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Appl Microbiol Biotechnol. 2012 Sep;95(5):1199-210. doi: 10.1007/s00253-011-3813-2. Epub 2011 Dec 24.

Synthesis of heparosan oligosaccharides by Pasteurella multocida PmHS2 single-action transferases.

Author information

1
Bioprocess Engineering Group, Wageningen University and Research Center, P.O. Box 8129, 6700 EV, Wageningen, the Netherlands.

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.

PMID:
22198719
PMCID:
PMC3418500
DOI:
10.1007/s00253-011-3813-2
[Indexed for MEDLINE]
Free PMC Article

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