High cell density cultivation of a recombinant E. coli strain expressing a key enzyme in bioengineered heparin production
Authors
Restaino, Odile Francesca
Bhaskar, Ujjwal
Paul, Priscilla
Li, Lingyun
De Rosa, Mario
Dordick, Jonathan S.
Linhardt, Robert J.
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
2013-05-01
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
Terms of Use
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Full Citation
High cell density cultivation of a recombinant E. coli strain expressing a key enzyme in bioengineered heparin production, O. F. Restaino, U. Bhaskar, P. Paul, L. Lin, M. De Rosa, J. S. Dordick, R. J. Linhardt, Applied Microbiology and Biotechnology, 97, 3893–3900, 2013.
Abstract
A bioengineered heparin, as a replacement for animal-derived heparin, is under development that relies on the fermentative production of heparosan by Escherichia coli K5 and its subsequent chemoenzymatic modification using biosynthetic enzymes. A critical enzyme in this pathway is the mammalian 6-O-sulfotransferase (6-OST-1) which specifically sulfonates the glucosamine residue in a heparin precursor. This mammalian enzyme, previously cloned and expressed in E. coli, is required in kilogram amounts if an industrial process for bioengineered heparin is to be established. In this study, high cell density cultivation techniques were exploited to obtain recombinant 6-OST-1. Physiological studies were performed in shake flasks to establish optimized growth and production conditions. Induction strategies were tested in fed-batch experiments to improve yield and productivity. High cell density cultivation in 7-l culture, together with a coupled inducer strategy using isopropyl β-D-1-thiogalactopyranoside and galactose, afforded 482 mg l(-1) of enzyme with a biomass yield of 16.2 mg gcdw (-1) and a productivity of 10.5 mg l(-1) h(-1).
Description
Applied Microbiology and Biotechnology, 97, 3893–3900
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.
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.
Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Springer
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Applied Microbiology and Biotechnology
https://harc.rpi.edu/
Rensselaer Polytechnic Institute, Troy, NY
Applied Microbiology and Biotechnology
https://harc.rpi.edu/
Access
A full text version is available in DSpace@RPI