Engineered artificial biosynthesis pathway enables simultaneous production and in-situ bio-dyeing of indigoids for textiles

Engineered artificial biosynthesis pathway enables simultaneous production and in-situ bio-dyeing of indigoids for textiles | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Engineered artificial biosynthesis pathway enables simultaneous production and in-situ bio-dyeing of indigoids for textiles Chenjie Zhu, Qi Na, Xiaowang Zhang, Linna Long, Siyu Qi, Zhiyu Zhang, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6670203/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Dec, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract The textile dyeing industry, empowered by synthetic chemistry, has shaped modern aesthetics through vibrant coloration technologies. Indigoid dyes, including indigo and Tyrian purple, embody this potential but are hindered by the toxic, hazardous chemo-synthesis and water-intensive chemo-dyeing processes. While biosynthesis and bio-dyeing efforts mitigate hazardous inputs, they remain constrained by disjointed synthesis-dyeing steps that lead to unsustainable water and energy demands. Also, the disadvantages of biosynthetic pathways including complexity, low atom economy, and inefficiency still remain. Here, we pioneer a one-pot, one-step approach for unifying biosynthesis and bio-dyeing of indigoid which circumvent water usage in the dyeing process. By combining alcohol dehydrogenase-catalyzed amino alcohol oxidation with flavin monooxygenase-driven indole oxygenation, we constructed a previously unreported artificial redox-neutral cascade biosynthetic pathway to produce indigoids. This innovation significantly simplified the catalytic process and achieved the highest reported atom economy and production efficiency of indigoids. Employing E. coli whole cell that generate biosynthetic bluish-violet indigoid dyes, we achieved biosynthesis and in situ dyeing of indigoids on various natural and synthetic fabrics. Crucially, this system successfully dyed synthetic nylon fibers with the historic purple hue—a milestone unattainable through conventional chemical methods. Physical sciences/Chemistry/Biosynthesis Biological sciences/Chemical biology/Biocatalysis Biological sciences/Biotechnology Biological sciences/Chemical biology/Synthetic biology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformation.pdf support information Cite Share Download PDF Status: Published Journal Publication published 26 Dec, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6670203","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":461138421,"identity":"b04c3e79-97c2-4d19-853c-88af2b46ea6d","order_by":0,"name":"Chenjie Zhu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA20lEQVRIiWNgGAWjYDACCRCuYGAGUmxAzEysljMMzDykaWFsY2AgXov87B7DD5bz7rDbszc/e8BQYZ3YwH72AF4tjHPOGEtIbnvGzMNzzNyA4Ux6YgNPXgJeLcwSOWYMktsOM/NI5LBJMLYdTmyQ4DHAq4UNrGUOUIv8G6CWf0Ro4QFraQDZwgPU0kCEFgmJtGIJiWNALWfSzCQSjqUbt/Hk4NciPyN542eJmsPJ7O2Hn0l8qLGW7Wc/g18LCDAD4yYZzEpggMQOQcD4gYHBjhiFo2AUjIJRMEIBACv5ODVljo8yAAAAAElFTkSuQmCC","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":true,"prefix":"","firstName":"Chenjie","middleName":"","lastName":"Zhu","suffix":""},{"id":461138422,"identity":"fb0428a0-f256-42b1-989a-88239897d7e1","order_by":1,"name":"Qi Na","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Qi","middleName":"","lastName":"Na","suffix":""},{"id":461138423,"identity":"59e72bcc-2351-4372-bb49-41487bfbe68e","order_by":2,"name":"Xiaowang Zhang","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Xiaowang","middleName":"","lastName":"Zhang","suffix":""},{"id":461138424,"identity":"a7dde0c0-bb27-4e0e-83a5-1e7bfd2462e6","order_by":3,"name":"Linna Long","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Linna","middleName":"","lastName":"Long","suffix":""},{"id":461138425,"identity":"49555197-7e4d-4a88-a7a3-9aa1dd5c6275","order_by":4,"name":"Siyu Qi","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Siyu","middleName":"","lastName":"Qi","suffix":""},{"id":461138426,"identity":"3a01bdcc-c1ae-4875-bbd4-c2ea149a31f4","order_by":5,"name":"Zhiyu Zhang","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Zhiyu","middleName":"","lastName":"Zhang","suffix":""},{"id":461138427,"identity":"513e9211-44b7-4a95-af67-a3a830edbf3b","order_by":6,"name":"Ruoyu Zhi","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Ruoyu","middleName":"","lastName":"Zhi","suffix":""},{"id":461138428,"identity":"4dace9c3-f403-4787-9cb7-0e75c90795db","order_by":7,"name":"Zhuotao