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Truncated echinococcus granulosus EG95 protein recombinant Pichia pastoris high-density fermentation process

A high-density fermentation technology for Echinococcus granulosus, applied in fermentation, microbial-based methods, biochemical equipment and methods, etc., can solve the problems of inability to proliferate in large quantities, limited antigen sources, and restrictions on wide application. Low expression cost, simple operation, and high protein expression

Inactive Publication Date: 2012-10-17
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0003] Studies have confirmed that the antigen secreted by Echinococcus hexacarpa has a good protective effect, but it cannot make it proliferate in vitro, and the source of antigen is limited, which limits its wide application in immune prevention

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  • Truncated echinococcus granulosus EG95 protein recombinant Pichia pastoris high-density fermentation process
  • Truncated echinococcus granulosus EG95 protein recombinant Pichia pastoris high-density fermentation process
  • Truncated echinococcus granulosus EG95 protein recombinant Pichia pastoris high-density fermentation process

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Embodiment Construction

[0014] The following are specific embodiments of the present invention. The recombinant yeast engineered bacteria monoclonal EG95-1 strain used in the following examples was constructed and identified by the inventor, and was preserved in the China Committee for Microorganism Culture Preservation on February 16, 2012. General Microorganism Center, the deposit number is CGMCC No5764.

[0015] 1. Working seed culture before fermentation

[0016] Pick a monoclonal EG95-1 strain from the streaked YPD plate (G418 concentration is 2mg / mL) and drop it into the first-grade seed YPD medium (200mL Erlenmeyer bottle with 15mL YPD) and cultivate it for about 24h to the cell concentration OD 600 When it reaches 5.0-6.0, the first-grade seed culture solution is obtained; then pour the above-mentioned culture solution into BMGY second-level seed medium (200mL culture solution in a 1000mL triangular bottle) and cultivate for about 18 hours until the cell concentration is OD 600 When it reach...

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Abstract

The invention discloses a truncated echinococcus granulosus EG95 protein tEG96 recombinant Pichia pastoris high-density fermentation process. The recombinant engineering yeast strain monoclonal EG95-1 employed by the invention is established and identified by the inventor, and is collected in China General Microbiological Culture Collection Center on February 16th, 2012 with a collection number of CGMCC No. 5764. The fermentation method provided by the invention comprises the steps that: a fermentation working seed solution is transferred into a BSM liquid culture medium; the pH value of the culture medium is regulated to 5.5, and enrichment is started; an aqueous solution containing glycerin and PTM 1 is supplemented during the enrichment process; and a solution containing a methanol solution of PTM 1 is separately added into the culture system in stages, such that protein induced expression is carried out.

Description

technical field [0001] The invention relates to a fermentation process of recombinant yeast, specifically, the invention relates to a high-density fermentation method of tEG95 recombinant Pichia pastoris of truncated Echinococcus granulosus EG95 protein. Background technique [0002] Echinococcosis, also known as hydatid disease, is a zoonotic parasitic disease that is extremely harmful to humans and animals. The disease is distributed worldwide, and my country is one of the countries with the highest incidence of echinococcosis. Cystic echinococcosis (CE) is the main disease in my country. CE is extremely harmful to humans and animals. People can be infected with the disease by eating insect eggs by mistake, and form space-occupying lesions in the liver, lungs and other organs. All kinds of animals (including humans) can die suddenly due to severe allergic reactions caused by vesicle rupture. The disease can lead to an annual direct economic loss of more than 3 billion y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12P21/02C12R1/84
Inventor 贾万忠娄忠子闫鸿斌李宏民倪兴维
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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