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Here, in vitro digestion-derived residues (RSs) from five high-amylose starches (HASs; four RS2 and one RS3) were evaluated for their effects on microbiota composition and SCFA production using a simulated colonic fermentation model inoculated with human fecal microbiota. RSs with more amorphous structures and lower short-range order generated the highest SCFA levels and promoted the growth of the primary RS degrader Bifidobacterium adolescentis. In contrast, RSs with higher resistance to digestion and retained B-type crystallinity favored propionate-associated taxa, including Veillonella spp. and Bacteroides spp., whereas less resistant RSs promoted butyrate-producing taxa such as Clostridium spp. These results suggest that structural features of RS are associated with differences in microbial composition and metabolic activity, providing insights into the design of HAS-based dietary fibers with targeted prebiotic functions.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06220","type":"journal-article","created":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T17:54:16Z","timestamp":1788112456000},"page":"27206-27217","source":"Crossref","is-referenced-by-count":0,"title":["Structure of\n                    <i>In Vitro<\/i>\n                    Digestion Residues\nfrom High-Amylose Starch Impacts Colonic Microbial Composition and\nMetabolic Activity"],"prefix":"10.1021","volume":"74","author":[{"given":"Yu","family":"Tian","sequence":"first","affiliation":[{"name":"Nanjing University of Finance and Economics , , ,","place":["Nanjing, China, 210023"]},{"name":"University of Copenhagen , , ,","place":["Frederiksberg C, Denmark, DK-1871"]},{"name":"Technical University of Denmark , , ,","place":["Kgs. Lyngby, Denmark, DK-2800"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Wang","sequence":"additional","affiliation":[{"name":"Technical University of Denmark , , ,","place":["Kgs. Lyngby, Denmark, DK-2800"]},{"name":"China Pharmaceutical University , , ,","place":["Nanjing, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinming","family":"Xu","sequence":"additional","affiliation":[{"name":"Leiden University , , ,","place":["Leiden, The Netherlands, 2333BE"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6580-2034","authenticated-orcid":true,"given":"Bekzod","family":"Khakimov","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , ,","place":["Frederiksberg C, Denmark, DK-1958"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9101-3525","authenticated-orcid":true,"given":"Xingxun","family":"Liu","sequence":"additional","affiliation":[{"name":"Nanjing University of Finance and Economics , , ,","place":["Nanjing, China, 210023"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lude","family":"Zhang","sequence":"additional","affiliation":[{"name":"Xi\u2019an Hengchuang Agricultural Technology Co., Ltd. , Weiyang District, , ,","place":["Xi\u2019an City, Shaanxi Province, China, 710016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuqing","family":"Xie","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , ,","place":["Frederiksberg C, Denmark, DK-1958"]},{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6265-0314","authenticated-orcid":true,"given":"Jacob J. K.","family":"Kirkensgaard","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , ,","place":["Frederiksberg C, Denmark, DK-1958"]},{"name":"University of Copenhagen , , ,","place":["Copenhagen, Denmark, DK-2200"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim H.","family":"Hebelstrup","sequence":"additional","affiliation":[{"name":"Aarhus University , , ,","place":["Slagelse, Denmark, 4200"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dennis S.","family":"Nielsen","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , ,","place":["Frederiksberg C, Denmark, DK-1958"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2993-8196","authenticated-orcid":true,"given":"Birte","family":"Svensson","sequence":"additional","affiliation":[{"name":"Technical University of Denmark , , ,","place":["Kgs. Lyngby, Denmark, DK-2800"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6476-9546","authenticated-orcid":true,"given":"Andreas","family":"Blennow","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,24]]},"reference":[{"issue":"3","key":"2026090311503703300_cit1","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1080\/19490976.2015.1134082","article-title":"Formation of Short Chain Fatty Acids by the Gut Microbiota\nand Their Impact on Human Metabolism","volume":"7","author":"Morrison","year":"2016","journal-title":"Gut Microbes"},{"issue":"1","key":"2026090311503703300_cit2","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1038\/s41579-020-0433-9","article-title":"Gut Microbiota\nin Human Metabolic Health and Disease","volume":"19","author":"Fan","year":"2021","journal-title":"Nat.