{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T17:39:12Z","timestamp":1793122752742,"version":"4.5.0"},"reference-count":54,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":3,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100011033","name":"Agencia Estatal de Investigaci\u00f3n","doi-asserted-by":"publisher","award":["TED2021\u2010129640B\u2010I00"],"award-info":[{"award-number":["TED2021\u2010129640B\u2010I00"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011033","name":"Agencia Estatal de Investigaci\u00f3n","doi-asserted-by":"publisher","award":["BIO2017\u201083533\u2010P"],"award-info":[{"award-number":["BIO2017\u201083533\u2010P"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011033","name":"Agencia Estatal de Investigaci\u00f3n","doi-asserted-by":"publisher","award":["PID2022\u2010140168NB\u2010I00"],"award-info":[{"award-number":["PID2022\u2010140168NB\u2010I00"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Mixed\u2010linkage \u03b2\u2010glucans (MLGs) are emerging as promising biopolymers with significant biotechnological potential due to their unique structural and rheological properties. In rhizobia, MLG biosynthesis is controlled by the second messenger cyclic di\u2010GMP (c\u2010di\u2010GMP) and mediated by the bicistronic operon\n                    <jats:italic>bgsBA<\/jats:italic>\n                    . However, the full composition of the biosynthetic machinery and strategies for enhanced production remain incompletely understood. In this study, we demonstrate that the outer membrane protein TolC is essential for MLG production in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Sinorhizobium meliloti<\/jats:italic>\n                    <\/jats:styled-content>\n                    . Genetic disruption of\n                    <jats:italic>tolC<\/jats:italic>\n                    abolished MLG synthesis, while its complementation restored production. We propose that TolC functions as part of a tripartite complex with BgsA and BgsB, enabling efficient polymer synthesis and\/or export. Furthermore, co\u2010overexpression of\n                    <jats:italic>tolC<\/jats:italic>\n                    ,\n                    <jats:italic>bgsBA<\/jats:italic>\n                    , and a constitutively active diguanylate cyclase (\n                    <jats:italic>pleD*<\/jats:italic>\n                    ) yielded an 8\u2010fold increase of MLG over a control plasmid without\n                    <jats:italic>tolC<\/jats:italic>\n                    , reaching up to ~10\u2009g\/L under bioreactor conditions. Additionally, this genetic module enabled de novo MLG production in otherwise non\u2010producer rhizobial hosts (e.g.,\n                    <jats:italic>Mesorhizobium japonicum<\/jats:italic>\n                    ), allowing bacterial chassis exchanges and highlighting its portability and potential for synthetic biology applications. Overall, our findings identify TolC as a key component of the MLG biosynthetic machinery and provide a robust platform for the scalable production of this valuable biopolymer.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.70424","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T06:24:54Z","timestamp":1785911094000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>TolC<\/scp>\n                    Is Required for a Mixed\u2010Linkage \u03b2\u2010Glucan (\n                    <scp>MLG<\/scp>\n                    ) Biosynthesis: Engineering Bacteria for\n                    <scp>MLG<\/scp>\n                    Overproduction"],"prefix":"10.1111","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8476-4431","authenticated-orcid":false,"given":"L.","family":"Ruiz\u2010S\u00e1ez","sequence":"first","affiliation":[{"name":"Department of Soil and Plant Microbiology Estaci\u00f3n Experimental del Zaid\u00edn, CSIC  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0408-5470","authenticated-orcid":false,"given":"P. J.","family":"Pacheco","sequence":"additional","affiliation":[{"name":"Department of Soil and Plant Microbiology Estaci\u00f3n Experimental del Zaid\u00edn, CSIC  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-2254-1141","authenticated-orcid":false,"given":"J.","family":"Peinado","sequence":"additional","affiliation":[{"name":"Department of Biology Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1284-4510","authenticated-orcid":false,"given":"J.","family":"Lloret","sequence":"additional","affiliation":[{"name":"Department of Biology Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9215-6518","authenticated-orcid":false,"given":"S.","family":"Mu\u00f1oz","sequence":"additional","affiliation":[{"name":"Department of Soil and Plant Microbiology Estaci\u00f3n Experimental del Zaid\u00edn, CSIC  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6404-8754","authenticated-orcid":false,"given":"J.","family":"Sanju\u00e1n","sequence":"additional","affiliation":[{"name":"Department of Soil and Plant Microbiology Estaci\u00f3n Experimental del Zaid\u00edn, CSIC  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9918-5671","authenticated-orcid":false,"given":"D.","family":"P\u00e9rez\u2010Mendoza","sequence":"additional","affiliation":[{"name":"Department of Soil and Plant Microbiology Estaci\u00f3n Experimental del Zaid\u00edn, CSIC  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,4]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2019.09.008"},{"key":"e_1_2_11_3_1","article-title":"Cellulose Synthesis in Cyanobacteria: Shared Pathways and Distinct Features With Bacteria and Plants","volume":"14","author":"An X.","year":"2025","journal-title":"Plants (Basel, Switzerland)"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2020.06.190"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.14624"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-84-1-188"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00303-23"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_11_9_1","doi-asserted-by":"crossref","unstructured":"Caseiro C. J. N. R.Dias C. M. G.deAndrade Fontes andP.Bule.2022. \u201cFrom Cancer Therapy to Winemaking: The Molecular Structure and Applications of \u03b2\u2010Glucans and \u03b2\u20101 3\u2010Glucanases.