[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-450215-105":59,"doc-detail-450215-en":134},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":127,"head_meta":129,"extra_data":131,"updated_unix":133},105,"en","engineering-galactoside-acetyltransferase-for-enhanced-hesperetin-7-o-glucoside-bioavailability","Engineering galactoside acetyltransferase for enhanced hesperetin-7-O-glucoside bioavailability","","Flavonoid glycosides show reduced bioavailability because glycosylated structures permeate cell membranes poorly. Enzymatic acetylation was used to enhance bioavailability: Hesperetin-7-O-glucoside (Hes-7-G) was acetylated by galactoside acetyltransferase (GAT), and Caco-2 apparent permeability increased by 69%, supporting therapeutic potential. GAT mutants were then engineered with computational and experimental approaches to improve acetylation efficiency and define the catalytic mechanism. Molecular dynamics identified key residues governing binding dynamics and proton transfer, while P148A elevated catalytic efficiency and interaction analysis clarified substrate regioselectivity, establishing an engineering framework and sustainable strategy for improving flavonoid pharmacokinetics.",{"@graph":69,"@context":126},[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":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/engineering-galactoside-acetyltransferase-for-enhanced-hesperetin-7-o-glucoside-bioavailability/450215/",{"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/engineering-galactoside-acetyltransferase-for-enhanced-hesperetin-7-o-glucoside-bioavailability/450215.png","ImageObject",300,407,{"name":92,"@type":93},"Asher","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-07","2026-09-30",true,{"@type":102,"interactionType":103,"userInteractionCount":39},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118,122],{"name":109,"@type":110,"acceptedAnswer":111},"Why was enzymatic acetylation applied to flavonoid glycosides in this study?","Question",{"text":112,"@type":113},"Flavonoid glycosides have compromised bioavailability due to low membrane permeability. Enzymatic acetylation was used to increase bioavailability by improving cellular transport properties.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What evidence shows that GAT improves Hes-7-G bioavailability?",{"text":117,"@type":113},"The study reports that Hes-7-O-glucoside was acetylated by GAT, and the apparent permeability (Papp) of the Caco-2 monolayer increased by 69%, indicating enhanced absorption potential.",{"name":119,"@type":110,"acceptedAnswer":120},"Which residues and steps were identified as key to GAT catalysis?",{"text":121,"@type":113},"Molecular dynamics indicated Tyr483 and Met127 control flavonoid binding via dynamic van der Waals interactions. His115 and Thr113 were linked to proton transfer, accounting for most catalytic activity (85–90%).",{"name":123,"@type":110,"acceptedAnswer":124},"How did the P148A mutation affect GAT catalytic performance?",{"text":125,"@type":113},"Rational substitution of Pro148 with alanine (P148A) increased flexibility in the cofactor binding ring and improved catalytic efficiency, increasing Kcat/KM by 21%.","https://schema.org",{"og:url":83,"og:type":128,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":130,"canonical":83},"index,follow",{"doc_id":132,"site_id":62},450215,1790812393,{"code":4,"msg":5,"data":135},{"doc_id":132,"user_id":136,"nickname":92,"user_avatar":137,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":138,"file_id":139,"file_url":140,"file_type":141,"file_size":142,"view_count":39,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":143,"language":144,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":145,"faqs":146,"seo_title":147,"seo_description":67,"update_tm":148,"read_time":149},687197207639,"https://ap-avatar.wpscdn.com/davatar_a8503ba1806abce46bf441b54a3ca4cd","Applied Microbiology and Biotechnology (2026) 110:2  \n[https://doi.org/10.1007/s00253-025-13661-5](https://doi.org/10.1007/s00253-025-13661-5)  \nEngineering galactoside acetyltransferase for enhanced hesperetin‑7‑O‑glucoside bioavailability  \nJia‑Xin Wang1 · Zi‑Feng Lin3 · Xin‑Yu Zheng1 · Jin‑Lin Zhou2 · Jia‑Jun Huang3 · Yu‑Jing Lu1,2  \nReceived: 2 April 2025 / Revised: 17 October 2025 / Accepted: 24 November 2025 © The Author(s) 2026  \nAbstract  \nFlavonoid glycosides exhibit compromised bioavailability due to low membrane permeability. To address this limitation, we acetylated flavonoids through enzymatic reactions to increase bioavailability. This study first reported that Hesperetin-7-Oglucoside (Hes-7-G) was acetylated by galactoside