De Novo Biosynthesis of Violacein Utilizing Saccharomyces cerevisiae as a Cell Factory

De Novo Biosynthesis of Violacein Utilizing Saccharomyces cerevisiae as a Cell Factory | 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 Research Article De Novo Biosynthesis of Violacein Utilizing Saccharomyces cerevisiae as a Cell Factory Yiheng Dong, Gengran Zhai, Keyao Ren, Jiale Chen, Kecheng Chen, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6768705/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Violacein is a naturally occurring purple pigment produced by microbes, commonly employed in synthetic biology as a readily detectable model compound. Despite its wide-ranging applications as an antimicrobial, antiviral, and antitumor compound, violacein production is hindered by low yield and strain instability, which are frequently affected by cultivation conditions. Furthermore, heterologous production remains limited to the laboratory scale. Consequently, rational design strategies for improving violacein biosynthesis have attracted increasing research interest. Results In this work, the violacein biosynthesis pathway was reconstructed in Saccharomyces cerevisiae ; however, no production of violacein was initially observed. Pathway analysis indicated that insufficient vioC expression contributed to the limited violacein output. By employing pathway engineering strategies—such as multicopy gene integration and overexpression—combined with fermentation medium optimization using the Box-Behnken design, violacein production in 250 mL shake flasks (with 100 mL culture volume) reached 143.89 mg/L, representing the highest yield reported in S. cerevisiae so far. Conclusions This study successfully demonstrates the potential of S. cerevisiae as a robust chassis for violacein biosynthesis via synthetic biology strategies. The combination of multicopy gene integration, targeted gene overexpression, and response surface optimization provides a scalable and safe platform for industrial production of violacein. Further improvement through metabolic engineering may bridge the gap toward full commercialization. De novo biosynthesis Multicopy integration Saccharomyces cerevisiae vio cluster Violacein Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryMaterial.docx Cite Share Download PDF Status: Posted 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-6768705","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":476743483,"identity":"09b2fb76-4afb-4f4b-8c8a-cb4221fb000a","order_by":0,"name":"Yiheng Dong","email":"","orcid":"","institution":"Zhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Sciences and Medicine, Zhejiang Sci-Tech 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Violacein","lastPublishedDoi":"10.21203/rs.3.rs-6768705/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6768705/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eViolacein is a naturally occurring purple pigment produced by microbes, commonly employed in synthetic biology as a readily detectable model compound. Despite its wide-ranging applications as an antimicrobial, antiviral, and antitumor compound, violacein production is hindered by low yield and strain instability, which are frequently affected by cultivation conditions. Furthermore, heterologous production remains limited to the laboratory scale. Consequently, rational design strategies for improving violacein biosynthesis have attracted increasing research interest.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eIn this work, the violacein biosynthesis pathway was reconstructed in \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e; however, no production of violacein was initially observed. Pathway analysis indicated that insufficient vioC expression contributed to the limited violacein output. By employing pathway engineering strategies\u0026mdash;such as multicopy gene integration and overexpression\u0026mdash;combined with fermentation medium optimization using the Box-Behnken design, violacein production in 250 mL shake flasks (with 100 mL culture volume) reached 143.89 mg/L, representing the highest yield reported in \u003cem\u003eS. cerevisiae\u003c/em\u003e so far.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eThis study successfully demonstrates the potential of S. cerevisiae as a robust chassis for violacein biosynthesis via synthetic biology strategies. The combination of multicopy gene integration, targeted gene overexpression, and response surface optimization provides a scalable and safe platform for industrial production of violacein. 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