{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T04:11:12Z","timestamp":1792123872308,"version":"4.4.0"},"reference-count":37,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T00:00:00Z","timestamp":1781568000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82574534"],"award-info":[{"award-number":["82574534"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["82574534"],"award-info":[{"award-number":["82574534"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Young Elite Scientists Sponsorship Program by CACM","award":["2022-QNRC2-B23"],"award-info":[{"award-number":["2022-QNRC2-B23"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Medicine food homology (MFH) plants are rich in nutrients and bioactive specialized metabolites, and their endophytic fungi mediate key biotransformation of host secondary metabolites. Licorice, a representative MFH herb, accumulates glycyrrhizin (GL) as its dominant bioactive triterpenoid saponin. Its hydrolysates glycyrrhetinic acid (GA) and glycyrrhetinic acid 3-O-mono-\u03b2-D-glucuronide (GAMG) show stronger bioactivity and bioavailability than GL. However, the enzymatic mechanisms of licorice-derived endophytic fungi-mediated GL biotransformation remain unclear. Here, nine licorice endophytic fungi were screened for significant GL-inducible \u03b2-glucuronidase activity. Four functional GH2 \u03b2-glucuronidase were obtained by prokaryotic and eukaryotic expression systems, and confirmed to catalyze glycyrrhizin biotransformation via two distinct hydrolytic pathways. Inoculation of these four strains into licorice markedly enhanced host glycyrrhizin accumulation. This study provides novel enzymatic resources for the efficient bioproduction of high-value glycyrrhizin derivatives, and proposes a green strategy to improve glycyrrhizin content in licorice, deepening the understanding of endophyte\u2013host metabolic crosstalk in medicinal herbs.<\/jats:p>","DOI":"10.3390\/ijms27125444","type":"journal-article","created":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T03:27:21Z","timestamp":1781666841000},"page":"5444","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Differential Biotransformation of Glycyrrhizin by Licorice-Derived Endophytic Fungi and Accumulation-Promoting Effects of Fungal Inoculation"],"prefix":"10.3390","volume":"27","author":[{"given":"Xin","family":"Zuo","sequence":"first","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangxi","family":"Ren","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5146-3812","authenticated-orcid":false,"given":"Dan","family":"Jiang","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunsheng","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"141111","DOI":"10.1016\/j.foodchem.2024.141111","article-title":"Medicine and food homology substances: A review of bioactive ingredients, pharmacological effects and applications","volume":"463","author":"Wang","year":"2025","journal-title":"Food Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"112439","DOI":"10.1016\/j.jep.2019.112439","article-title":"An \u201cessential herbal medicine\u201d-licorice: A review of phytochemicals and its effects in combination preparations","volume":"249","author":"Jiang","year":"2020","journal-title":"J. Ethnopharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"145332","DOI":"10.1016\/j.foodchem.2025.145332","article-title":"New insights of licorice (Glycyrrhiza genus) as a functional food","volume":"492","author":"Shao","year":"2025","journal-title":"Food Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"115686","DOI":"10.1016\/j.jep.2022.115686","article-title":"Chemical analysis of the Chinese herbal medicine licorice (Gan-Cao): An update review","volume":"299","author":"Shang","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1089\/jmf.2019.4459","article-title":"A review of the pharmacological efficacy and safety of licorice root from corroborative clinical trial findings","volume":"23","author":"Kwon","year":"2020","journal-title":"J. Med. Food"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zuo, J., Meng, T., Wang, Y., and Tang, W. (2023). A review of the antiviral activities of glycyrrhizic acid, glycyrrhetinic acid and glycyrrhetinic acid monoglucuronide. Pharmaceuticals, 