Recombinant Heparinase

Recombinant Heparinase

Recombinant Heparinase II and Heparinase III

Description

Recombinant, high-activity enzymes produced by expression in E.coli.  Low in endotoxin.

Sterilised by filtration (0.22uM)
Stabilised by 0.2% BSA (Sigma) and 10% glycerol

High stability enables shipping on a cold block, avoiding the higher costs of shipping on dry ice.

The package size for the Heparinase II and III enzymes is defined in International Units (IU) – one international unit is “the amount of enzyme that will liberate 1.0 μmole of product per minute from heparin (Hep II) or Heparan Sulphate (Hep III) substrate at 25º C” (Product is unsaturated saccharides)

Activity assay: 600 ug/ml heparin or heparan sulphate in 1ml 50mM sodium acetate/1mM calcium acetate pH 7.0 at 30C measuring rate of increase of absorption at 232nM

Specificity

Heparinase II (Hep II) degrades heparin and heparan sulfate. Low specificity enzyme attacks most linkages in heparin and HS but it does not effect complete depolymerisation of either GAG. It is very useful for analysis of disaccharide composition but its effectiveness is enhanced when used in combination with Heparinase I and Heparinase III. Heparinase II will cleave heparin and HS at N-unsubstituted glucosamine (GlcNH3+) residues; in contrast the GlcNH3+-GlcA (or IdoA) linkage is resistant to Heparinase I and Heparinase III. Heparinase I available as natural enzyme, see link Natural Enzymes

Heparinase III (Hep III/Heparitinase) EC 4.2.2.8 degrades heparan sulfate (HS). Catalyses the eliminative scission of glycosidic linkages between N-sulfated or N-acetylated glucosamine (GlcNSO3 or GlcNAc) and glucuronic acid. 

Heparinase III (Heparitinase) acts on Heparan Sulphate in regions of low sulfation and it has little activity against heparin. The preferred substrates for heparinase III are N-Acetylated or N-sulfated glucosamines linked to glucuronic acid. (i.e. GlcNAc/ GlcNSO3 1 – 4 GlcA). In consequence it acts primarily in the non-sulfated domains (NAc domains) and transition zones of HS whereas the highly sulfated S-domains are resistant to Heparinase III. The enzyme tolerates C-6 sulfation of amino sugars. Heparinase III has weak activity with disaccharides containing Iduronic Acid (IdoA) and its action is blocked if the iduronate residue is sulfated at C-2 (IdoA, 2SO3).