{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T22:37:44Z","timestamp":1792795064340,"version":"4.5.0"},"reference-count":105,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T00:00:00Z","timestamp":1744761600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009057","name":"Karl-Franzens-Universit\u00e4t Graz","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004955","name":"\u00d6sterreichische Forschungsf\u00f6rderungsgesellschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004955","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemSusChem"],"published-print":{"date-parts":[[2025,6,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Vanillin is one of the most important aroma compounds, naturally occurring in vanilla pods. Many routes to access natural vanillin from various renewables have been investigated, including a natural five\u2010step microbial transformation of eugenol to vanillin. Readily available eugenol was also the starting material for a chemical two\u2010step sequence to vanillin employed in the 19\n                    <jats:sup>th<\/jats:sup>\n                    century. Here we show that a two\u2010step sequence can also be realized using biocatalysts only and run it in one\u2010pot simultaneously. This was achieved by isomerizing the C=C double bond of eugenol by oxidation to coniferyl alcohol followed by oxidative C=C cleavage catalyzed by newly identified enzymes. Thus, two oxidative steps catalyzed by two different biocatalysts \u2010 one containing flavin and the other a non\u2010heme iron(II) cofactor \u2010 were successfully run simultaneously just requiring molecular oxygen as oxidant for each step. Using natural eugenol sources, e.\u2009g. clove oil, vanillin was obtained with 91\u2009% product formation. This study shows that natural pathways like the microbial transformation of eugenol to vanillin involving five steps can be shortened, hereto just two simultaneous steps, by exploiting and combining the repertoire of promiscuous enzymatic activities present in different organisms leading to new\u2010to\u2010nature cascades.\n                  <\/jats:p>","DOI":"10.1002\/cssc.202500387","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T04:23:56Z","timestamp":1742185436000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Transforming a Historical Chemical Synthetic Route for Vanillin Starting from Renewable Eugenol to a Cell\u2010Free Bi\u2010Enzymatic Cascade"],"prefix":"10.1002","volume":"18","author":[{"given":"Elisa","family":"Lanfranchi","sequence":"first","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valerio","family":"Ferrario","sequence":"additional","affiliation":[{"name":"Group Research BASF SE Carl-Bosch-Strasse 38  67056 Ludwigshafen am Rhein Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Somayyeh","family":"Gandomkar","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan E.","family":"Payer","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erna","family":"Zukic","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haris","family":"Rudalija","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Musi","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ines","family":"Gaberscek","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuliya","family":"Orel","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doreen","family":"Schachtschabel","sequence":"additional","affiliation":[{"name":"Group Research BASF SE Carl-Bosch-Strasse 38  67056 Ludwigshafen am Rhein Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Willrodt","sequence":"additional","affiliation":[{"name":"Group Research BASF SE Carl-Bosch-Strasse 38  67056 Ludwigshafen am Rhein Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Breuer","sequence":"additional","affiliation":[{"name":"Group Research BASF SE Carl-Bosch-Strasse 38  67056 Ludwigshafen am Rhein Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2151-6394","authenticated-orcid":false,"given":"Wolfgang","family":"Kroutil","sequence":"additional","affiliation":[{"name":"Austrian Center of Industrial Biotechnology (ACIB GmbH) c\/o Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"Group Research BASF SE Carl-Bosch-Strasse 38  67056 Ludwigshafen am Rhein Germany"},{"name":"Department of Chemistry University of Graz  Heinrichstra\u00dfe 28 8010 Graz Austria"},{"name":"BioTechMed Graz  8010 Graz Austria"},{"name":"Field of Excellence BioHealth University of Graz  8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,4,16]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2024.04.004"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.rser.2023.113905"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0031-9422(03)00149-3"},{"key":"e_1_2_8_4_1","unstructured":"Herzog von Sachsen Coburg und Gotha: Privileg Archiv Symrise Holzminden 1874."