{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:49:19Z","timestamp":1792003759176,"version":"4.4.0"},"reference-count":38,"publisher":"American Chemical Society (ACS)","issue":"40","license":[{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["P30 GM133893"],"award-info":[{"award-number":["P30 GM133893"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35 GM147557"],"award-info":[{"award-number":["R35 GM147557"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35 GM152049"],"award-info":[{"award-number":["R35 GM152049"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006132","name":"Office of Science","doi-asserted-by":"publisher","award":["DE-SC0012704"],"award-info":[{"award-number":["DE-SC0012704"]}],"id":[{"id":"10.13039\/100006132","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006206","name":"Biological and Environmental Research","doi-asserted-by":"publisher","award":["KP1605010"],"award-info":[{"award-number":["KP1605010"]}],"id":[{"id":"10.13039\/100006206","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100023581","name":"National Science Foundation Graduate Research Fellowship Program","doi-asserted-by":"publisher","award":["1937971"],"award-info":[{"award-number":["1937971"]}],"id":[{"id":"10.13039\/100023581","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Natural products have been paramount in inspiring novel chemistry as well as providing therapeutic agents and chemical tools for biological applications. A recently emerging class of natural products, the thio\/seleno-imidazoles (TSIs), contains C\u2013S or C\u2013Se bonds that are installed by nonheme iron sulfoxide and selenoxide synthases (NHISSs). These TSIs, which include ergothioneine, are ubiquitous, but little is known about the broader TSI chemical space or co-occurrence with other biosynthetic enzymes. Here, we use NHISSs as a bioinformatic hook to identify a widespread family of biosynthetic gene clusters that encode a combination of NHISS and ATP-grasp enzymes. This search strategy enabled the discovery of two 2-thiohistidine-containing dipeptide natural products, one of which we isolated from the original microbial host. Structural and biochemical analyses of the corresponding biosynthetic enzymes revealed a novel NHISS active site architecture and an ATP-grasp enzyme with unprecedented substrate selectivity for 2-thiohistidine. Together, these results expand the known enzymology and distribution of TSIs and establish 2-thiohistidine as a biosynthetically encoded building block, enabling future discovery and functional investigation into this new natural product family.<\/jats:p>","DOI":"10.1021\/jacs.6c03714","type":"journal-article","created":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T03:01:27Z","timestamp":1790910087000},"page":"42939-42948","source":"Crossref","is-referenced-by-count":0,"title":["Widespread Biosynthesis\nof Thiohistidine Dipeptides\nin Bacteria"],"prefix":"10.1021","volume":"148","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-6824-7796","authenticated-orcid":true,"given":"Marissa L.","family":"Abbott","sequence":"first","affiliation":[{"name":"Princeton University , , , ,","place":["Princeton, New Jersey, United States, 08544"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1077-2085","authenticated-orcid":true,"given":"Jacob T.","family":"Leeman","sequence":"additional","affiliation":[{"name":"Princeton University , , , ,","place":["Princeton, New Jersey, United States, 08544"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chase M.","family":"Kayrouz","sequence":"additional","affiliation":[{"name":"Princeton University , , , ,","place":["Princeton, New Jersey, United States, 08544"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0986-5142","authenticated-orcid":true,"given":"Kendra A.","family":"Ireland","sequence":"additional","affiliation":[{"name":"Emory University , , , ,","place":["Atlanta, Georgia, United States, 30322"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0258-8907","authenticated-orcid":true,"given":"Katherine M.","family":"Davis","sequence":"additional","affiliation":[{"name":"Emory University , , , ,","place":["Atlanta, Georgia, United States, 30322"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2707-4854","authenticated-orcid":true,"given":"Mohammad R.","family":"Seyedsayamdost","sequence":"additional","affiliation":[{"name":"Princeton University , , , ,","place":["Princeton, New Jersey, United States, 08544"]},{"name":"Princeton University , , , ,","place":["Princeton, New Jersey, United States, 08544"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,1]]},"reference":[{"key":"2026101414074151100_cit1","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1007\/s10295-013-1389-9","article-title":"Microbial genome mining for accelerated\nnatural products discovery: is a renaissance in the making?","volume":"41","author":"Bachmann","year":"2014","journal-title":"J. Ind. Microbiol. Biotechnol."