{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T16:51:21Z","timestamp":1791823881530,"version":"4.3.4"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"33","license":[{"start":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T00:00:00Z","timestamp":1786752000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T00:00:00Z","timestamp":1786752000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T00:00:00Z","timestamp":1786752000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"Department of Science and Technology of Sichuan Province","doi-asserted-by":"publisher","award":["2023ZHJY0022"],"award-info":[{"award-number":["2023ZHJY0022"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32272280"],"award-info":[{"award-number":["32272280"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,26]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>In this work, the protective mechanism of exopolysaccharide (EPS) on Lacticaseibacillus rhamnosus during freeze-drying was revealed through physiological, proteomic, and molecular dynamics simulation analyses. Physiological analysis suggested that the addition of EPS could enhance lactate dehydrogenase (LDH) and ATPase activities, improve cell membrane integrity, and raise the content of unsaturated fatty acids. Proteomic analysis revealed that EPS significantly upregulated the expression of LDH, regulated energy metabolism and nucleotide metabolism, and enhanced the repair ability of genetic material and the synthesis ability of proteins. Furthermore, molecular dynamics simulations indicated that EPS might form hydrogen bonds with the cell surface and inhibit ice crystal formation, thereby protecting cells from extreme freeze-drying conditions. The results presented in this study suggested that EPS might act as a physical barrier for probiotics, reducing the damage caused by freeze-drying, which would be conducive to the further application of EPS as a natural freeze-drying protective agent.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c07363","type":"journal-article","created":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T15:01:48Z","timestamp":1786806108000},"page":"26351-26363","source":"Crossref","is-referenced-by-count":1,"title":["Protective Mechanisms\nof Exopolysaccharide on\n                    <i>Lacticaseibacillus rhamnosus<\/i>\n                    During Freeze-Drying:\nInsights from Physiological Characteristics, Proteomic Analysis, and\nMolecular Dynamics Simulation"],"prefix":"10.1021","volume":"74","author":[{"given":"Min","family":"Zhang","sequence":"first","affiliation":[{"name":"Sichuan University , , ,","place":["Chengdu, China, 610065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jintao","family":"Xu","sequence":"additional","affiliation":[{"name":"Sichuan University , , ,","place":["Chengdu, China, 610065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huan","family":"Yang","sequence":"additional","affiliation":[{"name":"Sichuan University , , ,","place":["Chengdu, China, 610065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7964-7856","authenticated-orcid":true,"given":"Rongqing","family":"Zhou","sequence":"additional","affiliation":[{"name":"Sichuan University , , ,","place":["Chengdu, China, 610065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5896-3086","authenticated-orcid":true,"given":"Chongde","family":"Wu","sequence":"additional","affiliation":[{"name":"Sichuan University , , ,","place":["Chengdu, China, 610065"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,15]]},"reference":[{"issue":"1","key":"2026090104471745600_cit1","doi-asserted-by":"publisher","DOI":"10.1111\/1541-4337.13287","article-title":"Drying of probiotics to enhance the viability during preparation,\nstorage, food application, and digestion: A review","volume":"23","author":"Wang","year":"2024","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"issue":"9","key":"2026090104471745600_cit2","doi-asserted-by":"publisher","first-page":"7308","DOI":"10.3168\/jds.2021-21546","article-title":"Protective effects\nof whey protein hydrolysate on Bifidobacterium animalis ssp. lactis Probio-M8 during freeze-drying and\nstorage","volume":"105","author":"Wang","year":"2022","journal-title":"J. Dairy Sci."