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There are 10 serotypes of GBS, which are distinguished by their capsular polysaccharide (CPS) structure, with serotypes Ia, Ib, II, III, IV and V responsible for up to 99% of infections. Currently, there are no licensed vaccines against GBS. The most developed candidates are glycoconjugate vaccines, which can be highly effective but are also expensive to produce by existing approaches and unaffordable for many parts of the world. Biosynthesis of recombinant glycans and glycoconjugates in tractable strains of bacteria offers a low-cost alternative approach to current chemical conjugation methods. Methods: In this study, we apply combinatorial hierarchical DNA assembly to the heterologous biosynthesis of GBS III, IV and V CPSs in E. coli. Each gene was removed from its native regulation, paired with synthetic regulatory elements and rebuilt from the bottom up to generate libraries of reconstituted pathways. These pathways were screened for glycan biosynthesis using serotype-specific antisera. Results: We identified several configurations that successfully biosynthesised the GBS CPSs. Furthermore, we exploited the conserved nature of the GBS CPS biosynthesis loci and the flexibility of modular DNA assembly by constructing hybrid pathways from a minimal pool of glycosyltransferase genes. We show that transferase genes with homologous function can be used interchangeably between pathways, obviating the need to clone a complete locus for each new CPS assembly. Conclusions: In conclusion, we report the first demonstration of heterologous GBS CPS IV and V biosynthesis in E. coli, a key milestone towards the development of low-cost recombinant multivalent GBS glycoconjugate vaccines.<\/jats:p>","DOI":"10.3390\/vaccines13030279","type":"journal-article","created":{"date-parts":[[2025,3,6]],"date-time":"2025-03-06T06:02:13Z","timestamp":1741240933000},"page":"279","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Modular Combinatorial DNA Assembly of Group B Streptococcus Capsular Polysaccharide Biosynthesis Pathways to Expediate the Production of Novel Glycoconjugate Vaccines"],"prefix":"10.3390","volume":"13","author":[{"given":"Mark A.","family":"Harrison","sequence":"first","affiliation":[{"name":"Department of Infection Biology, London School of Hygiene & Tropical Medicine, London WC1E 7HT, 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 & Tropical Medicine, London WC1E 7HT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra","family":"Faulds-Pain","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John T.","family":"Heap","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brendan W.","family":"Wren","sequence":"additional","affiliation":[{"name":"Department of Infection Biology, London School of Hygiene & Tropical Medicine, London WC1E 7HT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4418-6957","authenticated-orcid":false,"given":"Ian J.","family":"Passmore","sequence":"additional","affiliation":[{"name":"Department of Infection Biology, London School of Hygiene & Tropical Medicine, London WC1E 7HT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"E3609","DOI":"10.1073\/pnas.1518311113","article-title":"Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies","volume":"113","author":"Chen","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","first-page":"51","article-title":"Purification and characterization of a Shigella conjugate vaccine, produced by glycoengineering Escherichia coli","volume":"26","author":"Ravenscroft","year":"2016","journal-title":"Glycobiology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4152","DOI":"10.1016\/j.vaccine.2016.06.067","article-title":"Immunogenicity and safety of a tetravalent E. coli O-antigen bioconjugate vaccine in animal models","volume":"34","author":"Serroyen","year":"2016","journal-title":"Vaccine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1038\/s41589-020-0595-9","article-title":"Engineering orthogonal human O-linked glycoprotein biosynthesis in bacteria","volume":"16","author":"Natarajan","year":"2020","journal-title":"Nat. Chem. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1016\/j.ymben.2018.04.014","article-title":"A flow cytometric approach to engineering Escherichia coli for improved eukaryotic protein glycosylation","volume":"47","author":"Glasscock","year":"2018","journal-title":"Metab. Eng."