Immobilized fusion enzyme and method for preparing glutathione by using immobilized fusion enzyme

Abstract

The invention relates to the field of polypeptide synthesis, and in particular relates to immobilized fusion enzyme and a method for preparing glutathione by using the immobilized fusion enzyme. According to the invention, a glutathione synthetase gene and a triphosadenine regenerative enzyme gene are respectively connected with a chitin protein domain gene; fusion enzyme is prepared by expression; by utilization of the fact that a chitin carrier can be bound to a chitin protein domain specifically and firmly, glutathione synthesis fusion enzyme and triphosadenine regenerative fusion enzyme are purified and immobilized through one step; the process is simple; and the space density of the immobilized enzyme is high. The immobilized fusion enzyme prepared by the invention is high in enzyme activity; continuous generation of glutathione can be realized; the batch and continuous process route of the immobilized fusion enzyme is adopted in the invention; glutathione up to 31 g / L can be generated; the concentration of a target product is high; no enzyme residue exists in product solution; and the later purification process is greatly simplified.

Description

technical field [0001] The invention relates to the field of polypeptide synthesis, in particular to an immobilized fusion enzyme and a method for preparing glutathione with it. Background technique [0002] Glutathione (GSH) is a natural tripeptide compound formed by linking L-glutamic acid, L-cysteine ​​and glycine, and its molecular formula is C 10 h 17 N 3 o 6 S, the molecular weight is 307.3. GSH widely exists in animals and plants, and plays an important antioxidant function in organisms. Since the active thiol-SH in GSH is easily oxidized and dehydrogenated, it can remove free radicals in the body; at the same time, GSH can also improve human immunity, maintain health, anti-aging, and slow down the aging cells. ; Coupled with broad-spectrum detoxification, therefore, glutathione is not only used as a safe drug additive, but also widely used as a base material for functional foods, as well as functions such as delaying aging, enhancing immunity, and anti-tumor. i...

AI Technical Summary

Problems solved by technology

[0005] Most of the existing technologies use free, unpurified glutathione synthase and ATP regeneration enzyme to produce glutathione. The liquid enzyme is used for one-time use and cannot be recycled, which greatly increases the production cost of the enzyme; At the same time, the mixed enzymes in the mixed enzyme liquid lead to the occurrence of many by-products, and the enzyme liquid itself is mixed with the product, which is not conducive to the purification of the final product; the fluctuation of each batch during the preparation of the mixed enzyme liquid in the early stage will affect the quality of the final product. Quality stability

Some technologies also involve immobilized enzymes. Most of the commonly used enzyme immobilization methods use enzyme purification to obtain free enzymes, and then fix them by adsorption, embedding, covalent bonding, etc. The process is cumbersome and the recovery rate of enzyme activity is low.

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