380616 Production of Chondroitin Sulfate with Escherichia coli K4

Wednesday, November 19, 2014: 3:15 PM
214 (Hilton Atlanta)
Wenqin He, Chemical & Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY, Robert J. Linhardt, Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY and Mattheos A.G. Koffas, Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY

Chondroitin sulfate is an important homopolymeric glycosaminoglycan consisting of repeating disaccharide units of N-acetyl-D-galactosamine and glucuronic acid with sulfation at various positions. It has been widely used for treating knee osteoarthrithis due to its anti-inflammatory and chondro-protective properties. The current method for manufacturing chondroitin sulfate relies solely on animal sourced tissues, which may be unreliable due to the risk of virus and/or prion contamination.  Given the importance of its use in medicine and as a dietary supplement, alternative production methods are necessary in order to provide stable and safe supply of chondroitin sulfate. The capsular polysaccharide of the pathogenic Escherichia coli K4 strain shares a similar structure to the chondroitin sulfate except for a β-linked fructose branching residue on the chondroitin backbone. The three genes, kfoA, kfoC and kfoF, which encode the major enzymes for the biosynthesis of the backbone of this chondroitin-like capsular polysaccharide in K4, were cloned into ePathbrick system vectors allowing a tunable gene copy number and promoter strength in a single plasmid, and then transformed into the non-pathogenic production strain E. coli BL21. A significant level of polysaccharide production was observed with an average yield of ~2.2 g/L chondroitin in shake flask. Functional expression of sulfotransferases has further allowed the production of a chondroitin sulfate derivative for the first time in E.coli.

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See more of this Session: Metabolic Engineering and Bioinformatics
See more of this Group/Topical: Food, Pharmaceutical & Bioengineering Division