{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T00:09:24Z","timestamp":1792109364274,"version":"4.4.0"},"reference-count":27,"publisher":"Wiley","issue":"19","license":[{"start":{"date-parts":[[2024,6,27]],"date-time":"2024-06-27T00:00:00Z","timestamp":1719446400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Fonden","doi-asserted-by":"publisher","award":["NNF17SA0031362"],"award-info":[{"award-number":["NNF17SA0031362"]}],"id":[{"id":"10.13039\/501100009708","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":[[2024,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Natural and pure <jats:italic>p<\/jats:italic>\u2010coumaric acid has valuable applications, and it can be produced via bioprocessing. However, fermentation processes have so far been unable to provide sufficient production metrics, while a biocatalytic process decoupling growth and production historically showed much promise. This biocatalytic process is revisited in order to tackle product inhibition of the key enzyme tyrosine ammonia lyase. In situ product removal is proposed as a possible solution, and a polymer\/salt aqueous two\u2010phase system is identified as a suitable system for extraction of <jats:italic>p<\/jats:italic>\u2010coumaric acid from an alkaline solution, with a partition coefficient of up to 13. However, a 10\u2009% salt solution was found to reduce tyrosine ammonia lyase activity by 19\u2009%, leading to the need for a more dilute system. The cloud points of two aqueous two\u2010phase systems at 40\u2009\u00b0C and pH\u200510 were found to be 3.8\u2009% salt and 9.5\u2009% polymer, and a 5\u2009% potassium phosphate and 12.5\u2009% poly(ethylene glycol\u2010<jats:italic>ran<\/jats:italic>\u2010propylene glycol) mW~2500 system was selected for in\u2005situ product removal. An immobilized tyrosine ammonia lyase biocatalyst in this aqueous two\u2010phase system produced up to 33\u2005g\/L <jats:italic>p<\/jats:italic>\u2010coumaric acid within 24\u2005hours, a 1.9\u2010fold improvement compared to biocatalysis without in\u2005situ product removal.<\/jats:p>","DOI":"10.1002\/cbic.202400178","type":"journal-article","created":{"date-parts":[[2024,5,14]],"date-time":"2024-05-14T08:46:45Z","timestamp":1715676405000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Biocatalysis with In\u2010Situ Product Removal Improves <i>p\u2010<\/i>Coumaric Acid Production"],"prefix":"10.1002","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4374-7641","authenticated-orcid":false,"given":"Alexander","family":"Virklund","sequence":"first","affiliation":[{"name":"Department of Chemical and Biochemical Engineering Technical University of Denmark  2800 Kgs. Lyngby Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6616-0187","authenticated-orcid":false,"given":"Alex Toftgaard","family":"Nielsen","sequence":"additional","affiliation":[{"name":"Novo Nordisk Foundation Center for Biosustainability Technical University of Denmark  2800 Kgs. Lyngby Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7976-2483","authenticated-orcid":false,"given":"John M.","family":"Woodley","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biochemical Engineering Technical University of Denmark  2800 Kgs. Lyngby Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,6,27]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07882-8"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodcont.2011.11.022"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.28292"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.61.080706.093248"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00405-15"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.202300111"},{"key":"e_1_2_9_7_1","unstructured":"A.\u2005A. Gatenby F.\u2005S. Sariaslani X.-S. Tang W.\u2005W. Qi T. Vannelli Bioproduction of Parahydroxycinnamic Acid 2002 US 6 368 837 B1."},{"key":"e_1_2_9_8_1","first-page":"2367","volume":"36","author":"Huo Y.","year":"2020","journal-title":"Shengwu Gongcheng Xuebao\/Chinese J. Biotechnol."},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1039\/D0CC00783H"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2014.07.003"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2021.125436"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jced.6b00361"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.seppur.2020.118170"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-816911-7.00005-0"},{"key":"e_1_2_9_15_1","volume-title":"Partitioning in Aqueous Two-Phase Systems: Theory, Methods, Uses, and Application to Biotechnology","author":"Walter H.","year":"1985"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19980705)59:1<73::AID-BIT10>3.0.CO;2-3"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC02691A"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02798461"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.1983.0011"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.260211002"},{"key":"e_1_2_9_21_1","first-page":"53","author":"Toorra J. L.","year":"2003","journal-title":"Immobil. Enzym. Cells"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp060379e"},{"key":"e_1_2_9_23_1","unstructured":"R. Dicosimo S.\u2005L. Haynie L.\u2005L. Huang F.\u2005S. Sariaslani T.\u2005K. Van Dyk R.\u2005J. Trotman Immobilized TAL Biocatalyst for Production of Para-Hydroxycinnamic Acid 2008 US 8 569 036 B2."},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/je700067a"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/jccs.201800265"},{"key":"e_1_2_9_26_1","volume":"2014","author":"Thomason L. C.","year":"2014","journal-title":"Curr. Protoc. Mol. Biol."},{"key":"e_1_2_9_27_1","doi-asserted-by":"crossref","unstructured":"C.\u2005T. Chung S.\u2005L. Niemela R.\u2005H. Miller One-Step Preparation of Competent Escherichia Coli: Transformation and Storage of Bacterial Cells in the Same Solution (Recombinant DNA) 1989.","DOI":"10.1073\/pnas.86.7.2172"}],"container-title":["ChemBioChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202400178","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T13:17:50Z","timestamp":1760275070000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cbic.202400178"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,27]]},"references-count":27,"journal-issue":{"issue":"19","published-print":{"date-parts":[[2024,10]]}},"alternative-id":["10.1002\/cbic.202400178"],"URL":"https:\/\/doi.org\/10.1002\/cbic.202400178","archive":["Portico"],"relation":{},"ISSN":["1439-4227","1439-7633"],"issn-type":[{"value":"1439-4227","type":"print"},{"value":"1439-7633","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,27]]},"assertion":[{"value":"2024-03-01","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-06-27","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202400178"}}