KR100302099B1 - High content biogermanium yeast and its manufacturing method - Google Patents

Abstract

The present invention relates to a method for producing a yeast containing a high content of biogerium by separating the process of producing a yeast cell and a process of introducing germanium ions into yeast cells, and a high content biogermanium yeast prepared therefrom. Method for producing a high content biogermanium yeast of the present invention is a yeast strain in a medium consisting of 0.3 to 0.5% peptone, 0.1 to 0.3% yeast extract, 3 to 5% glucose, 0.1 to 0.3% malt extract and water (rest) Incubating the cells at 25 to 35 ° C. for 24 to 48 hours to produce the cells and centrifuging the culture solution obtained in the above step to collect only the cells, and a solution in which the concentration of the germanium solution is 1000 to 5000 ppm is 0.5 to 2 (volume ratio). And mixing germanium ions into yeast cells. In the present invention, after culturing the yeast strain to isolate only the cells, the germanium solution was added to the cells to incubate the cells and germanium at the optimum pH, temperature and incubation time conditions by introducing germanium ions into the cells, high yield biogerium The yeast can be prepared and the germanium can be recovered, thereby limiting the problems of yeast cell production when producing germanium by adding germanium during yeast culture, and difficulty in recovering germanium ions remaining in the medium after the culture is completed. Very effective in solving.

Description

High content biogermanium yeast and preparation method thereof

The present invention relates to a method for preparing a high content biogerium yeast by introducing a large amount of germanium ions into yeast cells. More specifically, the present invention is to separate the process for producing yeast cells and the process for introducing germanium ions into yeast cells to produce a yeast containing a high content of biogerium and the high content biogermanium yeast produced thereby It is about.

Germanium is classified into two types, inorganic germanium and organic germanium. Of these, inorganic germanium is mainly present in minerals, and is a simple compound that is not bound to an organic compound. However, organic germanium is naturally present in microorganisms, mineral water, and medicinal plants, and organic germanium biosynthesized in cells using microorganisms is called 'biogerium', and is more effective than chemically synthesized organic germanium. safe.

Organo germanium provides intracellular oxygen supply (J. Orthomol. Medicin., 1: 145-148 (1986)), purification of blood (Medicin. Hypotheses., 26: 207-215 (1988)), NK cells And activation of marcrophage (Microbiol. Immunol., 29: 65-74 (1985)), induction of interferon secretion (Gantokagakutyoho, 9: 1976-1980), inhibition of cancer cell development and metastasis, neuralgia, osteoporosis It is known to be effective in high blood pressure, cholesterol lowering, hepatitis, and the like. In developed countries such as the United States and Japan, studies are being actively conducted to treat incurable diseases such as cancer and heart disease using organic germanium (see Anticancer Rec., 5). (5): 479-483 (1985).

As a method of obtaining such organic germanium, there is a method of extracting from natural ginseng, natural ganoderma, etc. and a method of chemically synthesizing a reaction between organic acid and germanium dioxide by a catalyst. However, the former is excessively consumed, the latter is not established as a safety or limited use of food or drugs, and biotechnologically mass production of biogermanium using yeast, solves all problems of organic germanium production Can be.

Yeast is a microorganism that has been important for baking and drinking since ancient times, and it has not only been used for human diet for thousands of years, but it is also a good source of nutrition by itself. It is attracting attention as a single cell protein that is a next generation protein source characterized by protein. In addition, yeast is the best source of vitamin B group as a single cell, and contains a large amount of enzymes that play an important role in metabolism in vivo, and is used as a health supplement.

Regarding the production of organic germanium using yeast, US Pat. No. 4,350,846 discloses a method for producing organic selenium by adding selenium to a culture medium of edible yeast, and Japanese Patent Laid-Open No. 53-130483 Although a yeast containing germanium and a method for producing the same are disclosed, these methods are a method of culturing yeast in a medium in which yeast is simply added with inorganic germanium, so that growth is generally inhibited under conditions in which microorganisms have added a large amount of inorganic germanium. In light of this, it is not possible to produce satisfactory amounts of organo germanium by the cultivation of yeast by this method. In addition, the applicant's Korean Patent No. 17638 discloses a yeast containing organic germanium and a method of manufacturing the same, but it is impossible to introduce germanium ions into the cells at high concentrations, which leads to restrictions on the production of high concentrations of cells and after fermentation ends. There was a problem that it is difficult to recover the germanium ions remaining in the balance.

Therefore, there is an urgent need in the art for a method for preparing biogerium yeast capable of introducing germanium ions into cells at high concentration and reusing the germanium ion solution.

Accordingly, the present inventors have made a result of diligent research to develop a method capable of recovering and reusing a germanium ion solution and producing a large amount of safe biogerium at a low cost and high content of biogerium yeast, and thus producing a yeast cell. By separating the process of introducing germanium ions into yeast cells and inducing high content of germanium ions into the cells, it was confirmed that a high content of biogermanium yeast could be prepared, and thus, the present invention was completed.

After all, the main object of the present invention is to provide a method for producing a high-content biogerium yeast by separating the process for producing yeast cells and the process for introducing germanium ions into yeast cells.

Another object of the present invention is to prepare a high content of the yeast cells produced by the production process of the process for producing yeast cells of the process and the process of inducing the introduction of germanium ions into the cells by separating, mixing and reacting in the germanium solution It is to provide biogermanium yeast.