},{"key":"2026101414074151100_cit2","doi-asserted-by":"publisher","first-page":"573","DOI":"10.1007\/s10295-016-1815-x","article-title":"Gifted\nmicrobes for genome mining and natural product discovery","volume":"44","author":"Baltz","year":"2017","journal-title":"J. Ind. Microbiol. Biotechnol."},{"key":"2026101414074151100_cit3","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1038\/nchem.269","article-title":"Natural products: Beyond\ngrind and find","volume":"1","author":"Miller","year":"2009","journal-title":"Nat. Chem."},{"key":"2026101414074151100_cit4","doi-asserted-by":"publisher","first-page":"18134","DOI":"10.1074\/jbc.C110.106377","article-title":"Identification\nof a novel selenium-containing compound,\nselenoneine, as the predominant chemical form of organic selenium\nin the blood of bluefin tuna","volume":"285","author":"Yamashita","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2026101414074151100_cit5","doi-asserted-by":"publisher","first-page":"821","DOI":"10.1021\/acschembio.6b00031","article-title":"Why Nature Chose\nSelenium","volume":"11","author":"Reich","year":"2016","journal-title":"ACS\nChem. Biol."},{"key":"2026101414074151100_cit6","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1038\/s41586-022-05174-2","article-title":"Biosynthesis\nof Selenium-Containing Small Molecules in Diverse Microorganisms","volume":"610","author":"Kayrouz","year":"2022","journal-title":"Nature"},{"key":"2026101414074151100_cit7","doi-asserted-by":"publisher","first-page":"1868","DOI":"10.1038\/s41557-024-01600-2","article-title":"Discovery\nof the Selenium-Containing Antioxidant Ovoselenol Derived from Convergent\nEvolution","volume":"16","author":"Kayrouz","year":"2024","journal-title":"Nat. Chem."},{"key":"2026101414074151100_cit8","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2024.102495","article-title":"Enzymatic\nstrategies for selenium incorporation into\nbiological molecules","volume":"81","author":"Kayrouz","year":"2024","journal-title":"Curr. Opin. Chem. Biol."},{"key":"2026101414074151100_cit9","doi-asserted-by":"publisher","first-page":"475","DOI":"10.1128\/jb.103.2.475-478.1970","article-title":"Biosynthesis\nof ergothioneine and hercynine by fungi and Actinomycetales","volume":"103","author":"Genghof","year":"1970","journal-title":"J. Bacteriol."},{"key":"2026101414074151100_cit10","doi-asserted-by":"publisher","first-page":"6632","DOI":"10.1021\/ja101721e","article-title":"In Vitro\nReconstitution of Mycobacterial Ergothioneine Biosynthesis","volume":"132","author":"Seebeck","year":"2010","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414074151100_cit11","doi-asserted-by":"publisher","first-page":"3309","DOI":"10.1021\/acs.biochem.8b00239","article-title":"Mini-Review: Ergothioneine and Ovothiol\nBiosyntheses, an Unprecedented Trans-Sulfur Strategy in Natural Product\nBiosynthesis","volume":"57","author":"Naowarojna","year":"2018","journal-title":"Biochemistry"},{"key":"2026101414074151100_cit12","doi-asserted-by":"publisher","first-page":"6906","DOI":"10.1021\/jacs.8b12596","article-title":"Structural\nand Mechanistic Basis for Anaerobic Ergothioneine Biosynthesis","volume":"141","author":"Leisinger","year":"2019","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414074151100_cit13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.sbi.2020.04.002","article-title":"Structural basis of ergothioneine\nbiosynthesis","volume":"65","author":"Stampfli","year":"2020","journal-title":"Curr. Opin. Struct. Biol."},{"key":"2026101414074151100_cit14","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.cbpa.2020.06.007","article-title":"The catalytic mechanism\nof sulfoxide synthases","volume":"59","author":"Stampfli","year":"2020","journal-title":"Curr. Opin. Chem. Biol."