{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T02:38:56Z","timestamp":1792463936606,"version":"4.5.0"},"reference-count":47,"publisher":"American Chemical Society (ACS)","issue":"34","license":[{"start":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T00:00:00Z","timestamp":1787443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["DI 1536\/13-1"],"award-info":[{"award-number":["DI 1536\/13-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["GR 1861\/13-1"],"award-info":[{"award-number":["GR 1861\/13-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Type I terpene synthases generate complex polycyclic scaffolds through carbocation cascades. However, how closely related enzymes convert a common C25 precursor to distinct sesterterpene frameworks remains unresolved. Here, we combine high-resolution crystal structures with systematic mutagenesis of four bacterial sesterterpene synthases to define the structural basis for pathway divergence. These analyses show how local active-site interactions within a conserved fold redirect carbocation trajectories and thereby control product formation. The structures reveal a preorganized binding mode that positions the substrate in a product-like conformation and directs the cyclization cascade. Comparative structural analyses further support distinct carbocation trajectories involving either centralized cation hubs or sequential rearrangement pathways with the exact pathway being enzyme-dependent. Structure-guided mutagenesis targets these features, alters product profiles, and enables the formation of new terpene scaffolds. Together, crystallographic data and mutagenesis demonstrate that closely related enzymes with the same overall fold can follow distinct carbocation trajectories and reveal how local architectural changes control the cyclization outcome.<\/jats:p>","DOI":"10.1021\/jacs.6c06677","type":"journal-article","created":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T12:25:02Z","timestamp":1787919902000},"page":"36648-36661","source":"Crossref","is-referenced-by-count":1,"title":["Local Active-Site Architecture\nDirects Divergent Carbocation\nCascades in Sesterterpene Synthases"],"prefix":"10.1021","volume":"148","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2584-896X","authenticated-orcid":true,"given":"Philip","family":"Troycke","sequence":"first","affiliation":[{"name":"Technical University Munich , , ,","place":["Garching, Germany, 85748"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6406-7269","authenticated-orcid":true,"given":"Heng","family":"Li","sequence":"additional","affiliation":[{"name":"Rheinische Friedrich-Wilhelms-University Bonn , , Gerhard-Domagk-Stra\u00dfe 1 , ,","place":["Bonn, Germany, 53121"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kexin","family":"Yang","sequence":"additional","affiliation":[{"name":"Rheinische Friedrich-Wilhelms-University Bonn , , Gerhard-Domagk-Stra\u00dfe 1 , ,","place":["Bonn, Germany, 53121"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0102-0631","authenticated-orcid":true,"given":"Jeroen S.","family":"Dickschat","sequence":"additional","affiliation":[{"name":"Rheinische Friedrich-Wilhelms-University Bonn , , Gerhard-Domagk-Stra\u00dfe 1 , ,","place":["Bonn, Germany, 53121"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1660-340X","authenticated-orcid":true,"given":"Michael","family":"Groll","sequence":"additional","affiliation":[{"name":"Technical University Munich , , ,","place":["Garching, Germany, 85748"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,23]]},"reference":[{"key":"2026090312105775600_cit1","doi-asserted-by":"publisher","first-page":"10040","DOI":"10.1002\/anie.201702363","article-title":"Importance\nof Inherent Substrate Reactivity in Enzyme-Promoted Carbocation Cyclization\/Rearrangements","volume":"56","author":"Tantillo","year":"2017","journal-title":"Angew. 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Biophys."},{"key":"2026090312105775600_cit8","doi-asserted-by":"publisher","first-page":"7652","DOI":"10.1002\/anie.201403648","article-title":"Induced Fit Mechanism in Class I\nTerpene Cyclases","volume":"53","author":"Baer","year":"2014","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit9","doi-asserted-by":"publisher","first-page":"13543","DOI":"10.1073\/pnas.231313098","article-title":"Structure\nof trichodiene synthase\nfrom Fusarium sporotrichioides provides mechanistic\ninferences on the terpene cyclization cascade","volume":"98","author":"Rynkiewicz","year":"2001","journal-title":"Proc. Natl. Acad. Sci. 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Soc."},{"key":"2026090312105775600_cit30","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202209785","article-title":"Crystal Structure Based\nMutagenesis of Cattleyene Synthase Leads to the Generation of Rearranged\nPolycyclic Diterpenes","volume":"61","author":"Xing","year":"2022","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit31","doi-asserted-by":"publisher","first-page":"664","DOI":"10.1038\/s41589-022-01026-2","article-title":"Ancient\nplant-like\nterpene biosynthesis in corals","volume":"18","author":"Burkhardt","year":"2022","journal-title":"Nat. Chem. Biol."},{"key":"2026090312105775600_cit32","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2408064121","article-title":"Engineering\nsubstrate channeling in a bifunctional terpene synthase","volume":"121","author":"Wenger","year":"2024","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090312105775600_cit33","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-39706-9","article-title":"Molecular insights into the catalytic\npromiscuity of a bacterial diterpene synthase","volume":"14","author":"Li","year":"2023","journal-title":"Nat. Commun."