Engineering in vivo sulfation in Escherichia coli for the complete biosynthesis of sulfated glycosaminoglycans

Authors
Badri, Abinaya
ORCID
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Other Contributors
Koffas, Mattheos A. G.
Linhardt, Robert J.
Dordick, Jonathan S.
Barquera, Blanca L.
Issue Date
2020-08
Keywords
Chemical engineering
Degree
PhD
Terms of Use
This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.
Full Citation
Abstract
The PAPS-accumulating E. coli developed was then further engineered to produce CS in the second part of the work. Here, we focus on synthesis of 4-O-sulfated CS (CS-A). Assembly of metabolic steps involved in the biosynthesis of all required precursors enabled in vivo CS production for the very first time in a microbial system. We also investigate the effects of manipulating PAPS levels, sulfotransferase activity, induction conditions and GAG export on the sulfation levels of the product. Our studies indicate that CS-A with different sulfation levels can be produced microbially. Moreover, we also show achievement of high 4-O-sulfation levels (~55%) comparable to animal-sourced CS-A (~70%). The microbial CS-A obtained from our engineered strain also has a distinct advantage since it only consists of 4-O-sulfated CS-A unlike animal CS-A (which contain other CS types like 6-O-sulfated CS). Overall, this is the first demonstration of a simple, one-step microbial production of a sulfated GAG and marks an important milestone in the animal-free production of these molecules.
Description
August 2020
School of Engineering
Department
Dept. of Chemical and Biological Engineering
Publisher
Rensselaer Polytechnic Institute, Troy, NY
Relationships
Rensselaer Theses and Dissertations Online Collection
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