{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T18:33:05Z","timestamp":1786732385167,"version":"build-2736575974"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T00:00:00Z","timestamp":1657670400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010198","name":"Ministerio de Asuntos Econ\u00f3micos y Transformaci\u00f3n Digital, Gobierno de Espa\u00f1a","doi-asserted-by":"publisher","award":["FPU 13\/00831"],"award-info":[{"award-number":["FPU 13\/00831"]}],"id":[{"id":"10.13039\/501100010198","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["BIO2014-56752-R"],"award-info":[{"award-number":["BIO2014-56752-R"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["BIO2017-82462-R"],"award-info":[{"award-number":["BIO2017-82462-R"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["PID2020-113062RB-100"],"award-info":[{"award-number":["PID2020-113062RB-100"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006382","name":"Universidad de Oviedo","doi-asserted-by":"publisher","award":["PAPI-20-GR-2010-0019"],"award-info":[{"award-number":["PAPI-20-GR-2010-0019"]}],"id":[{"id":"10.13039\/501100006382","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100011941","name":"Gobierno del Principado de Asturias","doi-asserted-by":"publisher","award":["FC-15-GRUPIN14-014"],"award-info":[{"award-number":["FC-15-GRUPIN14-014"]}],"id":[{"id":"10.13039\/100011941","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,8,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Largimycins A1 and A2 are key members of a recently identified family of hybrid nonribosomal peptide polyketides belonging to the scarcely represented group of antitumor leinamycins. They are encoded by the gene cluster lrg of Streptomyces argillaceus. This cluster contains a halogenase gene and two sets of genes for the biosynthesis and incorporation of \u03b2 branches at C3 and C9. Noticeably, largimycins A1 and A2 are nonhalogenated compounds and only contain a \u03b2 branch at C3. By generating mutants in those genes and characterizing chemically their accumulated compounds, we could confirm the existence of a chlorination step at C19, the introduction of an acetyl-derived olefinic exomethylene group at C9, and a propionyl-derived \u03b2 branch at C3 in the biosynthesis pathway. Since the olefinic exomethylene group and the chlorine atom are absent in the final products, those biosynthetic steps can be considered cryptic in the overall pathway but essential to generating keto and epoxide functionalities at C9 and C18\/C19, respectively. We propose that chlorination at C19 is utilized as an activation strategy that creates the precursor halohydrin to finally yield the epoxy functionality at C18\/C19. This represents a novel strategy to create such functionalities and extends the small number of natural product biosynthetic pathways that include a cryptic chlorination step.<\/jats:p>","DOI":"10.1021\/acschembio.2c00416","type":"journal-article","created":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T11:49:33Z","timestamp":1657712973000},"page":"2320-2331","source":"Crossref","is-referenced-by-count":12,"title":["Biosynthesis\nof Largimycins in\n                    <i>Streptomyces\nargillaceus<\/i>\n                    Involves Transient \u03b2\u2011Alkylation\nand Cryptic Halogenation Steps Unprecedented in the Leinamycin Family"],"prefix":"10.1021","volume":"17","author":[{"given":"Adriana","family":"Becerril","sequence":"first","affiliation":[{"name":"Universidad de Oviedo , , ,","place":["Oviedo, Spain, 33006"]},{"name":"Instituto de Investigacio\u0301n Sanitaria de Asturias (ISPA) , ,","place":["Oviedo, Spain, 33011"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4556-688X","authenticated-orcid":true,"given":"Ignacio","family":"Pe\u0301rez-Victoria","sequence":"additional","affiliation":[{"name":"Fundacio\u0301n MEDINA, Centro de Excelencia en Investigacio\u0301n de Medicamentos Innovadores en Andaluci\u0301a , Armilla , ,","place":["Granada, Spain, 18016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesu\u0301s M.","family":"Marti\u0301n","sequence":"additional","affiliation":[{"name":"Fundacio\u0301n MEDINA, Centro de Excelencia en Investigacio\u0301n de Medicamentos Innovadores en Andaluci\u0301a , Armilla , ,","place":["Granada, Spain, 18016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1607-5106","authenticated-orcid":true,"given":"Fernando","family":"Reyes","sequence":"additional","affiliation":[{"name":"Fundacio\u0301n MEDINA, Centro de Excelencia en Investigacio\u0301n de Medicamentos Innovadores en Andaluci\u0301a , Armilla , ,","place":["Granada, Spain, 18016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose A.","family":"Salas","sequence":"additional","affiliation":[{"name":"Universidad de Oviedo , , ,","place":["Oviedo, Spain, 33006"]},{"name":"Instituto de Investigacio\u0301n Sanitaria de Asturias (ISPA) , ,","place":["Oviedo, Spain, 33011"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2729-841X","authenticated-orcid":true,"given":"Carmen","family":"Me\u0301ndez","sequence":"additional","affiliation":[{"name":"Universidad de Oviedo , , ,","place":["Oviedo, Spain, 33006"]},{"name":"Instituto de Investigacio\u0301n Sanitaria de Asturias (ISPA) , ,","place":["Oviedo, Spain, 33011"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2022,7,13]]},"reference":[{"key":"2026081413093701500_cit1","doi-asserted-by":"publisher","first-page":"4688","DOI":"10.1002\/anie.200806121","article-title":"The biosynthetic\nlogic of polyketide diversity","volume":"48","author":"Hertweck","year":"2009","journal-title":"Angew. Chem.,\nInt. Ed. Engl."