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In recent years, it has become apparent that the so\u2010called non\u2010pylori <jats:italic>Helicobacters<\/jats:italic>, normally infecting pigs, cats, and dogs, may also be involved in human pathology via zoonotic transmission. Indeed, more than 30 species of non\u2010pylori <jats:italic>Helicobacters<\/jats:italic> have been identified thus far. One such organism is <jats:italic>Helicobacter canadensis<\/jats:italic>, an emerging pathogen whose genome sequence was published in 2009. Given our long\u2010standing interest in the biosynthesis of <jats:italic>N<\/jats:italic>\u2010formylated sugars found in the O\u2010antigens of some Gram\u2010negative bacteria, we were curious as to whether <jats:italic>H. canadensis<\/jats:italic> produces such unusual carbohydrates. Here, we demonstrate using both biochemical and structural techniques that the proteins encoded by the <jats:italic>HCAN_0198<\/jats:italic>, <jats:italic>HCAN_0204<\/jats:italic>, and <jats:italic>HCAN_0200<\/jats:italic> genes in <jats:italic>H. canadensis,<\/jats:italic> correspond to a 3,4\u2010ketoisomerase, a pyridoxal 5\u2032\u2010phosphate aminotransferase, and an <jats:italic>N<\/jats:italic>\u2010formyltransferase, respectively. For this investigation, five high\u2010resolution X\u2010ray structures were determined and the kinetic parameters for the isomerase and the <jats:italic>N<\/jats:italic>\u2010formyltransferase were measured. Based on these data, we suggest that the unusual sugar, 3\u2010formamido\u20103,6\u2010dideoxy\u2010<jats:sc>d<\/jats:sc>\u2010glucose, will most likely be found in the O\u2010antigen of <jats:italic>H. canadensis<\/jats:italic>. Whether <jats:italic>N<\/jats:italic>\u2010formylated sugars found in the O\u2010antigen contribute to virulence is presently unclear, but it is intriguing that they have been observed in such pathogens as <jats:italic>Francisella tularensis<\/jats:italic>, <jats:italic>Mycobacterium tuberculosis<\/jats:italic>, and <jats:italic>Brucella melitensis<\/jats:italic>.<\/jats:p>","DOI":"10.1002\/pro.4169","type":"journal-article","created":{"date-parts":[[2021,8,11]],"date-time":"2021-08-11T13:09:29Z","timestamp":1628687369000},"page":"2144-2160","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Investigation of the enzymes required for the biosynthesis of an unusual formylated sugar in the emerging human pathogen <scp><i>Helicobacter canadensis<\/i><\/scp>"],"prefix":"10.1002","volume":"30","author":[{"given":"Colton J.","family":"Heisdorf","sequence":"first","affiliation":[{"name":"Department of Biochemistry University of Wisconsin  Madison Wisconsin USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William A.","family":"Griffiths","sequence":"additional","affiliation":[{"name":"Department of Biochemistry University of Wisconsin  Madison Wisconsin USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James B.","family":"Thoden","sequence":"additional","affiliation":[{"name":"Department of Biochemistry University of Wisconsin  Madison Wisconsin USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6214-3638","authenticated-orcid":false,"given":"Hazel M.","family":"Holden","sequence":"additional","affiliation":[{"name":"Department of Biochemistry University of Wisconsin  Madison Wisconsin USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,8,31]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(84)91816-6"},{"key":"e_1_2_9_3_1","first-page":"397","article-title":"Transfer of Campylobacter pylori and Campylobacter mustelae to Helicobacter gen. 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