},{"key":"e_1_2_8_5_1","unstructured":"Kaiserliches Patentamt: Patentschrift Nr. DE27992\u2005C. Haarmann & Reimer in Holzminden Germany 1884."},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/cber.187400701193"},{"key":"e_1_2_8_7_1","unstructured":"B.\u2005B. Kuhse Vanillin \u2013 Historie und Schulrelevanz: Die Geschichte einer regionalen Riechstoffindustrie und deren Verwendung in einem praxisorientierten Chemieunterricht Cuvillier Verlag G\u00f6ttingen 2010."},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed054p776"},{"key":"e_1_2_8_9_1","unstructured":"Kaiserliches Patentamt: Patentschrift Nr.\u200593938. Haarmann & Reimer in Holzminden Verfahren zur Darstellung von Vanillin aus i-Eugenol order Eugenol Germany 1897."},{"key":"e_1_2_8_10_1","unstructured":"Kaiseliches Patentamt: Patenschrift Nr.\u200557568. Haarmann & Reimer in Holzminden Verfahren zur Darstellung von monomolecularen S\u00e4urederivative des Isoeugenol Germany 1891."},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed074p1055"},{"key":"e_1_2_8_12_1","first-page":"40","volume":"8","author":"Gallage N. J.","year":"2015","journal-title":"Mol. Plant Pathol."},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1039\/D1GC04416H"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssuschemeng.5b01344"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.indcrop.2023.117372"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.procbio.2024.12.012"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom14111413"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.3390\/biochem4040017"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-023-02144-9"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.apcata.2019.117117"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13399-019-00418-0"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-013-5303-1"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-6-13"},{"key":"e_1_2_8_24_1","unstructured":"M. Gelo-Pujic A. Amory (Rhodia Operations) WO2016001203 2016."},{"key":"e_1_2_8_25_1","unstructured":"G. Kindel J.-M. Hilmer E.-E. Gross (Symrise) EP2772142A1 2014."},{"key":"e_1_2_8_26_1","unstructured":"A. Muheim B. M\u00fcller T. M\u00fcnch M. Wetli (Givaudan SA) EP0885968B1 2006."},{"key":"e_1_2_8_27_1","unstructured":"T. Zougmore R. Djillali L. Garel (Rhodia Operations) WO2022194771\u2005A1 2022."},{"key":"e_1_2_8_28_1","unstructured":"HSF Biotech. can be found under https:\/\/de.hsfbiotech.com\/natenfla-natural-flavors\/natural-vanillin-powder.html (accessed: 19.\u200902.\u20092025)."},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ces.2022.118049"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep09993"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-9-84"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002030050772"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1080\/14786419.2022.2066101"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.3c08723"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.8.2595-2607.1997"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002530051598"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.69.11.6569-6576.2003"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2019.08.007"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.apcata.2020.117934"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2024.06.012"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.2c00168"},{"key":"e_1_2_8_42_1","author":"De Vitis V.","year":"2023","journal-title":"ChemBioChem"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssuschemeng.2c06534"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-020-10433-1"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b08177"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbt.2015.03.002"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201402215"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00233-24"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-34912-3"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.202300868"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202402546"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202316760"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202314566"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1039\/D3GC04191C"},{"key":"e_1_2_8_55_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202301178"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202300906"},{"key":"e_1_2_8_57_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07506-w"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacsau.4c00026"},{"key":"e_1_2_8_59_1","doi-asserted-by":"publisher","DOI":"10.1002\/ejoc.202301236"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202217713"},{"key":"e_1_2_8_61_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202309012"},{"key":"e_1_2_8_62_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202300334"},{"key":"e_1_2_8_63_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.3c00555"},{"key":"e_1_2_8_64_1","volume":"38","author":"See W. W. L.","year":"2022","journal-title":"Curr. Opin. Green Sustain. Chem."