A Novel Microbial Dissolution - Reprecipitation Pathway for Biosintering of Limestone for Biocement Production

A Novel Microbial Dissolution - Reprecipitation Pathway for Biosintering of Limestone for Biocement Production | 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 Novel Microbial Dissolution - Reprecipitation Pathway for Biosintering of Limestone for Biocement Production Raja Murugan, Anant Aishwarya Dubey, Navdeep K Dhami, Abhijit Mukherjee This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7268390/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Dec, 2025 Read the published version in Applied Microbiology and Biotechnology → Version 1 posted You are reading this latest preprint version Abstract In nature, remarkable geological structures, such as stromatolites, thrombolites, and beachrocks, are formed through biocementation, a process involving the successive dissolution and reprecipitation of CaCO 3 . This research demonstrates mimicking natural cement via bio-sintering of limestone, a process that involves successive dissolution and reprecipitation of limestone facilitated by bacteria under ambient environmental conditions. When the bacterium Acetobacter aceti (ATCC 15973) was introduced into a mixture of ethanol and limestone powder, the pH dropped rapidly, leading to the dissolution of limestone into calcium acetate. After ethanol was fully consumed, the pH gradually increased due to acetate oxidation, causing biocement crystals to precipitate. All reaction rates were measured, and the products characterized through Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy, Quantitative X-ray Diffraction, and a Particle Size Analyzer. The detailed characterization indicates that the precipitate is mainly calcite and that the dissolved calcium carbonate sinters microbially. This pathway offers new opportunities in biocementation research by using limestone as a direct calcium source, reducing the overall carbon footprint, and eliminating the need for urea or other chemical additives. It provides a foundation for developing sustainable, low-carbon cementing materials suitable for next-generation construction applications. Biocement Biosintering CaCO3 dissolution-reprecipitation Ethanol and Acetate oxidation Limestone as Calcium Source Acetobacter aceti Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryFile.docx Cite Share Download PDF Status: Published Journal Publication published 23 Dec, 2025 Read the published version in Applied Microbiology and Biotechnology → 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. 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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-7268390","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":508011303,"identity":"08851680-ce18-44b5-9707-27a5b809f473","order_by":0,"name":"Raja Murugan","email":"","orcid":"","institution":"Curtin University","correspondingAuthor":false,"prefix":"","firstName":"Raja","middleName":"","lastName":"Murugan","suffix":""},{"id":508011305,"identity":"b0a54ca3-051b-4739-98fc-55ebcd1d31ac","order_by":1,"name":"Anant Aishwarya Dubey","email":"","orcid":"","institution":"Curtin University","correspondingAuthor":false,"prefix":"","firstName":"Anant","middleName":"Aishwarya","lastName":"Dubey","suffix":""},{"id":508011306,"identity":"d2ad1ce8-c689-419e-a5db-39d005009a6d","order_by":2,"name":"Navdeep K Dhami","email":"","orcid":"","institution":"Curtin University","correspondingAuthor":false,"prefix":"","firstName":"Navdeep","middleName":"K","lastName":"Dhami","suffix":""},{"id":508011307,"identity":"0c681b50-7074-4bd4-ba4a-7da56203dfab","order_by":3,"name":"Abhijit Mukherjee","email":"data:image/png;base64,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","orcid":"","institution":"Curtin University","correspondingAuthor":true,"prefix":"","firstName":"Abhijit","middleName":"","lastName":"Mukherjee","suffix":""}],"badges":[],"createdAt":"2025-08-01 07:23:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7268390/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7268390/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00253-025-13658-0","type":"published","date":"2025-12-23T15:56:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":99172210,"identity":"d586382a-c901-4dc5-b771-15b9eff4a03a","added_by":"auto","created_at":"2025-12-29 16:00:15","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":746668,"visible":true,"origin":"","legend":"","description":"","filename":"ANovelMicrobialDissolutionReprecipitationPathwayforBiosinteringofLimestoneforBiocementProduction.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7268390/v1_covered_45f1a874-593f-4006-bf8e-244766a8ccd4.pdf"},{"id":90407939,"identity":"798a097b-9b5d-4e0f-a25f-0fd0fa136c81","added_by":"auto","created_at":"2025-09-02 11:33:55","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":72625,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFile.docx","url":"https://assets-eu.researchsquare.com/files/rs-7268390/v1/8ed5ed0241d7c1ed71512357.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"A Novel Microbial Dissolution - 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This research demonstrates mimicking natural cement via bio-sintering of limestone, a process that involves successive dissolution and reprecipitation of limestone facilitated by bacteria under ambient environmental conditions. When the bacterium \u003cem\u003eAcetobacter aceti\u003c/em\u003e (ATCC 15973) was introduced into a mixture of ethanol and limestone powder, the pH dropped rapidly, leading to the dissolution of limestone into calcium acetate. After ethanol was fully consumed, the pH gradually increased due to acetate oxidation, causing biocement crystals to precipitate. All reaction rates were measured, and the products characterized through Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy, Quantitative X-ray Diffraction, and a Particle Size Analyzer. The detailed characterization indicates that the precipitate is mainly calcite and that the dissolved calcium carbonate sinters microbially. This pathway offers new opportunities in biocementation research by using limestone as a direct calcium source, reducing the overall carbon footprint, and eliminating the need for urea or other chemical additives. It provides a foundation for developing sustainable, low-carbon cementing materials suitable for next-generation construction applications.\u003c/p\u003e","manuscriptTitle":"A Novel Microbial Dissolution - Reprecipitation Pathway for Biosintering of Limestone for Biocement Production","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-02 11:33:51","doi":"10.21203/rs.3.rs-7268390/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","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}}],"origin":"","ownerIdentity":"7cd7bb51-2903-44f1-948a-68cc4069a267","owner":[],"postedDate":"September 2nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-29T15:58:50+00:00","versionOfRecord":{"articleIdentity":"rs-7268390","link":"https://doi.org/10.1007/s00253-025-13658-0","journal":{"identity":"applied-microbiology-and-biotechnology","isVorOnly":false,"title":"Applied Microbiology and Biotechnology"},"publishedOn":"2025-12-23 15:56:56","publishedOnDateReadable":"December 23rd, 2025"},"versionCreatedAt":"2025-09-02 11:33:51","video":"","vorDoi":"10.1007/s00253-025-13658-0","vorDoiUrl":"https://doi.org/10.1007/s00253-025-13658-0","workflowStages":[]},"version":"v1","identity":"rs-7268390","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7268390","identity":"rs-7268390","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}