Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 419449
Title Synthesis of heparosan oligosaccharides by Pasteurella multocida PmHS2 single-action transferases
Author(s) Chavaroche, A.A.E.; Broek, L.A.M. van den; Boeriu, C.G.; Eggink, G.
Source Applied Microbiology and Biotechnology 95 (2012)5. - ISSN 0175-7598 - p. 1199 - 1210.
Department(s) Bioprocess Engineering
FBR Sustainable Chemistry & Technology
FBR Bioconversion
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) heparan-sulfate polymerization - molecular-weight heparin - chemoenzymatic synthesis - hyaluronic-acid - capsular polysaccharide - glycosaminoglycans - identification - synthase - glycosyltransferase - biosynthesis
Abstract Pasteurella multocida heparosan synthase PmHS2 is a dual action glycosyltransferase that catalyzes the polymerization of heparosan polymers in a non-processive manner. The two PmHS2 single-action transferases, obtained previously by site-directed mutagenesis, have been immobilized on Ni(II)-nitrilotriacetic acid agarose during the purification step. A detailed study of the polymerization process in the presence of non-equal amounts of PmHS2 single-action transferases revealed that the glucuronyl transferase (PmHS2-GlcUA+) is the limiting catalyst in the polymerization process. Using experimental design, it was determined that the N-acetylglucosaminyl transferase (PmHS2-GlcNAc+) plays an important role in the control of heparosan chain elongation depending on the number of heparosan chains and the UDP-sugar concentrations present in the reaction mixture. Furthermore, for the first time, the synthesis of heparosan oligosaccharides alternately using PmHS2-GlcUA+ and PmHS2-GlcNAc+ is reported. It was shown that the synthesis of heparosan oligosaccharides by PmHS2 single-action transferases do not require the presence of template molecules in the reaction mixture.
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