\nRev. Microbiol."},{"issue":"1","key":"2026090311503703300_cit3","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1038\/s44324-025-00066-1","article-title":"Mechanisms and Implications of the\nGut Microbial Modulation of Intestinal\nMetabolic Processes","volume":"3","author":"Jyoti","year":"2025","journal-title":"Npj Metab. Health Dis."},{"issue":"23","key":"2026090311503703300_cit4","doi-asserted-by":"publisher","first-page":"3838","DOI":"10.1016\/j.jmb.2014.07.028","article-title":"A Perspective on\nthe Complexity of Dietary Fiber Structures\nand Their Potential Effect on the Gut Microbiota","volume":"426","author":"Hamaker","year":"2014","journal-title":"J. Mol. Biol."},{"issue":"4","key":"2026090311503703300_cit5","doi-asserted-by":"publisher","first-page":"2613","DOI":"10.1021\/acs.jafc.4c08872","article-title":"Increased Crystallite\nStability Enhances\nGut Microbial Fermentability of Type 5 Resistant Starch","volume":"73","author":"Wang","year":"2025","journal-title":"J. Agric. Food Chem."},{"issue":"15","key":"2026090311503703300_cit6","doi-asserted-by":"publisher","first-page":"2509","DOI":"10.1080\/10408398.2021.1976722","article-title":"Align Resistant Starch\nStructures from Plant-Based Foods with Human\nGut Microbiome for Personalized Health Promotion","volume":"63","author":"Li","year":"2023","journal-title":"Crit. Rev. Food Sci. Nutr."},{"issue":"1","key":"2026090311503703300_cit7","doi-asserted-by":"publisher","first-page":"1926842","DOI":"10.1080\/19490976.2021.1926842","article-title":"Resistant Starch, Microbiome, and\nPrecision Modulation","volume":"13","author":"Dobranowski","year":"2021","journal-title":"Gut Microbes"},{"issue":"2","key":"2026090311503703300_cit8","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1038\/nrmicro1817","article-title":"Polysaccharide\nUtilization by Gut Bacteria: Potential for New Insights from Genomic\nAnalysis","volume":"6","author":"Flint","year":"2008","journal-title":"Nat. Rev. Microbiol."},{"key":"2026090311503703300_cit9","doi-asserted-by":"publisher","first-page":"640253","DOI":"10.3389\/fmicb.2021.640253","article-title":"In Vitro Fermentation Reveals Changes\nin Butyrate Production Dependent on Resistant Starch Source and Microbiome\nComposition","volume":"12","author":"Teichmann","year":"2021","journal-title":"Front. Microbiol."},{"issue":"9","key":"2026090311503703300_cit10","doi-asserted-by":"publisher","first-page":"2009","DOI":"10.1007\/s10068-024-01621-0","article-title":"Important\nRoles of Ruminococcaceae in the Human Intestine for\nResistant Starch Utilization","volume":"33","author":"Kim","year":"2024","journal-title":"Food Sci. Biotechnol."},{"key":"2026090311503703300_cit11","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.copbio.2019.10.008","article-title":"Resistant Starch: Impact on the Gut Microbiome and\nHealth","volume":"61","author":"DeMartino","year":"2020","journal-title":"Curr. Opin. Biotechnol."},{"key":"2026090311503703300_cit12","doi-asserted-by":"publisher","first-page":"100388","DOI":"10.1016\/j.bcdf.2023.100388","article-title":"The Prebiotic Potential of RS-3 Preparations for Pre-\nand Post-Weaning Piglets","volume":"30","author":"Klostermann","year":"2023","journal-title":"Bioact. Carbohydr.\nDiet. Fibre"},{"issue":"1","key":"2026090311503703300_cit13","doi-asserted-by":"publisher","first-page":"3502","DOI":"10.1038\/s41467-024-47594-w","article-title":"The\nGut Commensal Blautia Maintains Colonic Mucus Function under Low-Fiber\nConsumption through Secretion of Short-Chain Fatty Acids","volume":"15","author":"Holmberg","year":"2024","journal-title":"Nat. Commun."},{"issue":"2","key":"2026090311503703300_cit14","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.tim.2019.09.004","article-title":"Starch\nDigestion by Gut Bacteria: Crowdsourcing for Carbs","volume":"28","author":"Cerqueira","year":"2020","journal-title":"Trends Microbiol."