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Although\n                      <jats:italic toggle=\"yes\">Bacillus<\/jats:italic>\n                      lipopeptides are known to exhibit antibacterial activity against\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      , the specific compounds responsible for the inhibition of ETEC in probiotic-relevant\n                      <jats:italic toggle=\"yes\">Bacillus<\/jats:italic>\n                      strains remain undefined. Here, we sought to identify extracellular metabolite(s) associated with the anti-ETEC activity of\n                      <jats:italic toggle=\"yes\">Bacillus subtilis<\/jats:italic>\n                      DB9011. DB9011 culture supernatant strongly inhibited the growth of ETEC and other enteric pathogens, including\n                      <jats:italic toggle=\"yes\">Salmonella<\/jats:italic>\n                      Typhimurium. Bioactivity-guided fractionation of the culture supernatant, followed by liquid chromatography\u2013quadrupole time-of-flight mass spectrometry, identified C17 fengycin B as a major active metabolite enriched in inhibitory fractions, but absent from the non-inhibitory type strain NBRC13719\u1d40. A fengycin-containing fraction enriched in C17 fengycin B exhibited concentration-dependent antibacterial activity against ETEC and enhanced growth inhibition when combined with colistin, whereas incomplete inhibition at native supernatant-equivalent concentrations indicated that additional metabolites contribute to the overall antibacterial phenotype. While fengycins have been primarily recognized for antifungal activity and, more recently, for effects on gram-negative bacteria, our findings identify C17 fengycin B as a major contributor to the ETEC-inhibitory phenotype of\n                      <jats:italic toggle=\"yes\">B. subtilis<\/jats:italic>\n                      DB9011. These findings provide mechanistic insights into how probiotic-relevant\n                      <jats:italic toggle=\"yes\">Bacillus<\/jats:italic>\n                      strains inhibit enteric pathogens; nonetheless, further studies are required to evaluate\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      relevance and functional performance under intestinal conditions.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Antimicrobial resistance poses a major threat to livestock production, where antibiotics remain widely used to control post-weaning diarrhea caused by enterotoxigenic\n                        <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                        (ETEC). Supplementation with\n                        <jats:italic toggle=\"yes\">Bacillus subtilis<\/jats:italic>\n                        DB9011 is known to reduce pathogenic\n                        <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                        colonization in piglets; however, the metabolites responsible for this protective effect remain unidentified. Here, using a bioactivity-guided fractionation approach, we identify C17 fengycin B as a major contributing metabolite associated with the anti-ETEC activity of DB9011 and show enhanced growth inhibition during co-treatment with colistin. These findings clarify the functional contribution of\n                        <jats:italic toggle=\"yes\">Bacillus<\/jats:italic>\n                        -derived metabolites to pathogen inhibition and establish a mechanistic basis for strain-specific antimicrobial activity.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/aem.00276-26","type":"journal-article","created":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T13:04:40Z","timestamp":1780578280000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Role of C17 fengycin B from\n                    <i>Bacillus subtilis<\/i>\n                    DB9011 in enterotoxigenic\n                    <i>Escherichia coli<\/i>\n                    inhibition"],"prefix":"10.1128","volume":"92","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-6247-7513","authenticated-orcid":false,"given":"Yuki","family":"Indo","sequence":"first","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]},{"name":"SDS Biotech K.K., Tsukuba Research & Technology Center","place":["Tsukuba, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kosuke","family":"Shimizu","sequence":"additional","affiliation":[{"name":"SDS Biotech K.K., Tsukuba Research & Technology Center","place":["Tsukuba, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayane","family":"Sato","sequence":"additional","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fu","family":"Namai","sequence":"additional","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]},{"name":"Livestock Immunology Unit, International Education and Research Centre for Food and Agricultural Immunology (CFAI), Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weichen","family":"Gong","sequence":"additional","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]},{"name":"Livestock Immunology Unit, International Education and Research Centre for Food and Agricultural Immunology (CFAI), Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio","family":"Villena","sequence":"additional","affiliation":[{"name":"Division of Animal Immunology and Omics, CFAI, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chie","family":"Hikita","sequence":"additional","affiliation":[{"name":"SDS Biotech K.K., Tsukuba Research & Technology Center","place":["Tsukuba, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6483-0669","authenticated-orcid":true,"given":"Keita","family":"Nishiyama","sequence":"additional","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]},{"name":"Livestock Immunology Unit, International Education and Research Centre for Food and Agricultural Immunology (CFAI), Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4586-5553","authenticated-orcid":false,"given":"Haruki","family":"Kitazawa","sequence":"additional","affiliation":[{"name":"Laboratory of Animal Food Function, Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]},{"name":"Livestock Immunology Unit, International Education and Research Centre for Food and Agricultural Immunology (CFAI), Graduate School of Agricultural Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","unstructured":"United Nations. 2025. 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