},{"key":"2026081413093701500_cit2","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1016\/S0076-6879(09)04608-4","article-title":"Type I polyketide synthases\nthat require discrete acyltransferases","volume":"459","author":"Cheng","year":"2009","journal-title":"Methods\nEnzymol."},{"key":"2026081413093701500_cit3","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1039\/C5NP00125K","article-title":"Biosynthesis of polyketides by trans-AT polyketide synthases","volume":"33","author":"Helfrich","year":"2016","journal-title":"Nat. Prod. Rep."},{"key":"2026081413093701500_cit4","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1039\/b807243d","article-title":"Isoprenoid-like\nalkylations in polyketide biosynthesis","volume":"25","author":"Calderone","year":"2008","journal-title":"Nat.\nProd. Rep."},{"key":"2026081413093701500_cit5","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1039\/D0NP00045K","article-title":"Polyketide \u03b2\nbranching: diversity, mechanism and selectivity","volume":"38","author":"Walker","year":"2021","journal-title":"Nat. Prod. Rep."},{"key":"2026081413093701500_cit6","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1038\/nature07870","article-title":"Metamorphic enzyme assembly in polyketide diversification","volume":"459","author":"Gu","year":"2009","journal-title":"Nature."},{"key":"2026081413093701500_cit7","doi-asserted-by":"publisher","first-page":"6900","DOI":"10.1021\/ja9012134","article-title":"Bifunctional acyltransferase\/decarboxylase\nLnmK as the missing link for beta-alkylation in polyketide biosynthesis","volume":"131","author":"Liu","year":"2009","journal-title":"J. Am. Chem. 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Microbiol."},{"key":"2026081413093701500_cit10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0198145","article-title":"Uncovering\nproduction of specialized metabolites by Streptomyces argillaceus: Activation of cryptic biosynthesis gene clusters using nutritional\nand genetic approaches","volume":"13","author":"Becerril","year":"2018","journal-title":"PLoS One."},{"key":"2026081413093701500_cit11","doi-asserted-by":"publisher","first-page":"1541","DOI":"10.1021\/acschembio.0c00160","article-title":"Discovery\nof Cryptic Largimycins in Streptomyces Reveals Novel\nBiosynthetic Avenues Enriching the Structural Diversity of the Leinamycin\nFamily","volume":"15","author":"Becerril","year":"2020","journal-title":"ACS Chem. Biol."},{"key":"2026081413093701500_cit12","doi-asserted-by":"publisher","first-page":"E11131","DOI":"10.1073\/pnas.1716245115","article-title":"Discovery\nof the leinamycin family of natural products by mining actinobacterial\ngenomes","volume":"114","author":"Pan","year":"2017","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026081413093701500_cit13","doi-asserted-by":"publisher","first-page":"8977","DOI":"10.1073\/pnas.0603148103","article-title":"Convergence\nof isoprene and polyketide biosynthetic machinery: isoprenyl-S-carrier\nproteins in the pksX pathway of Bacillus subtilis","volume":"103","author":"Calderone","year":"2006","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026081413093701500_cit14","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1038\/nchembio.1342","article-title":"A conserved motif flags\nacyl carrier proteins for \u03b2 branching in polyketide synthesis","volume":"9","author":"Haines","year":"2013","journal-title":"Nat. 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Chem."},{"key":"2026081413093701500_cit17","doi-asserted-by":"publisher","first-page":"1760","DOI":"10.1039\/D1NP00010A","article-title":"Cryptic halogenation\nreactions in natural product biosynthesis","volume":"38","author":"Adak","year":"2021","journal-title":"Nat.\nProd. Rep."},{"key":"2026081413093701500_cit18","doi-asserted-by":"publisher","first-page":"5619","DOI":"10.1021\/acs.chemrev.6b00571","article-title":"Enzymatic Halogenation\nand Dehalogenation\nReactions: Pervasive and Mechanistically Diverse","volume":"117","author":"Agarwal","year":"2017","journal-title":"Chem. Rev."},{"key":"2026081413093701500_cit19","doi-asserted-by":"publisher","first-page":"11009","DOI":"10.1002\/anie.201804351","article-title":"Total synthesis establishes the biosynthesis\npathway to the naphterpin and marinone natural products","volume":"57","author":"Murray","year":"2018","journal-title":"Angew. Chem., Int. Ed. 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Tu\u0308 6176: Characterization\nof the Nataxazole Biosynthesis\nPathway","volume":"16","author":"Cano-Prieto","year":"2015","journal-title":"Chembiochem"},{"key":"2026081413093701500_cit33","doi-asserted-by":"publisher","first-page":"167","DOI":"10.1128\/AEM.72.1.167-177.2006","article-title":"Deoxysugar\ntransfer during chromomycin A3 biosynthesis in Streptomyces\ngriseus subsp. griseus: new derivatives with antitumor activity","volume":"72","author":"Mene\u0301ndez","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"2026081413093701500_cit34","doi-asserted-by":"publisher","first-page":"4929","DOI":"10.1128\/JB.180.18.4929-4937.1998","article-title":"Identification of two genes from Streptomyces\nargillaceus encoding glycosyltransferases involved\nin transfer of a disaccharide during biosynthesis of the antitumor\ndrug mithramycin","volume":"180","author":"Fernandez","year":"1998","journal-title":"J. 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