},{"issue":"10","key":"2026090311503703300_cit15","doi-asserted-by":"publisher","first-page":"3209","DOI":"10.1021\/acs.jafc.0c08187","article-title":"RS5 Produced More Butyric\nAcid through\nRegulating the Microbial Community of Human Gut Microbiota","volume":"69","author":"Qin","year":"2021","journal-title":"J. Agric. Food Chem."},{"issue":"1","key":"2026090311503703300_cit16","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1016\/j.cell.2013.12.016","article-title":"Microbiota-Generated Metabolites Promote Metabolic\nBenefits via Gut-Brain Neural Circuits","volume":"156","author":"De Vadder","year":"2014","journal-title":"Cell"},{"issue":"3","key":"2026090311503703300_cit17","doi-asserted-by":"publisher","first-page":"424","DOI":"10.1038\/ijo.2014.153","article-title":"The Short\nChain Fatty Acid Propionate Stimulates GLP-1 and PYY Secretion via\nFree Fatty Acid Receptor 2 in Rodents","volume":"39","author":"Psichas","year":"2015","journal-title":"Int.\nJ. Obes."},{"issue":"7480","key":"2026090311503703300_cit18","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1038\/nature12726","article-title":"Metabolites Produced\nby Commensal Bacteria Promote Peripheral Regulatory T-Cell Generation","volume":"504","author":"Arpaia","year":"2013","journal-title":"Nature"},{"issue":"3","key":"2026090311503703300_cit19","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1016\/j.chom.2020.01.006","article-title":"Precision Microbiome Modulation with Discrete Dietary\nFiber Structures Directs Short-Chain Fatty Acid Production","volume":"27","author":"Deehan","year":"2020","journal-title":"Cell Host Microbe"},{"issue":"1","key":"2026090311503703300_cit20","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02566-18","article-title":"Dynamics of Human\nGut Microbiota and Short-Chain Fatty\nAcids in Response to Dietary Interventions with Three Fermentable\nFibers","volume":"10","author":"Baxter","year":"2019","journal-title":"mBio"},{"issue":"1","key":"2026090311503703300_cit21","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1039\/D3FO01763J","article-title":"Presence of Digestible\nStarch Impacts in Vitro Fermentation of Resistant Starch","volume":"15","author":"Klostermann","year":"2024","journal-title":"Food Funct."},{"issue":"3","key":"2026090311503703300_cit22","doi-asserted-by":"publisher","first-page":"56","DOI":"10.3390\/agronomy7030056","article-title":"Understanding\nStarch Structure: Recent Progress","volume":"7","author":"Bertoft","year":"2017","journal-title":"Agronomy"},{"issue":"23","key":"2026090311503703300_cit23","doi-asserted-by":"publisher","first-page":"6412","DOI":"10.1080\/10408398.2022.2031101","article-title":"New Definition of Resistant\nStarch Types from the Gut Microbiota\nPerspectives \u2013 a Review","volume":"63","author":"Li","year":"2023","journal-title":"Crit. Rev. Food\nSci. Nutr."},{"key":"2026090311503703300_cit24","doi-asserted-by":"publisher","first-page":"108621","DOI":"10.1016\/j.foodhyd.2023.108621","article-title":"Interfacial Enzyme Kinetics Reveals Degradation Mechanisms\nbehind Resistant Starch","volume":"140","author":"Tian","year":"2023","journal-title":"Food Hydrocolloids"},{"key":"2026090311503703300_cit25","doi-asserted-by":"publisher","first-page":"109593","DOI":"10.1016\/j.foodhyd.2023.109593","article-title":"Characterization\nof Different High\nAmylose Starch Granules. Part II: Structure Evolution during Digestion\nand Distinct Digestion Mechanisms","volume":"149","author":"Tian","year":"2024","journal-title":"Food Hydrocolloids"},{"issue":"11","key":"2026090311503703300_cit26","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1038\/s43016-020-00159-8","article-title":"A Natural Mutation\nin Pisum Sativum L. (Pea) Alters Starch Assembly\nand Improves Glucose Homeostasis\nin Humans","volume":"1","author":"Petropoulou","year":"2020","journal-title":"Nat. Food"},{"issue":"1","key":"2026090311503703300_cit27","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1186\/1471-2229-12-223","article-title":"Concerted\nSuppression of All Starch Branching Enzyme Genes in Barley Produces\nAmylose-Only Starch Granules","volume":"12","author":"Carciofi","year":"2012","journal-title":"BMC Plant Biol."