{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:43:11Z","timestamp":1792784591199,"version":"4.5.0"},"reference-count":37,"publisher":"American Chemical Society (ACS)","issue":"37","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"name":"National Natural Science Foundation of China (NSFC)","award":["32172204"],"award-info":[{"award-number":["32172204"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China (NSFC)","award":["32672951"],"award-info":[{"award-number":["32672951"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\u03b1-Amylase inhibitors often exhibit markedly inconsistent activities across substrate systems, yet the molecular basis remains poorly understood. Here, using epigallocatechin gallate (EGCG) as a model inhibitor, we compared corn starch, a continuous \u03b1-1,4-glucan, with the small, structurally defined chromogenic substrate GalG2CNP, demonstrating that substrate structural organization, rather than the inhibitor itself, governs apparent inhibitory outcomes. Their distinct enzyme affinities produced opposing responses: low-affinity GalG2CNP was readily inhibited by EGCG, whereas high-affinity starch resisted inhibition even with excess inhibitor. Multimodal biophysics, molecular docking, and molecular dynamics simulations confirmed that EGCG occupies the \u03b1-amylase active pocket without substantially perturbing enzyme structure. Ternary system analyses further revealed that high-affinity starch displaced prebound EGCG, whereas GalG2CNP induced only partial re-equilibration. We therefore propose a substrate-affinity-driven tug-of-war model and a two-tier screening strategy, using artificial substrates for primary screening and starch for physiological verification, providing a mechanistic framework for more reliably evaluating candidate dietary \u03b1-amylase inhibitors.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c11481","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T16:01:50Z","timestamp":1788883310000},"page":"29607-29623","source":"Crossref","is-referenced-by-count":0,"title":["Substrate \u03b1-1,4-Glucan\nStructural Organization\nDetermines EGCG Inhibition of \u03b1-Amylase through an Enzyme-Substrate-Inhibitor\nTug-of-War"],"prefix":"10.1021","volume":"74","author":[{"given":"Chunming","family":"Zou","sequence":"first","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jifan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liping","family":"Qin","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Puying","family":"Chen","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1097-4365","authenticated-orcid":true,"given":"Yutang","family":"Wang","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuebo","family":"Liu","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4814-6485","authenticated-orcid":true,"given":"Lijun","family":"Sun","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"issue":"6A","key":"2026092310085257600_cit1","doi-asserted-by":"publisher","first-page":"4H","DOI":"10.1016\/S0002-9149(01)01830-6","article-title":"Pathophysiologic\nMechanisms of Postprandial Hyperglycemia","volume":"88","author":"Gavin","year":"2001","journal-title":"Am.\nJ. 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Nutr."},{"key":"2026092310085257600_cit4","doi-asserted-by":"publisher","first-page":"137749","DOI":"10.1016\/j.foodchem.2023.137749","article-title":"The Inhibition Mechanism of Bound Polyphenols Extracted from Mung\nBean Coat Dietary Fiber on Porcine Pancreatic \u03b1-Amylase: Kinetic,\nSpectroscopic, Differential Scanning Calorimetric and Molecular Docking","volume":"436","author":"Sun","year":"2024","journal-title":"Food Chem."},{"key":"2026092310085257600_cit5","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1016\/j.tifs.2020.08.003","article-title":"Inhibition of \u03b1-Amylase by\nPolyphenolic Compounds: Substrate Digestion, Binding Interactions\nand Nutritional Intervention","volume":"104","author":"Sun","year":"2020","journal-title":"Trends Food Sci.\nTechnol."},{"key":"2026092310085257600_cit6","doi-asserted-by":"publisher","first-page":"133047","DOI":"10.1016\/j.foodchem.2022.133047","article-title":"Enzymic Catalyzing\nAffinity to Substrate Affects Inhibitor-Enzyme\nBinding Interactions: Inhibition Behaviors of EGCG against Starch\nDigestion by Individual and Co-Existing \u03b1-Amylase and Amyloglucosidase","volume":"388","author":"Zhu","year":"2022","journal-title":"Food Chem."},{"key":"2026092310085257600_cit7","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1016\/j.jff.2015.10.003","article-title":"Inhibition of Human \u03b1-Amylase\nby Dietary Polyphenols","volume":"19","author":"Nyambe-Silavwe","year":"2015","journal-title":"J. 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Nutr. Food Res."