Heparinase enzymes

Heparinase Enzymes

Heparinase I, Heparinase II and Heparinase III - isolated from natural strains of Flavobacterium heparinum. 

For structural and disaccharide analysis of heparin and/or heparan sulphate.

Suitable for USP Chemical Tests / ⟨207⟩ 1,6-Anhydro Derivative for Enoxaparin Sodium.

Please email [email protected] for any questions, for a quote or to place and order

The multistep preparation method gives baseline chromatographic separation between Heparinase I, Heparinase II and Heparinase III.

The enzymes are stabilised with 0.1-0.2% BSA (Sigma A4378), 0.22uM-sterile filtered and delivered worldwide as frozen solutions.

International Units: The package size for the Heparinase enzymes is defined in International Units (IU) – one international unit is “the amount of enzyme that will form 1.0 μmole of unsaturated uronic acid per minute from a heparin or heparan sulphate substrate”. 

Heparinase I (Hep I) EC 4.2.2.7

Degrades Heparin and the sulphated (S)-domains of HS by specific scission of the linkage between N-sulfated glucosamine (GlcN.SO3, +/-6-OSO3) and 2-O-sulfated iduronic acid (IdoA,2SO3); the elimination reaction yields di-and oligosaccharides with unsaturated, end-chain 2-O-sulphated uronic acids that absorb at 232nM. 

Linkage Cleaved by Heparinase I   

 -4 GlcN.SO3 (+/-6SO3) 1 – 4 IdoA,2SO3 1-

Heparinase II (Hep II) EC number not assigned

Degrades Heparin and Heparan Sulphate

Catalyses the eliminative scission of the glycosidic linkages between N-sulfated or N-acetylated glucosamine and glucuronic or iduronic acid. This is a broad activity enzyme that tolerates O-sulfation of the uronic acid and glucosamine residues. It will also cleave amino sugar – uronic acid linkages where the amine is unsubstituted ie GlcNH3+ – GlcA/IdoA; these linkages are resistant to Hep I and Hep III. 

Linkage cleaved by Hep II

 -4 GlcN.R (+/-6SO3) 1 – 4 GlcA/IdoA (+/-2SO3) 1-

 R is Acetyl (Ac) or SO3

Heparinase III (Hep III/Heparitinase) EC 4.2.2.8 

Heparinase III (Hep III) degrades Heparan Sulphate (HS) in regions of low and intermediate sulfation and has very limited activity against heparin. The primary substrates for Hep III are N-acetylated or N-sulfated glucosamines linked to glucuronic acid (i.e. GlcNAc/GlcNSO3 1 – 4 GlcA). Hep III has weak activity on disaccharides containing Iduronic Acid (IdoA) and its action is blocked if the iduronate residue is sulfated at C-2 (IdoA, 2SO3). The highly sulphated regions in HS (the S-domains) are resistant to Hep III.

Linkage in HS cleaved by Hep III

-4 GlcN.R(+/-6SO3) 1-4 GlcA 1-

 R is Acetyl (Ac) or SO3