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5403","DOI":"10.1038\/s41467-019-13283-2","article-title":"Cytoplasmic glycoengineering enables biosynthesis of nanoscale glycoprotein assemblies","volume":"10","author":"Tytgat","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1080\/14760584.2020.1775077","article-title":"Improving protein glycan coupling technology (PGCT) for glycoconjugate vaccine production","volume":"19","author":"Dow","year":"2020","journal-title":"Expert Rev. Vaccines"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1038\/s41541-019-0110-z","article-title":"Recent advances in the production of recombinant glycoconjugate vaccines","volume":"4","author":"Kay","year":"2019","journal-title":"npj Vaccines"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e17","DOI":"10.1093\/nar\/gky1182","article-title":"Start-Stop Assembly: A functionally scarless DNA assembly system optimized for metabolic engineering","volume":"47","author":"Taylor","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1038\/nbt.3063","article-title":"Functional optimization of gene clusters by combinatorial design and assembly","volume":"32","author":"Smanski","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7085","DOI":"10.1073\/pnas.1120788109","article-title":"Refactoring the nitrogen fixation gene cluster from Klebsiella oxytoca","volume":"109","author":"Temme","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2446","DOI":"10.1038\/s41467-020-16175-y","article-title":"Application of combinatorial optimization strategies in synthetic biology","volume":"11","author":"Naseri","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1093\/glycob\/cwac082","article-title":"A combinatorial DNA assembly approach to biosynthesis of N-linked glycans in E. coli","volume":"33","author":"Passmore","year":"2023","journal-title":"Glycobiology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S200","DOI":"10.1093\/cid\/cix664","article-title":"Estimates of the Burden of Group B Streptococcal Disease Worldwide for Pregnant Women, Stillbirths, and Children","volume":"65","author":"Seale","year":"2017","journal-title":"Clin. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e807","DOI":"10.1016\/S2214-109X(22)00093-6","article-title":"Group B streptococcus infection during pregnancy and infancy: Estimates of regional and global burden","volume":"10","author":"Procter","year":"2022","journal-title":"Lancet Glob. Health"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7307","DOI":"10.1016\/j.vaccine.2016.12.029","article-title":"WHO consultation on group B Streptococcus vaccine development: Report from a meeting held on 27\u201328 April 2016","volume":"37","author":"Kobayashi","year":"2019","journal-title":"Vaccine"},{"key":"ref_17","unstructured":"(2017). WHO Preferred Product Characteristics for Group B Streptococcus Vaccines, World Health Organization."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6930","DOI":"10.1016\/j.vaccine.2020.08.056","article-title":"Safety and immunogenicity of an investigational maternal trivalent group B streptococcus vaccine in pregnant women and their infants: Results from a randomized placebo-controlled phase II trial","volume":"38","author":"Swamy","year":"2020","journal-title":"Vaccine"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Navarro-Torn\u00e9, A., Curcio, D., Mo\u00efsi, J.C., and Jodar, L. (2021). Burden of invasive group B Streptococcus disease in non-pregnant adults: A systematic review and meta-analysis. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0258030"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1002\/aah.10132","article-title":"Prevention and Control of Streptococcosis in Tilapia Culture: A Systematic Review","volume":"33","author":"Maulu","year":"2021","journal-title":"J. Aquat. Anim. Health"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6682","DOI":"10.1016\/j.vaccine.2020.08.052","article-title":"Systematic review of Group B Streptococcal capsular types, sequence types and surface proteins as potential vaccine candidates","volume":"38","author":"Paul","year":"2020","journal-title":"Vaccine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1056\/NEJMoa2116045","article-title":"Potential for Maternally Administered Vaccine for Infant Group B Streptococcus","volume":"389","author":"Madhi","year":"2023","journal-title":"N. Engl. J. Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Martin, P., and Alaimo, C. (2022). The Ongoing Journey of a Shigella Bioconjugate Vaccine. Vaccines, 10.","DOI":"10.3390\/vaccines10020212"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1038\/s41541-024-00885-1","article-title":"A randomized phase 1\/2a trial of ExPEC10V vaccine in adults with a history of UTI","volume":"9","author":"Fierro","year":"2024","journal-title":"npj Vaccines"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3111","DOI":"10.1021\/acsinfecdis.1c00415","article-title":"Development and Immunogenicity of a Prototype Multivalent Group B Streptococcus Bioconjugate Vaccine","volume":"7","author":"Duke","year":"2021","journal-title":"ACS Infect. Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3096","DOI":"10.1128\/IAI.73.5.3096-3103.2005","article-title":"Structural and genetic diversity of group B streptococcus capsular polysaccharides","volume":"73","author":"Cieslewicz","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_27","unstructured":"Bachmann, B.J. (1996). Derivations and Genotypes of Some Mutant Derivatives of Escherichia coli K-12, ASM Press."