Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris

Authors
He, Wenqin
Zhu, Yuanyuan
Shirke, Abhijeet
Sun, Xiaojun
Liu, Jian
Gross, Richard A.
Koffas, Mattheos A.G.
Linhardt, Robert J.
Li, Ming
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
2017-09-01
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
Terms of Use
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Full Citation
Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris, W. He, Y. Zhu, A. Shirke, X. Sun, J. Liu, R. A. Gross, M. A. G. Koffas, R. J. Linhardt, M. Li, Applied Microbiology and Biotechnology, 101, 6919–6928, 2017.
Abstract
Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase, Homo sapiens C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both Escherichia coli and Pichia pastoris in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.
Description
Applied Microbiology and Biotechnology, 101, 6919–6928
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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
Applied Microbiology and Biotechnology
https://harc.rpi.edu/
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