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Culture medium for expressing O type foot-and-mouth disease inclusion body engineering bacteria

A technology of inclusion bodies and engineering bacteria, which is applied in the field of culture medium for expressing O-type foot-and-mouth disease inclusion body engineering bacteria, and achieves the effects of high viable bacterial count, clear composition, and prevention of the decrease of viable bacterial count.

Inactive Publication Date: 2019-03-01
天津威特生物医药有限责任公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem of high-density fermentation and cultivation of engineering bacteria, the present invention proposes a culture medium expressing O-type foot-and-mouth disease inclusion body engineering bacteria

Method used

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  • Culture medium for expressing O type foot-and-mouth disease inclusion body engineering bacteria
  • Culture medium for expressing O type foot-and-mouth disease inclusion body engineering bacteria
  • Culture medium for expressing O type foot-and-mouth disease inclusion body engineering bacteria

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Experimental program
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Effect test

Embodiment 1

[0016] Research experiment on the large-scale fermentation process of the culture medium formulated according to the formula of the present invention

[0017] (1) Fermentation adopts fed-batch mode, 50L fermenter (Worixin), prepares 20L basal medium, sterilizes the fermenter in situ and prepares it for later use, prepares 5L of feed culture, and sterilizes it for later use.

[0018] The basal medium was prepared according to the following formula: yeast extract 4g / L, tryptone 2.5g / L, magnesium sulfate heptahydrate 1g / L, cobalt chloride hexahydrate 0.2g / L, manganese chloride tetrahydrate 0.15g / L, Potassium dihydrogen phosphate 0.45g / L, dipotassium hydrogen phosphate 1.45g / L, calcium pantothenate 0.25g / L, biotin 0.1g / L, folic acid 0.1g / L, the medium is adjusted to the appropriate pH with alkaline solution such as NaOH value.

[0019] The feed medium was prepared according to the following formula: yeast extract 40g / L, tryptone 25g / L, potassium dihydrogen phosphate 4.5g / L, dipot...

Embodiment 2

[0027] When cultivating Escherichia coli engineering bacteria with variable index flow and feeding strategy, glucose solution and ammonia water were used to adjust the pH value of the experiment.

[0028] 1. Fermentation parameter control

[0029] Both tanks A and B are 2L basal medium + 1L feed medium, and the strategy of adding feed medium is the same. The difference is that when the sugar in the fermentation tank is not enough, tank A adds a certain amount of glucose solution, and then The pH is regulated by 25% ammonia water; the pH is regulated by 60% glucose solution in tank B, and the specific fermentation parameters are shown in Table 1 and Table 2:

[0030] Table 1 Fermentation control parameters of Tank A

[0031]

[0032] Table 2 B tank fermentation control parameters

[0033]

[0034] Note: 1. Both tank A and tank B are 2L basal medium + 1L feed medium.

[0035] 2. The strategy of fed-batch feed medium in tank A and tank B is exactly the same.

[0036] 3....

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Abstract

The invention discloses a culture medium for expressing O type foot-and-mouth disease inclusion body engineering bacteria, and belongs to the technical field of biology. The culture medium comprises abasal culture medium and a material supplementation culture medium and contains no a quick-acting carbon source, a carbon source is slowly supplemented through feeding 60 percent of glucose, the generation of acetic acid can be reduced, and decrement of a viable count caused by pH (Potential of Hydrogen) regulation by ammonia water is also prevented from being generated. The material supplementation culture medium of the culture medium is capable of synthesizing required amino acids through bacteria, different kinds of amino acids can be reasonably added, and waste of overnutrition can be avoided. The culture medium disclosed by the invention is capable of remarkably increasing the engineering fermentation density of escherichia coli, meanwhile, high viable count under high density can beensured, and the expression quantity of mycoprotein per unit cannot be reduced; the total expression quantity of target protein can be increased, the production efficiency can be increased, and the production cost can be reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for expressing O-type foot-and-mouth disease inclusion body engineering bacteria. Background technique [0002] With the development of biopharmaceuticals, the engineering bacteria obtained by cloning Escherichia coli using recombinant DNA technology produces more and more biopharmaceuticals, which is the main way for biotechnology to obtain biopharmaceutical products. [0003] Generally, the yield of biopharmaceuticals is closely related to the fermentation density of Escherichia coli, and the yield of Escherichia coli is directly related to the yield of biopharmaceuticals. Therefore, under the condition that the protein expression efficiency of bacteria is equivalent, the fermentation density of E. coli directly affects the efficiency and cost of biopharmaceuticals. Especially under the same purification technology conditions, some scholars continue to...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12R1/19
CPCC12P21/02
Inventor 秦天达扶星星黄银君温建波王科牟克斌武志宇刘斌李金杰李佳霞邢向茹陈晓宇岑静刘瑞宫士越史效茹
Owner 天津威特生物医药有限责任公司
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