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Recently, ADP-d<\/jats:sc>-glycero<\/jats:italic>-\u03b2<\/jats:italic>-d<\/jats:sc>-manno<\/jats:italic>-heptose (ADP-d<\/jats:sc>,d<\/jats:sc>-manno<\/jats:italic>-heptose) and its C-6\u2032\u2032 epimer, ADP-l<\/jats:sc>-glycero<\/jats:italic>-\u03b2<\/jats:italic>-d<\/jats:sc>-manno<\/jats:italic>-heptose (ADP-l<\/jats:sc>,d<\/jats:sc>-manno<\/jats:italic>-heptose), were identified as potent pathogen-associated molecular patterns (PAMPs) that can trigger robust innate immune responses. Although the production of ADP-d<\/jats:sc>,d<\/jats:sc>-manno<\/jats:italic>-heptose has been studied in several different pathogenic Gram-negative bacteria, current knowledge of ADP-\u03b2<\/jats:italic>-d<\/jats:sc>-manno<\/jats:italic>-heptose biosynthesis in Vibrio<\/jats:italic> strains remains limited. Here, we characterized the biosynthetic enzymes of ADP-d<\/jats:sc>,d<\/jats:sc>-manno<\/jats:italic>-heptose and the epimerase that converts it to ADP-l<\/jats:sc>,d<\/jats:sc>-manno<\/jats:italic>-heptose from Vibrio cholerae<\/jats:italic> (the causative agent of pandemic cholera) and Vibrio parahaemolyticus<\/jats:italic> (non-cholera pathogen causing vibriosis with clinical manifestations of gastroenteritis and wound infections) in comparison with their isozymes from Escherichia coli<\/jats:italic>. Moreover, we discovered that \u03b2<\/jats:italic>-d<\/jats:sc>-mannose 1-phosphate, but not \u03b1<\/jats:italic>-d<\/jats:sc>-mannose 1-phosphate, could be activated to its ADP form by the nucleotidyltransferase domains of bifunctional kinase\/nucleotidyltransferases HldEVC<\/jats:sub> (from V. cholerae<\/jats:italic>) and HldEVP<\/jats:sub> (from V. parahaemolyticus<\/jats:italic>). Kinetic analyses of the nucleotidyltransferase domains of HldEVC<\/jats:sub> and HldEVP<\/jats:sub> together with the E. coli<\/jats:italic>\u2013derived HldEEC<\/jats:sub> were thus carried out using \u03b2<\/jats:italic>-d<\/jats:sc>-mannose 1-phosphate as a mimic sugar substrate. Overall, our works suggest that V. cholerae<\/jats:italic> and V. parahaemolyticus<\/jats:italic> are capable of synthesizing ADP-\u03b2<\/jats:italic>-d<\/jats:sc>-manno<\/jats:italic>-heptoses and lay a foundation for further physiological function explorations on manno<\/jats:italic>-heptose metabolism in Vibrio<\/jats:italic> strains.<\/jats:p>\n <\/jats:sec>\n \n Key points<\/jats:title>\n \n \u2022 Vibrio strains adopt the same biosynthetic pathway as E. coli in synthesizing ADP-\u03b2-<\/jats:italic>\n d<\/jats:sc>\n -manno-heptoses.<\/jats:italic>\n <\/jats:p>\n \n \u2022 HldEs from two Vibrio strains and E. coli could activate \u03b2-<\/jats:italic>\n d<\/jats:sc>\n -mannose 1-phosphate to ADP-\u03b2-<\/jats:italic>\n d<\/jats:sc>\n -mannose.<\/jats:italic>\n <\/jats:p>\n \n \u2022 Comparable nucleotidyltransfer efficiencies were observed in the kinetic studies of HldEs.<\/jats:italic>\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1007\/s00253-024-13108-3","type":"journal-article","created":{"date-parts":[[2024,3,18]],"date-time":"2024-03-18T09:02:20Z","timestamp":1710752540000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Characterization of the ADP-\u03b2-d-manno-heptose biosynthetic enzymes from two pathogenic Vibrio strains"],"prefix":"10.1007","volume":"108","author":[{"given":"Zhaoxiang","family":"Shi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yue","family":"Tang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhenyi","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zijian","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingming","family":"Jia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9862-182X","authenticated-orcid":false,"given":"Yihua","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,3,18]]},"reference":[{"issue":"8","key":"13108_CR1","doi-asserted-by":"publisher","first-page":"2385","DOI":"10.4049\/jimmunol.1801012","volume":"201","author":"IA Adekoya","year":"2018","unstructured":"Adekoya IA, Guo CX, Gray-Owen SD, Cox AD, Sauvageau J (2018) d-glycero-\u03b2-d-manno-heptose 1-phosphate and d-glycero-\u03b2-d-manno-heptose 1,7-biphosphate are both innate immune agonists. 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