Expression and secretion of glycosylated heparin biosynthetic enzymes using Komagataella pastoris
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Authors
Englaender, Jacob A.
Zhu, Yuanyuan
Shirke, Abhijit N.
Lin, Lei
Liu, Xinyue
Zhang, Fuming
Gross, Richard A.
Koffas, Mattheos A.G.
Linhardt, Robert J.
Issue Date
2017-04-01
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
Heparin, an anticoagulant drug, is biosynthesized in selected animal cells. The heparin biosynthetic enzymes mainly consist of sulfotransferases and all are integral transmembrane glycoproteins. These enzymes are generally produced in engineered Escherichia coli as without their transmembrane domains as non-glycosylated fusion proteins. In this study, we used the yeast, Komagataella pastoris, to prepare four sulfotransferases involved in heparin biosynthesis as glycoproteins. While the yields of these yeast-expressed enzymes were considerably lower than E. coli-expressed enzymes, these enzymes were secreted into the fermentation media simplifying their purification and were endotoxin free. The activities of these sulfotransferases, expressed as glycoproteins in yeast, were compared to the bacterially expressed proteins. The yeast-expressed sulfotransferase glycoproteins showed improved kinetic properties than the bacterially expressed proteins.
Description
Applied Microbiology and Biotechnology, 101, 2843–2851
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.
Full Citation
Expression and secretion of glycosylated heparin biosynthetic enzymes using Komagataella pastoris, J. A. Englaender, Y. Zhu, A.N. Shirke, L. Lin, X. Liu, F. Zhang, R.A. Gross, M.A.G. Koffas, R. J. Linhardt, Applied Microbiology and Biotechnology, 101, 2843–2851, 2017
Publisher
Terms of Use
Journal
Volume
Issue
PubMed ID
DOI
ISSN
14320614
1757598
1757598