Molecular Insights into Cadaverine–Alkaline Phosphatase Interactions: Combined Spectroscopic and Computational Approaches

Molecular Insights into Cadaverine–Alkaline Phosphatase Interactions: Combined Spectroscopic and Computational Approaches | 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 Molecular Insights into Cadaverine–Alkaline Phosphatase Interactions: Combined Spectroscopic and Computational Approaches Nasim Babaknejad, Behzad Shareghi, Ali Akbar Saboury, Raheleh jahanbazi goujani This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7552310/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract The relevance of enzyme stabilization is increasing with the growing number of enzyme applications. The lack of proper stability of enzymes in processing conditions is one of the significant problems associated with using enzymes in industrial biotechnology. Natural organic molecules, such as osmolytes, offer several advantages, including increased solubility in hydrophobic substrates. In this study, we examined the effect of cadaverine as an osmolyte on the stability and kinetics of bovine alkaline phosphatase (BALP). The results revealed that cadaverine binds to the enzyme and alters its absorption spectra. Additionally, cadaverine lowers the fluorescence spectrum of the enzyme through a static quenching mechanism. According to fluorescence and docking measurements, cadaverine interacts with BALP through hydrogen bonds and van der Waals interactions. The circular dichroism spectroscopy study showed that the presence of cadaverine alters the structure of BALP. Cadaverine also enhances BALP activity by increasing Km and Vmax. Molecular simulation results further supported cadaverine binding to BALP and its impact on the structure. Overall, these findings suggest that cadaverine has the potential to stabilize BALP. Biological sciences/Biochemistry Biological sciences/Biological techniques Biological sciences/Biophysics Biological sciences/Biotechnology Biological sciences/Chemical biology Physical sciences/Chemistry BALP Cadaverine Activity Stability Molecular simulation Full Text Additional Declarations No competing interests reported. Supplementary Files suplementury.docx Cite Share Download PDF Status: Published Journal Publication published 27 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 29 Oct, 2025 Reviews received at journal 28 Oct, 2025 Reviewers agreed at journal 09 Oct, 2025 Reviews received at journal 09 Oct, 2025 Reviewers agreed at journal 09 Oct, 2025 Reviewers agreed at journal 09 Oct, 2025 Reviewers invited by journal 09 Oct, 2025 Editor invited by journal 03 Oct, 2025 Editor assigned by journal 10 Sep, 2025 Submission checks completed at journal 09 Sep, 2025 First submitted to journal 06 Sep, 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. 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-7552310","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":527364046,"identity":"8e752d0b-456c-4c6a-973c-2710bc3edd4a","order_by":0,"name":"Nasim Babaknejad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYFACHgZmhgMMDIwNDIYPQFw+UrQYG4C4bERrAQIzCRBJUIs5A+/BzwVn7sgxtx/eVvk1x06GjYH54aMbeLRYNvAlS8+48cyYsSet7LbstmSgw9iMjXPwaDE4wGMgzfPhcGJjQ47ZbcltzEAtPGzSBLQY/wZqqW/sf2NWLLmtnigtZtI8Nw4nMM7IMWP8uO0wEVoO86VZzzhz2LBxxrNiacZtx3nYmAn55Xjv4dsFxw7LG/Ynb/z4c1u1PT9788PH+LQAIwUCDBuAbB5kEYJAHogZfxCrehSMglEwCkYUAAC1K0it5giuYQAAAABJRU5ErkJggg==","orcid":"","institution":"Shahid Ashrafi Esfahani University","correspondingAuthor":true,"prefix":"","firstName":"Nasim","middleName":"","lastName":"Babaknejad","suffix":""},{"id":527364047,"identity":"82f8ff63-d5ab-4dc5-a764-1672ebbbf8c9","order_by":1,"name":"Behzad Shareghi","email":"","orcid":"","institution":"University of Shahrekord","correspondingAuthor":false,"prefix":"","firstName":"Behzad","middleName":"","lastName":"Shareghi","suffix":""},{"id":527364049,"identity":"bc43c140-8f68-4891-8ff7-25378cb742e9","order_by":2,"name":"Ali Akbar Saboury","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"prefix":"","firstName":"Ali","middleName":"Akbar","lastName":"Saboury","suffix":""},{"id":527364050,"identity":"4d2812cf-c549-4617-b563-317e6df38e79","order_by":3,"name":"Raheleh jahanbazi goujani","email":"","orcid":"","institution":"Islamic Azad University","correspondingAuthor":false,"prefix":"","firstName":"Raheleh","middleName":"jahanbazi","lastName":"goujani","suffix":""}],"badges":[],"createdAt":"2025-09-06 