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A kind of method using oxygen carrier to improve glutathione titer in Saccharomyces cerevisiae fermentation broth

A technology of Saccharomyces cerevisiae and glutathione, which is applied in the field of microbial fermentation, can solve the problems of aggravated feedback inhibition, low productivity, and long fermentation production cycle, and achieve the effects of increasing the yield of expression products, promoting high-density fermentation, and shortening the fermentation cycle

Active Publication Date: 2015-09-02
安徽正兴生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the glutathione produced by fermentation exists in the cells of microorganisms, so while increasing the content of glutathione, the product reduced glutathione can synthesize γ-L-glutamyl-L-cysteine Feedback inhibition of the enzyme is also aggravated, limiting further increases in glutathione production
In addition, the cell permeability of the bacteria during the fermentation process is not good, making it difficult for the added substrate amino acid to enter the cell for the synthesis of glutathione, resulting in a low yield of glutathione relative to the added amino acid , and the fermentation production cycle is long, the productivity is low, and it is not conducive to the separation and extraction of products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] This embodiment includes the following steps:

[0023] (1) Seed preparation: Take Saccharomyces cerevisiae and insert it into the seed medium, cultivate it for 8 hours at 30°C with a stirring speed of 150-200rpm, and then transfer it to the second-generation seed medium under the conditions of pH 6.5-7, temperature 30 Cultivate for 10-15 hours at ℃, and prepare secondary seeds for later use;

[0024] The seed medium is: yeast extract 5-10g / L, peptone 10-20g / L, glucose 5-20g / L, pH=7, the pH value is adjusted by ammonia water;

[0025] In this example, Saccharomyces cerevisiae was purchased from the market, and the Saccharomyces cerevisiae was preserved by China Industrial Microorganism Collection Center, and its number was CICC1898;

[0026] (2) Fermentation culture: Inoculate the obtained seed culture liquid into the fermentation medium, 30°C, 200-300rpm, add sterile oxygen carrier n-hexane to the fermentation medium, the amount of oxygen carrier added is the volume of t...

Embodiment 2

[0028] In this example, the mixture of oxygen carrier n-dodecane, n-hexane and soybean oil is added in an amount of 0.5% of the volume of the fermentation medium, and the glutathione in the final fermentation liquid is 2.1g / L. Other embodiments and Example 1 same. Example 3

Embodiment 3

[0029] This embodiment includes the following steps:

[0030] (1) Seed preparation: Take Saccharomyces cerevisiae and insert it into the seed medium, cultivate it for 8 hours at 35°C with a stirring speed of 150-200rpm, then transfer it to the second-generation seed medium, and add sterile oxygen carrier p-12 at the same time The addition of alkane and oxygen carrier is 8% of the volume of the seed medium, pH=7, and the temperature is 35° C. for 10 to 15 hours to prepare secondary seeds for later use;

[0031] (2) Fermentation culture: Inoculate the obtained seed culture solution into the fermentation medium, add sterile oxygen carrier n-dodecane into the fermentation tank at 35°C, 200-300rpm, and the amount of oxygen carrier added is the fermentation medium Constant temperature culture at 8% volume, control dissolved oxygen 30-50%, add glucose 500g / L to maintain cell growth, adjust pH value to 7 with ammonia water, cell density OD600nm=120, add precursor amino acid L-glutamic...

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PUM

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Abstract

The invention discloses a method for increasing the valence of glutathione in saccharomyces cerevisiae fermentation liquor by utilizing an oxygen carrier. The method comprises the following steps: (1) preparation of seeds, namely inoculating saccharomyces cerevisiae in a seed medium for culturing so as to obtain a seed culture solution; and (2) fermentation culture, namely inoculating the seed culture solution obtained in step (1) in a fermentation medium, then adding an aseptic oxygen carrier of which the added amount is 0.5-10 percent of the volume of the fermentation medium, and fermenting to obtain the glutathione. According to the method disclosed by the invention, by adding the proper oxygen carrier into the medium without addition of other substances and extra energy consumption, the concentration of dissolved oxygen and the permeability of cell membranes in the medium are greatly increased; in the process, the carrier disclosed by the invention acts as the oxygen carrier, and the permeability of the cell membranes is improved as well, so the aim of obviously increasing the yield of an expression product can be successfully achieved, the fermentation period is shortened and the production efficiency is improved.

Description

technical field [0001] The invention relates to a microbial fermentation method, in particular to a method for increasing glutathione titer in brewer's yeast fermentation liquid by using an oxygen carrier. Background technique [0002] Glutathione (GSH) is a tripeptide compound composed of L-glutamic acid, L-cysteine ​​and glycine. It is an important physiologically active substance in organisms, especially for maintaining the redox environment in organisms. play a very important role. Glutathione is a small molecular peptide substance containing sulfhydryl groups, which has two important antioxidant effects and integrated detoxification effects. The sulfhydryl group on cysteine ​​in the molecular structure of glutathione is its active group, so glutathione is often abbreviated as GSH, and it is easy to react with heavy metal salts such as iodoacetic acid, mustard gas (a poisonous gas), lead, mercury, arsenic, etc. Complexation, and has an integrated detoxification effect....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/865
Inventor 邵千飞汪德林崔秀云黄绪民
Owner 安徽正兴生物医药科技有限公司