Method for producing tagatose by immobilizing multiple enzymes in artificial oil bodies

Abstract

The invention is an immobilized multienzyme based on artificial oil body and its application in the preparation of tagatose. The present invention uses artificial oil bodies to mix the fusion protein of the expressed target protease-oleosome protein with the oil body, and ultrasonically treats the fusion protein to anchor the fusion protein to the surface of the oil body through the specific hydrophobicity of human protein to form an artificial oil containing the target protease body, so that the purification and immobilization of the enzyme can be completed simultaneously. The invention provides an immobilized multi-enzyme that can be used in the production of tagatose. For the first time, artificial oil bodies are used as the substrate of the immobilized enzyme, which significantly improves the stability of the immobilized enzyme and reduces the production cost of the current enzymatic method for preparing tagatose. , and the preparation process is simple.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for expressing enzyme proteins and preparing immobilized multi-enzymes for producing tagatose. Background technique [0002] At present, the mainstream production method of tagatose is to produce pure tagatose through steps such as galactose isomerization, desalination, decolorization, separation, concentration, and crystallization. However, this method also has defects, galactose cannot be completely converted into tagatose, and the final product is a mixture of galactose and tagatose, resulting in complex separation process of tagatose, low conversion rate, high separation cost, and half of the raw material Lactose is not cheap and ultimately tagatose is expensive to produce. (Rhimi M, Aghajari N, Juy M, Chouayekh H, Maguin E, Haser R, Bejar S: Rational design of Bacillus stearothermophilus US100l-arabinose isomerase: Potential applications for d-tagatose production...

AI Technical Summary

Problems solved by technology

However, this new synthesis route involves multiple enzyme molecules, which need to undergo cumbersome extraction and purification before they can be used in catalytic reactions, resulting in high production costs for enzymes; and biological enzymes are water-soluble molecules that dissolve in water after the catalytic reaction. Sexual enzyme molecules are difficult to recycle and reuse, resulting in waste of enzymes

These factors lead to high production costs of the new pathway of tagatose synthesis. It is urgent to develop a multi-enzyme immobilization method to immobilize the multi-enzyme in the new pathway of tagatose synthesis so that it can be reused and reduce production costs.

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