17:08:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7552310/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7552310/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-33248-4","type":"published","date":"2026-01-27T15:58:47+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":94157664,"identity":"30a1544f-1271-4664-8d94-d8f6e6d55ce0","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2215244,"visible":true,"origin":"","legend":"","description":"","filename":"TheeffectofcadaverineonthestructureCopy.docx","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/52fa8dcab60a2f62f39f3557.docx"},{"id":94157660,"identity":"ac1bf46b-7ced-48e8-8d6e-8052ae8f4ff4","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5852,"visible":true,"origin":"","legend":"","description":"","filename":"90c4e773fa2248598f90649e58a33712.json","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/9492c15181c7e9b79bec982e.json"},{"id":94157669,"identity":"933955f4-d824-421f-bf7e-a07d748d21ff","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":179204,"visible":true,"origin":"","legend":"","description":"","filename":"suplementury.docx","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/78e35539d996c4e5c9aa72e1.docx"},{"id":94157662,"identity":"bafd0955-fb90-4a63-9050-a137f32cdfce","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"xml","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":111188,"visible":true,"origin":"","legend":"","description":"","filename":"90c4e773fa2248598f90649e58a337121enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/0343dd6b7ce78bf2d046a16f.xml"},{"id":94158200,"identity":"18ddfb10-b684-4734-bd9e-d60ba016ff3d","added_by":"auto","created_at":"2025-10-23 03:24:10","extension":"jpeg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":487519,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/7255dc9a61fdbf48eeacea8a.jpeg"},{"id":94158203,"identity":"5750d3cd-d613-4031-86ea-8452df9a7c1b","added_by":"auto","created_at":"2025-10-23 03:24:10","extension":"emf","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9556100,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.emf","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/68ac48b1b889c96ee26ebade.emf"},{"id":94157665,"identity":"eeee3d34-34f5-4538-8211-b7e8f0c9b4d4","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":89579,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/aa95a2348c43408aa186e0f4.jpeg"},{"id":94157663,"identity":"14a8e7de-8622-4e52-b90a-7b3665d753aa","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"jpeg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":106786,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/7123ac6da1ae73cd4be998c1.jpeg"},{"id":94157661,"identity":"ea104916-c8a8-4350-9920-f8802c645c66","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"jpeg","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":64471,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/381240e9bf9671d7252f4e3b.jpeg"},{"id":94158201,"identity":"d4a53a66-c018-4b02-a459-b9ce63e7224d","added_by":"auto","created_at":"2025-10-23 03:24:10","extension":"png","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":113129,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/5160993d10ff92f4a7d0d868.png"},{"id":94157670,"identity":"f69d8e48-4877-4dab-b5e2-50748ad33caf","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":392139,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/611718a6b886119aa0e13e3d.png"},{"id":94157673,"identity":"cc43b092-0a1b-4f42-8246-3da314b25240","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"png","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":26955,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/075f8825647dd7a7c873da7e.png"},{"id":94158202,"identity":"c8bfda1f-05fc-43f6-8d08-d92d2c1ea075","added_by":"auto","created_at":"2025-10-23 03:24:10","extension":"png","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":34821,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/cbba531b2d6cf107fcc02b8c.png"},{"id":94157667,"identity":"0eef3541-0102-4775-a6f6-e93e84c4368b","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"png","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":19614,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/960fe10fcb85a2a91a83cc0c.png"},{"id":94157675,"identity":"e5216ab4-7971-4e7a-ad22-9d51fa64fb99","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"xml","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":111131,"visible":true,"origin":"","legend":"","description":"","filename":"90c4e773fa2248598f90649e58a337121structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/aff221b158805df8ee2eb38c.xml"},{"id":94157676,"identity":"a31f0e1f-f290-4a19-af29-807164d078e5","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":124261,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/9d99895be479b420861f0591.html"},{"id":101690529,"identity":"bf1a8dcc-4ff1-47fc-a121-954a9c302c90","added_by":"auto","created_at":"2026-02-02 16:04:49","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":813535,"visible":true,"origin":"","legend":"","description":"","filename":"TheeffectofcadaverineonthestructureCopy.