Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of recombinant Escherichia coli shake flask culture medium and the method for improving the expression amount of target protein

A technology for recombinant Escherichia coli and protein expression, which is applied in the biological field, can solve the problems of low yield of target protein research needs, inability to scale up large-scale fermentation production needs, and insufficient nutrients in the medium, so as to reduce production costs and shorten the production cycle , The effect of reducing equipment investment

Active Publication Date: 2021-10-08
海南省产品质量监督检验所
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the culture medium for shake flask expression is LB medium (containing 10 grams of peptone per liter, 5 grams of yeast extract, and 10 grams of sodium chloride). This medium is not rich in nutrition and the culture conditions are relatively simple. There are few bacteria and the low yield of target protein cannot meet the needs of follow-up research, nor can it be scaled up to the needs of large-scale fermentation production

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of recombinant Escherichia coli shake flask culture medium and the method for improving the expression amount of target protein
  • A kind of recombinant Escherichia coli shake flask culture medium and the method for improving the expression amount of target protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A recombinant Escherichia coli shake flask culture medium, comprising the following preparation method: weighing the following components: 20g of soybean peptone, 10g of yeast powder, 2.5g of ammonium sulfate, 2.3g of potassium dihydrogen phosphate, 11.84g of disodium hydrogen phosphate, and 3g of ginkgo polysaccharide , Potassium magnesium sulfate 1g, sodium naphthalene acetate 12g, add purified water to dissolve and make up to 900mL, then add 0.1mL antifoaming agent, after preparation, sterilize at 121°C for 30 minutes to obtain.

[0026] The method for improving the expression of recombinant protein by applying the described recombinant Escherichia coli shake flask culture medium comprises the following steps:

[0027] a. Take 50 mL of LB culture solution, add 50 uL of 35 mg / mL kanamycin sulfate solution, inoculate the bacteria containing the recombinant protein into the LB culture solution, and culture overnight at 37°C and 50 rpm after inoculation;

[0028] b. Take...

Embodiment 2

[0032] A recombinant Escherichia coli shake flask culture medium is characterized in that it comprises the following preparation method: weighing the following ingredients: 15g of soybean peptone, 15g of yeast powder, 2g of ammonium sulfate, 3g of potassium dihydrogen phosphate, 10g of disodium hydrogen phosphate, ginkgo polysaccharide 2g, potassium magnesium sulfate 1g, sodium naphthalene acetate 12g, add water to dissolve and make up to 900mL, then add 0.3mL antifoaming agent, after the preparation, sterilize at 121°C for 45 minutes to obtain the product.

[0033] The method for improving the expression of recombinant protein by applying the described recombinant Escherichia coli shake flask culture medium comprises the following steps:

[0034] a. Take 50 mL of LB culture solution, add 50 uL of 30 mg / mL kanamycin sulfate solution, inoculate the bacteria containing the recombinant protein into the LB culture solution, and culture overnight at 37°C and 70 rpm after inoculation...

Embodiment 3

[0039]A recombinant Escherichia coli shake flask culture medium, comprising the following preparation method: weighing the following ingredients: 30g of soybean peptone, 8g of yeast powder, 4g of ammonium sulfate, 1g of potassium dihydrogen phosphate, 15g of disodium hydrogen phosphate, 3g of ginkgo polysaccharide, potassium sulfate Add 2g of magnesium and 10g of sodium naphthaleneacetate, dissolve in water and make up to 900mL, then add 0.1-0.3mL of antifoaming agent, and sterilize at 121°C for 45 minutes after preparation.

[0040] The method for improving the expression of recombinant protein by applying the described recombinant Escherichia coli shake flask culture medium comprises the following steps:

[0041] a. Take 50 mL of LB culture solution, add 50 μL of 50 mg / mL kanamycin sulfate solution, inoculate the bacteria containing the recombinant protein into the LB culture solution, and culture overnight at 37°C and 70 rpm after inoculation;

[0042] b. Take 100mL of 20% ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a recombinant Escherichia coli shake flask culture medium, comprising the following components: soybean peptone, yeast powder, ammonium sulfate, potassium dihydrogen phosphate, disodium hydrogen phosphate, ginkgo polysaccharide, potassium magnesium sulfate, and sodium naphthalene acetate. The strain culture conditions and culture methods in the shake flask culture were also optimized, and a shake flask expression method capable of high-density growth of the bacteria and efficient expression of the target protein was established. On the premise of increasing the amount of bacteria per unit volume, the content of total protein per unit volume is increased, thereby increasing the output of the target product, and finally improving the specific productivity of the product (the output of the product per unit volume and unit time) can not only reduce the culture volume , Strengthening downstream separation and extraction can also shorten the production cycle, reduce equipment investment and thus reduce production costs, and can greatly improve the competitiveness in the market.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant Escherichia coli shake flask culture medium and a method for increasing the expression level of a target protein. Background technique [0002] The prokaryotic expression system has the characteristics of short expression period, high expression level, simple operation, easy cultivation, and low cost. Therefore, it is usually used as an expression system for exogenous genes in DNA recombination technology, and the E. coli expression system is currently the most mature technology. Prokaryotic expression system. [0003] For the expression of recombinant bacteria for the purpose of producing foreign proteins, it is hoped to obtain as much target protein as possible during the expression process, improve the volume production efficiency of the product, and increase the yield of the final product. In the process of expressing foreign proteins, recombinant engineering bacte...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/00C12R1/19
CPCC12N1/20C12P21/00
Inventor 赵宏亮杨晓红杨景豪毛海梅吴毓炜
Owner 海南省产品质量监督检验所