Polysaccharide Sequence Influences the Specificity and Catalytic Activity of Glucuronyl C5-Epimerase

Authors
Vaidyanathan, Deepika
Ke, Xia
Yu, Yanlei
Linhardt, Robert J.
Dordick, Jonathan S.
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
2020-07-14
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
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Full Citation
Polysaccharide Sequence Influences the Specificity and Catalytic Activity of Glucuronyl C5-Epimerase, Deepika Vaidyanathan, Xia Ke, Yanlei Yu, Robert J. Linhardt, Jonathan S. Dordick, Biochemistry, 59, 2576-2584, 2020.
Abstract
Heparin is a widely used biotherapeutic produced from animal tissues. However, it might be possible to produce a bioengineered version using a multienzyme process, relying on the isolation of the E. coli K5 capsule heparosan and its chemical conversion to N-sulfoheparosan, NSH. Glucuronyl C5-epimerase, the first enzyme that acts on NSH, catalyzes the reversible conversion of glucuronic acid (GlcA) to iduronic acid (IdoA). Using full-length NSH, containing different amounts of N-acetylglucosamine (GlcNAc) residues, we demonstrate that C5-epimerase specificity relates to polysaccharide sequence, particularly the location of GlcNAc residues within the chain. We leveraged the deuterium exchange and the novel β-glucuronidase heparanase BP, which cleaves at the GlcA residue. Liquid chromatography–mass spectrometry and gel permeation chromatography of partial/complete heparanase BP digestion products from various NSH substrates treated with C5-epimerase provide information on C5-epimerase activity and action pattern. This study provides insight into optimizing the large-scale production of bioengineered heparin.
Description
Biochemistry, 59, 2576-2584
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Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Biochemistry
https://harc.rpi.edu/
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https://login.libproxy.rpi.edu/login?url=https://doi.org/10.1021/acs.biochem.0c00419