acetyltransferase (GAT), and the apparent permeability (Papp) of the Caco-2 monolayer was increased by 69%, indicating the acetylated Hes-7-G application potential to improve bioavailability. Subsequently, we designed GAT mutants through comprehensive computational and experimental methods to improve the acetylation efficiency and elucidate the catalytic mechanism. Molecular Dynamics (MD) simulations found that Tyr483 and Met127 are key residues that control flavonoid binding through dynamic van der Waals interactions, while His115 and Thr113 mediated proton transfer accounts for 85–90% of the catalytic activity. Rational substitution of Pro148 with alanine (P148A) increased the flexibility of the cofactor binding ring and increased the catalytic efficiency (Kcat/KM) by 21% . Average noncovalent interaction (aNCI) analysis revealed that regional selectivity in the glucose portion was controlled by hydrophobic interactions with Tyr483 and hydrogen bonding with Gly125, and rhamnose substitution caused spatial conflict. This work deciphered the structure-activity relationship of GAT, established a framework for protein engineering, and highlighted enzyme-driven acetylation as a sustainable strategy for optimizing flavonoid pharmacokinetics.  \nKey points  \n• Engineered acetyltransferase enhances flavonoid glycoside absorption.  \n• P148A mutation improves catalytic efficiency.  \n• Insight into the catalytic mechanism of GAT by flavonoid glycoside substrates.  \nKeywords Flavonoid glycoside · Acetyltransferase engineering · Bioavailability optimization · MD simulation · ANCI analysis  \nIntroduction  \nFlavonoids, a diverse family of polyphenolic secondary metabolites ubiquitously present in plants, have garnered considerable attention for their extensive biological activities, including antioxidant, anti-inflammatory, and anticancer effects (Maleki et al. 2019 ; Tungmunnithum et al. 2018 ; Agati et al. 2012 ; Khachatoorian et al. 2012) . Hesperetin-7-O-glucoside (Hes-7-G), a compound found in flavonoids, has drawn our attention due to its ability to modulate the metabolism of microbial flora and exhibit potent antiinflammatory properties (Wu et al. 2021b, 2021a, 2022) . However, the clinical translation of flavonoid glycosides is hindered by their inherently low bioavailability, which can be attributed to three primary limitations: poor membrane permeability of glycosylated structures, inefficient transport  \nacross biological barriers, and susceptibility to phase II metabolic transformations in intestinal epithelia (Thilakarathna and Rupasinghe 2013) .  \nHowever, conventional chemical acetylation strategies that rely on pyridine catalysts and elevated temperatures encounter significant challenges in achieving regioselectivity control and ensuring environmental compatibility (Xiao et al. 2010 ; Biely et al. 2014 ; Araújo et al. 2017 ; Wang et al. 2022) . In this context, enzymatic acylation has emerged asa green alternative, offering precise regiospecificity and sustainable production methods (Liu and Kong 2018) . Galactoside acetyltransferase (GAT), a versatile acyltransferase capable of multi-site modification on glucosides, has drawn increasing interest. In spite of this, the current research mainl","cbCaigPnL8oZNA3e","https://ap.wps.com/l/cbCaigPnL8oZNA3e","pdf",1908278,15,"English","# Abstract\n## Key points\n## Keywords\n## Introduction","[{\"question\":\"Why was enzymatic acetylation applied to flavonoid glycosides in this study?\",\"answer\":\"Flavonoid glycosides have compromised bioavailability due to low membrane permeability. Enzymatic acetylation was used to increase bioavailability by improving cellular transport properties.\"},{\"question\":\"What evidence shows that GAT improves Hes-7-G bioavailability?\",\"answer\":\"The study reports that Hes-7-O-glucoside was acetylated by GAT, and the apparent permeability (Papp) of the Caco-2 monolayer increased by 69%, indicating enhanced absorption potential.\"},{\"question\":\"Which residues and steps were identified as key to GAT catalysis?\",\"answer\":\"Molecular dynamics indicated Tyr483 and Met127 control flavonoid binding via dynamic van der Waals interactions. His115 and Thr113 were linked to proton transfer, accounting for most catalytic activity (85–90%).\"},{\"question\":\"How did the P148A mutation affect GAT catalytic performance?\",\"answer\":\"Rational substitution of Pro148 with alanine (P148A) increased flexibility in the cofactor binding ring and improved catalytic efficiency, increasing Kcat/KM by 21%.\"}]","Engineering galactoside acetyltransferase for enhanced hesperetin-7-O-glucoside bioavailability | PDF",1790732477,38]