{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T13:28:59Z","timestamp":1787232539553,"version":"build-2736575974"},"reference-count":53,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM129325"],"award-info":[{"award-number":["GM129325"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P20 GM121310"],"award-info":[{"award-number":["P20 GM121310"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["2020-67014-32496"],"award-info":[{"award-number":["2020-67014-32496"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["WYO-630-22"],"award-info":[{"award-number":["WYO-630-22"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Wyoming School of Computing Graduate Scholarship"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2026,8,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Listeria monocytogenes (Lm<\/jats:italic>\n                      ) is a major foodborne pathogen capable of persisting on fresh produce and in food-processing environments. Although exopolysaccharides (EPSs) are recognized as central components of resilient bacterial biofilms,\n                      <jats:italic toggle=\"yes\">Lm<\/jats:italic>\n                      biofilms have long been considered to be EPS-less or EPS-poor. We previously identified a listerial EPS, Pss, whose biosynthesis is encoded by the highly conserved\n                      <jats:italic toggle=\"yes\">pss<\/jats:italic>\n                      operon and is stimulated by elevated c-di-GMP levels. However, the prevalence of Pss in wild-type strains remained unclear, in part, due to the lack of a specific detection probe. Here, we engineered a genetically encoded fluorescent probe, PssZ*-mS, based on a catalytically inactive variant (E72Q) of the Pss hydrolase PssZ. Structural modeling predicted that the E72Q substitution abolishes catalysis without disrupting Pss binding. PssZ*-mS bound purified Pss with high affinity (\n                      <jats:italic toggle=\"yes\">K<\/jats:italic>\n                      <jats:sub>d<\/jats:sub>\n                      , 50 nM) and showed no detectable interaction with other tested EPSs, including cellulose, pectin, maltodextrin, or alginate. Using PssZ*-mS, we demonstrate that the reference wild-type strain EGDe produces substantial Pss under standard conditions in liquid minimal glucose-containing medium and on diverse abiotic surfaces relevant to food environments. A survey of\n                      <jats:italic toggle=\"yes\">Lm<\/jats:italic>\n                      isolates revealed that Pss production is common, though not universal, among\n                      <jats:italic toggle=\"yes\">Lm<\/jats:italic>\n                      strains and is also present in non-\n                      <jats:italic toggle=\"yes\">monocytogenes Listeria<\/jats:italic>\n                      species. Notably, three genetically distinct strains from the infamous 2011 whole-cantaloupe listeriosis outbreak synthesize Pss. These findings challenge the prevailing view of listerial biofilms and identify Pss as a widespread component of\n                      <jats:italic toggle=\"yes\">Lm<\/jats:italic>\n                      biofilms, underscoring the need for targeted anti-Pss strategies to enhance food safety.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic toggle=\"yes\">Listeria monocytogenes<\/jats:italic>\n                        persists on fresh produce and in food-processing environments despite rigorous sanitation, yet the molecular basis of this resilience remains poorly understood. We developed a specific fluorescent probe for the conserved Pss exopolysaccharide (EPS) and show that Pss is a common component of listerial biofilms. These findings challenge the long-standing view that\n                        <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                        forms primarily EPS-less or EPS-poor biofilms and identify Pss as an important biofilm component. By enabling direct visualization of Pss, this work provides a new tool for studying listerial biofilms and opens avenues for targeted antibiofilm strategies to improve food safety and public health.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00153-26","type":"journal-article","created":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T16:36:56Z","timestamp":1784047016000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["The conserved Pss exopolysaccharide is a common component of\n                    <i>Listeria<\/i>\n                    biofilms"],"prefix":"10.1128","volume":"208","author":[{"given":"Aili","family":"Wang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01485tq96","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Biology, University of Wyoming","place":["Laramie, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed M.","family":"Elbakush","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01485tq96","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Biology, University of Wyoming","place":["Laramie, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oliver","family":"Trunschke","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01485tq96","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Biology, University of Wyoming","place":["Laramie, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4077-3932","authenticated-orcid":true,"given":"Mark","family":"Gomelsky","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01485tq96","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Biology, University of Wyoming","place":["Laramie, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2171"},{"key":"e_1_3_2_3_2","unstructured":"Centers for Disease Control and Prevention (CDC). 2024. 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