},{"key":"2026090104471745600_cit3","doi-asserted-by":"publisher","first-page":"100145","DOI":"10.1016\/j.fochms.2022.100145","article-title":"Effects of galactosyltransferase\non EPS biosynthesis\nand freeze-drying resistance of Lactobacillus acidophilus NCFM","volume":"5","author":"Kong","year":"2022","journal-title":"Food Chem. Mol. Sci."},{"key":"2026090104471745600_cit4","doi-asserted-by":"publisher","first-page":"104425","DOI":"10.1016\/j.tifs.2024.104425","article-title":"Research progress on improving the freeze-drying resistance\nof probiotics:\nA review","volume":"147","author":"Ge","year":"2024","journal-title":"Trends Food Sci. Technol."},{"key":"2026090104471745600_cit5","doi-asserted-by":"publisher","first-page":"108463","DOI":"10.1016\/j.lwt.2019.108463","article-title":"Protective\neffect of polysaccharides from Pholiota nameko on Lactobacillus casei ATCC 334 subjected\nto freeze-drying","volume":"115","author":"Haiping","year":"2019","journal-title":"LWT- Food Sci. Technol."},{"key":"2026090104471745600_cit6","doi-asserted-by":"publisher","first-page":"108496","DOI":"10.1016\/j.lwt.2019.108496","article-title":"Protective effects of \u03b2-glucan extracted from\nspent brewer yeast during freeze-drying, storage and exposure to simulated\ngastrointestinal conditions of probiotic lactobacilli","volume":"116","author":"da Silva Guedes","year":"2019","journal-title":"LWT - Food Sci. Technol."},{"issue":"3","key":"2026090104471745600_cit7","doi-asserted-by":"publisher","first-page":"2606","DOI":"10.3168\/jds.2020-19110","article-title":"Polysaccharides can improve the survival of Lactiplantibacillus plantarum subjected to freeze-drying","volume":"104","author":"Wang","year":"2021","journal-title":"J. Dairy Sci."},{"issue":"4","key":"2026090104471745600_cit8","doi-asserted-by":"publisher","first-page":"3066","DOI":"10.3168\/jds.2019-17685","article-title":"Influence of freezing\ntemperature before freeze-drying on the viability of various Lactobacillus plantarum strains","volume":"103","author":"Wang","year":"2020","journal-title":"J.\nDairy Sci."},{"issue":"7","key":"2026090104471745600_cit9","doi-asserted-by":"publisher","first-page":"308","DOI":"10.3390\/md22070308","article-title":"Protective effects of Laminaria japonica polysaccharide\ncomposite microcapsules on the survival of Lactobacillus plantarum during simulated gastrointestinal digestion and heat treatment","volume":"22","author":"Guo","year":"2024","journal-title":"Mar. Drugs"},{"key":"2026090104471745600_cit10","doi-asserted-by":"publisher","first-page":"111083","DOI":"10.1016\/j.foodres.2022.111083","article-title":"Structural characterization\nand bioactivity\nof novel exopolysaccharides produced by Tetragenococcus halophilus","volume":"155","author":"Zhang","year":"2022","journal-title":"Food Res. Int."},{"key":"2026090104471745600_cit11","doi-asserted-by":"publisher","first-page":"133304","DOI":"10.1016\/j.foodchem.2022.133304","article-title":"A novel exopolysaccharide\nproduced by Zygosaccharomyces rouxii with cryoprotective and freeze-drying protective activities","volume":"392","author":"Zhang","year":"2022","journal-title":"Food Chem."},{"issue":"6","key":"2026090104471745600_cit12","doi-asserted-by":"publisher","first-page":"3559","DOI":"10.1002\/jsfa.13240","article-title":"Improvement\nof the viability of Tetragenococcus halophilus under\nacidic stress by forming the biofilm cell structure based on RNA-Seq\nand iTRAQ analyses","volume":"104","author":"Yao","year":"2024","journal-title":"J. Sci. Food Agric."