High density fermentation method of recombinant porcine interferon alpha 1 (rPoIFN alpha 1) gene engineering bacteria

Abstract

The invention discloses a high density fermentation method of recombinant porcine interferon alpha 1(rPoIFN alpha 1) gene engineering bacteria, and belongs to the technical field of fermentation engineering. The high density fermentation method of the recombinant porcine interferon alpha 1 (rPoIFN alpha 1) gene engineering bacteria is as follows: recombinant escherichia coli BL21 / pET-32a-rPoIFN alpha 1 which can construct and express rPoIFN alpha 1 by itself is used as a production bacteria strain for fermentation at 37 DEG C for 2-3h, inducer isopropyl-beta-d-thiogalactoside (IPTG) with the concentration of 50 g / L-100 g / L is added into each liter fermentation solution by one time, fermentation is continued at 32 DEG C for 4-5 h, and then the fermentation process is completed. The high density fermentation method can realize the high-efficiency expression of the rPoIFN alpha 1, an objective protein expression amount accounts for more than 40% of the total bacteria protein, the potency can reach above 1*10<6>IU / ml, the protein is a soluble protein, denaturation and renaturation problems of inclusion body expression can be avoided, the production cycle is shortened, and the high density fermentation method provides a solid foundation for industrial production of the rPoIFN alpha 1.

Description

technical field [0001] The invention mainly relates to a high-density fermentation method of recombinant porcine interferon α1 genetically engineered bacteria, which belongs to the technical field of fermentation process. Background technique [0002] Interferon (IFN) is a protein with broad-spectrum anti-virus, anti-tumor and immunoregulatory effects induced by viral infection. It mainly inhibits the growth, reproduction and development of viruses by inhibiting viral gene transcription and degrading viral RNA. Antitumor activity. Interferon is divided into three types: α, β, and γ according to the different production cells, biochemical characteristics, and the role it plays in the body's immunity. The mechanism by which α-interferon can exert broad-spectrum and high-efficiency antiviral effects in vivo and in vitro is mainly to inhibit the synthesis of viral proteins, and can selectively act on infected cells without any drug residue after use. At present, more than ...

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Problems solved by technology

[0004] At present, the large-scale fermentation production technology of genetically engineered recombinant porcine interferon α has the following problems: 1) Most of the expressed porcine interferon α protein exists in the form of inactive inclusion bodies, and the step of denaturation and renaturation must be added in the purification process. 2) The expression level of genetically engineered recombinant porcine interferon α is low, and the potency is not high enough to meet the requirements of industrialization

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