},{"key":"2026101414074151100_cit15","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1016\/j.bbadis.2011.09.017","article-title":"Ergothioneine;\nAntioxidant Potential, Physiological\nFunction and Role in Disease","volume":"1822","author":"Cheah","year":"2012","journal-title":"Biochim. Biophys.\nActa, Mol. Basis Dis."},{"key":"2026101414074151100_cit16","doi-asserted-by":"publisher","first-page":"144","DOI":"10.4331\/wjbc.v1.i5.144","article-title":"Discovery\nof the Strong Antioxidant Selenoneine in Tuna and Selenium Redox Metabolism","volume":"1","author":"Yamashita","year":"2010","journal-title":"World J. Biol. Chem."},{"key":"2026101414074151100_cit17","doi-asserted-by":"publisher","first-page":"4526","DOI":"10.1016\/j.cell.2022.10.008","article-title":"A Microbial Transporter of the Dietary\nAntioxidant Ergothioneine","volume":"185","author":"Dumitrescu","year":"2022","journal-title":"Cell"},{"key":"2026101414074151100_cit18","doi-asserted-by":"publisher","first-page":"5256","DOI":"10.1073\/pnas.0408624102","article-title":"Discovery\nof the ergothioneine transporter","volume":"102","author":"Gru\u0308ndemann","year":"2005","journal-title":"Proc. Natl.\nAcad. Sci. U.S.A."},{"key":"2026101414074151100_cit19","doi-asserted-by":"publisher","first-page":"5275","DOI":"10.1021\/jacs.8b13023","article-title":"An Alternative Active Site Architecture for O2 Activation in the Ergothioneine Biosynthetic EgtB from Chloracidobacterium thermophilum","volume":"141","author":"Stampfli","year":"2019","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414074151100_cit20","doi-asserted-by":"publisher","DOI":"10.1155\/2019\/9593467","article-title":"A Computational\nInvestigation of\nthe Binding of the Selenium Analogues of Ergothioneine and Ovothiol\nto Cu(I) and Cu(II) and the Effect of Binding on the Redox Potential\nof the Cu(II)\/Cu(I) Redox Couple","volume":"2019","author":"Wiebe","year":"2019","journal-title":"J. Chem."},{"key":"2026101414074151100_cit21","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202400439","article-title":"5-Thiohistidine\nN-Acetyltransferase from Proteiniphilum Saccharofermentans","volume":"26","author":"Liao","year":"2025","journal-title":"ChemBioChem"},{"key":"2026101414074151100_cit22","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/j.bioorg.2011.08.004","article-title":"The ATP-Grasp Enzymes","volume":"39","author":"Fawaz","year":"2011","journal-title":"Bioorg. Chem."},{"key":"2026101414074151100_cit23","doi-asserted-by":"publisher","first-page":"2022","DOI":"10.1111\/j.1742-4658.2012.08588.x","article-title":"The major facilitator superfamily\n(MFS) revisited","volume":"279","author":"Reddy","year":"2012","journal-title":"FEBS J."},{"key":"2026101414074151100_cit24","doi-asserted-by":"publisher","first-page":"1757","DOI":"10.1021\/ja109378e","article-title":"Identification and\nCharacterization of the First Ovothiol\nBiosynthetic Enzyme","volume":"133","author":"Braunshausen","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414074151100_cit25","doi-asserted-by":"publisher","first-page":"15417","DOI":"10.1021\/acscatal.3c04026","article-title":"Biochemical and Structural Characterization of OvoATh2: A Mononuclear\nNonheme Iron Enzyme from Hydrogenimonas Thermophila for Ovothiol Biosynthesis","volume":"13","author":"Wang","year":"2023","journal-title":"ACS Catal."},{"key":"2026101414074151100_cit26","doi-asserted-by":"publisher","first-page":"2013","DOI":"10.1016\/j.str.2024.08.006","article-title":"Structural Insights into the Convergent\nEvolution of\nSulfoxide Synthase EgtB-IV, an Ergothioneine-Biosynthetic Homolog\nof Ovothiol Synthase OvoA","volume":"32","author":"Ireland","year":"2024","journal-title":"Structure"},{"key":"2026101414074151100_cit27","doi-asserted-by":"publisher","first-page":"2821","DOI":"10.1002\/anie.201410045","article-title":"Structure of the Sulfoxide Synthase\nEgtB from the Ergothioneine Biosynthetic Pathway","volume":"54","author":"Goncharenko","year":"2015","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026101414074151100_cit28","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2023.128428","article-title":"Structural\nInsights into a Novel Nonheme Iron-Dependent Oxygenase in