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

A technology of fermentation process and engineering bacteria, applied in fermentation, microbial-based methods, organic chemistry, etc., to achieve the effects of increased expression, simple production process, and high recovery rate

Inactive Publication Date: 2010-03-17
SHANXI LIFEGEN
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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 opt

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

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Embodiment Construction

[0023] The present invention is based on the work of predecessors, constructs engineering bacteria containing Kringle 5 gene, studies a fermentation process of recombinant engineering bacteria containing Kringle 5 gene and a method of purifying Kringle by affinity chromatography and cation exchange chromatography 5 protein method.

[0024] The fermentation process of the engineering bacterium containing Kringle 5 gene of the present invention is---designed 15 kinds of different fermentation mediums, under shake flask culture mode, by measuring the specific growth rate and the specific growth rate of engineering bacteria in 15 kinds of mediums The expression level of the target protein kringle5 determined the medium suitable for the growth of engineering bacteria and the expression of the target protein; using the optimized medium, through orthogonal experiments, the best fermentation process was determined; according to the optimized conditions of shake flask culture , carried...

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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...

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Application Information

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IPC IPC(8): C12P21/06C07K1/22C07K1/18C07K14/435C12R1/19
Inventor 边六交党红艳杨晓燕陈超
Owner SHANXI LIFEGEN
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