Spontaneous fermentation of raisin water to form wine

Spontaneous fermentation of raisin water to form wine | 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 Spontaneous fermentation of raisin water to form wine Wataru Hashimoto, Mamoru Hio, Kensuke Murata, Ryuichi Takase, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4771016/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 Yeast obtained from the water containing raisins has been conventionally used for breadmaking. It is assumed that people consumed naturally fermented raisin water as wine in ancient times. In this study, we demonstrated spontaneous fermentation by yeast that colonizes raisins. Raisin water samples containing Saccharomyces and non-Saccharomyces species underwent alcohol fermentation. As the fermentation progressed, the diversity and abundance of Aspergillus decreased. A significant difference in β diversity was detected between the microbiomes of raw grapes and raisins, with raisins containing a higher ratio of Saccharomycetaceae. The yeast colonizing raisin fermented alcohol more efficiently than those on raw grapes. Outdoor-dried raisins harbored a distinct eukaryotic microbiome that successfully fermented alcohol, indicating that these yeasts migrate from the environment. Therefore, our data suggest that sun-drying grapes can be utilized for food preservation and making alcoholic beverages, validating the ancient practice of wine-making from raisins due to spontaneous fermentation. eukaryotic microbiome grape non-Saccharomyces raisin Saccharomyces spontaneous fermentation Full Text Additional Declarations There is NO Competing Interest. Tables are available in the Supplementary Files section. Supplementary Files 240622Hio2024RaisinTable1.docx Table 1. Effects of temperatures and raisin contents on fermentation. 240622Hio2024RaisinTable2.docx Table 2. Fermentation tests using raisins made from different grape varieties. 240622Hio2024SupplementaryTable1.pdf Supplementary Table 1. Yeasts isolated from raisins. 240622Hio2024SupplementaryTable2.pdf Supplementary Table 2. Yeasts isolated from grapes. 240622Hio2024SupplementaryTable3.pdf Supplementary Table 3. Detailed information of BioProject PRJDB17448. Hio202420240520SupplementaryFigures1.pdf Supplementary Figure 1. Spontaneous fermentation of raisin-containing water by raisin-colonized S. cerevisiae. Hio202420240520SupplementaryFigures2.pdf Supplementary Figure 2. Images of raisin water (n = 3, #1–3) related to Figure 1. Hio202420240520SupplementaryFigures3.pdf Supplementary Figure 3. Ethanol production and ethanol tolerance of raisin-isolated yeasts. Hio202420240520SupplementaryFigures4.pdf Supplementary Figure 4. Effects of raisin contents on ethanol production and glucose consumption. Hio202420240520SupplementaryFigures5.pdf Supplementary Figure 5. Effects of incubation temperatures on ethanol production. Hio202420240520SupplementaryFigures6.pdf Supplementary Figure 6. Difference of the fermentation across raisin products. Hio202420240520SupplementaryFigures7.pdf Supplementary Figure 7. Comparison of the abundance ratio of microorganisms between grapes (n = 7) and raisins (n = 8) (related to Figure 3). Hio202420240520SupplementaryFigures8.pdf Supplementary Figure 8. Comparison of α diversity between grapes and raisins (related to Figure 3). Hio202420240520SupplementaryFigures9.pdf Supplementary Figure 9. Glucose consumption and growth of isolated yeasts from grapes and raisins in 10% glucose-containing YNB medium. Hio202420240520SupplementaryFigures10.pdf Hio202420240520SupplementaryFigures11.pdf Supplementary Figure 11. Ethanol production and glucose consumption of raisin water made from laboratory-made raisins. 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. 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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-4771016","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":338321702,"identity":"44fd05d9-3652-46ff-bd43-11f38d4547fc","order_by":0,"name":"Wataru Hashimoto","email":"data:image/png;base64,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","orcid":"","institution":"Kyoto University","correspondingAuthor":true,"prefix":"","firstName":"Wataru","middleName":"","lastName":"Hashimoto","suffix":""},{"id":338321703,"identity":"a428c662-8c3e-4dfd-8211-f784c87dd4c8","order_by":1,"name":"Mamoru Hio","email":"","orcid":"","institution":"Kyoto University","correspondingAuthor":false,"prefix":"","firstName":"Mamoru","middleName":"","lastName":"Hio","suffix":""},{"id":338321704,"identity":"e551202e-72a8-4a4d-91a8-a6a9a67e5e32","order_by":2,"name":"Kensuke Murata","email":"","orcid":"","institution":"Kyoto University","correspondingAuthor":false,"prefix":"","firstName":"Kensuke","middleName":"","lastName":"Murata","suffix":""},{"id":338321705,"identity":"c3a0bf86-6df5-44cd-a186-ae26066845bd","order_by":3,"name":"Ryuichi