Isolation and optimisation of chitin-degrading bacteria from bird faecal waste and its chitinase activity

Isolation and optimisation of chitin-degrading bacteria from bird faecal waste and its chitinase activity | 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 Isolation and optimisation of chitin-degrading bacteria from bird faecal waste and its chitinase activity Mayavati Satish Patil, Aniket Bhanudas Fasate, Samadhan Sanjabrao Deshmukh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9475559/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 Chitin-degrading microorganisms play a vital role in nutrient cycling and have significant biotechnological potential. In this study, chitinolytic bacteria were isolated from bird faecal samples using chitin-containing selective media. Six isolates were obtained, among which strain A3 exhibited the highest chitin-degrading activity, as indicated by a prominent clear zone on chitin agar. Morphological, biochemical, and phylogenetic analyses identified the isolate as Enterobacter cloacae . Quantitative estimation of enzyme activity revealed that E. cloacae produced maximum chitinase activity of 5.61 U/mL at 48 h and pH 5.5, indicating that enzyme production is strongly influenced by incubation time and acidic conditions. Medium components affecting chitinase production were screened using the Plackett–Burman design. Chitin, yeast extract, and CaCl₂ were identified as the most significant factors influencing enzyme yield. Further optimisation using the central composite design demonstrated good agreement between actual and predicted chitinase activities, although some deviations were observed at extreme factor levels. The response surface model showed strong fitting (R² = 0.914), but a lower predicted R² indicated limited predictive ability beyond the experimental range. Phylogenetic analysis confirmed the close evolutionary proximity of the isolate to known Enterobacter cloacae strains, validating its taxonomic classification. Chitinase chitin Plackett–Burman Full Text Supplementary Files finalresized920x300.jpg 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-9475559","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":627751827,"identity":"316919bd-69f1-457d-8052-3ad72b64320b","order_by":0,"name":"Mayavati Satish Patil","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA50lEQVRIiWNgGAWjYJCCAw+ABBsD8wEgJSFDhAZmhgMJYC1sIEqChygtDAlgBo8BmCSowZz9/MEDiTl1dn38Zz6/ulFjwcPAfvjoBnxaLHuSGQ4kbjuc3CaRu8065xjQYTxpaTfwaTE4ANZyIJlNgnebcQ4bUIsEjxl+Lecfg7TUJbPxn3lmnPOPGC03wLYw27Ex5DA/zm0jQovljMcGIL8ksEmkmTHn9knwsBHyizl/4uMPH7fV2cv3H378OedbnRw/++Fj+B0GpRMbgLEpAWKx4VOOrMUeiJk/EFI9CkbBKBgFIxMAAH9QSJl4R7bdAAAAAElFTkSuQmCC","orcid":"","institution":"Government Institute Of Science","correspondingAuthor":true,"prefix":"","firstName":"Mayavati","middleName":"Satish","lastName":"Patil","suffix":""},{"id":627751828,"identity":"9e9b4f17-8163-4ee9-9088-e5dab48bbd91","order_by":1,"name":"Aniket Bhanudas Fasate","email":"","orcid":"","institution":"Institute of Science","correspondingAuthor":false,"prefix":"","firstName":"Aniket","middleName":"Bhanudas","lastName":"Fasate","suffix":""},{"id":627751829,"identity":"4905df9c-a100-4649-850d-fe2a766c3f1a","order_by":2,"name":"Samadhan Sanjabrao Deshmukh","email":"","orcid":"","institution":"Institute of Science","correspondingAuthor":false,"prefix":"","firstName":"Samadhan","middleName":"Sanjabrao","lastName":"Deshmukh","suffix":""}],"badges":[],"createdAt":"2026-04-20 18:15:57","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9475559/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9475559/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108492121,"identity":"8af5e9ec-8a72-4aaf-a87b-f90ea5f1cda1","added_by":"auto","created_at":"2026-05-05 09:56:54","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":807818,"visible":true,"origin":"","legend":"","description":"","filename":"Chitinase.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9475559/v1_covered_9f1646dc-725a-473c-a2e7-53d919321a8d.pdf"},{"id":108199886,"identity":"2a9e5596-6ae6-41a1-82cf-d9f8c1af381c","added_by":"auto","created_at":"2026-04-30 11:41:21","extension":"jpg","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":83416,"visible":true,"origin":"","legend":"","description":"","filename":"finalresized920x300.jpg","url":"https://assets-eu.researchsquare.com/files/rs-9475559/v1/b2edeb9783e69c48389ef548.jpg"}],"financialInterests":"","formattedTitle":"Isolation and optimisation of chitin-degrading bacteria from bird faecal waste and its chitinase activity","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":"Chitinase, chitin, Plackett–Burman ","lastPublishedDoi":"10.21203/rs.3.rs-9475559/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9475559/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChitin-degrading microorganisms play a vital role in nutrient cycling and have significant biotechnological potential. In this study, chitinolytic bacteria were isolated from bird faecal samples using chitin-containing selective media. Six isolates were obtained, among which strain A3 exhibited the highest chitin-degrading activity, as indicated by a prominent clear zone on chitin agar. Morphological, biochemical, and phylogenetic analyses identified the isolate as \u003cem\u003eEnterobacter cloacae\u003c/em\u003e. Quantitative estimation of enzyme activity revealed that \u003cem\u003eE. cloacae\u003c/em\u003e produced maximum chitinase activity of 5.61 U/mL at 48 h and pH 5.5, indicating that enzyme production is strongly influenced by incubation time and acidic conditions. Medium components affecting chitinase production were screened using the Plackett–Burman design. Chitin, yeast extract, and CaCl₂ were identified as the most significant factors influencing enzyme yield. 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