(366d) Engineering Designer Biocatalysts and Pathways for Microbial Glycobiology

Carbohydrates add a level of diversity across all forms of life that is unparalleled by the information content of nucleic acids and proteins. The lack of a simple template to translate a glycan code into defined sugar structures contributes to this complexity and impedes efforts aimed at the production of biologically important glycans and glycoconjugates. With the discovery of glycoprotein synthesis in bacteria and functional transfer of glycosylation pathways between species, Escherichia coli cells have become a tractable host for understanding glycosylation and the underlying glycan code of living cells. Moreover, efforts to manipulate the pathways from sugar nucleotides to glycolipids to glycoproteins have transformed E. coli into a living factory for scalable, bottom-up production of complex glycoconjugates by design. Here, I will discuss our efforts to develop E. coli for the biosynthesis of a diverse array of glycan structures, which can be used to tailor the activity, stability, half-life, and shelf-life of a diverse array of protein targets ranging from biopharmaceuticals to industrial enzymes. I will also discuss our efforts to unify protein glycosylation in E. coli with the advanced techniques of protein engineering such as chimeragenesis, protein-fragment complementation, and directed evolution. The result is a powerful new platform for engineering the enzymes, pathways, end-products, and genomes of glycoengineered bacteria for creating the next generation of biotechnology-derived products.