{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T17:03:32Z","timestamp":1783962212092,"version":"3.55.0"},"reference-count":42,"publisher":"Wiley","issue":"16","license":[{"start":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:00:00Z","timestamp":1690243200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100018774","name":"Bundesministerium f\u00fcr Klimaschutz, Umwelt, Energie, Mobilit\u00e4t, Innovation und Technologie","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100018774","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012416","name":"Bundesministerium f\u00fcr Digitalisierung und Wirtschaftsstandort","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100012416","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007945","name":"Steirische Wirtschaftsf\u00f6rderungsgesellschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007945","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011035","name":"Standortagentur Tirol","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100011035","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015053","name":"Amt der N\u00d6 Landesregierung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2023,8,15]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Biocatalytic decarboxylation of hydroxycinnamic acids yields phenolic styrenes, which are important precursors for antioxidants, epoxy coatings, adhesives and other polymeric materials. <jats:italic>Bacillus subtilis<\/jats:italic> decarboxylase (BsPAD) is a cofactor\u2010independent enzyme that catalyzes the cleavage of carbon dioxide from <jats:italic>p<\/jats:italic>\u2010coumaric\u2010, caffeic\u2010, and ferulic acid with high catalytic efficiency. Real\u2010time spectroscopic assays for decarboxylase reactions remove the necessity of extensive sample workup, which is required for HPLC, mass spectrometry, gas chromatography, or NMR methods. This work presents two robust and sensitive assays based on photometry and fluorimetry that allow decarboxylation reactions to be followed with high sensitivity while avoiding product extraction and long analysis times. Optimized assay procedures were used to measure BsPAD activity in cell lysates and to determine the kinetic constants (<jats:italic>K<\/jats:italic><jats:sub>M<\/jats:sub> and <jats:italic>V<\/jats:italic><jats:sub>max<\/jats:sub>) of the purified enzyme for <jats:italic>p<\/jats:italic>\u2010coumaric\u2010, caffeic\u2010 and ferulic acid. Substrate inhibition was shown for caffeic acid.<\/jats:p>","DOI":"10.1002\/cbic.202300207","type":"journal-article","created":{"date-parts":[[2023,5,16]],"date-time":"2023-05-16T10:05:07Z","timestamp":1684231507000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Spectrophotometric and Fluorimetric High\u2010Throughput Assays for Phenolic Acid Decarboxylase"],"prefix":"10.1002","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9625-8177","authenticated-orcid":false,"given":"Henrik","family":"Terholsen","sequence":"first","affiliation":[{"name":"Institute of Biochemistry Department of Biotechnology and Enzyme Catalysis University of Greifswald  Felix-Hausdorff-Stra\u00dfe 4 17487 Greifswald Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5283-8436","authenticated-orcid":false,"given":"Kamela","family":"Myrtollari","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biotechnology Graz University of Technology  Petersgasse 14 8010 Graz Austria"},{"name":"Henkel AG &amp; Co. KGaA Adhesive Research  Henkelstra\u00dfe 67 40191 D\u00fcsseldorf Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirna","family":"Larva","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biotechnology Graz University of Technology  Petersgasse 14 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8286-1589","authenticated-orcid":false,"given":"Christina","family":"M\u00f6ller","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry Department of Biotechnology and Enzyme Catalysis University of Greifswald  Felix-Hausdorff-Stra\u00dfe 4 17487 Greifswald Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3394-7503","authenticated-orcid":false,"given":"Andreas","family":"Taden","sequence":"additional","affiliation":[{"name":"Henkel AG &amp; Co. KGaA Adhesive Research  Henkelstra\u00dfe 67 40191 D\u00fcsseldorf Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2853-3525","authenticated-orcid":false,"given":"Robert","family":"Kourist","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biotechnology Graz University of Technology  Petersgasse 14 8010 Graz Austria"},{"name":"acib \u2013 Austrian Centre of Industrial Biotechnology  Petersgasse 14 8010 Graz Austria"},{"name":"BioTechMed-Graz  Mozartgasse 12\/II 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0685-2696","authenticated-orcid":false,"given":"Uwe T.","family":"Bornscheuer","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry Department of Biotechnology and Enzyme Catalysis University of Greifswald  Felix-Hausdorff-Stra\u00dfe 4 17487 Greifswald Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3856-3170","authenticated-orcid":false,"given":"Daniel","family":"Kracher","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biotechnology Graz University of Technology  Petersgasse 14 8010 Graz Austria"},{"name":"BioTechMed-Graz  Mozartgasse 12\/II 8010 Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,7,25]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13068-019-1610-9"},{"key":"e_1_2_8_2_1","first-page":"e01123","volume":"84","author":"Santamar\u00eda L.","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-09-14-0292-FI"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00475-14"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2009.03.025"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-813167-1.00005-0"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.66.12.5322-5328.2000"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcatb.2006.09.001"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2020.583617"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodres.2015.06.046"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.eurpolymj.2020.109534"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/ma961129p"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-009-2026-4"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/jctb.5590"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-012-4358-8"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssuschemeng.9b03455"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2012.04.051"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2018.07.021"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1039\/c3gc40748a"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.01798"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-014-6313-3"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1039\/D2GC01629J"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b08177"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssuschemeng.0c03489"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.1.326-332.1995"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2021.107821"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0016262"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10295-010-0709-6"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2016.10.131"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.5344\/ajev.1993.44.1.76"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1997.tb10256.x"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.4.1466-1471.1998"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2014.12.064"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201900571"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.66.8.3368-3375.2000"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb5008103"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-07110-w"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.oprd.6b00362"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1039\/c2cy20015e"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1186\/2191-0855-2-4"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules26216500"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-890-0:571"}],"container-title":["ChemBioChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202300207","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T11:38:45Z","timestamp":1760269125000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cbic.202300207"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,25]]},"references-count":42,"journal-issue":{"issue":"16","published-print":{"date-parts":[[2023,8,15]]}},"alternative-id":["10.1002\/cbic.202300207"],"URL":"https:\/\/doi.org\/10.1002\/cbic.202300207","archive":["Portico"],"relation":{},"ISSN":["1439-4227","1439-7633"],"issn-type":[{"value":"1439-4227","type":"print"},{"value":"1439-7633","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,25]]},"assertion":[{"value":"2023-03-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-07-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202300207"}}