Combinatorial one-pot chemoenzymatic synthesis of heparin

Ujjwal Bhaskar, Guoyun Li, Li Fu, Akihiro Onishi, Mathew Suflita, Jonathan S. Dordick, Robert J. Linhardt

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

Contamination in heparin batches during early 2008 has resulted in a significant effort to develop a safer bioengineered heparin using bacterial capsular polysaccharide heparosan and recombinant enzymes derived from the heparin/heparan sulfate biosynthetic pathway. This requires controlled chemical N-deacetylation/N-sulfonation of heparosan followed by epimerization of most of its glucuronic acid residues to iduronic acid and O-sulfation of the C2 position of iduronic acid and the C3 and C6 positions of the glucosamine residues. A combinatorial study of multi-enzyme, one-pot, in vitro biocatalytic synthesis, carried out in tandem with sensitive analytical techniques, reveals controlled structural changes leading to heparin products similar to animal-derived heparin active pharmaceutical ingredients. Liquid chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy analysis confirms an abundance of heparin's characteristic trisulfated disaccharide, as well as 3-O-sulfo containing residues critical for heparin binding to antithrombin III and its anticoagulant activity. The bioengineered heparins prepared using this simplified one-pot chemoenzymatic synthesis also show in vitro anticoagulant activity.

Original languageEnglish
Pages (from-to)399-407
Number of pages9
JournalCarbohydrate Polymers
Volume122
DOIs
StatePublished - 20 May 2015
Externally publishedYes

Keywords

  • Bioengineered heparin
  • Liquid chromatography-mass spectrometry
  • Nuclear magnetic resonance spectroscopy
  • One-pot synthesis
  • United States Pharmacopeia

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