},{"issue":"2","key":"2026090104471745600_cit13","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1007\/s00253-011-3757-6","article-title":"A combined physiological\nand proteomic approach to reveal lactic-acid-induced\nalterations in Lactobacillus casei Zhang and its\nmutant with enhanced lactic acid tolerance","volume":"93","author":"Wu","year":"2012","journal-title":"Appl. Microbiol. Biotechnol."},{"issue":"9","key":"2026090104471745600_cit14","doi-asserted-by":"publisher","first-page":"1681","DOI":"10.4014\/jmb.1702.02060","article-title":"Effect of exogenous proline on metabolic\nresponse of Tetragenococcus halophilus under salt\nstress","volume":"27","author":"He","year":"2017","journal-title":"J. Microbiol Biotechn."},{"issue":"D1","key":"2026090104471745600_cit15","doi-asserted-by":"publisher","first-page":"D1211","DOI":"10.1093\/nar\/gky869","article-title":"iProX: an integrated proteome resource","volume":"47","author":"Ma","year":"2019","journal-title":"Nucleic\nAcids Res."},{"issue":"D1","key":"2026090104471745600_cit16","doi-asserted-by":"publisher","first-page":"D1522","DOI":"10.1093\/nar\/gkab1081","article-title":"iProX in 2021: connecting\nproteomics data sharing with big data","volume":"50","author":"Chen","year":"2022","journal-title":"Nucleic\nAcids Res."},{"key":"2026090104471745600_cit17","doi-asserted-by":"publisher","first-page":"139263","DOI":"10.1016\/j.ijbiomac.2024.139263","article-title":"Understanding the mechanism of saccharides\ntype and concentration affecting texture of freeze-dried pectin-CMC\ncryogels through experiment and molecular dynamic simulation","volume":"292","author":"Ma","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"2026090104471745600_cit18","doi-asserted-by":"publisher","first-page":"109915","DOI":"10.1016\/j.foodhyd.2024.109915","article-title":"Elucidate the molecular basis of\nampholytic chitosan as a high-performance cryoprotectant to myosin\ndenaturation: The importance of saccharide charges","volume":"152","author":"Zhang","year":"2024","journal-title":"Food Hydrocolloids"},{"key":"2026090104471745600_cit19","doi-asserted-by":"publisher","first-page":"122312","DOI":"10.1016\/j.carbpol.2024.122312","article-title":"Higher molecular weight\npectin inhibits ice crystal growth and its effect on the microstructural\nand physical properties of pectin cryogels","volume":"340","author":"Ma","year":"2024","journal-title":"Carbohydr. Polym."},{"issue":"1","key":"2026090104471745600_cit20","doi-asserted-by":"publisher","first-page":"1323","DOI":"10.1038\/s41467-021-21717-z","article-title":"The atomistic\ndetails of the ice recrystallisation inhibition activity of PVA","volume":"12","author":"Bachtiger","year":"2021","journal-title":"Nat. Commun."},{"key":"2026090104471745600_cit21","doi-asserted-by":"publisher","first-page":"109579","DOI":"10.1016\/j.foodhyd.2023.109579","article-title":"Retarding ice recrystallization\nby tamarind seed polysaccharide: Investigation in ice cream mixes\nand insights from molecular dynamics simulation","volume":"149","author":"Sun","year":"2024","journal-title":"Food Hydrocoll."},{"key":"2026090104471745600_cit22","doi-asserted-by":"publisher","first-page":"129468","DOI":"10.1016\/j.ijbiomac.2024.129468","article-title":"Sodium carboxymethyl\ncelluloses as a cryoprotectant for survival improvement of lactic\nacid bacterial strains subjected to freeze-drying","volume":"260","author":"Liu","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"issue":"8","key":"2026090104471745600_cit23","doi-asserted-by":"publisher","first-page":"1724","DOI":"10.1021\/acs.jafc.6b05508","article-title":"Effect of drying methods on protein and DNA conformation\nchanges in Lactobacillus rhamnosus GG cells by Fourier\ntransform infrared spectroscopy","volume":"65","author":"Hlaing","year":"2017","journal-title":"J. Agr. Food\nChem."},{"key":"2026090104471745600_cit24","doi-asserted-by":"publisher","first-page":"109519","DOI":"10.1016\/j.foodhyd.2023.109519","article-title":"Stabilising and functional effects\nof Spirulina (Arthrospira platensis) protein isolate\non encapsulated Lacticaseibacillus rhamnosus GG during\nprocessing, storage\nand gastrointestinal digestion","volume":"149","author":"Fortuin","year":"2024","journal-title":"Food Hydrocoll."