16.","DOI":"10.3390\/ph16050641"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2155","DOI":"10.1038\/s41401-021-00651-2","article-title":"Pharmacokinetics-based identification of pseudoaldosterogenic compounds originating from Glycyrrhiza uralensis roots (Gancao) after dosing LianhuaQingwen capsule","volume":"42","author":"Lan","year":"2021","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Shinu, P., Gupta, G.L., Sharma, M., Khan, S., Goyal, M., Nair, A.B., Kumar, M., Soliman, W.E., Rahman, A., and Attimarad, M. (2023). Pharmacological features of 18\u03b2-Glycyrrhetinic Acid: A pentacyclic triterpenoid of therapeutic potential. Plants, 12.","DOI":"10.3390\/plants12051086"},{"key":"ref_9","first-page":"1346","article-title":"Mechanisms of fungal endophytes in plant protection against pathogens","volume":"4","author":"Gao","year":"2010","journal-title":"Afr. J. Microbiol. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"eaau6389","DOI":"10.1126\/science.aau6389","article-title":"A specialized metabolic network selectively modulates Arabidopsis root microbiota","volume":"364","author":"Huang","year":"2019","journal-title":"Science"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1111\/nph.70254","article-title":"Beyond defense: Microbial modifications of plant specialized metabolites alter and expand their ecological functions","volume":"247","author":"Unger","year":"2025","journal-title":"New Phytol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"You, C., Qin, D., Wang, Y., Lan, W., Li, Y., Yu, B., Peng, Y., Xu, J., and Dong, J. (2021). Plant triterpenoids regulate endophyte community to promote medicinal plant Schisandra sphenanthera growth and metabolites accumulation. J. Fungi, 7.","DOI":"10.3390\/jof7100788"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/j.jgg.2023.10.004","article-title":"Two diversities meet in the rhizosphere: Root specialized metabolites and microbiome","volume":"51","author":"Wang","year":"2024","journal-title":"Genet. Genom."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Jia, M., Chen, L., Xin, H.L., Zheng, C.J., Rahman, K., Han, T., and Qin, L.P. (2016). A friendly relationship between endophytic fungi and medicinal plants: A systematic review. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.00906"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1074\/jbc.M117.801910","article-title":"Structure-guided engineering of the substrate specificity of a fungal \u03b2-glucuronidase toward triterpenoid saponins","volume":"293","author":"Lv","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"16286","DOI":"10.1021\/acs.jafc.2c06859","article-title":"Tetramerization of GH2 \u03b2-Glucuronidases is essential for catalyzing the hydrolysis of the large substrate glycyrrhizin","volume":"70","author":"Wang","year":"2002","journal-title":"J. Agric. Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"9193","DOI":"10.1007\/s00253-018-9285-x","article-title":"Characteristics and molecular determinants of a highly selective and efficient glycyrrhizin-hydrolyzing \u03b2-glucuronidase from Staphylococcus pasteuri 3I10","volume":"102","author":"Wei","year":"2018","journal-title":"Appl. Microbiol. Biot."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Liu, Y., Huangfu, J., Qi, F., Kaleem, I., Wenwen, E., and Li, C. (2012). Effects of a non-conservative sequence on the properties of \u03b2-glucuronidase from Aspergillus terreus Li-20. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030998"},{"key":"ref_19","first-page":"792","article-title":"Purification and characterization of two novel \u03b2-D-glucuronidases converting glycyrrhizin to 18\u03b2-glycyrrhetinic acid-3-O-\u03b2-D-glucuronide from Streptococcus LJ-22","volume":"15","author":"Park","year":"2005","journal-title":"J. Microbiol. Biotech."