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Here we identify an \u03b11,3-fucosyltransferase from Akkermansia muciniphila (AkkFT) that functions as a lactose-selective \u03b11,3-fucosylation module for HMO synthesis. AkkFT preferentially acts on glycans containing a lactose unit and shows no detectable conversion of type-1 or type-2 N-acetyllactosamine acceptors under standard conditions, contrasting with commonly used bacterial \u03b11,3\/4-fucosyltransferases. This lactose-anchored selectivity enables late-stage \u03b11,3-fucosylation across neutral and sialylated HMO acceptors, including efficient formation of lacto-N-neofucopentaose V and the blood group B antigen. Notably, AkkFT converts disialyl-lacto-N-tetraose to disialyl-lacto-N-fucopentaose V under conditions where Helicobacter pylori \u03b11,3-fucosyltransferases show no detectable activity. AkkFT also converts free 2\u2032-fucosyllactose to lactodifucotetraose on a preparative scale and was implemented in an engineered Escherichia coli strain for whole-cell 3-fucosyllactose production. Overall, AkkFT expands the repertoire of selectivity-defined biosynthetic parts for fucosylated HMO production and demonstrates how enzyme discovery can complement engineered microbial platforms for glycan biomanufacturing.<\/jats:p>","DOI":"10.1021\/acssynbio.6c00427","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T23:02:21Z","timestamp":1790204541000},"page":"3966-3974","source":"Crossref","is-referenced-by-count":0,"title":["A Lactose-Selective \u03b11,3-Fucosyltransferase from\n                    <i>Akkermansia muciniphila<\/i>\n                    Expands the Biosynthetic Toolkit for Fucosylated Human Milk Oligosaccharides"],"prefix":"10.1021","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8821-2371","authenticated-orcid":false,"given":"Cheng-Huan","family":"Liu","sequence":"first","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3332-2739","authenticated-orcid":false,"given":"Kai-Eng","family":"Ooi","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9134-3255","authenticated-orcid":false,"given":"Mei-Ting","family":"Lin","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yung-Jen","family":"Cheng","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8993-0256","authenticated-orcid":true,"given":"Guan-Chun","family":"Wu","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan-Yu","family":"Hsieh","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3199-5434","authenticated-orcid":true,"given":"Hsiang-En","family":"Chen","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1856-9815","authenticated-orcid":false,"given":"Po-Ting","family":"Chen","sequence":"additional","affiliation":[{"name":"Southern Taiwan University of Science and Technology , , 1, Nantai Street, Yungkang District , ,","place":["Tainan, Taiwan, 710301"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2103-6419","authenticated-orcid":true,"given":"Ching-Ching","family":"Yu","sequence":"additional","affiliation":[{"name":"National Tsing Hua University , , 101, Section 2, Kuang-Fu Road , ,","place":["Hsinchu, Taiwan, 300044"]},{"name":"Institute of Biological Chemistry, Academia Sinica , 128, Academia Road Sec. 2, Nankang , ,","place":["Taipei, Taiwan, 115201"]},{"name":"Kaohsiung Medical University , , 100, Shiquan first Road , ,","place":["Kaohsiung, Taiwan, 807378"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,23]]},"reference":[{"key":"2026101610072636500_cit1","doi-asserted-by":"publisher","first-page":"7767","DOI":"10.1039\/d4ob01006j","article-title":"Recent Progress in Chemoenzymatic Synthesis of Human Glycans","volume":"22","author":"Ma","year":"2024","journal-title":"Org. 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