A biosynthetic route for polysialylating proteins in Escherichia coli
| Autor: | Manuela Mally, Ivan Hang, Jörg Schneider, Markus Aebi, Michael Wetter, Fabian Müller, Christoph Rutschmann, Simona Russo, Michael Steffen, Chia-Wei Lin, Matthias Zuppiger, Timothy G. Keys, Amirreza Faridmoayer |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Glycosylation Green Fluorescent Proteins Bioengineering medicine.disease_cause Applied Microbiology and Biotechnology law.invention Glycoengineering N-glycosyltransferase Polysialic acid DARPin Stealth polymer 03 medical and health sciences 0302 clinical medicine law Glycosyltransferase medicine Escherichia coli biology Immunogenicity Recombinant Proteins 030104 developmental biology Biochemistry Cytoplasm biology.protein Recombinant DNA Sialic Acids Target protein 030217 neurology & neurosurgery Biotechnology Protein Modification Translational |
| Zdroj: | Metabolic Engineering, 44 |
| ISSN: | 1096-7184 |
| Popis: | Polysialic acid (polySia) is a posttranslational modification found on only a handful of proteins in the central nervous and immune systems. The addition of polySia to therapeutic proteins improves pharmacokinetics and reduces immunogenicity. To date, polysialylation of therapeutic proteins has only been achieved in vitro by chemical or chemoenzymatic strategies. In this work, we develop a biosynthetic pathway for site-specific polysialylation of recombinant proteins in the cytoplasm of Escherichia coli. The pathway takes advantage of a bacterial cytoplasmic polypeptide-glycosyltransferase to establish a site-specific primer on the target protein. The glucose primer is extended by glycosyltransferases derived from lipooligosaccharide, lipopolysaccharide and capsular polysaccharide biosynthesis from different bacterial species to synthesize long chain polySia. We demonstrate the new biosynthetic route by modifying green fluorescent proteins and a therapeutic DARPin (designed ankyrin repeat protein). |
| Databáze: | OpenAIRE |
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