},{"key":"e_1_2_8_65_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41557-022-00931-2"},{"key":"e_1_2_8_66_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201703353"},{"key":"e_1_2_8_67_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.7b00033"},{"key":"e_1_2_8_68_1","doi-asserted-by":"publisher","DOI":"10.1039\/C4CC08752F"},{"key":"e_1_2_8_69_1","doi-asserted-by":"publisher","DOI":"10.1039\/D3CS00595J"},{"key":"e_1_2_8_70_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M809060200"},{"key":"e_1_2_8_71_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0510144103"},{"key":"e_1_2_8_72_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-313X.2008.03412.x"},{"key":"e_1_2_8_73_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000993"},{"key":"e_1_2_8_74_1","doi-asserted-by":"crossref","unstructured":"T.\u2005A. Ewing G. Gygli M.\u2005W. Fraaije W.\u2005J.\u2005H. van\u2005Berkel inThe Enzymes Vol. 47(Eds.: P. Chaiyen F. Tamanoi) Academic Press 2020 pp. 87\u2013116.","DOI":"10.1016\/bs.enz.2020.05.003"},{"key":"e_1_2_8_75_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.3c01418"},{"key":"e_1_2_8_76_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41929-020-00556-z"},{"key":"e_1_2_8_77_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pisc.2016.04.106"},{"key":"e_1_2_8_78_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1235"},{"key":"e_1_2_8_79_1","doi-asserted-by":"crossref","unstructured":"C. Martin C. Binda M.\u2005W. Fraaije A. Mattevi inThe Enzymes Vol. 47(Eds.: P. Chaiyen F. Tamanoi) Academic Press 2020 pp. 63\u201386.","DOI":"10.1016\/bs.enz.2020.05.002"},{"key":"e_1_2_8_80_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201600148"},{"key":"e_1_2_8_81_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2007.05767.x"},{"key":"e_1_2_8_82_1","first-page":"1","volume":"168","author":"Zhou H.","year":"2021","journal-title":"Biochem. Eng. J."},{"key":"e_1_2_8_83_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-016-0472-0"},{"key":"e_1_2_8_84_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.202304172"},{"key":"e_1_2_8_85_1","doi-asserted-by":"publisher","DOI":"10.1002\/ffj.3739"},{"key":"e_1_2_8_86_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.procbio.2018.05.013"},{"key":"e_1_2_8_87_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-007-9602-4"},{"key":"e_1_2_8_88_1","doi-asserted-by":"publisher","DOI":"10.3390\/catal11101199"},{"key":"e_1_2_8_89_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12010-020-03478-5"},{"key":"e_1_2_8_90_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.120715"},{"key":"e_1_2_8_91_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiosc.2020.04.012"},{"key":"e_1_2_8_92_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12858-018-0098-4"},{"key":"e_1_2_8_93_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(05)80360-2"},{"key":"e_1_2_8_94_1","unstructured":"The UniProt Consortium Nucleic Acids Res.2023 51 D523-D531."},{"key":"e_1_2_8_95_1","volume":"24","author":"Yeung W.","year":"2023","journal-title":"Brief. Bioinform."},{"key":"e_1_2_8_96_1","doi-asserted-by":"publisher","DOI":"10.1002\/cite.202200176"},{"key":"e_1_2_8_97_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25034-3"},{"key":"e_1_2_8_98_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.oprd.5b00278"},{"key":"e_1_2_8_99_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1818"},{"key":"e_1_2_8_100_1","doi-asserted-by":"publisher","DOI":"10.1214\/aoms\/1177699147"},{"key":"e_1_2_8_101_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(05)80360-2"},{"key":"e_1_2_8_102_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg500"},{"key":"e_1_2_8_103_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btm226"},{"key":"e_1_2_8_104_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetlet.2023.154665"},{"key":"e_1_2_8_105_1","doi-asserted-by":"publisher","DOI":"10.1002\/ejoc.202201112"}],"container-title":["ChemSusChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cssc.202500387","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T17:37:59Z","timestamp":1761154679000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cssc.202500387"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,16]]},"references-count":105,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2025,6,2]]}},"alternative-id":["10.1002\/cssc.202500387"],"URL":"https:\/\/doi.org\/10.1002\/cssc.202500387","archive":["Portico"],"relation":{},"ISSN":["1864-5631","1864-564X"],"issn-type":[{"value":"1864-5631","type":"print"},{"value":"1864-564X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,16]]},"assertion":[{"value":"2025-02-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-16","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202500387"}}