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A kind of process of producing glutathione by fermentation of Saccharomyces cerevisiae

A technology of Saccharomyces cerevisiae and glutathione, applied in fermentation, microorganism-based methods, peptides, etc., can solve the problems of consumption of glutathione to protect cells, decrease of glutathione production, and inhibition of bacterial growth, etc. The effect of maintaining long-term oxidative stimulation, promoting the accumulation of GSH, and alleviating the effect of yeast growth

Active Publication Date: 2019-11-08
盐城仁越生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Glucose feeding can significantly increase the cell density in the fermentation broth, but in the later stage of fermentation, factors such as nitrogen source and growth factors limit the biomass accumulation
The study on the effect of oxidative stimulation on the production of glutathione by brewer's yeast fermentation found that adding KMnO to the fermentation broth 4 or H 2 o 2 , can improve the production of glutathione to a certain extent, but the addition of a large amount of instant oxidants will not only inhibit the growth of bacteria, but also consume intracellular glutathione to protect cells, which will lead to a rapid decline in glutathione production

Method used

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  • A kind of process of producing glutathione by fermentation of Saccharomyces cerevisiae

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Batch fermentation experiment using O-media medium

[0036] This embodiment adopts the traditional culture method:

[0037] First, the yeast culture was inoculated into a fermenter (10 L) containing traditional O-media medium at a volume ratio of 5% for fermentation.

[0038] Fermentation conditions: culture temperature 30°C, initial pH 6.0, dissolved oxygen concentration 30%;

[0039] Counting from the time of fermentation, after 120 hours of fermentation, the wet weight of Saccharomyces cerevisiae cells in the fermentation broth was detected to reach 45.9 g / L, and the accumulation of GSH reached 0.47 g / L.

[0040] Determination method of GSH: Chromatographic conditions: GSH is determined by HPLC method, high performance liquid chromatography (Shimadzu LC10A), reversed-phase C18 chromatographic column (Hanbon Sci. & Tech). Phosphate buffer solution and methanol were mixed at a volume ratio of 9:1 as the mobile phase, the flow rate was 1 mL / min, and the dete...

Embodiment 2

[0041] Example 2 Batch fermentation test using improved O-media medium

[0042] On the basis of Example 1, the traditional O-media medium in the fermenter was replaced with an improved O-media medium, and other fermentation conditions were the same as in Example 1.

[0043] After 120 h of fermentation, the wet weight of Saccharomyces cerevisiae cells in the fermentation broth reached 41.5 g / L, and the GSH accumulation reached 0.83 g / L.

Embodiment 3

[0044] Embodiment 3 molasses fed-batch experiment

[0045] Real-time monitoring of the carbon source concentration in the fermented liquid during the fermentation process of Example 2 found that when the fermentation was carried out for 16 hours, the glucose was exhausted. Therefore, on the basis of the experiment in Example 2, 16 hours after the start of fermentation, the carbon source was added to the The molasses was fed into the fermentation broth at a constant speed for 72 hours (that is, the feeding was stopped 88 hours after the fermentation started), and the other fermentation conditions were the same as in Example 2.

[0046] The experiment was divided into three groups. The feeding amount of molasses was 0.5 g / L / h, 1.5 g / L / h and 2.0 g / L / h respectively, and they were numbered 1-3 in sequence. Yeast cell wet weight and GSH accumulation, the results are shown in Table 1:

[0047] Table 1 Results of molasses fed-batch experiment

[0048] serial number Cell ...

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Abstract

The invention discloses a technology for producing glutathione by fermenting saccharomyces cerevisiae and belongs to the field of industrial microbial fermentation. The technology for producing glutathione by fermenting saccharomyces cerevisiae comprises the following steps: utilizing an improved O-media fermentation medium to ferment for 16h, and then adding molasses and corn liquor at constant speed of 1.5g / L / h and 0.4g / L / h; after the cell density reaches a certain degree, adding a certain amount of KMnO4 slow-released grains and sodium citrate into the fermentation liquor; fermenting for 120h so that the cell wet weight in the fermentation liquor reaches up to 121.4g / L and the GSH accumulated volume reaches up to 5.78g / L. According to the invention, the cell density of the fermentation liquor is promoted through the improvement for the O-media fermentation medium and the flow adding of the molasses and corn liquor; the oxidative stress for yeast cells is maintained and meanwhile the influence thereof on the yeast growth is relieved through the addition of oxidized stimulant KMnO4 slow-released grains; intracellular ATP level is improved through the addition of the energy auxiliary matter sodium citrate, and finally, the accumulated volume of glutathione is obviously improved and the production cost is lowered.

Description

technical field [0001] The invention relates to the field of industrial microbial fermentation, in particular to a fermentation process for glutathione production by Saccharomyces cerevisiae. Background technique [0002] Glutathione (GSH), a tripeptide antioxidant widely present in eukaryotic and prokaryotic cells, consists of L-glutamic acid, L-cysteine ​​and glycine. Due to the presence of reactive sulfhydryl groups, it maintains the redox potential of cells and prevents the damaging effects of reactive oxygen species (ROS) on cells. In addition, it can also protect cells from ultraviolet rays, heavy metals and various exogenous substances. Therefore, it is often used clinically for the prevention and treatment of anti-radiation, tumor, oxidation and aging, and is widely used in the fields of medicine, health products, food and cosmetics. [0003] GSH can be obtained through extraction, chemical, enzymatic and fermentation methods, with enzymatic and fermentation method...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/865
CPCC07K5/0215C12P21/02
Inventor 陈海龙陈思宇韦平和
Owner 盐城仁越生物科技有限公司
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