\u201d23:3156.","DOI":"10.3390\/ijms23063156"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells10030510"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-21-7-0947"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.27099-0"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2004.09.007"},{"key":"e_1_2_11_14_1","doi-asserted-by":"crossref","unstructured":"Flores A. J.Zaera L.Ruiz\u2010Saez et\u00a0al.2026.\u201cCrystal Structure of Alternating Mixed\u2010Linkage \u03b2\u2010Glucan by X\u2010Ray Diffraction.\u201dPreprint SSRN.https:\/\/dx.doi.org\/10.2139\/ssrn.6170295.","DOI":"10.2139\/ssrn.6170295"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2011.03.008"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2109"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1093\/femsml\/uqad026"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2022.104024"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.73.011303.074104"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.d01-1880.x"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2020-0232"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0040460"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1039\/C4NP00086B"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13977"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-3617-6_9"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2004.12.001"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2006.05539.x"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-019-0313-3"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jenvman.2014.05.010"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780470920541.ch1"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0091645"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-09290-2"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2011.07849.x"},{"key":"e_1_2_11_34_1","first-page":"E757","article-title":"Novel Mixed\u2010Linkage Beta\u2010Glucan Activated by c\u2010di\u2010GMP in \n                     Sinorhizobium meliloti","volume":"112","author":"P\u00e9rez\u2010Mendoza D.","year":"2015","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"e_1_2_11_35_1","doi-asserted-by":"publisher","DOI":"10.3390\/biology11091364"},{"key":"e_1_2_11_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2015.12.004"},{"key":"e_1_2_11_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2026.02.009"},{"key":"e_1_2_11_38_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12866-014-0298-z"},{"key":"e_1_2_11_39_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.21.7471-7480.2005"},{"key":"e_1_2_11_40_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.67.3.389"},{"key":"e_1_2_11_41_1","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.00043-12"},{"key":"e_1_2_11_42_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2005.04697.x"},{"key":"e_1_2_11_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/325279a0"},{"key":"e_1_2_11_44_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpmb.38"},{"key":"e_1_2_11_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(94)90324-7"},{"key":"e_1_2_11_46_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00795-15"},{"key":"e_1_2_11_47_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1702435114"},{"key":"e_1_2_11_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1183-784"},{"key":"e_1_2_11_49_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/9.1.31"},{"key":"e_1_2_11_50_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.17.4416-4425.1998"},{"key":"e_1_2_11_51_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1399004714001850"},{"key":"e_1_2_11_52_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.711762"},{"key":"e_1_2_11_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2012.10.001"},{"key":"e_1_2_11_54_1","doi-asserted-by":"publisher","DOI":"10.1038\/s44259-025-00078-3"},{"key":"e_1_2_11_55_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2008.10.010"}],"container-title":["Microbial Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70424","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/1751-7915.70424","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70424","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T17:30:49Z","timestamp":1793122249000},"score":1,"resource":{"primary":{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/10.1111\/1751-7915.70424"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8]]},"references-count":54,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2026,8]]}},"alternative-id":["10.1111\/1751-7915.70424"],"URL":"https:\/\/doi.org\/10.1111\/1751-7915.70424","archive":["Portico"],"relation":{},"ISSN":["1751-7915","1751-7915"],"issn-type":[{"value":"1751-7915","type":"print"},{"value":"1751-7915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8]]},"assertion":[{"value":"2026-04-30","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-27","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-04","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70424"}}