Skip Navigation


Glycobiology Advance Access originally published online on July 14, 2004
Glycobiology 2004 14(12):1217-1228; doi:10.1093/glycob/cwh129
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Data Supplement
Right arrow All Versions of this Article:
14/12/1217    most recent
cwh129v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (2)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Saribas, A. S.
Right arrow Articles by Wang, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Saribas, A. S.
Right arrow Articles by Wang, J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Glycobiology vol. 14 no. 12 © Oxford University Press 2004; all rights reserved.

Production of N-sulfated polysaccharides using yeast-expressed N-deacetylase/N-sulfotransferase-1 (NDST-1)

A. Sami Saribas1, Ali Mobasseri, Pavlo Pristatsky, Xi Chen2, Roger Barthelson3, David Hakes and Jin Wang

Neose Technologies, Inc., 102 Witmer Road, Horsham, PA 19044

Received on February 4, 2004; revised on July 6, 2004; accepted on July 7, 2004

Heparan sulfate/heparin N-deacetylase/N-sulfotransferase-1 (NDST-1) is a critical enzyme involved in heparan sulfate/heparin biosynthesis. This dual-function enzyme modifies the GlcNAc-GlcA disaccharide repeating sugar backbone to make N-sulfated heparosan. N-sulfation is an absolute requirement for the subsequent epimerization and O-sulfation steps in heparan sulfate/heparin biosynthesis. We have expressed rat liver (r) NDST-1 in Saccharomyces cerevisiae as a soluble protein. The yeast-expressed enzyme has both N-deacetylase and N-sulfotransferase activities. N-acetyl heparosan, isolated from Escherichia coli K5 polysaccharide, de-N-sulfated heparin (DNSH) and completely desulfated N-acetylated heparan sulfate (CDSNAcHS) are all good substrates for the rNDST-1. However, N-desulfated, N-acetylated heparin (NDSNAcH) is a poor substrate. The rNDST-1 was partially purified on heparin Sepharose CL-6B. Purified rNDST-1 requires Mn2+ for its enzymatic activity, can utilize PAPS regenerated in vitro by the PAPS cycle (PAP plus para-nitrophenylsulfate in the presence of arylsulfotransferase IV), and with the addition of exogenous PAPS is capable of producing 60–65% N-sulfated heparosan from E. coli K5 polysaccharide or Pasteurella multocida polysaccharide.

2 Present address: Department of Chemistry, University of California-Davis, Davis, CA 95616

3 Present address: Department of Plant Sciences, University of Arizona, Tucson, AZ 85721

1 To whom correspondence should be addressed; e-mail: ssaribas{at}neose.com


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
J. Chen, F. Y. Avci, E. M. Munoz, L. M. McDowell, M. Chen, L. C. Pedersen, L. Zhang, R. J. Linhardt, and J. Liu
Enzymatic Redesigning of Biologically Active Heparan Sulfate
J. Biol. Chem., December 30, 2005; 280(52): 42817 - 42825.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.