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1_covered_2c4aa8ae-aa62-4872-a844-7f15068e18bd.pdf"},{"id":94157666,"identity":"88bad9be-95ce-420e-8746-a66d4775fa23","added_by":"auto","created_at":"2025-10-23 03:16:10","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":179204,"visible":true,"origin":"","legend":"","description":"","filename":"suplementury.docx","url":"https://assets-eu.researchsquare.com/files/rs-7552310/v1/85acc431c8dfcb024adf4903.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Molecular Insights into Cadaverine–Alkaline Phosphatase Interactions: Combined Spectroscopic and Computational Approaches","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"BALP, Cadaverine, Activity, Stability, Molecular simulation","lastPublishedDoi":"10.21203/rs.3.rs-7552310/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7552310/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe relevance of enzyme stabilization is increasing with the growing number of enzyme applications. The lack of proper stability of enzymes in processing conditions is one of the significant problems associated with using enzymes in industrial biotechnology. Natural organic molecules, such as osmolytes, offer several advantages, including increased solubility in hydrophobic substrates. In this study, we examined the effect of cadaverine as an osmolyte on the stability and kinetics of bovine alkaline phosphatase (BALP). The results revealed that cadaverine binds to the enzyme and alters its absorption spectra. Additionally, cadaverine lowers the fluorescence spectrum of the enzyme through a static quenching mechanism. According to fluorescence and docking measurements, cadaverine interacts with BALP through hydrogen bonds and van der Waals interactions. The circular dichroism spectroscopy study showed that the presence of cadaverine alters the structure of BALP. Cadaverine also enhances BALP activity by increasing Km and Vmax. Molecular simulation results further supported cadaverine binding to BALP and its impact on the structure. Overall, these findings suggest that cadaverine has the potential to stabilize BALP.\u003c/p\u003e","manuscriptTitle":"Molecular Insights into Cadaverine–Alkaline Phosphatase Interactions: Combined Spectroscopic and Computational Approaches","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-23 03:16:05","doi":"10.21203/rs.3.rs-7552310/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-29T11:47:58+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-28T09:46:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"95735049021409361989683914410841196846","date":"2025-10-10T00:31:22+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-09T20:14:09+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"325562530327715257655021514203363304048","date":"2025-10-09T19:55:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"251972419450368517290857798804801766351","date":"2025-10-09T19:42:57+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-09T19:37:28+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-10-03T06:39:23+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-10T06:19:28+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-09T12:45:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-09-06T17:02:59+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5b75531a-4beb-4514-8f6a-d30e3476df52","owner":[],"postedDate":"October 23rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":56056694,"name":"Biological sciences/Biochemistry"},{"id":56056695,"name":"Biological sciences/Biological techniques"},{"id":56056696,"name":"Biological sciences/Biophysics"},{"id":56056697,"name":"Biological sciences/Biotechnology"},{"id":56056698,"name":"Biological sciences/Chemical biology"},{"id":56056699,"name":"Physical sciences/Chemistry"}],"tags":[],"updatedAt":"2026-02-02T16:01:20+00:00","versionOfRecord":{"articleIdentity":"rs-7552310","link":"https://doi.org/10.1038/s41598-025-33248-4","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-01-27 15:58:47","publishedOnDateReadable":"January 27th, 2026"},"versionCreatedAt":"2025-10-23 03:16:05","video":"","vorDoi":"10.1038/s41598-025-33248-4","vorDoiUrl":"https://doi.org/10.1038/s41598-025-33248-4","workflowStages":[]},"version":"v1","identity":"rs-7552310","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7552310","identity":"rs-7552310","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}