},{"key":"2026092310085257600_cit25","doi-asserted-by":"publisher","first-page":"146815","DOI":"10.1016\/j.foodchem.2025.146815","article-title":"Ligand-Enzyme\nThermodynamic Bindings Provide an Efficient and Microscale\nEvaluating Approach for \u03b1-Amylase Inhibition of Polyphenols","volume":"496","author":"Zhang","year":"2025","journal-title":"Food Chem."},{"issue":"8","key":"2026092310085257600_cit26","doi-asserted-by":"publisher","first-page":"4173","DOI":"10.1039\/C8FO00562A","article-title":"Inhibition\nof \u03b1-Glucosidase and \u03b1-Amylase\nby Flavonoid Glycosides from Lu\u2019an GuaPian Tea: Molecular Docking\nand Interaction Mechanism","volume":"9","author":"Hua","year":"2018","journal-title":"Food Funct."},{"issue":"13","key":"2026092310085257600_cit27","doi-asserted-by":"publisher","first-page":"2174","DOI":"10.3390\/foods14132174","article-title":"Effect of\nSugarcane Polyphenol Extract on \u03b1-Amylase Inhibition\nand Mechanism Exploration","volume":"14","author":"Wang","year":"2025","journal-title":"Foods"},{"issue":"14","key":"2026092310085257600_cit28","doi-asserted-by":"publisher","first-page":"4526","DOI":"10.3390\/molecules27144526","article-title":"Molecular\nDocking and Dynamics Simulation of Natural Compounds from Betel Leaves\n(Piper betle L.) for Investigating the Potential Inhibition of Alpha-Amylase\nand Alpha-Glucosidase of Type 2 Diabetes","volume":"27","author":"Ahmed","year":"2022","journal-title":"Molecules"},{"issue":"12","key":"2026092310085257600_cit29","doi-asserted-by":"publisher","first-page":"10195","DOI":"10.3390\/ijms241210195","article-title":"Assessments\nof Alpha-Amylase Inhibitory Potential of Tagetes Flavonoids through\nin Vitro, Molecular Docking, and Molecular Dynamics Simulation Studies","volume":"24","author":"Mohamed","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"2026092310085257600_cit30","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1016\/j.abb.2019.02.001","article-title":"Atomic Force\nMicroscopy for Single Molecule Characterization of Protein Aggregation","volume":"664","author":"Ruggeri","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"issue":"1","key":"2026092310085257600_cit31","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1515\/amylase-2018-0002","article-title":"Inhibition\nStudies on \u03b1-Amylase Using Isothermal Titration Calorimetry","volume":"2","author":"Lehoczki","year":"2018","journal-title":"Amylase"},{"issue":"6","key":"2026092310085257600_cit32","first-page":"3622","article-title":"Mutual\nInteractions\nbetween \u03b1-Amylase and Starch Substrates with Different Structures","volume":"13","author":"Zhou","year":"2022","journal-title":"Food Funct."},{"key":"2026092310085257600_cit33","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.foodhyd.2017.12.011","article-title":"Soluble Polysaccharides Reduce Binding\nand Inhibitory Activity of Tea Polyphenols against Porcine Pancreatic\n\u03b1-Amylase","volume":"79","author":"Sun","year":"2018","journal-title":"Food Hydrocolloids"},{"key":"2026092310085257600_cit34","doi-asserted-by":"crossref","DOI":"10.1002\/9781118540398","volume-title":"Evaluation of Enzyme\nInhibitors in Drug Discovery: a Guide for Medicinal Chemists and Pharmacologists","author":"Copeland","year":"2013","edition":"2nd"},{"key":"2026092310085257600_cit35","doi-asserted-by":"publisher","first-page":"114504","DOI":"10.1016\/j.foodres.2024.114504","article-title":"Structure\u2013Function\nRelationships in (Poly)phenol\u2013Enzyme Binding: Direct Inhibition\nof Human Salivary and Pancreatic \u03b1-Amylases","volume":"188","author":"Visvanathan","year":"2024","journal-title":"Food Res. Int."},{"key":"2026092310085257600_cit36","doi-asserted-by":"publisher","first-page":"17931","DOI":"10.1021\/acs.jmedchem.4c02052","article-title":"Dose\u2013Response Curves and the Determination of\nIC50 and EC50 Values","volume":"67","author":"Srinivasan","year":"2024","journal-title":"J. Med. Chem."},{"issue":"12","key":"2026092310085257600_cit37","first-page":"5396","article-title":"Maltoheptaoside\nHydrolysis with Chromatographic Detection and Starch Hydrolysis with\nReducing Sugar Analysis: Comparison of Assays Allows Assessment of\nthe Roles of Direct \u03b1-Amylase Inhibition and Starch Complexation","volume":"12","author":"Barber","year":"2021","journal-title":"Food Funct."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c11481","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/37\/29607\/68450059\/acs.jafc.6c11481.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/37\/29607\/68450059\/acs.jafc.6c11481.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T14:43:13Z","timestamp":1790174593000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/37\/29607\/5426442\/Substrate-1-4-Glucan-Structural-Organization"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":37,"journal-issue":{"issue":"37","published-online":{"date-parts":[[2026,9,8]]},"published-print":{"date-parts":[[2026,9,23]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c11481","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]}}}