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    High cell density cultivation of a recombinant E. coli strain expressing a key enzyme in bioengineered heparin production

    Author
    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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    HIGH CELL DENSITY CULTIVATION OF A RECOMBINANT E. COLI STRAIN.pdf (1.911Mb)
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    Date Issued
    2013-05-01
    Subject
    Biology; Chemistry and chemical biology; Chemical and biological engineering; Biomedical engineering
    Degree
    Terms of Use
    In Copyright : this Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). https://rightsstatements.org/page/InC/1.0/;
    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.
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    URI
    https://hdl.handle.net/20.500.13015/5301; https://doi.org/10.1007/s00253-012-4682-z
    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.
    Department
    The Linhardt Research Labs.; 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/;
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    A full text version is available in DSpace@RPI;
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