Selenoneine\nBiosynthesis","volume":"256","author":"Liu","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"2026101414074151100_cit29","doi-asserted-by":"publisher","first-page":"4604","DOI":"10.1021\/jacs.7b13628","article-title":"Use of a Tyrosine Analogue\nto Modulate the Two Activities of a Nonheme Iron Enzyme OvoA in Ovothiol\nBiosynthesis, Cysteine Oxidation versus Oxidative C-S Bond Formation","volume":"140","author":"Chen","year":"2018","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414074151100_cit30","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1021\/acscatal.8b03903","article-title":"Mechanistic\nStudies of a Nonheme Iron Enzyme OvoA in Ovothiol Biosynthesis Using\na Tyrosine Analogue, 2-Amino-3-(4-Hydroxy-3-(Methoxyl) Phenyl) Propanoic\nAcid (MeOTyr)","volume":"9","author":"Chen","year":"2019","journal-title":"ACS Catal."},{"key":"2026101414074151100_cit31","doi-asserted-by":"publisher","first-page":"3589","DOI":"10.1039\/D1SC05479A","article-title":"OvoAMtht from\nMethyloversatilis Thermotolerans\nOvothiol Biosynthesis Is a Bifunction Enzyme: Thiol Oxygenase and\nSulfoxide Synthase Activities","volume":"13","author":"Cheng","year":"2022","journal-title":"Chem. Sci."},{"key":"2026101414074151100_cit32","doi-asserted-by":"publisher","first-page":"1945","DOI":"10.1039\/C5CC07772A","article-title":"Conversion of a\nNon-Heme Iron-Dependent Sulfoxide Synthase\ninto a Thiol Dioxygenase by a Single Point Mutation","volume":"52","author":"Goncharenko","year":"2016","journal-title":"Chem. Commun."},{"key":"2026101414074151100_cit33","doi-asserted-by":"publisher","first-page":"4854","DOI":"10.1021\/ol402275t","article-title":"Regioselectivity of\nthe Oxidative C-S Bond Formation in Ergothioneine and Ovothiol Biosyntheses","volume":"15","author":"Song","year":"2013","journal-title":"Org. Lett."},{"key":"2026101414074151100_cit34","doi-asserted-by":"publisher","first-page":"1847","DOI":"10.1002\/prot.24311","article-title":"Structure\nand Function of Escherichia Coli RimK,\nan ATP-Grasp Fold, l-Glutamyl Ligase Enzyme","volume":"81","author":"Zhao","year":"2013","journal-title":"Proteins: Struct., Funct., Bioinf."},{"key":"2026101414074151100_cit35","doi-asserted-by":"publisher","first-page":"376","DOI":"10.1002\/prot.20103","article-title":"YbdK Is a Carboxylate-Amine\nLigase\nwith a \u03b3-Glutamyl: Cysteine Ligase Activity: Crystal Structure\nand Enzymatic Assays","volume":"56","author":"Lehmann","year":"2004","journal-title":"Proteins: Struct., Funct.,\nGenet."},{"key":"2026101414074151100_cit36","doi-asserted-by":"publisher","DOI":"10.1002\/psc.3339","article-title":"Ergothioneine in a peptide: Substitution\nof histidine\nwith 2-thiohistidine in bioactive peptides","volume":"27","author":"Jenny","year":"2021","journal-title":"J. Pept. Sci."},{"key":"2026101414074151100_cit37","doi-asserted-by":"publisher","first-page":"3309","DOI":"10.1021\/acs.biochem.8b00239","article-title":"Mini-review:\nErgothioneine and Ovothiol\nBiosyntheses, an Unprecedented Trans-Sulfur Strategy in Natural Product\nBiosynthesis","volume":"57","author":"Naowarojna","year":"2018","journal-title":"Biochemistry"},{"key":"2026101414074151100_cit38","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202304989","article-title":"Structure-Function\nAnalysis of the S-Glycosylation Reaction in the Biosynthesis of Lincosamide\nAntibiotics","volume":"62","author":"Mori","year":"2023","journal-title":"Angew. Chem., Int. Ed."}],"container-title":["Journal of the American\nChemical Society"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacs.6c03714","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/40\/42939\/68731387\/jacs.6c03714.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/40\/42939\/68731387\/jacs.6c03714.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:17:58Z","timestamp":1792001878000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jacsat\/article\/148\/40\/42939\/5443653\/Widespread-Biosynthesis-of-Thiohistidine"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,1]]},"references-count":38,"journal-issue":{"issue":"40","published-online":{"date-parts":[[2026,10,1]]},"published-print":{"date-parts":[[2026,10,14]]}},"URL":"https:\/\/doi.org\/10.1021\/jacs.6c03714","relation":{},"ISSN":["0002-7863","1520-5126"],"issn-type":[{"value":"0002-7863","type":"print"},{"value":"1520-5126","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,1]]}}}