},{"key":"2026090312105775600_cit34","doi-asserted-by":"publisher","first-page":"6209","DOI":"10.1021\/acscatal.5c01540","article-title":"A Hydrophobic Tunnel and a Gatekeeper\nat the Active Site Entrance in Two Type I Terpene Synthases","volume":"15","author":"Li","year":"2025","journal-title":"ACS Catal."},{"key":"2026090312105775600_cit35","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-60537-3","article-title":"Structure of bifunctional variediene\nsynthase yields\nunique insight on biosynthetic diterpene assembly and cyclization","volume":"16","author":"Wenger","year":"2025","journal-title":"Nat. Commun."},{"key":"2026090312105775600_cit36","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202405140","article-title":"Structural\nInsights into the Terpene\nCyclization Domains of Two Fungal Sesterterpene Synthases and Enzymatic\nEngineering for Sesterterpene Diversification","volume":"63","author":"Xu","year":"2024","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit37","doi-asserted-by":"publisher","first-page":"6981","DOI":"10.1021\/jacs.5c17651","article-title":"Structural Insights into Three Bifunctional Sesterterpene Synthases\nand Product Profile Investigation by Domain Swapping and Active Site\nMutation","volume":"148","author":"Lei","year":"2026","journal-title":"J. Am. Chem. Soc."},{"key":"2026090312105775600_cit38","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202215688","article-title":"Mechanistic\nCharacterisation and\nEngineering of Sesterviolene Synthase from Streptomyces violens","volume":"62","author":"Gu","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit39","doi-asserted-by":"publisher","first-page":"1386","DOI":"10.3762\/bjoc.19.100","article-title":"Functional Characterisation of Twelve\nTerpene Synthases from Actinobacteria","volume":"19","author":"Chhalodia","year":"2023","journal-title":"Beilstein\nJ. Org. Chem."},{"key":"2026090312105775600_cit40","doi-asserted-by":"publisher","first-page":"19961","DOI":"10.1002\/anie.202010084","article-title":"The Biosynthetic\nGene Cluster for Sestermobaraenes\n\u2013 Discovery of a Geranylfarnesyl Diphosphate Synthase and a\nMultiproduct Sesterterpene Synthase from Streptomyces mobaraensis","volume":"59","author":"Hou","year":"2020","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit41","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202313789","article-title":"Subrutilane\n\u2013 A Hexacyclic Sesterterpene from Streptomyces subrutilus","volume":"62","author":"Gu","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit42","doi-asserted-by":"publisher","first-page":"2441","DOI":"10.3762\/bjoc.17.161","article-title":"Targeting Active Site Residues and Structural Anchoring\nPositions in Terpene Synthases","volume":"17","author":"Hou","year":"2021","journal-title":"Beilstein J.\nOrg. Chem."},{"key":"2026090312105775600_cit43","doi-asserted-by":"publisher","first-page":"6696","DOI":"10.1021\/acs.orglett.7b03418","article-title":"Focused Genome\nMining of Structurally Related Sesterterpenes: Enzymatic Formation\nof Enantiomeric and Diastereomeric Products","volume":"19","author":"Narita","year":"2017","journal-title":"Org. Lett."},{"key":"2026090312105775600_cit44","doi-asserted-by":"publisher","first-page":"10053","DOI":"10.1002\/chem.201702766","article-title":"Mechanistic\nCharacterization of two Chimeric Sesterterpene Synthases from Penicillium","volume":"23","author":"Mitsuhashi","year":"2017","journal-title":"Chem. \u2013 Eur. J."},{"key":"2026090312105775600_cit45","doi-asserted-by":"publisher","first-page":"1658","DOI":"10.1002\/anie.201509263","article-title":"An Unusual Chimeric\nDiterpene Synthase from Emericella variecolor and\nIts Functional Conversion into a Sesterterpene Synthase by Domain\nSwapping","volume":"55","author":"Qin","year":"2016","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090312105775600_cit46","doi-asserted-by":"publisher","first-page":"25468","DOI":"10.1002\/anie.202110842","article-title":"A Cryptic Plant Terpene Cyclase Producing\nUnconventional 18- and\n14-Membered Macrocyclic C25 and C20 Terpenoids\nwith Immunosuppressive Activity","volume":"60","author":"Chen","year":"2021","journal-title":"Angew. Chem.,\nInt. Ed."},{"key":"2026090312105775600_cit47","doi-asserted-by":"publisher","first-page":"12723","DOI":"10.1021\/acscatal.3c03740","article-title":"Turning\na Sesquiterpene Synthase into a Di- and Sesterterpene\nSynthase","volume":"13","author":"Xu","year":"2023","journal-title":"ACS Catal."}],"container-title":["Journal of the American\nChemical Society"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacs.6c06677","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/34\/36648\/67243956\/jacs.6c06677.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/34\/36648\/67243956\/jacs.6c06677.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T16:20:35Z","timestamp":1788452435000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jacsat\/article\/148\/34\/36648\/5312811\/Local-Active-Site-Architecture-Directs-Divergent"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,23]]},"references-count":47,"journal-issue":{"issue":"34","published-online":{"date-parts":[[2026,8,23]]},"published-print":{"date-parts":[[2026,9,2]]}},"URL":"https:\/\/doi.org\/10.1021\/jacs.6c06677","relation":{},"ISSN":["0002-7863","1520-5126"],"issn-type":[{"value":"0002-7863","type":"print"},{"value":"1520-5126","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,23]]}}}