A heparanase and its preparation method and application

WJS HPSE, prepared by isolating from the white jade snail and using an insect baculovirus expression system, solves the problems of insufficient thermostability and substrate specificity of existing heparan sulfate enzymes, achieving efficient cleavage of HS and heparin sugar chains, and is suitable for sugar chain structure analysis and low molecular weight heparin preparation.

CN120924522BActive Publication Date: 2026-04-28JIANGXI NORMAL UNIV
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Patent Information

Application Number
CN202511045766.6
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2025-07-29
Publication Date
2026-04-28
Estimated Expiration
2045-07-29

AI Technical Summary

Technical Problem

Existing heparan sulfate enzymes are insufficient in terms of thermal stability and substrate specificity, making it difficult to meet industrial requirements. Furthermore, existing heparin lyases also lack sufficient thermal stability, hindering their ability to cleave substrate structures and limiting the precision analysis of complex glycan structures.

Method used

Using heparan sulfate enzyme (WJS HPSE) derived from the white jade snail, the coding sequence of WJS HPSE was isolated from the white jade snail using gene cloning technology. Recombinant expression was achieved using an insect baculovirus expression system, and high-purity active enzyme was obtained by affinity chromatography purification. WJS HPSE exhibited the best catalytic activity at pH 4.0 and 57℃, and can efficiently and specifically cleave the β-1,4 glycosidic bond between IdoA2S or GlcA and GlcNAc.

Benefits of technology

WJS HPSE exhibits significantly superior thermal stability compared to human HPSE and possesses the ability to efficiently and specifically cleave HS and heparin glycan chains. It is suitable for applications such as HS glycan structure analysis and low molecular weight heparin preparation, providing a novel tool enzyme for glycan structure analysis and targeted preparation of pharmacological oligosaccharide fragments.

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Abstract

The application belongs to the technical field of bioengineering technology, and particularly relates to a heparanase and a preparation method and application thereof. The amino acid sequence of the heparanase (HPSE) is shown as SEQ ID NO. 2. The heparanase (WJS HPSE) provided in the application can specifically degrade HS glycan substrates, which are white jade snail polysaccharide (WJS PS) containing IdoA2S-GlcNAc disaccharide repeating units, heparin or HS. In addition, the heparanase (WJS HPSE) provided in the application has high thermal stability, has optimal enzyme activity at 57-67 DEG C and pH=4.0, significantly improves the stability and reuse efficiency in a catalytic process, and thus improves the application value of the HPSE.
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