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S-equol, an exclusive gut bacterial metabolite, is associated with health benefits such as reduced blood pressure, cardiovascular disease prevention, and protection against hormone-related cancers due to its estrogen-mimicking structure and antioxidant properties. However, the limited availability of S-equol-producing bacteria has hindered its production and utilization. This study investigates the isolation and characterization of S-equol-producing microbes from albino Wistar rats and explores the impact of dietary interventions on S-equol production. Preliminary tests showed that both dietary groups excreted more S-equol in feces than urine, with rats on fermented soy feed showing higher S-equol levels due to the presence of daidzein, a precursor. In this study, we isolated four anaerobic S-equol-producing bacteria \u2014 MG1 (PX459562), MG2 (PX459563), MG3 (PX459564), and MG4 (PX459565) from the intestine and feces of albino Wistar rats. High-Performance Thin-Layer Chromatography (HPTLC) and High-Performance Liquid Chromatography (HPLC) confirmed the presence of S-equol, with concentrations ranging from 5.90 to 7.56\u00a0\u00b5g\/g of fermented soybean across different strains. Phylogenetic analysis revealed that the isolates belonged to the Enterobacteriaceae and Enterococcaceae families, identifying MG1 as\n                      <jats:italic>C. freundii<\/jats:italic>\n                      strain ATCC 8090, MG2 as\n                      <jats:italic>Escherichia fergusonii<\/jats:italic>\n                      strain NBRC 102419, and both MG3 and MG4 as\n                      <jats:italic>Enterococcus faecalis<\/jats:italic>\n                      strain NBRC 100480. Our findings underscore the significant role of gut microbiota in metabolizing daidzein into S-equol, highlighting the potential for utilizing these bacterial strains in functional food development and therapeutic applications. While the pathogenic nature of\n                      <jats:italic>E. fergusonii<\/jats:italic>\n                      (MG2) precludes its therapeutic use, strains MG1, MG3, and MG4, which match common commensal bacteria, show promise for commercial S-equol production and may serve as valuable resources for further investigation and utilization in promoting health and preventing associated diseases.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Dietary intervention modulates gut microbiota in albino Wistar rats.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Soybean fermentation enables efficient conversion of daidzin to bioactive S-equol.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Novel S-equol\u2013producing microbes were isolated and identified.<\/jats:italic>\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1007\/s00253-026-13759-4","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T07:55:18Z","timestamp":1772438118000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["S-equol producing bacteria: isolation and identification from Albino Wistar rat gut microbiota"],"prefix":"10.1007","volume":"110","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8087-2789","authenticated-orcid":false,"given":"Megha","family":"Gangwar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanaa Ismael","family":"Abduljabbar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5146-4743","authenticated-orcid":false,"given":"Jalaluddin","family":"Khan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Sarwar","family":"Alam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kahksha","family":"Ahmed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sameena","family":"Naaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5110-2945","authenticated-orcid":false,"given":"Bibhu Prasad","family":"Panda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,3,2]]},"reference":[{"issue":"1","key":"13759_CR1","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1089\/jwh.2011.2753","volume":"21","author":"T Aso","year":"2012","unstructured":"Aso T, Uchiyama S, Matsumura Y, Taguchi M, Nozaki M, Takamatsu K, Ishizuka B, Kubota T, Mizunuma H, Ohta H (2012) A natural S-equol supplement alleviates hot flushes and other menopausal symptoms in equol nonproducing postmenopausal Japanese women. 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