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Using Limosilactobacillus fermentum in combination with Saccharomyces cerevisiae, optimal activity occurred after 48 h at 30 °C, with antioxidant activity, α-amylase inhibition, and α-glucosidase inhibition reported. Fermented milk reduced pro-inflammatory cytotoxins and ROS in LPS-induced RAW 267.4 macrophage cells, while peptide fractions were separated by RP-HPLC and identified via BIOPEP database comparison. Molecular docking showed strong binding to human α-amylase and α-glucosidase, and further in vivo validation was recommended.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/exploring-the-molecular-interaction-of-bioactive-peptides-with-antioxidative-antidiabetic-and-anti-inflammatory-attributes-from-sheep-milk-fermented-with-lactobacillus-and-yeast/459783/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/exploring-the-molecular-interaction-of-bioactive-peptides-with-antioxidative-antidiabetic-and-anti-inflammatory-attributes-from-sheep-milk-fermented-with-lactobacillus-and-yeast/459783.png","ImageObject",300,407,{"name":92,"@type":93},"Rizky","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-31","2026-09-30",true,{"@type":102,"interactionType":103,"userInteractionCount":29},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"Which microorganisms were used to ferment sheep milk in this study?","Question",{"text":112,"@type":113},"Fermentation used Limosilactobacillus fermentum (KGL4, MTCC 25515) combined with Saccharomyces cerevisiae (WBS2A, MG101828).","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What fermentation conditions produced the best bioactivity?",{"text":117,"@type":113},"Optimal bioactivity was observed after 48 hours of fermentation at 30 °C.",{"name":119,"@type":110,"acceptedAnswer":120},"How were the generated peptides separated and identified?",{"text":121,"@type":113},"Ultrafiltered water-soluble extracts were fractionated by reverse-phase HPLC, and identified sequences were assessed by comparison with the BIOPEP database to confirm antidiabetic and antioxidative activities.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},459783,1790813036,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":29,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":46,"language":139,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":140,"faqs":141,"seo_title":142,"seo_description":67,"update_tm":143,"read_time":31,"preview_page_count":4},962085564807,"https://ap-avatar.wpscdn.com/davatar_6f874abed73319feea01a86fa6f0fab8","[www. nature.com/scientificreports](www. nature.com/scientificreports)  \nOPEN  \nExploring the molecular interaction of bioactive peptides with antioxidative, antidiabetic and anti-inflammatory attributes from sheep milk fermented with lactobacillus and yeast  \nPrashantkumar Natubhai Padhiyar1, Pooja M. Mankad2, AmarA. Sakure3, Arka Bhattacharya4, Kanthi Kiran Kondepudi4, Brij Pal Singh5, Souparno Paul6, Preetam Sarkar7, Ashish Patel8, Shaikh Adil9􀀍 & Subrota Hati1􀀍  \nSheep milk is a rich source of proteins with potential to generate bioactive peptides through microbial fermentation. However, limited studies have focused on the Panchali sheep breed of India and the multifunctional health properties of its fermented products. This study investigated the generation of bioactive peptides with antidiabetic, antioxidative, and anti-inflammatory properties through the fermentation of sheep milk using a potent LAB, Limosilactobacillus fermentum (KGL4, MTCC 25515), in combination with theyeast strain Saccharomyces cerevisiae (WBS2A, MG101828). Optimal bioactivity was observed after 48 h of fermentation at 30 °C, with antioxidant activity reaching 40.08%, α-amylase inhibition at 72.48%, and α-glucosidase inhibition at 75.96% . Proteolytic activity was assessed at varying inoculum concentrations (1.5, 2 and 2.5%) and incubation time (0, 12, 24, 36 and 48 h), with the highest activity (6.74 mg/mL) observed at a 2.5% inoculum after 48 h. Additionally, fermented milk effectively lowered the production of pro-inflammatory cytotoxins and ROS in LPS-induced RAW 267.4 macrophage cells, indicating its anti-inflammatory properties. Protein profiling through SDS-PAGE revealed bands between 15 and 70 kDa, while 2D electrophoresis identified 39 distinct protein spots associated with peptide production. Reverse-phase HPLC of ultrafiltered water-soluble extracts (WSEs) was used to separate peptide fractions, and identified sequences were assessed through comparison with BIOPEP database to confirm antidiabetic and antioxidative activities. The most potent selected peptide showed strong binding affinity and stable interaction within the active pockets of human α-amylase and α-glucosidase, as well as high HADDOCK and HPEPDOCK scores in molecular docking analyses. These findings indicate that fermented sheep milk is a promising source of multifunctional bioactive peptides with potential applications in functional foods targeting diabetes and inflammation. Further in vivo studies are warranted to validate physiological relevance, bioavailability, and health benefits.  \n[www. nature.com/scientificreports/](www. nature.com/scientificreports/)  \nKeywords Antidiabetic, Anti-inflammatory, Antioxidative, Bioactive properties, Fermentation, Sheep milk  \nAbbreviations  \nABTS 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)  \nALA α-linolenic acid  \nATR Attenuated total reflection  \nCLA Conjugated linoleic acids  \nCLSM Confocal laser scanning microscopy  \nDCFDA 2’,7’-dichlorofluorescin diacetate DMSO Dimethyl sulfoxide  \nDNSA 3,5-dinitrosalicylic acid  \nELISA Enzyme-linked immunosorbent assay  \nEMS Enhanced MS  \nEPI Enhanced product ion  \nESI Electrospray ionization  \nFAO Food and agriculture organization  \nFBS Fetal bovine serum  \nFITC Fluorescein isothiocyanate FSM Fermented sheep milk  \nFTIR Fourier-transform infrared spectroscopy  \nIDA Information-dependent acquisition  \nIL-1β Interleukin 1 beta  \nIL-6 Interleukin 6  \nIPG Immobilized pH gradient  \nLAB Lactic acid bacteria  \nLC Liquid chromatography  \nLPS Lipopolysaccharide  \nMPL Milk polar lipids  \nMS Mass spectrometry  \nMTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide OPA o-phthaldialdehyde  \nP-NPG p-nitrophenyl-α-D-glucopyranoside  \nRP-HPLC Reverse phase high-performance liquid chromatography SDS-PAGE Sodium dodecyl sulfate–polyacrylamide gel electrophoresis TFA Trifluoroacetic acid  \nTNF-α Tumor necrosis factor alpha  \nWSE Water-soluble extracts  \n1Department of Dairy Microbiology, SMC ","cbCaiaJJmZCRnwnd","https://ap.wps.com/l/cbCaiaJJmZCRnwnd","pdf",6777524,"English","# Abstract\n# Study Rationale\n# Fermentation Conditions and Bioactivity Results\n# Proteolytic Activity and Protein Profiling\n# Peptide Separation and Identification\n# Molecular Docking and Mechanistic Insights\n# Conclusions and Future Work","[{\"question\":\"Which microorganisms were used to ferment sheep milk in this study?\",\"answer\":\"Fermentation used Limosilactobacillus fermentum (KGL4, MTCC 25515) combined with Saccharomyces cerevisiae (WBS2A, MG101828).\"},{\"question\":\"What fermentation conditions produced the best bioactivity?\",\"answer\":\"Optimal bioactivity was observed after 48 hours of fermentation at 30 °C.\"},{\"question\":\"How were the generated peptides separated and identified?\",\"answer\":\"Ultrafiltered water-soluble extracts were fractionated by reverse-phase HPLC, and identified sequences were assessed by comparison with the BIOPEP database to confirm antidiabetic and antioxidative activities.\"}]","Exploring the molecular interaction of bioactive peptides with antioxidative, antidiabetic and anti-inflammatory attributes from sheep milk fermented with lactobacillus and yeast | PDF",1790756893]