},{"issue":"8","key":"2026090104471745600_cit25","doi-asserted-by":"publisher","first-page":"R152","DOI":"10.1111\/j.1750-3841.2011.02347.x","article-title":"Role of glassy state\non stabilities of freeze-dried probiotics","volume":"76","author":"Santivarangkna","year":"2011","journal-title":"J.\nFood Sci."},{"key":"2026090104471745600_cit26","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/j.foodres.2015.03.016","article-title":"Membrane fluidity-related\nadaptive response mechanisms of foodborne bacterial pathogens under\nenvironmental stresses","volume":"72","author":"Yoon","year":"2015","journal-title":"Food Res. Int."},{"key":"2026090104471745600_cit27","doi-asserted-by":"publisher","first-page":"126849","DOI":"10.1016\/j.foodchem.2020.126849","article-title":"Effects of buffer salts\non the freeze-drying survival rate of Lactobacillus plantarum LIP-1 based on transcriptome and proteome analyses","volume":"326","author":"Jingjing","year":"2020","journal-title":"Food Chem."},{"key":"2026090104471745600_cit28","doi-asserted-by":"publisher","first-page":"110694","DOI":"10.1016\/j.foodres.2021.110694","article-title":"Effects of different\ninitial pH values on freeze-drying resistance\nof Lactiplantibacillus plantarum LIP-1 based on transcriptomics\nand proteomics","volume":"149","author":"E","year":"2021","journal-title":"Food Res. Int."},{"issue":"6","key":"2026090104471745600_cit29","doi-asserted-by":"publisher","first-page":"2355","DOI":"10.1016\/j.fshw.2023.03.039","article-title":"The protective\neffect of oleic acid on Lactiplantibacillus plantarum during freeze-drying","volume":"12","author":"Xia","year":"2023","journal-title":"Food Sci. Hum. Wellness"},{"issue":"4","key":"2026090104471745600_cit30","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1038\/s41929-023-01103-2","article-title":"A non-carboxylative route for the efficient synthesis\nof central metabolite malonyl-CoA and its derived products","volume":"7","author":"Li","year":"2024","journal-title":"Nat. Catal."},{"issue":"2","key":"2026090104471745600_cit31","doi-asserted-by":"publisher","first-page":"253","DOI":"10.20892\/j.issn.2095-3941.2019.0284","article-title":"ABC transporters\nand the hallmarks of cancer: roles in cancer aggressiveness beyond\nmultidrug resistance","volume":"17","author":"Muriithi","year":"2020","journal-title":"Cancer Biol. Med."},{"key":"2026090104471745600_cit32","doi-asserted-by":"publisher","first-page":"138259","DOI":"10.1016\/j.foodchem.2023.138259","article-title":"Molecular\nmodelling studies reveal cryoprotective mechanism\nof L-Proline during the frozen storage of shrimp (Litopenaeus vannamei)","volume":"441","author":"Zhang","year":"2024","journal-title":"Food Chem."},{"issue":"7","key":"2026090104471745600_cit33","doi-asserted-by":"publisher","first-page":"3877","DOI":"10.1021\/acs.biomac.3c00958","article-title":"Antifreeze polysaccharides from wheat\nbran: the structural characterization and antifreeze mechanism","volume":"25","author":"Yang","year":"2024","journal-title":"Biomacromolecules"},{"key":"2026090104471745600_cit34","doi-asserted-by":"publisher","first-page":"117568","DOI":"10.1016\/j.lwt.2025.117568","article-title":"Tea polysaccharide\u2019s\nrole in inhibited protein alterations of Hypophthalmichthys molitrix and in-depth molecular interaction mechanism during\nfluctuated frozen storage","volume":"220","author":"Walayat","year":"2025","journal-title":"LWT - Food Sci. Technol."},{"key":"2026090104471745600_cit35","doi-asserted-by":"publisher","first-page":"111828","DOI":"10.1016\/j.foodhyd.2025.111828","article-title":"Effect of pomegranate\npeel polysaccharides on gelation properties\nand freeze-thaw stability of grass carp surimi and mechanism","volume":"171","author":"Wang","year":"2026","journal-title":"Food Hydrocoll."