Heparinase

Heparinase (Heparinase I) degrades heparin to oligosaccharide or unsaturated disaccharide. Heparinase can be used in the preparation of low molecular weight heparin (LMWH)^[1].

Product Information

Heparinase has a theoretical molecular weight of 42.5 kDa and an isoelectric point of 9.3-9.5. It selectively cleaves (through an elimination mechanism) highly sulfated polysaccharide chains containing 1-4 linkages between hexosamine and O-sulfated iduronic acid residues. The reaction produces oligosaccharide products (primarily disaccharides) containing unsaturated uronic acids, detectable by UV spectroscopy at 232 nm.

Dissolution
The enzyme is typically dissolved at a concentration of 1 mg/mL in 20 mM Tris-HCl (pH 7.5), 50 mM NaCl, 4 mM CaCl2, and 0.01% BSA.

Reaction Conditions

1. Stable pH range: 4.0-9.0, optimal pH: 7.0-7.5

2. Stable temperature range: 20-37℃, optimal temperature: 30℃

3. Heparinase is calcium-dependent and exhibits maximum activity at 1-5 mM CaCl2. Adding CaCl2 to the reaction system is recommended.

Note: The incubation time and enzyme concentration need to be determined based on practical experience.

References:
[1]. Liu CY, et al. Cloning, expression, and characterization of a novel heparinase I from Bacteroides eggerthii. Prep Biochem Biotechnol. 2020;50(5):477-485.

产品信息
肝素酶理论分子量 42.5 kda，等电点 9.3-9.5，可以选择性地（通过消除机制）切割高度硫酸化的多糖链，该多糖链含有 1-4 个连接，连接点位于己糖胺和 O-硫酸化艾杜糖醛酸残基之间。该反应产生含有不饱和糖醛酸的寡糖产物（主要是二糖），可通过 232 nm 的紫外光谱检测。

溶解
该酶通常以 1 mg/mL 的浓度溶解于 20 mM Tris-HCl (pH 7.5)、50 mM NaCl、4 mM CaCl2 、0.01% BSA 中。

反应条件
1. pH 稳定范围 4.0-9.0 ，最适 pH 7.0-7.5
2. 温度稳定范围20-37℃，最适温度 30℃
3. 肝素酶具有钙依赖性，在 1-5 mM CaCl₂ 条件下具有最大活性，反应体系中建议加入 CaCl₂。
注：酶切底物需要根据实际经验确定孵育时间及酶浓度。