Thioredoxin mutant, preparation method thereof and application of thioredoxin mutant in production of recombinant fusion proteins

Abstract

The invention discloses a thioredoxin mutant, a preparation method thereof and application of the thioredoxin mutant in the production of recombinant fusion proteins, and belongs to the technical field of biology. The thioredoxin mutant is a non-natural protein dissoluble at a high temperature. The thioredoxin mutant is still dissoluble even when staying at a high temperature for long time. The thioredoxin mutant has both thermal stability and solubility expression in host bacteria, that is to say, a recombinant fusion protein generated by expression can be released quickly; desired reagents can be extracted quickly, experimental equipment has low price, and the thioredoxin mutant is applicable to industrial large-scale production, and has good solubility stability and degeneration resistance at a high temperature.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to thioredoxin mutants, these mutations lead to changes in thermal stability, solubility and net charge properties of thioredoxin mutants, a preparation method of thioredoxin mutants and the use of the sulfur Application of redoxin mutants in the production of recombinant fusion proteins. Background technique [0002] Many peptides and proteins with biological activity have potential uses and commercial value in the fields of modern agriculture, biochemical industry, medicine and health, beauty care and food care, but their application is often limited by industrial production conditions, such as heat, organic Variability under solvent and various chemical operating conditions. However, proteins that are stable under these conditions are widely used in industrial production. [0003] The three-dimensional structure of a protein is critical to its function. In fact, the diversity of p...

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Problems solved by technology

Except for the high-pressure homogenization that can be used for large-scale industrial production, the physical methods are difficult to meet the needs of large-scale production. However, the equipment used in the high-pressure homogenization method is expensive, and the processing time is usually more than 10 hours, which is not conducive to pressure-sensitive or protease degradation. The production of recombinant polypeptides or proteins; the second is chemical cell destruction, mainly using solvents and surfactants to extract cells. Loss of activity, but also cause environmental pollution

The third is biological cell lysis, which mainly uses some compound lysozyme for cell lysis. However, enzymatic cell lysis takes a long time and is generally incomplete, and the yield of recombinant polypeptide or protein is not high. At the same time, the lysozyme added from an external source makes the subsequent purification process Complicated, resulting in lower yields, in addition, there will be potential hidden dangers to the quality control of finished products

[0009] Therefore, the release and extraction of recombinant polypeptides or proteins (referred to as recombinant fusion proteins) heterologously expressed by bacterial hosts in the cell-breaking methods currently used in the art cannot meet the industrial requirements of high efficiency, low cost and large-scale production.

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