Takase","email":"","orcid":"","institution":"Kyoto University","correspondingAuthor":false,"prefix":"","firstName":"Ryuichi","middleName":"","lastName":"Takase","suffix":""},{"id":338321706,"identity":"90331c75-d393-407e-89aa-b2a984182667","order_by":4,"name":"Kohei Ogura","email":"","orcid":"","institution":"Kyoto University","correspondingAuthor":false,"prefix":"","firstName":"Kohei","middleName":"","lastName":"Ogura","suffix":""}],"badges":[],"createdAt":"2024-07-20 03:30:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4771016/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4771016/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":66401347,"identity":"d027a666-0631-463d-b819-c2cdac20a88d","added_by":"auto","created_at":"2024-10-11 11:18:33","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1381654,"visible":true,"origin":"","legend":"","description":"","filename":"CommBio240720Hio2024RaisinManuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1_covered_bf67eb67-48e6-4d17-bbba-b9afcec4208d.pdf"},{"id":64054979,"identity":"71b86d09-d4c5-43ef-8722-5340c29511ec","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":28787,"visible":true,"origin":"","legend":"\u003cp\u003eTable 1. Effects of temperatures and raisin contents on fermentation.\u003c/p\u003e","description":"","filename":"240622Hio2024RaisinTable1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/ed2e2915de9246e40de2c85f.docx"},{"id":64054980,"identity":"257b6c92-e788-4e91-8364-773b837f8c11","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":33193,"visible":true,"origin":"","legend":"\u003cp\u003eTable 2. Fermentation tests using raisins made from different grape varieties.\u003c/p\u003e","description":"","filename":"240622Hio2024RaisinTable2.docx","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/045cd38293e126fbfd558670.docx"},{"id":64054983,"identity":"63b7eb62-9bd4-4c41-a7ba-8ef1aa07857e","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":99062,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Table 1. Yeasts isolated from raisins.\u003c/p\u003e","description":"","filename":"240622Hio2024SupplementaryTable1.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/270d982727f074cd459f5252.pdf"},{"id":64055192,"identity":"9e5bed10-aea5-4d92-a6ca-3d9e55d971aa","added_by":"auto","created_at":"2024-09-05 18:29:52","extension":"pdf","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":101142,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Table 2. Yeasts isolated from grapes.\u003c/p\u003e","description":"","filename":"240622Hio2024SupplementaryTable2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/50821575ced75cedb6e4a050.pdf"},{"id":64054981,"identity":"05f99e46-4857-40be-83bb-a445a78422ff","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":46123,"visible":true,"origin":"","legend":"Supplementary Table 3. Detailed information of BioProject PRJDB17448.","description":"","filename":"240622Hio2024SupplementaryTable3.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/1739d77b43c2fe3f7a58b113.pdf"},{"id":64054985,"identity":"c44e8228-2ade-40fe-a575-4fd3cf157758","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":3314307,"visible":true,"origin":"","legend":"Supplementary Figure 1. Spontaneous fermentation of raisin-containing water by raisin-colonized S. cerevisiae.","description":"","filename":"Hio202420240520SupplementaryFigures1.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/ad799c7dc7af33c57dd646f6.pdf"},{"id":64054987,"identity":"fced5c05-e25e-43a8-9ebf-493022e3f76c","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":4869538,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Figure 2. Images of raisin water (n = 3, #1–3) related to Figure 1.\u003c/p\u003e","description":"","filename":"Hio202420240520SupplementaryFigures2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/bfb306d6ffdc79dad9ae290f.pdf"},{"id":64055196,"identity":"d13ffd30-d50e-4ae0-b739-22eee1b1e78f","added_by":"auto","created_at":"2024-09-05 18:29:53","extension":"pdf","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":589995,"visible":true,"origin":"","legend":"Supplementary Figure 3. Ethanol production and ethanol tolerance of raisin-isolated yeasts.","description":"","filename":"Hio202420240520SupplementaryFigures3.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/404eec76e5c0c085661bae08.pdf"},{"id":64054993,"identity":"54204afb-65dc-463f-93b1-3837ed172449","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":2243265,"visible":true,"origin":"","legend":"Supplementary Figure 4. Effects of raisin contents on ethanol production and glucose consumption.","description":"","filename":"Hio202420240520SupplementaryFigures4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/d097e5ac928e6f8ad4f62f79.pdf"},{"id":64054989,"identity":"b43fbb4a-5514-4cb9-93df-ed52526c2f83","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":2187792,"visible":true,"origin":"","legend":"Supplementary Figure 5. Effects of incubation temperatures on ethanol production.","description":"","filename":"Hio202420240520SupplementaryFigures5.