Combinatorial one-pot chemoenzymatic synthesis of heparin. (20th May 2015)

• Record Type: Journal Article
• Authors: Bhaskar, Ujjwal; Li, Guoyun; Fu, Li; Onishi, Akihiro; Suflita, Mathew; Dordick, Jonathan S.; Linhardt, Robert J.
• Is Part Of: Carbohydrate polymers. Volume 122(2015)
• Page Start: 399
• Page End: 407
• Publication Date: 2015-05-20
• Subjects: Bioengineered heparin -- One-pot synthesis -- Liquid chromatography–mass spectrometry -- United States Pharmacopeia -- Nuclear magnetic resonance spectroscopy; Polysaccharides -- Periodicals; Polysaccharides -- Periodicals; Polysaccharides -- Périodiques; Electronic journals; 547.78
• DOI: 10.1016/j.carbpol.2014.10.054
• Languages: English
• ISSNs: 0144-8617

Abstract

Highlights: A simple one pot method for heparin synthesis. Bioengineered heparin with activity similar to USP heparin. 2- O -sulfation must occur before 6- O -sulfation. 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.