{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T15:52:32Z","timestamp":1762271552317,"version":"build-2065373602"},"reference-count":35,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2024,6,10]],"date-time":"2024-06-10T00:00:00Z","timestamp":1717977600000},"content-version":"vor","delay-in-days":9,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"publisher","award":["BB\/M01925X\/1"],"award-info":[{"award-number":["BB\/M01925X\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2024,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The application of bacterial oligosaccharyltransferases (OSTs) such as the\n                    <jats:italic>Campylobacter jejuni<\/jats:italic>\n                    PglB for glycoengineering has attracted considerable interest in glycoengineering and glycoconjugate vaccine development. However, PglB has limited specificity for glycans that can be transferred to candidate proteins, which along with other factors is dependent on the reducing end sugar of glycans. In this study, we developed a cell\u2010free glycosylation assay that offers the speed and simplicity of a \u2018yes\u2019 or \u2018no\u2019 determination. Using the assay, we tested the activity of eleven PglBs from\n                    <jats:italic>Campylobacter<\/jats:italic>\n                    species and more distantly related bacteria. The following assorted glycans with diverse reducing end sugars were tested for transfer, including\n                    <jats:italic>Streptococcus pneumoniae<\/jats:italic>\n                    capsule serotype 4,\n                    <jats:italic>Salmonella<\/jats:italic>\n                    enterica serovar Typhimurium O antigen (B1),\n                    <jats:italic>Francisella tularensis<\/jats:italic>\n                    O antigen,\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    O9 antigen and\n                    <jats:italic>Campylobacter jejuni<\/jats:italic>\n                    heptasaccharide. Interestingly, while PglBs from the same genus showed high activity, whereas divergent PglBs differed in their transfer of glycans to an acceptor protein. Notably for glycoengineering purposes,\n                    <jats:italic>Campylobacter hepaticus<\/jats:italic>\n                    and\n                    <jats:italic>Campylobacter subantarcticus<\/jats:italic>\n                    PglBs showed high glycosylation efficiency, with\n                    <jats:italic>C. hepaticus<\/jats:italic>\n                    PglB potentially being useful for glycoconjugate vaccine production. This study demonstrates the versatility of the cell\u2010free assay in rapidly assessing an OST to couple glycan\/carrier protein combinations and lays the foundation for future screening of PglBs by linking amino acid similarity to glycosyltransferase activity.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.14480","type":"journal-article","created":{"date-parts":[[2024,6,10]],"date-time":"2024-06-10T23:46:05Z","timestamp":1718063165000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Investigation into the efficiency of diverse\n                    <i>N<\/i>\n                    \u2010linking oligosaccharyltransferases for glycoengineering using a standardised cell\u2010free assay"],"prefix":"10.1111","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3478-597X","authenticated-orcid":false,"given":"Burhan","family":"Lehri","sequence":"first","affiliation":[{"name":"Department of Infection Biology London School of Hygiene and Tropical Medicine  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1783-6889","authenticated-orcid":false,"given":"Elizabeth","family":"Atkins","sequence":"additional","affiliation":[{"name":"Department of Infection Biology London School of Hygiene and Tropical Medicine  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7042-8609","authenticated-orcid":false,"given":"Timothy A.","family":"Scott","sequence":"additional","affiliation":[{"name":"Department of Medicine, Cambridge Institute of Therapeutic Immunology &amp; Infectious Disease (CITIID), Jeffrey Cheah Biomedical Centre University of Cambridge  Cambridge UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3802-3059","authenticated-orcid":false,"given":"Sherif","family":"Abouelhadid","sequence":"additional","affiliation":[{"name":"Department of Infection Biology London School of Hygiene and Tropical Medicine  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6140-9489","authenticated-orcid":false,"given":"Brendan W.","family":"Wren","sequence":"additional","affiliation":[{"name":"Department of Infection Biology London School of Hygiene and Tropical Medicine  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4538-4007","authenticated-orcid":false,"given":"Jon","family":"Cuccui","sequence":"additional","affiliation":[{"name":"Department of Infection Biology London School of Hygiene and Tropical Medicine  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,6,10]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwx067"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1155\/2010\/148178"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1080\/14760584.2020.1775077"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-313X.2006.02842.x"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1099\/00207713-43-4-631"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M807113200"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1558-5646.1985.tb00420.x"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2014.00381"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.445940"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijs.0.000048"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(17)30108-1"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1098\/rsob.140227"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05110-x"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00211-10"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwx110"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.thromres.2012.09.011"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-019-0110-z"},{"key":"e_1_2_10_19_1","first-page":"1192","article-title":"A minimal sequon sufficient for O\u2010linked glycosylation by the versatile oligosaccharyltransferase PglS","volume":"31","author":"Knoot C.J.","year":"2021","journal-title":"Glycobiology"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.2217\/fmb.09.10"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz239"},{"key":"e_1_2_10_22_1","unstructured":"LimmaTech Biologics AG. 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