Culture medium and culture method for high-density culture of edwardsiella tarda

A high-density cultivation and culture medium technology, applied in the field of bioengineering, can solve problems such as a large number of bacteria

Inactive Publication Date: 2021-06-15
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when using this attenuated strain to prepare vaccines on a large scale, a large amount of bacteria is needed, so it is urgent to find a method for high-density fermentation to produce this strain

Method used

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  • Culture medium and culture method for high-density culture of edwardsiella tarda
  • Culture medium and culture method for high-density culture of edwardsiella tarda
  • Culture medium and culture method for high-density culture of edwardsiella tarda

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Embodiment 1, tryptone soy broth (TSB) batch fermentation

[0107] In this example, the medium used is a commercial medium TSB for fermentation of Edwardsiella tarda.

[0108] 1. Culture medium preparation

[0109] The formula of TSB medium is: casein trypsin digest 17g / L, soybean powder papain digest 3g / L, sodium chloride 5g / L, dipotassium hydrogen phosphate 2.5g / L, glucose 2.5g / L, and finally pH7.3±0.2.

[0110] 2. Fermentation culture

[0111] Pick a single colony and inoculate it in TSB medium for cultivation (primary fermentation) for 16 hours to obtain a first-level seed solution, and take the above-mentioned seed solution according to the inoculum size of 4% and insert it into TSB medium for 16 hours to obtain a second-level seed solution. The seed liquid is taken and inserted into the medium prepared above according to the inoculum amount of 10% to carry out fermentation culture (tertiary fermentation). The initial pH is 6.8, and ammonia water is used to con...

Embodiment 2

[0114] Embodiment 2, process optimization: optimize culture medium and carry out common fermentation of Edwardsiella tarda

[0115] After repeated research, screening and experimental analysis, the inventor obtained a medium that is beneficial to the cultivation of Edwardsiella tarda, and fermented with this medium.

[0116] 1. Culture medium preparation

[0117] The present inventors comprehensively analyzed the utilization of carbon sources (including glucose, sucrose, starch, etc.) by Edwardsiella tarda, and selected glycerol as the carbon source.

[0118] The present inventor comprehensively analyzed the utilization of nitrogen sources (including peptone, corn flour, yeast powder, beef extract, etc.) by Edwardsiella tarda, and selected peptone and yeast powder as nitrogen sources.

[0119] The optimized fermentation medium is: peptone 20g / L, yeast powder 12g / L, potassium dihydrogen phosphate 3g / L, glycerin 7.5g / L, disodium hydrogen phosphate dodecahydrate 8.2g / L, magnesiu...

Embodiment 3

[0126] Embodiment 3, process optimization: optimize culture medium and carry out process optimization of Edwardsiella tarda reaction process

[0127] 1. Culture medium preparation

[0128] Weigh 40g of peptone, 24g of yeast powder, 6g of potassium dihydrogen phosphate, 15g of glycerin, 16.4g of disodium hydrogen phosphate dodecahydrate, and dissolve it with some deionized water; Mix to 2L; add defoamer 1ml after filling; sterilize.

[0129] 2. Preparation of feeding materials

[0130] Weigh 40g of peptone, 24g of yeast powder, and 15g of glycerol, dissolve in deionized water, dilute to 1L, and sterilize separately.

[0131] 3. Fermentation culture

[0132] Pick a single colony and inoculate it in TSB medium for 16 hours to obtain a first-level seed solution, take the above-mentioned seed solution and insert it into TSB medium for 16 hours to obtain a second-level seed solution according to the inoculation amount of 4%, and take the seed solution according to 10% inoculation...

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Abstract

The invention provides a culture medium and a culture method for high-density culture of edwardsiella tarda. The invention discloses a novel fermentation strategy of the edwardsiella tarda, which can realize high-density and long-period fermentation. According to the method, the characteristics of the edwardsiella tarda are closely combined, a proper supplementing strategy is further adopted in the fermentation process while the culture medium is optimized, the growth density of the edwardsiella tarda can be effectively improved, and the duration of the fermentation period is promoted. The method provided by the invention has the advantages of controllable conditions, simple process control and low cost, and can effectively improve the yield of the edwardsiella tarda in single fermentation.

Description

technical field [0001] The invention belongs to the field of bioengineering, and relates to a microbial culture medium and a microbial fermentation process; more specifically, the invention relates to a culture medium and a culture process for high-density cultivation of Edwardsiella tarda. Background technique [0002] In view of the continuous growth of the world population and the depletion of natural fishery resources, aquaculture, as a traditional industry, has developed rapidly and effectively in modern times, and has highlighted its important position in society, economy and people's lives. In 2002, China's aquatic products accounted for 71% of the world's total output and 49.8% of the total output value, ranking first in the world. According to FAO statistics, in 2004 China's total aquatic products reached 47.5 million tons. However, due to the limitation of water and soil resources, it is difficult to continue to develop by increasing the production area by increas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 刘琴蔡孟浩王擎王启要刘晓红张元兴
Owner EAST CHINA UNIV OF SCI & TECH
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