},{"issue":"27","key":"2026090311503703300_cit28","doi-asserted-by":"publisher","first-page":"8568","DOI":"10.1080\/10408398.2022.2056871","article-title":"High-Amylose Starch:\nStructure, Functionality and Applications","volume":"63","author":"Zhong","year":"2023","journal-title":"Crit.\nRev. Food Sci. Nutr."},{"key":"2026090311503703300_cit29","doi-asserted-by":"publisher","first-page":"105820","DOI":"10.1016\/j.foodhyd.2020.105820","article-title":"High-Amylose Wheat\nand Maize Starches Have Distinctly\nDifferent Granule Organization and Annealing Behaviour: A Key Role\nfor Chain Mobility","volume":"105","author":"Li","year":"2020","journal-title":"Food Hydrocolloids"},{"issue":"2","key":"2026090311503703300_cit30","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1111\/1541-4337.12416","article-title":"High-Amylose\nStarches to Bridge the\n\u201cFiber Gap\u201d: Development, Structure, and Nutritional\nFunctionality","volume":"18","author":"Li","year":"2019","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"2026090311503703300_cit31","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.4268","article-title":"CoMiniGut\u2500a Small Volume in Vitro Colon Model for the Screening of Gut Microbial\nFermentation Processes","volume":"6","author":"Wiese","year":"2018","journal-title":"PeerJ"},{"issue":"5","key":"2026090311503703300_cit32","doi-asserted-by":"publisher","first-page":"875","DOI":"10.1080\/10408398.2014.922043","article-title":"Mechanisms\nof Starch Digestion by \u03b1-Amylase\u2500Structural Basis for\nKinetic Properties","volume":"57","author":"Dhital","year":"2017","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"2026090311503703300_cit33","doi-asserted-by":"publisher","first-page":"119327","DOI":"10.1016\/j.carbpol.2022.119327","article-title":"Different Genetic Strategies to Generate High Amylose\nStarch Mutants by Engineering the Starch Biosynthetic Pathways","volume":"287","author":"Zhong","year":"2022","journal-title":"Carbohydr. Polym."},{"key":"2026090311503703300_cit34","doi-asserted-by":"publisher","first-page":"109286","DOI":"10.1016\/j.foodhyd.2023.109286","article-title":"Characterization of\nDifferent High Amylose Starch Granules. Part I: Multi-Scale Structures\nand Relationships to Thermal Properties","volume":"146","author":"Tian","year":"2024","journal-title":"Food\nHydrocolloids"},{"issue":"6","key":"2026090311503703300_cit35","doi-asserted-by":"publisher","first-page":"1113","DOI":"10.1039\/C3FO60702J","article-title":"A Standardised Static in Vitro Digestion\nMethod Suitable for Food \u2013 an International Consensus","volume":"5","author":"Minekus","year":"2014","journal-title":"Food Funct."},{"key":"2026090311503703300_cit36","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1016\/j.carbpol.2016.06.016","article-title":"Compact\nStructure and Proteins of Pasta Retard in Vitro Digestive\nEvolution of Branched Starch Molecular Structure","volume":"152","author":"Zou","year":"2016","journal-title":"Carbohydr. Polym."},{"issue":"3","key":"2026090311503703300_cit37","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1093\/ajcn\/69.3.448","article-title":"Rapidly\nAvailable Glucose in Foods: An in Vitro Measurement\nThat Reflects the Glycemic Response","volume":"69","author":"Englyst","year":"1999","journal-title":"Am. J.\nClin. Nutr."},{"issue":"12","key":"2026090311503703300_cit38","doi-asserted-by":"publisher","first-page":"266","DOI":"10.1007\/s10965-018-1651-y","article-title":"Study of the Structural\nOrder of Native Starch Granules Using Combined FTIR and XRD Analysis","volume":"25","author":"Pozo","year":"2018","journal-title":"J. Polym. Res."},{"key":"2026090311503703300_cit39","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.carbpol.2015.11.066","article-title":"Infrared Spectroscopy\nas a Tool to\nCharacterise Starch Ordered Structure\u2500a Joint FTIR\u2013ATR,\nNMR, XRD and DSC Study","volume":"139","author":"Warren","year":"2016","journal-title":"Carbohydr. Polym."},{"issue":"8","key":"2026090311503703300_cit40","doi-asserted-by":"publisher","first-page":"3885","DOI":"10.1021\/acs.jafc.2c08338","article-title":"Soluble, Diferuloylated\nCorn Bran Glucuronoarabinoxylans Modulate the Human Gut Microbiota In Vitro","volume":"71","author":"Lin","year":"2023","journal-title":"J. Agric. 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