},{"key":"2026090104471745600_cit36","doi-asserted-by":"publisher","first-page":"134824","DOI":"10.1016\/j.foodchem.2022.134824","article-title":"Molecular\ndynamics simulation of the interaction of food proteins\nwith small molecules","volume":"405","author":"Hu","year":"2023","journal-title":"Food Chem."},{"key":"2026090104471745600_cit37","doi-asserted-by":"publisher","first-page":"103180","DOI":"10.1016\/j.fbio.2023.103180","article-title":"Scope of bacterial\nsurface effector molecules beyond probiotics","volume":"56","author":"Bhawal","year":"2023","journal-title":"Food Biosci."},{"key":"2026090104471745600_cit38","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1016\/B978-0-12-818607-7.00024-4","article-title":"Cell surface protein\nenrichment for biomarker and drug target discovery using mass spectrometry-based\nproteomics","volume-title":"Proteomic And Metabolomic Approaches\nTo Biomarker Discovery","author":"Ye","year":"2020"},{"key":"2026090104471745600_cit39","doi-asserted-by":"publisher","first-page":"105164","DOI":"10.1016\/j.tifs.2025.105164","article-title":"Revolutionizing\ncryopreservation: Bio-based cryoprotectants,\nmechanisms, and advanced applications for sustainable pan-food systems","volume":"163","author":"Liu","year":"2025","journal-title":"Trends Food Sci. Technol."},{"key":"2026090104471745600_cit40","doi-asserted-by":"publisher","first-page":"137352","DOI":"10.1016\/j.foodchem.2023.137352","article-title":"Interaction\nmechanism between surface layer protein and yeast mannan: Insights\nfrom multi-spectroscopic and molecular dynamics simulation analyses","volume":"433","author":"Fu","year":"2024","journal-title":"Food Chem."},{"key":"2026090104471745600_cit41","doi-asserted-by":"publisher","first-page":"109393","DOI":"10.1016\/j.lwt.2020.109393","article-title":"Trehalose and alginate oligosaccharides\naffect the stability of myosin in whiteleg shrimp (Litopenaeus vannamei): The water-replacement mechanism confirmed by molecular\ndynamic simulation","volume":"127","author":"Zhang","year":"2020","journal-title":"LWT - Food Sci. Technol."},{"key":"2026090104471745600_cit42","doi-asserted-by":"publisher","first-page":"143212","DOI":"10.1016\/j.foodchem.2025.143212","article-title":"Rational design\nof phospholipase D to enhance thermostability and catalytic activity\nfor efficient biocatalytic phosphatidylserine synthesis","volume":"474","author":"Zhang","year":"2025","journal-title":"Food Chem."},{"key":"2026090104471745600_cit43","doi-asserted-by":"publisher","first-page":"118224","DOI":"10.1016\/j.lwt.2025.118224","article-title":"Rosmarinic\nacid improved the binding pattern and gel properties of low sodium\nmyofibrillar protein and Pediococcus acidilactici exopolypolysaccharide","volume":"229","author":"Wang","year":"2025","journal-title":"LWT - Food Sci. Technol."},{"key":"2026090104471745600_cit44","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1016\/j.foodchem.2018.11.087","article-title":"Insights into ice-growth\ninhibition by trehalose and alginate oligosaccharides in peeled Pacific\nwhite shrimp (Litopenaeus vannamei) during frozen\nstorage","volume":"278","author":"Zhang","year":"2019","journal-title":"Food Chem."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c07363","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/33\/26351\/66604527\/acs.jafc.6c07363.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/33\/26351\/66604527\/acs.jafc.6c07363.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T08:47:34Z","timestamp":1788252454000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/33\/26351\/5259303\/Protective-Mechanisms-of-Exopolysaccharide-on"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,15]]},"references-count":44,"journal-issue":{"issue":"33","published-online":{"date-parts":[[2026,8,15]]},"published-print":{"date-parts":[[2026,8,26]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c07363","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,15]]}}}