A new yeast strain for valorisation of vinasse, a rum distillery waste product

A new yeast strain for valorisation of vinasse, a rum distillery waste product | 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 A new yeast strain for valorisation of vinasse, a rum distillery waste product Brigita Simonaviciene, Ayokunle Araoyinbo, Juwayria Ali, Jamie McGowan, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6593393/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Jul, 2025 Read the published version in Biotechnology for Biofuels and Bioproducts → Version 1 posted 12 You are reading this latest preprint version Abstract Background: Waste valorisation refers to processes of reusing or recycling waste materials to create valuable products. In the Rum distillery industry, the primary waste byproducts include bagasse, a solid waste made up of sugar cane residue and vinasse, a thick and acidic liquid. Although vinasse has been repurposed in agricultural fields, it has also contributed to both soil and ocean pollution. Despite several potential solutions having been suggested, an effective and environmentally safe use for vinasse has yet to be found. Results: The valorisation of vinasse for biofuel production was explored by assessing its potential as a growth medium for lipid production by non-conventional yeasts. The oleaginous yeast strain Yarrowia lipolytica, known for its lipid production capabilities, was initially tested on vinasse but required further adaptation and optimization. To circumvent this, we isolated a novel yeast strain from old vinasse waste, named V1, which demonstrated strong growth potential. The growth conditions of V1, including temperature and acidity, were characterized, and its potential for bioengineering was evaluated. This strain exhibited resistance to highly acidic pH levels and higher temperatures when cultivated on YPV, an artificial laboratory medium designed to mimic the acidity and glycerol content of vinasse. Whole genome sequencing (WGS) identified V1 as Pichia kudriavzevii. We demonstrated that V1 could be transformed with Yarrowia lipolytica vectors using the classical yeast heat shock protocol, thus enabling potential genetic engineering. Lastly, lipid content in V1 was analysed in different conditions, confirming the strain's potential for biofuel production. Conclusions: Pichia kudriavzevii is not a traditional yeast, but its ability to adapt and grow under extreme pH and higher temperature conditions makes it a promising candidate for rum industry waste management applications. This strain could potentially be utilised to convert vinasse and other food waste products into valuable biofuels. Although further research is required to engineer and optimize this novel strain for vinasse cultivation, our findings highlight its great potential as a micro-factory in rum-producing regions and high locations where agricultural waste is in need of valorisation solutions. Vinasse Pichia kudriavzevii Yarrowia lipolytica Biofuel rum distillery Full Text Additional Declarations No competing interests reported. Supplementary Files SupplTable1Microbiome.pdf SupplTable2OrthologsSNPSanalysisoriginal1.xlsx SupplTable3SimonavicieneV128vsYL.xlsx SupplTable4SimonavicieneV128vsV137.xlsx Cite Share Download PDF Status: Published Journal Publication published 18 Jul, 2025 Read the published version in Biotechnology for Biofuels and Bioproducts → Version 1 posted Editorial decision: Revision requested 22 May, 2025 Reviews received at journal 14 May, 2025 Reviewers agreed at journal 13 May, 2025 Reviews received at journal 12 May, 2025 Reviewers agreed at journal 12 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers invited by journal 07 May, 2025 Editor assigned by journal 06 May, 2025 Submission checks completed at journal 06 May, 2025 First submitted to journal 05 May, 2025 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. 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Bioproducts","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Vinasse, Pichia kudriavzevii, Yarrowia lipolytica, Biofuel, rum distillery","lastPublishedDoi":"10.21203/rs.3.rs-6593393/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6593393/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground: Waste valorisation refers to processes of reusing or recycling waste materials to create valuable products. In the Rum distillery industry, the primary waste byproducts include bagasse, a solid waste made up of sugar cane residue and vinasse, a thick and acidic liquid. Although vinasse has been repurposed in agricultural fields, it has also contributed to both soil and ocean pollution. Despite several potential solutions having been suggested, an effective and environmentally safe use for vinasse has yet to be found.\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults:\u0026nbsp;The valorisation of vinasse for biofuel production was explored by assessing its potential as a growth medium for lipid production by non-conventional yeasts. The oleaginous yeast strain Yarrowia lipolytica, known for its lipid production capabilities, was initially tested on vinasse but required further adaptation and optimization. To circumvent this, we isolated a novel yeast strain from old vinasse waste, named V1, which demonstrated strong growth potential. The growth conditions of V1, including temperature and acidity, were characterized, and its potential for bioengineering was evaluated. This strain exhibited resistance to highly acidic pH levels and higher temperatures when cultivated on YPV, an artificial laboratory medium designed to mimic the acidity and glycerol content of vinasse. Whole genome sequencing (WGS) identified V1 as Pichia kudriavzevii. We demonstrated that V1 could be transformed with\u0026nbsp; Yarrowia lipolytica vectors using the classical yeast heat shock protocol, thus enabling potential genetic engineering. Lastly, lipid content in V1 was analysed in different conditions, confirming the strain's potential for biofuel production.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConclusions:\u0026nbsp;Pichia kudriavzevii is not a traditional yeast, but its ability to adapt and grow under extreme pH and higher temperature conditions makes it a promising candidate for rum industry waste management applications. This strain could potentially be utilised to convert vinasse and other food waste products into valuable biofuels. Although further research is required to engineer and optimize this novel strain for vinasse cultivation, our findings highlight its great potential as a micro-factory in rum-producing regions and high locations where agricultural waste is in need of valorisation solutions.\u003c/p\u003e","manuscriptTitle":"A new yeast strain for valorisation of vinasse, a rum distillery waste product","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-12 08:45:14","doi":"10.21203/rs.3.rs-6593393/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-22T07:30:51+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-14T20:06:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"4458035109505278017469007438441347734","date":"2025-05-13T04:37:16+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-12T19:44:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"54822368684043413317154993355662442726","date":"2025-05-12T11:00:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"72927286637024243438565728616997975071","date":"2025-05-07T21:42:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"118244873881769138844899904730506425537","date":"2025-05-07T17:19:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"47541943773279543076155699707267547007","date":"2025-05-07T09:08:39+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-05-07T08:35:35+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-05-06T15:07:29+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-05-06T04:24:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biotechnology for Biofuels and Bioproducts","date":"2025-05-05T10:03:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biotechnology-for-biofuels-and-bioproducts","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bbio","sideBox":"Learn more about [Biotechnology for Biofuels](http://biotechnologyforbiofuels.biomedcentral.com/)","snPcode":"13068","submissionUrl":"https://submission.nature.com/new-submission/13068/3","title":"Biotechnology for Biofuels and Bioproducts","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fd9ef569-84ff-4394-8c49-83a366082e48","owner":[],"postedDate":"May 12th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-08-07T07:21:12+00:00","versionOfRecord":{"articleIdentity":"rs-6593393","link":"https://doi.org/10.1186/s13068-025-02671-0","journal":{"identity":"biotechnology-for-biofuels-and-bioproducts","isVorOnly":false,"title":"Biotechnology for Biofuels and Bioproducts"},"publishedOn":"2025-07-18 16:05:44","publishedOnDateReadable":"July 18th, 2025"},"versionCreatedAt":"2025-05-12 08:45:14","video":"","vorDoi":"10.1186/s13068-025-02671-0","vorDoiUrl":"https://doi.org/10.1186/s13068-025-02671-0","workflowStages":[]},"version":"v1","identity":"rs-6593393","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6593393","identity":"rs-6593393","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}