{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T11:26:26Z","timestamp":1788348386438,"version":"build-2803163510"},"reference-count":50,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2002,10,1]],"date-time":"2002-10-01T00:00:00Z","timestamp":1033430400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2002,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The expression of heterologous bacterial glycosyltransferases is of interest for potential application in the emerging field of carbohydrate engineering in gram-positive organisms. To assess the feasibility of using enzymes from gram-negative bacteria, the functional expression of the genes\n                    <jats:italic>wbaP<\/jats:italic>\n                    (formerly\n                    <jats:italic>rfbP<\/jats:italic>\n                    ),\n                    <jats:italic>wecA<\/jats:italic>\n                    (formerly\n                    <jats:italic>rfe<\/jats:italic>\n                    ), and\n                    <jats:italic>wbbO<\/jats:italic>\n                    (formerly\n                    <jats:italic>rfbF<\/jats:italic>\n                    ) from enterobacterial lipopolysaccharide O-polysaccharide biosynthesis pathways was examined in\n                    <jats:italic>Bacillus subtilis<\/jats:italic>\n                    . WbaP and WecA are initiation enzymes for O-polysaccharide formation, catalyzing the transfer of galactosyl 1-phosphate from UDP-galactose and\n                    <jats:italic>N<\/jats:italic>\n                    -acetylglucosaminyl 1-phosphate from UDP-\n                    <jats:italic>N<\/jats:italic>\n                    -acetylglucosamine, respectively, to undecaprenylphosphate. The WecA product (undecaprenylpyrophosphoryl GlcNAc) is used as an acceptor to which the bifunctional\n                    <jats:italic>wbbO<\/jats:italic>\n                    gene product sequentially adds a galactopyranose and a galactofuranose residue from the corresponding UDP sugars to form a lipid-linked trisaccharide. Genes were cloned into the shuttle vectors pRB374 and pAW10. In\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    hosts, the genes were effectively transcribed under the\n                    <jats:italic>vegII<\/jats:italic>\n                    promoter control of pRB374, but the plasmids were susceptible to rearrangements and deletion. In contrast, pAW10-based constructs, in which genes were cloned downstream of the\n                    <jats:italic>tet<\/jats:italic>\n                    resistance cassette, were stable but yielded lower levels of enzyme activity. In vitro glycosyltransferase assays were performed in\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    and\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    , using membrane preparations as sources of enzymes and endogenous undecaprenylphosphate as an acceptor. Incorporation of radioactivity from UDP-\u03b1-\n                    <jats:sc>d<\/jats:sc>\n                    -\n                    <jats:sup>14<\/jats:sup>\n                    C-sugar into reaction products verified the functionality of WbaP, WecA, and WbbO in either host. Enzyme activities in\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    varied between 20 and 75% of those measured in\n                    <jats:italic>E. coli.<\/jats:italic>\n                  <\/jats:p>","DOI":"10.1128\/aem.68.10.4722-4730.2002","type":"journal-article","created":{"date-parts":[[2002,9,25]],"date-time":"2002-09-25T14:02:42Z","timestamp":1032962562000},"page":"4722-4730","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":27,"title":["Functional Expression of Enterobacterial O-Polysaccharide Biosynthesis Enzymes in\n                    <i>Bacillus subtilis<\/i>"],"prefix":"10.1128","volume":"68","author":[{"given":"Christina","family":"Scha\u0308ffer","sequence":"first","affiliation":[{"name":"Zentrum fu\u0308r Ultrastrukturforschung and Ludwig Boltzmann-Institut fu\u0308r Molekulare Nanotechnologie, Universita\u0308t fu\u0308r Bodenkultur Wien, A-1180 Vienna, Austria"},{"name":"Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Wugeditsch","sequence":"additional","affiliation":[{"name":"Zentrum fu\u0308r Ultrastrukturforschung and Ludwig Boltzmann-Institut fu\u0308r Molekulare Nanotechnologie, Universita\u0308t fu\u0308r Bodenkultur Wien, A-1180 Vienna, Austria"},{"name":"Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Messner","sequence":"additional","affiliation":[{"name":"Zentrum fu\u0308r Ultrastrukturforschung and Ludwig Boltzmann-Institut fu\u0308r Molekulare Nanotechnologie, Universita\u0308t fu\u0308r Bodenkultur Wien, A-1180 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chris","family":"Whitfield","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(00)99871-6"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.176.22.7079-7084.1994"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.5631930.x"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(92)90048-T"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M106481200"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.14.4614-4621.1992"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.177.19.5411-5418.1995"},{"key":"e_1_3_2_9_2","first-page":"1321","volume":"5","year":"1999","unstructured":"Drummelsmith, J., and C. 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