Tan","email":"","orcid":"https://orcid.org/0000-0001-6314-8422","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Zhuotao","middleName":"","lastName":"Tan","suffix":""},{"id":461138429,"identity":"a9ebca5f-8ad8-49a6-8486-15af66f9b4b1","order_by":8,"name":"Hanjie Ying","email":"","orcid":"","institution":"Nanjing Tech University","correspondingAuthor":false,"prefix":"","firstName":"Hanjie","middleName":"","lastName":"Ying","suffix":""}],"badges":[],"createdAt":"2025-05-15 07:57:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6670203/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6670203/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41467-025-67935-7","type":"published","date":"2025-12-26T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":101551995,"identity":"5abf891c-b560-4230-8486-3c37c21dec5c","added_by":"auto","created_at":"2026-01-31 08:06:31","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1767921,"visible":true,"origin":"","legend":"Article File","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6670203/v1_covered_08ab7727-a378-49a4-9a13-fd49c2d8fe32.pdf"},{"id":86625721,"identity":"6810acdc-8ae0-479b-9f6f-0f53648ad304","added_by":"auto","created_at":"2025-07-14 05:13:10","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":9552427,"visible":true,"origin":"","legend":"support information","description":"","filename":"SupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6670203/v1/854018a83ffe064038e3e514.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Engineered artificial biosynthesis pathway enables simultaneous production and in-situ bio-dyeing of indigoids for textiles","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6670203/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6670203/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The textile dyeing industry, empowered by synthetic chemistry, has shaped modern aesthetics through vibrant coloration technologies. Indigoid dyes, including indigo and Tyrian purple, embody this potential but are hindered by the toxic, hazardous chemo-synthesis and water-intensive chemo-dyeing processes. While biosynthesis and bio-dyeing efforts mitigate hazardous inputs, they remain constrained by disjointed synthesis-dyeing steps that lead to unsustainable water and energy demands. Also, the disadvantages of biosynthetic pathways including complexity, low atom economy, and inefficiency still remain. Here, we pioneer a one-pot, one-step approach for unifying biosynthesis and bio-dyeing of indigoid which circumvent water usage in the dyeing process. By combining alcohol dehydrogenase-catalyzed amino alcohol oxidation with flavin monooxygenase-driven indole oxygenation, we constructed a previously unreported artificial redox-neutral cascade biosynthetic pathway to produce indigoids. This innovation significantly simplified the catalytic process and achieved the highest reported atom economy and production efficiency of indigoids. Employing E. coli whole cell that generate biosynthetic bluish-violet indigoid dyes, we achieved biosynthesis and in situ dyeing of indigoids on various natural and synthetic fabrics. Crucially, this system successfully dyed synthetic nylon fibers with the historic purple hue—a milestone unattainable through conventional chemical methods.","manuscriptTitle":"Engineered artificial biosynthesis pathway enables simultaneous production and in-situ bio-dyeing of indigoids for textiles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-14 05:13:05","doi":"10.21203/rs.3.rs-6670203/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"c6357885-a661-4e25-af70-c35b3505acce","owner":[],"postedDate":"July 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":48984999,"name":"Physical sciences/Chemistry/Biosynthesis"},{"id":48985000,"name":"Biological sciences/Chemical biology/Biocatalysis"},{"id":48985001,"name":"Biological sciences/Biotechnology"},{"id":48985002,"name":"Biological sciences/Chemical biology/Synthetic biology"}],"tags":[],"updatedAt":"2026-01-31T08:06:25+00:00","versionOfRecord":{"articleIdentity":"rs-6670203","link":"https://doi.org/10.1038/s41467-025-67935-7","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2025-12-26 05:00:00","publishedOnDateReadable":"December 26th, 2025"},"versionCreatedAt":"2025-07-14 05:13:05","video":"","vorDoi":"10.1038/s41467-025-67935-7","vorDoiUrl":"https://doi.org/10.1038/s41467-025-67935-7","workflowStages":[]},"version":"v1","identity":"rs-6670203","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6670203","identity":"rs-6670203","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}