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e00755-18","DOI":"10.1128\/AEM.00755-18","article-title":"A novel \u03b2-Glucuronidase from Talaromyces pinophilus Li-93 precisely hydrolyzes glycyrrhizin into glycyrrhetinic acid 3-o-mono-\u03b2-d-glucuronide","volume":"84","author":"Xu","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"140484","DOI":"10.1016\/j.ijbiomac.2025.140484","article-title":"Licorice endophytes activate glycyrrhizin synthesis metabolic flux through feedback of \u03b2-glucuronidase conversion activity","volume":"302","author":"Zuo","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"bar020","DOI":"10.1093\/database\/bar020","article-title":"Curation of characterized glycoside hydrolases of fungal origin","volume":"2011","author":"Murphy","year":"2011","journal-title":"Database"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.procbio.2023.03.035","article-title":"\u03b2-glucosidase: An overview on immobilization and some aspects of structure, function, applications and cost","volume":"130","author":"Boscolo","year":"2023","journal-title":"Process Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.molcatb.2003.11.011","article-title":"Biochemical and catalytic properties of two intracellular \u03b2-glucosidases from the fungus Penicillium decumbens active on flavonoid glucosides","volume":"27","author":"Mamma","year":"2004","journal-title":"Mol. Catal. B Enzym."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Zhu, H., Zhang, R., Huang, Z., and Zhou, J. (2023). Progress in the conversion of ginsenoside Rb1 into minor ginsenosides using \u03b2-Glucosidases. Foods, 12.","DOI":"10.3390\/foods12020397"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zhang, X., Wang, Y., Zhang, T., Yuan, Z., and Wei, Y. (2024). Efficient biotransformation of icariin to baohuoside I using two novel GH1 \u03b2-Glucosidases. Molecules, 29.","DOI":"10.3390\/molecules29225280"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"175","DOI":"10.2323\/jgam.2023.02.002","article-title":"Bacillus velezensis S141, a soybean growth-promoting bacterium, hydrolyzes isoflavone glycosides into aglycones","volume":"69","author":"Kondo","year":"2023","journal-title":"J. Gen. Appl. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"782","DOI":"10.1016\/j.ijbiomac.2020.05.047","article-title":"Purification and characterization of a novel \u03b2-glucuronidase precisely converts glycyrrhizin to glycyrrhetinic acid 3-O-mono-\u03b2-D-glucuronide from plant endophytic Chaetomium globosum DX-THS3","volume":"159","author":"Zhang","year":"2020","journal-title":"Int. J. Biol. Macromo."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/j.procbio.2012.12.008","article-title":"Purification and characterization of a highly selective glycyrrhizin-hydrolyzing \u03b2-glucuronidase from Penicillium purpurogenum Li-3","volume":"48","author":"Zou","year":"2013","journal-title":"Process Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s13007-018-0343-2","article-title":"Improving agroinfiltration-based transient gene expression in Nicotiana benthamiana","volume":"14","author":"Norkunas","year":"2018","journal-title":"Plant Methods"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"289","DOI":"10.18388\/abp.2006_3341","article-title":"Optimization of transient Agrobacterium-mediated gene expression system in leaves of Nicotiana benthamiana","volume":"53","author":"Wydro","year":"2006","journal-title":"Acta Biochim. Pol."},{"key":"ref_32","first-page":"51204","article-title":"Optimization and utilization of Agrobacterium-mediated transient protein production in Nicotiana","volume":"19","author":"Shamloul","year":"2014","journal-title":"Jove-J. Vis. Exp."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1446","DOI":"10.1038\/s41467-023-37133-4","article-title":"Revealing evolution of tropane alkaloid biosynthesis by analyzing two genomes in the Solanaceae family","volume":"14","author":"Zhang","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2289","DOI":"10.1039\/C4MB00223G","article-title":"Multi-well fungal co-culture for de novo metabolite-induction in time-series studies based on untargeted metabolomics","volume":"10","author":"Bertrand","year":"2014","journal-title":"Mol. Biosyst."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1093\/nar\/gkt1178","article-title":"The carbohydrate-active enzymes database (CAZy) in 2013","volume":"42","author":"Lombard","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of biomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/12\/5444\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T04:40:13Z","timestamp":1781844013000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/12\/5444"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,16]]},"references-count":37,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["ijms27125444"],"URL":"https:\/\/doi.org\/10.3390\/ijms27125444","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,16]]}}}