Increased 3’-phosphoadenosine-5’-phosphosulfate levels in engineered Escherichia coli cell lysate facilitates in vitro synthesis of chondroitin sulfate A

Loading...
Thumbnail Image
Authors
Badri, A.
Williams, A.
Xia, K.
Linhardt, Robert J.
Koffas, M.A.G.
Issue Date
2019
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Research Projects
Organizational Units
Journal Issue
Alternative Title
Abstract
Chondroitin sulfates (CSs) are linear glycosaminoglycans that have important applications in the medical and food industries. Engineering bacteria for the microbial production of CS will facilitate a one-step, scalable production with good control over sulfation levels and positions in contrast to extraction from animal sources. To achieve this goal, Escherichia coli (E. coli) is engineered in this study using traditional metabolic engineering approaches to accumulate 3'-phosphoadenosine-5'-phosphosulfate (PAPS), the universal sulfate donor. PAPS is one of the least-explored components required for the biosynthesis of CS. The resulting engineered E. coli strain shows an ≈1000-fold increase in intracellular PAPS concentrations. This study also reports, for the first time, in vitro biotransformation of CS using PAPS, chondroitin, and chondroitin-4-sulfotransferase (C4ST), all synthesized from different engineered E. coli strains. A 10.4-fold increase is observed in the amount of CS produced by biotransformation by employing PAPS from the engineered PAPS-accumulating strain. The data from the biotransformation experiments also help evaluate the reaction components that need improved production to achieve a one-step microbial synthesis of CS. This will provide a new platform to produce CS.
Description
Biotechnology Journal, 14, 1800436
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Increased 3’-phosphoadenosine-5’-phosphosulfate levels in engineered Escherichia coli cell lysate facilitates in vitro synthesis of chondroitin sulfate A, A. Badri, A. Williams, K. Xia, R. J. Linhardt, M. A.G. Koffas, Biotechnology Journal, 14, 1800436, 2019.
Publisher
Wiley
Terms of Use
Journal
Volume
Issue
PubMed ID
DOI
ISSN
EISSN