"Ryu","sequence":"additional","affiliation":[{"name":"Amway Research and Development, Ada, MI 49355, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eun Sung","family":"Jung","sequence":"additional","affiliation":[{"name":"HEM Pharma Inc., Suwon 16229, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Plaza-Diaz, J., Herrera-Quintana, L., Olivares-Arancibia, J., and V\u00e1zquez-Lorente, H. (2026). Personalized nutrition through the gut microbiome in metabolic syndrome and related comorbidities. Nutrients, 18.","DOI":"10.3390\/nu18020290"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Santhiravel, S., Bekhit, A.E.D.A., Mendis, E., Jacobs, J.L., Dunshea, F.R., Rajapakse, N., and Ponnampalam, E.N. (2022). The impact of plant phytochemicals on the gut microbiota of humans for a balanced life. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23158124"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lippolis, T., Cofano, M., Caponio, G.R., De Nunzio, V., and Notarnicola, M. (2023). Bioaccessibility and bioavailability of diet polyphenols and their modulation of gut microbiota. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24043813"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Kang, S., Lim, Y., Kim, Y.J., Jung, E.S., Suh, D.H., Lee, C.H., Park, E., Hong, J., Velliquette, R.A., and Kwon, O. (2019). Multivitamin and mineral supplementation containing phytonutrients scavenges reactive oxygen species in healthy subjects: A randomized, double-blinded, placebo-controlled trial. Nutrients, 11.","DOI":"10.3390\/nu11010101"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bedu-Ferrari, C., Biscarrat, P., Langella, P., and Cherbuy, C. (2022). Prebiotics and the human gut microbiota: From breakdown mechanisms to the impact on metabolic health. Nutrients, 14.","DOI":"10.3390\/nu14102096"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1600240","DOI":"10.1002\/mnfr.201600240","article-title":"Probiotics, gut microbiota, and their influence on host health and disease","volume":"61","author":"Delgado","year":"2017","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2305716","DOI":"10.1080\/19490976.2024.2305716","article-title":"Exploring substrate\u2013microbe interactions: A metabiotic approach toward developing targeted synbiotic compositions","volume":"16","author":"Speckmann","year":"2024","journal-title":"Gut Microbes"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Kezer, G., Paramithiotis, S., Khwaldia, K., Harahap, I.A., \u010cagalj, M., \u0160imat, V., Smaoui, S., Elfalleh, W., Ozogul, F., and Esatbeyoglu, T. (2025). A comprehensive overview of the effects of probiotics, prebiotics and synbiotics on the gut\u2013brain axis. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1651965"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.1016\/j.cell.2016.05.041","article-title":"From dietary fiber to host physiology: Short-chain fatty acids as key bacterial metabolites","volume":"165","author":"Koh","year":"2016","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2325","DOI":"10.1194\/jlr.R036012","article-title":"The role of short-chain fatty acids in the interplay between diet, gut microbiota, and host energy metabolism","volume":"54","author":"Groen","year":"2013","journal-title":"J. Lipid Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1111\/j.1753-4887.2011.00388.x","article-title":"Propionate as a health-promoting microbial metabolite in the human gut","volume":"69","author":"Hosseini","year":"2011","journal-title":"Nutr. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Salvi, P.S., and Cowles, R.A. (2021). Butyrate and the intestinal epithelium: Modulation of proliferation and inflammation in homeostasis and disease. Cells, 10.","DOI":"10.3390\/cells10071775"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e3","DOI":"10.1017\/gmb.2022.3","article-title":"Microbial lactate utilisation and the stability of the gut microbiome","volume":"3","author":"Louis","year":"2022","journal-title":"Gut Microbiome"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1017\/S0029665113000025","article-title":"Inter-individual differences in response to dietary intervention: Integrating omics platforms towards personalised dietary recommendations","volume":"72","author":"Lampe","year":"2013","journal-title":"Proc. Nutr. Soc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"G403","DOI":"10.1152\/ajpgi.00087.2025","article-title":"Emerging advances in intestinal models for in vitro preclinical research","volume":"329","author":"Adesina","year":"2025","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1038\/s41522-026-00978-w","article-title":"A colon mimetic screening approach reveals Lactobacillus fermentum as a microbiome-based therapy for COPD","volume":"12","author":"Kim","year":"2026","journal-title":"npj Biofilms Microbiomes"},{"key":"ref_17","unstructured":"Park, S., and Ji, Y. (2022). Method for Screening Personalized Intestinal Environment-Improving Material and Composition Therefor. (US 11237172 B2), U.S. Patent."},{"key":"ref_18","unstructured":"Park, S., and Ji, Y. (2020). PMAS Method for Screening Personalized Intestinal Environment-Improving Material. (10-2124474 B1), KR Patent."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"311","DOI":"10.3934\/microbiol.2024016","article-title":"Why does increased microbial fermentation in the human colon shift toward butyrate?","volume":"10","author":"Flint","year":"2024","journal-title":"AIMS Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.ijfoodmicro.2016.10.019","article-title":"Lactate- and acetate-based cross-feeding interactions between selected strains of lactobacilli, bifidobacteria and colon bacteria in the presence of inulin-type fructans","volume":"241","author":"Moens","year":"2017","journal-title":"Int. J. Food Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1038\/s41579-025-01183-w","article-title":"Gut microbiota-derived short-chain fatty acids and their role in human health and disease","volume":"23","author":"Mukhopadhya","year":"2025","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5810","DOI":"10.1128\/AEM.70.10.5810-5817.2004","article-title":"Lactate-utilizing bacteria isolated from human feces that produce butyrate as a major fermentation product","volume":"70","author":"Duncan","year":"2004","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s12934-019-1085-1","article-title":"Cell factories converting lactate and acetate to butyrate: Clostridium butyricum and microbial communities from dark fermentation bioreactors","volume":"18","author":"Detman","year":"2019","journal-title":"Microb. Cell Fact."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Honda, S., Eguchi, H., Okino, Y., and Wang, D.S. (2025). The probiotic strain Clostridium butyricum TO-A produces butyrate by utilizing lactate and acetate. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26072951"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1111\/j.1574-6941.2011.01085.x","article-title":"Kinetic modelling of lactate utilization and butyrate production by key human colonic bacterial species","volume":"76","author":"Laroche","year":"2011","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1093\/advances\/nmz049","article-title":"The role of the gut microbiome in predicting response to diet and the development of precision nutrition models. Part II: Results","volume":"10","author":"Hughes","year":"2019","journal-title":"Adv. Nutr."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Parada Venegas, D., De la Fuente, M.K., Landskron, G., Gonz\u00e1lez, M.J., Quera, R., Dijkstra, G., Harmsen, H.J.M., Faber, K.N., and Hermoso, M.A. (2019). Short chain fatty acids (SCFAs)-mediated gut epithelial and immune regulation and its relevance for inflammatory bowel diseases. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.01486"}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/15\/13\/1006\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T13:45:52Z","timestamp":1782395152000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/15\/13\/1006"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,25]]},"references-count":27,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["biology15131006"],"URL":"https:\/\/doi.org\/10.3390\/biology15131006","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,25]]}}}