{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:19:19Z","timestamp":1792804759411,"version":"4.5.0"},"reference-count":50,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2022,9,16]],"date-time":"2022-09-16T00:00:00Z","timestamp":1663286400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Irish Department of Agriculture Food and the Marine","award":["13\/F\/463"],"award-info":[{"award-number":["13\/F\/463"]}],"id":[{"id":"https:\/\/ror.org\/008gjgb19","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Irish Department of Agriculture Food and the Marine","award":["12\/RI\/2346"],"award-info":[{"award-number":["12\/RI\/2346"]}],"id":[{"id":"https:\/\/ror.org\/008gjgb19","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Science Foundation Ireland","award":["13\/F\/463"],"award-info":[{"award-number":["13\/F\/463"]}],"id":[{"id":"https:\/\/ror.org\/0271asj38","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Science Foundation Ireland","award":["12\/RI\/2346"],"award-info":[{"award-number":["12\/RI\/2346"]}],"id":[{"id":"https:\/\/ror.org\/0271asj38","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The naturally occurring sulphur-containing histidine derivative, ergothioneine (EGT), exhibits potent antioxidant properties and has been proposed to confer human health benefits. Although it is only produced by select fungi and prokaryotes, likely to protect against environmental stress, the GRAS organism Saccharomyces cerevisiae does not produce EGT naturally. Herein, it is demonstrated that the recombinant expression of a single gene, Aspergillus fumigatus egtA, in S. cerevisiae results in EgtA protein presence which unexpectedly confers complete EGT biosynthetic capacity. Both High Performance Liquid Chromatography (HPLC) and LC\u2013mass spectrometry (MS) analysis were deployed to detect and confirm EGT production in S. cerevisiae. The localisation and quantification of the resultant EGT revealed a significantly (p &lt; 0.0001) larger quantity of EGT was extracellularly present in culture supernatants than intracellularly accumulated in 96 h yeast cultures. Methionine addition to cultures improved EGT production. The additional expression of two candidate cysteine desulfurases from A. fumigatus was thought to be required to complete EGT biosynthesis, namely AFUA_2G13295 and AFUA_3G14240, termed egt2a and egt2b in this study. However, the co-expression of egtA and egt2a in S. cerevisiae resulted in a significant decrease in the observed EGT levels (p &lt; 0.05). The AlphaFold prediction of A. fumigatus EgtA 3-Dimensional structure illuminates the bidomain structure of the enzyme and the opposing locations of both active sites. Overall, we clearly show that recombinant S. cerevisiae can biosynthesise and secrete EGT in an EgtA-dependent manner which presents a facile means of producing EGT for biotechnological and biomedical use.<\/jats:p>","DOI":"10.3390\/ijms231810832","type":"journal-article","created":{"date-parts":[[2022,9,18]],"date-time":"2022-09-18T23:39:22Z","timestamp":1663544362000},"page":"10832","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["A Single Aspergillus fumigatus Gene Enables Ergothioneine Biosynthesis and Secretion by Saccharomyces cerevisiae"],"prefix":"10.3390","volume":"23","author":[{"given":"Sean","family":"Doyle","sequence":"first","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daragh D.","family":"Cuskelly","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"},{"name":"Kerry Group, Naas, Co., W91 W923 Kildare, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9534-1742","authenticated-orcid":false,"given":"Niall","family":"Conlon","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"},{"name":"Pfizer, Grangecastle, Co., D22 V8F8 Dublin, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7345-6998","authenticated-orcid":false,"given":"David A.","family":"Fitzpatrick","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ciara B.","family":"Gilmartin","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophia H.","family":"Dix","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gary W.","family":"Jones","sequence":"additional","affiliation":[{"name":"Department of Biology, Maynooth University, Maynooth, Co., W23 F2K8 Kildare, Ireland"},{"name":"Centre for Biomedical Science Research, School of Health, Leeds Beckett University, Leeds LS1 3HE, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6632","DOI":"10.1021\/ja101721e","article-title":"In vitro reconstitution of Mycobacterial ergothioneine biosynthesis","volume":"132","author":"Seebeck","year":"2010","journal-title":"J. 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