},{"key":"ref_28","unstructured":"Chandran, S., and George, K.W. (2020). Design and Implementation of Multi-protein Expression Constructs and Combinatorial Libraries using Start-Stop Assembly. DNA Cloning and Assembly: Methods and Protocols, Springer."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2615","DOI":"10.1038\/s41467-019-10224-x","article-title":"COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors","volume":"10","author":"Naseri","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"6620","DOI":"10.1093\/nar\/gkv466","article-title":"Versatile genetic assembly system (VEGAS) to assemble pathways for expression in S.\u2009cerevisiae","volume":"43","author":"Mitchell","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Valenzuela-Ortega, M., and French, C. (2021). Joint universal modular plasmids (JUMP): A flexible vector platform for synthetic biology. Synth. Biol., 6.","DOI":"10.1093\/synbio\/ysab003"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Jones, J.A., Vernacchio, V.R., Lachance, D.M., Lebovich, M., Fu, L., Shirke, A.N., Schultz, V.L., Cress, B., Linhardt, R.J., and Koffas, M.A.G. (2015). ePathOptimize: A Combinatorial Approach for Transcriptional Balancing of Metabolic Pathways. Sci. Rep., 5.","DOI":"10.1038\/srep11301"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6199","DOI":"10.1016\/j.vaccine.2020.07.050","article-title":"A review of the costs of delivering maternal immunisation during pregnancy","volume":"38","author":"Procter","year":"2020","journal-title":"Vaccine"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"S143","DOI":"10.1093\/cid\/cix654","article-title":"Intrapartum Antibiotic Chemoprophylaxis Policies for the Prevention of Group B Streptococcal Disease Worldwide: Systematic Review","volume":"65","author":"Turner","year":"2017","journal-title":"Clin. Infect. Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2067","DOI":"10.1038\/s41390-022-02375-3","article-title":"Evolving antibiotic resistance in Group B Streptococci causing invasive infant disease: 1970\u20132021","volume":"93","author":"Sabroske","year":"2023","journal-title":"Pediatr. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1093\/infdis\/jiz062","article-title":"A Novel Hexavalent Capsular Polysaccharide Conjugate Vaccine (GBS6) for the Prevention of Neonatal Group B Streptococcal Infections by Maternal Immunization","volume":"220","author":"Buurman","year":"2019","journal-title":"J. Infect. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2079","DOI":"10.1093\/cid\/ciy838","article-title":"A Phase 2, Randomized, Control Trial of Group B Streptococcus (GBS) Type III Capsular Polysaccharide-tetanus Toxoid (GBS III-TT) Vaccine to Prevent Vaginal Colonization With GBS III","volume":"68","author":"Hillier","year":"2019","journal-title":"Clin. Infect. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3769","DOI":"10.1016\/j.vaccine.2004.03.012","article-title":"Efficacy of Streptococcus agalactiae (group B) vaccine in tilapia (Oreochromis niloticus) by intraperitoneal and bath immersion administration","volume":"22","author":"Evans","year":"2004","journal-title":"Vaccine"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1016\/j.fsi.2019.11.044","article-title":"An effective live attenuated vaccine against Streptococcus agalactiae infection in farmed Nile tilapia (Oreochromis niloticus)","volume":"98","author":"Zhang","year":"2020","journal-title":"Fish Shellfish Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.ejar.2024.09.006","article-title":"Implementation of formalin-inactivated Streptococcus agalactiae vaccine in Nile tilapia Oreochromis niloticus broodstock: Efficient vaccination regime, antibody response, and immune-related genes","volume":"50","author":"Piamsomboon","year":"2024","journal-title":"Egypt. J. Aquat. Res."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Bashir, S., Phuoc, N.N., Herath, T., Basit, A., Zadoks, R.N., and Murdan, S. (2023). An oral pH-responsive Streptococcus agalactiae vaccine formulation provides protective immunity to pathogen challenge in tilapia: A proof-of-concept study. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0278277"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2371","DOI":"10.1080\/22221751.2022.2122585","article-title":"Development and evaluation of a multi-epitope subunit vaccine against group B Streptococcus infection","volume":"11","author":"Zhang","year":"2022","journal-title":"Emerg. Microbes Infect."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/3\/279\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:48:15Z","timestamp":1760028495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/3\/279"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,6]]},"references-count":42,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["vaccines13030279"],"URL":"https:\/\/doi.org\/10.3390\/vaccines13030279","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,6]]}}}