Kringle 5 gene-containing engineered bacterium fermentation process and kringle 5 protein purification process

Abstract

The invention relates a kringle 5 gene-containing engineered bacterium fermentation process and a kingle 5 protein purification process. In the processes, optimal media suitable for the growth of recombinant engineering bacteria and the expression of a kingle 5 protein are determined, the fermentation conditions of the recombinant bacteria are optimized by an orthogonal experiment method, the fermented kringle 5 protein is purified by a two-step method of metal affinity chromatography and cation-exchange chromatography in high, medium and low chromatographs of a device which has gradient elution and can display ultraviolet ray and conductivity parameters at the same time, and an chicken chorioallantoic membrane angiogenesis inhibition test for activity detection is performed. The processescan obvious improve the expression of kringle and conveniently and quickly purify kringle 5 protein to ensure the kringle 5 meets required standards in both purity and biological activity. The processes are simple in production process and high in yield and can be linearly amplified for industrial large-scale production.

Description

technical field [0001] The invention belongs to the technology of biology and medicine development. It adopts gene recombination technology to express angiogenesis inhibitor Kringle 5 in Escherichia coli, optimizes the fermentation process of engineering bacteria through orthogonal experiments, and adopts affinity chromatography and cation exchange chromatography. Two-step purification of the target protein. Background technique [0002] Folkman et al. first proposed in 1972 that the growth of tumors depends on the formation of blood vessels. Although small tumors can obtain nutrients from surrounding tissues through osmosis, their further growth depends on the formation of new blood vessels to obtain sufficient nutrients. Angiogenesis not only provides essential nutrition for tumor growth, but also provides an important way for its diffusion and metastasis. Many experiments have proved that tumor growth and migration depend on angiogenesis. Inhibiting tumor growth by inhib...

AI Technical Summary

Problems solved by technology

At present, in terms of optimizing the fermentation conditions of genetically engineered bacteria, there are few studies on the optimization of the medium, and the optimization of the fermentation conditions is mainly carried out through single-factor experiments. The experimental scheme and results still need to be further improved.

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