High-expression and high-activity bacteroides thetaiotaomicron heparinase I fusion protein, and encoding gene and application thereof

Abstract

The invention relates to a high-expression and high-activity bacteroides thetaiotaomicron heparinase I fusion protein. The fusion protein has the amino acid sequence shown in SEQ ID NO.1. After the fusion protein is fused with an SUMO tag, the soluble expression of a target protein is increased, the soluble expression reaches 95% or more and the specific enzyme activity is increased by 48.9%. Theenzyme activity of the bacteroides thetaiotaomicron heparinase I fusion protein in a 5L fermentation tank can reach 3.412*10<5> IU / L, the expression quantity can reach 3.4 g / L, and the foundation is laid for application development of the bacteroides thetaiotaomicron heparinase I.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a high-expression and high-activity Bacteroides polymorpha heparanase I fusion protein and its coding gene and application. Background technique [0002] Heparin is a specific polydisperse mixed sulfated polysaccharide, which is widely distributed in mammalian tissues and binds to proteins in the form of covalent bonds. At present, commercial heparin is mainly extracted from bovine lung and pig small intestinal mucosa. It has a complex structure and has a variety of important biological functions. It is generally used clinically for the treatment of thrombosis, cardiovascular and other diseases. Low molecular weight heparin (Low molecular heparin, LMWH) is a small segment of heparin produced by cleavage of heparin by certain physical and chemical methods. The ability to bind to proteins or cells is reduced, but the anticoagulant activity is significantly increased. C...

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Problems solved by technology

[0006] The object of the present invention is to overcome the deficiencies in the prior art, overcome the shortcoming of current low yield and low activity of heparanase I, and provide a high expression and high activity Bacteroides polymorpha heparanase I fusion protein and its coding gene and Application, after the fusion protein is fused with the SUMO tag, the soluble expression of the target protein is improved, and the soluble expression reaches more than 95%, and the specific enzyme activity is increased by 48.9%; Up to 3.412×10 5 IU / L, the expression level can reach 3.4g / L, laying the foundation for the application and development of Bacteroides polymorpha heparanase I

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