Engineering in vivo sulfation in Escherichia coli for the complete biosynthesis of sulfated glycosaminoglycans
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Authors
Badri, Abinaya
Issue Date
2020-08
Type
Electronic thesis
Thesis
Thesis
Language
ENG
Keywords
Chemical engineering
Alternative Title
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
School of Engineering
Full Citation
Publisher
Rensselaer Polytechnic Institute, Troy, NY