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/71c1560b2eaadc96cbd43fe3.pdf"},{"id":64054994,"identity":"37acf7ab-42c7-4efc-af2f-d62ca05b2519","added_by":"auto","created_at":"2024-09-05 18:21:53","extension":"pdf","order_by":11,"title":"","display":"","copyAsset":false,"role":"supplement","size":6447655,"visible":true,"origin":"","legend":"Supplementary Figure 6. Difference of the fermentation across raisin products.","description":"","filename":"Hio202420240520SupplementaryFigures6.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/f3c48978346782e685ce9f14.pdf"},{"id":64054984,"identity":"73e34db7-e64f-4af5-addf-4a86754e958f","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":12,"title":"","display":"","copyAsset":false,"role":"supplement","size":401586,"visible":true,"origin":"","legend":"Supplementary Figure 7. Comparison of the abundance ratio of microorganisms between grapes (n = 7) and raisins (n = 8) (related to Figure 3).","description":"","filename":"Hio202420240520SupplementaryFigures7.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/44a3599d9578e43b7ae23ce7.pdf"},{"id":64055193,"identity":"a406002d-21f6-4d7b-acdc-b7568af3261e","added_by":"auto","created_at":"2024-09-05 18:29:52","extension":"pdf","order_by":13,"title":"","display":"","copyAsset":false,"role":"supplement","size":117111,"visible":true,"origin":"","legend":"Supplementary Figure 8. Comparison of \u0026#x03B1; diversity between grapes and raisins (related to Figure 3).","description":"","filename":"Hio202420240520SupplementaryFigures8.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/12f5e2d9e53f2c0c43e699c5.pdf"},{"id":64054992,"identity":"b64d80d6-6064-47bf-9018-5234b5ed613e","added_by":"auto","created_at":"2024-09-05 18:21:52","extension":"pdf","order_by":14,"title":"","display":"","copyAsset":false,"role":"supplement","size":530240,"visible":true,"origin":"","legend":"Supplementary Figure 9. Glucose consumption and growth of isolated yeasts from grapes and raisins in 10% glucose-containing YNB medium.","description":"","filename":"Hio202420240520SupplementaryFigures9.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/ad0739798a38c3ceb4cdc787.pdf"},{"id":64055195,"identity":"7e03b7a3-cd26-424a-b6d9-c163ace3426b","added_by":"auto","created_at":"2024-09-05 18:29:52","extension":"pdf","order_by":15,"title":"","display":"","copyAsset":false,"role":"supplement","size":527147,"visible":true,"origin":"","legend":"","description":"","filename":"Hio202420240520SupplementaryFigures10.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/62b37a494af83dad89f4a34e.pdf"},{"id":64055194,"identity":"831e24e3-2a56-410b-abed-c696a3fc5f07","added_by":"auto","created_at":"2024-09-05 18:29:52","extension":"pdf","order_by":16,"title":"","display":"","copyAsset":false,"role":"supplement","size":174480,"visible":true,"origin":"","legend":"Supplementary Figure 11. Ethanol production and glucose consumption of raisin water made from laboratory-made raisins.","description":"","filename":"Hio202420240520SupplementaryFigures11.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4771016/v1/9dbc2bb96068dc7e7bc91d8d.pdf"}],"financialInterests":"\u003cp\u003eThere is \u003cstrong\u003eNO\u003c/strong\u003e Competing Interest.\u003c/p\u003e\n\u003cp\u003eTables are available in the Supplementary Files section.\u003c/p\u003e","formattedTitle":"Spontaneous fermentation of raisin water to form wine","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"eukaryotic microbiome, grape, non-Saccharomyces, raisin, Saccharomyces, spontaneous fermentation","lastPublishedDoi":"10.21203/rs.3.rs-4771016/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4771016/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Yeast obtained from the water containing raisins has been conventionally used for breadmaking. It is assumed that people consumed naturally fermented raisin water as wine in ancient times. In this study, we demonstrated spontaneous fermentation by yeast that colonizes raisins. Raisin water samples containing Saccharomyces and non-Saccharomyces species underwent alcohol fermentation. As the fermentation progressed, the diversity and abundance of Aspergillus decreased. A significant difference in β diversity was detected between the microbiomes of raw grapes and raisins, with raisins containing a higher ratio of Saccharomycetaceae. The yeast colonizing raisin fermented alcohol more efficiently than those on raw grapes. Outdoor-dried raisins harbored a distinct eukaryotic microbiome that successfully fermented alcohol, indicating that these yeasts migrate from the environment. 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