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Method for crudely extracting saccharomyces cerevisiae glutathione

A technology of glutathione and brewer's yeast, which is applied in the field of glutathione extraction and brewer's yeast glutathione extraction, can solve the problems of mercury content easily exceeding the standard, high cost, and large pollution, so as to improve the extraction rate , reduce the amount of use, improve the effect of purity

Inactive Publication Date: 2010-05-19
JIANGNAN UNIV
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Problems solved by technology

The methods for separating and extracting GSH from fermented liquid include copper salt method, two-phase method, metal chelate affinity chromatography (Wang Hui, Feng Wanxiang, Separation and Purification of Glutathione with Mercury Resin, Journal of East China University of Science and Technology, 1996, 22 (6)), ion exchange resin separation method (Haruhiko Maki Hideki Fukuda The Separation of glutathione and glutamic acidusing a simulated noving-bed absorber system, J Fermend, 1987, 65 (1): 61-70), wherein the copper salt method is A more traditional method, the production process requires a large amount of H 2 S, the pollution is relatively large, the process is complex, and the residual copper ions also affect the application of GSH in the field of medicine and food; the small-scale aqueous two-phase separation method has the advantages of less investment and less pollution, but it is difficult to industrialize; metal chelation affinity And chromatography generally requires mercury-containing resins, which are sensitive to acid and alkali and have poor stability. The advantages of separation and easy industrialization are becoming a popular technology for the separation of GSH in the later stage
[0004] At present, most of the high-purity GSH products needed in China rely on imports. This is because the current domestic GSH separation and purification technology is relatively backward, resulting in low output and high cost. The product quality is difficult to meet the requirements of the pharmaceutical and food industries in terms of purity and concentration. Therefore, The production of high-purity GSH products that meet the requirements of food and pharmaceutical grades needs to be solved urgently

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  • Method for crudely extracting saccharomyces cerevisiae glutathione
  • Method for crudely extracting saccharomyces cerevisiae glutathione
  • Method for crudely extracting saccharomyces cerevisiae glutathione

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Experimental program
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Embodiment 1

[0030] The optimization of embodiment 1 solid-liquid ratio:

[0031] Take 10ml of deionized water each, according to the ratio of material to liquid (dry yeast g / ml of deionized water) 1:6, 1:7, 1:8, 1:10, 1:12.5, 1:20, 1:40 Add dry beer yeast, mix well, adjust the pH to 3.5 with phosphoric acid, stir and heat at 69-72°C for 14min, put in the refrigerator and cool to room temperature and centrifuge (6000r / min, 12min), measure the supernatant in 9 different conditions The concentration of GSH was used to calculate the extraction yield of GSH. See figure 1 As shown, the experimental results show that the maximum GSH extraction yield is 0.5013% when the solid-liquid ratio is 1:40, followed by 0.4766% when the ratio is 1:8. The extraction rate of the former is 5.18% higher than that of the latter, but the amount of deionized water used is 5 times that of the latter. In actual production, considering the comprehensive reduction of labor intensity, water saving and energy saving, ...

Embodiment 2

[0032] The optimization of embodiment 2 extraction temperature:

[0033] Take 10ml of deionized water, add 1.25g of brewer's dry yeast, mix well, adjust the pH to 3.5 with phosphoric acid, and set the pH in 5 different temperature ranges (49-52, 59-62, 69-72, 79-82, 89- 92,92-95°C), stirred and heated for 12min, placed in the refrigerator to cool to room temperature and centrifuged (6000r / min, 12min), measured the concentration of GSH in the supernatant under 5 different conditions, and calculated the extraction yield. See figure 2 As shown, the experimental results show that as the temperature increases, the GSH extraction first increases and then decreases, and the maximum is 0.48% at 69-72°C. When the temperature is low, the permeability of the cell membrane is poor, and GSH is not completely released from the membrane; if the temperature is too high, the permeability of the cell membrane is too high, and the release of oxidative substances in the cell will also lead to t...

Embodiment 3

[0034] The optimization of embodiment 3 extraction pH:

[0035] Take 10ml of deionized water, add 1.25g of beer dry yeast, mix well, adjust pH to 2.3, 3.0, 3.5, 4.0, 5.0 with phosphoric acid respectively, stir and heat for 12min at 69-72°C, place in refrigerator to cool to room temperature Centrifuge (6000r / min, 12min), measure the concentration of GSH in the supernatant under 4 different conditions, and calculate the extraction yield. See image 3 As shown, the experimental results show that with the increase of pH, the GSH extraction first increases and then decreases, and the maximum value is 0.483% at pH 3.5. When the pH is low, the permeability of the cell membrane is increased, or even destroyed, and the oxidative substances in the membrane are released, which will lead to the oxidation of GSH; when the pH is high, the oxidation of GSH is also increased, so pH 3.5 is selected as the best extraction pH.

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Abstract

The invention relates to a method for crudely extracting saccharomyces cerevisiae glutathione, which comprises the following steps: (1) adjusting the pH value of saccharomyces cerevisiae solution by using an acetic acid-hydrochloric acid system to between 3.3 and 3.5; (2) stirring and heating the saccharomyces cerevisiae solution to between 69 and 72 DEG C, preserving the heat for certain time and extracting, and then cooling the solution to the room temperature; and (3) centrifugalizing the solution to remove saccharomyces, and adjusting the pH of the filtered solution to between 2.0 and 2.5 by using concentrated acid, wherein preferably, the material-liquid ratio of the gram of the dried saccharomyces cerevisiae in the saccharomyces cerevisiae solution to the milliliter of deionized water is 1 to 7.5 to 1 to10; the pH is adjusted to between 3.5 and 3.7 by adding acetic acid, and the pH is adjusted to between 3.3 and 3.5 by adding hydrochloric acid; the preservation time is 12 to 17 minutes, and the cooling step adopts ice-water bath; the centrifugal rotating speed is 5,000 to 6,000rpm, and the time is 10 to 15 minutes; and the concentrated acid is concentrated phosphoric acid. The method also comprises step (4) of performing micro-filtration and ultra-filtration. The method has the advantages of skilful design, high extracting yield, high extraction purity, mild condition, easy operation, low cost, light environment pollution, and suitability for industrialized large-scale production.

Description

technical field [0001] The invention relates to the technical field of glutathione extraction, in particular to the technical field of brewer's yeast glutathione extraction, in particular to a crude extraction method of brewer's yeast glutathione. Background technique [0002] Glutathione (GSH) is a natural active tripeptide (L-γ-glutamyl-L-cysteinyl-glycine) with important physiological functions (Meister A, Anderson M.Glutathione[J].A NNU REV Biochem, 1983, 52: 711-760), easily oxidized to oxidized glutathione (GSSG) in the air, has the functions of anti-oxidation, detoxification, and flavor improvement, and is widely used in clinical medicine, food industry, cosmetic industry, etc. The field has a wide range of uses (Izawa S, Inoue Y, Kimura A. Oxidative stress respnse in yeast: effect of glutathione on adaptation to hydrogen peroxide stress in Saccharomycescerevisiae[J]. FEBS). Recent studies have found that GSH can also inhibit AIDS (Liu Nayu, Research and Application ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/037C07K1/34C07K1/14C12R1/865
Inventor 杨海麟张玉然辛瑜张玲夏小乐仝艳军王武
Owner JIANGNAN UNIV
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