The present invention is a method for producing a large amount of high concentration of biogermanium, culturing Saccharomyces cerevisiae (KCTC-1199) strain was distributed by the KRIBB Gene Bank (KCTC) After separating only the cells, the germanium solution was added to the cells to incubate the cells and germanium under a constant condition to introduce a large amount of germanium ions into the cells to produce a high content biogerium yeast. In general, the growth of microorganisms is limited in the environment in which high concentrations of minerals are present. Therefore, the production of biogermanium by adding germanium during yeast culture is restricted to the production of yeast cells and also remains in the medium after the culture is completed. Since it is difficult to recover the germanium ions, separating the process of cultivating yeast and producing a cell and the process of introducing germanium ions into the cell can produce a high yield of biogerium yeast, as well as recovering germanium. It is very effective in solving these problems.

Hereinafter, the method for preparing a high content biogermanium yeast of the present invention will be described in detail by dividing step.

Step 1: Production of Cells

Yeast strains were cultured for 24 to 48 hours at 25 to 35 ° C. in a medium consisting of 0.3 to 0.5% of peptone, 0.1 to 0.3% of yeast extract, 3 to 5% of glucose, 0.1 to 0.3% of malt extract, and water (rest). To produce. At this time, it is particularly preferable to culture Saccharomyces cerevisiae (KCTC-1199) strain, which was distributed by the KRIBB Gene Bank (KCTC).

Second Step: Introducing Germanium Ion into Cells

Centrifugation of the culture solution obtained in the above step to collect only the yeast cells, and mixed with a solution containing germanium at a concentration of 1000 to 5000ppm, preferably 3000 to 5000ppm in a ratio (volume ratio) of 1: 0.5 to 2, When the germanium-containing culture solution is placed in a reaction tank maintained at pH 5 to 7 and 30 to 40 ° C. and incubated for 15 to 25 hours, high content biogermanium yeast can be produced. Subsequently, the cultured cells are recovered by centrifugation, washed several times to remove inorganic germanium from the surface of the cells and dried, thereby preparing a high content biogermanium dry yeast. In addition, the germanium-containing culture solution can be reused by filtering with a 0.2 ㎛ membrane filter.

Hereinafter, the present invention will be described in detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

Example 1 Determination of Optimal Concentration Ratio by Nutrients

As a protein source, Saccharomyces cerevisiae, obtained from the KRIBB Gene Bank (KCTC) by varying the concentration of nutrients using peptone as a carbohydrate source, glucose and yeast extract as a source of vitamins and trace elements Saccharomyces cerevisiae (KCTC-1199) strains were cultured at 30 ° C. for 48 hours, showing the highest amount of glucose in a medium containing 3.0 wt% glucose, 0.3 wt% yeast extract, 0.5 wt% peptone and water (rest). Cell production was achieved (see Table 1).

Production of Yeast Cells According to Nutrient Ratio (%, ㎎ / ㎖, Hydration) Glucose (%) peptone(%) Yeast Extract (%) Cell Production (Yeast) 1.0 0.1 0.1 41.6 2.0 0.1 0.1 44.5 3.0 0.1 0.1 49.8 4.0 0.1 0.1 41.4 5.0 0.1 0.1 52.2 3.0 0.2 0.1 50.9 3.0 0.3 0.1 56.7 3.0 0.4 0.1 53.3 3.0 0.5 0.1 53.6 3.0 0.5 0.2 53.5 3.0 0.5 0.3 67.4 3.0 0.5 0.4 58.8 3.0 0.5 0.5 53.3

Example 2: Changes in Germanium Content in Yeast Cells According to Germanium Concentration

The cells were recovered by centrifuging the culture broth after the yeast culture, and the germanium solution was added to the cells at a ratio of 1 to 1 by volume (volume ratio), followed by incubation at pH 6.0 and temperature at 30 ° C. for 20 hours. As the solution concentration was higher, more germanium ions were introduced into the yeast cells, and a very high germanium content was shown at 3000 to 5000 ppm (see Table 2). Meanwhile, in order to quantify germanium introduced into yeast cells, 30 ml of nitric acid was added to 1 g of dry bacteria, and then completely decomposed by heating. After calibrating the pH to 6.0, 1 ml of sample, 1 ml of phenylfluorine solution and 1 ml of cyclohexane were added. After mixing and leaving at 30 ° C. for 30 hours, the absorbance was measured at 525 nm to determine the germanium content.

Comparison of Germanium Content in Cells According to Germanium Concentration Germanium solution concentration (ppm) Germanium content in cells (ppm) 1,000 789 2,000 1504 3,000 2551 4,000 3690 5,000 4575

Example 3: Changes in Germanium Content in Yeast Cells According to the Mixing Ratio of Yeast Cells and Germanium Solutions

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After the incubation was completed, only the yeast cells were recovered and the germanium solution (5000 ppm) was mixed therein at various ratios. The pH was 6.0 and the incubation temperature was incubated at 30 ° C. for 20 hours. As shown in Table 3 below, the cells and germanium were incubated. The mixing ratio of the solution showed a high germanium content at 1: 0.5 to 2 (volume ratio).

Comparison of Germanium Content in Cells According to the Mixing Ratio (volume Ratio) of Yeast Cells and Germanium Solutions Mixing ratio (cells: germanium solution) Germanium content in yeast cells (ppm) 1:05 5215 1: 1 4698 1: 2 2672 1: 3 1340

Example 4 Optimal Conditions for Maximizing Germanium Ion Inflow into Yeast Cells

After the completion of the yeast culture, the yeast cells and the germanium solution (5000 ppm) recovered by centrifugation are mixed in a ratio of 1: 1 (volume ratio), incubated in the reactor for 20 hours, and a large amount of germanium ions can be introduced into the cells. As a result of experimenting with the optimum conditions, it was shown that the yeast inflows maximum germanium ions into the cells at a pH of 5 to 7, a reaction temperature of 30 to 40 ℃ (see Table 4).

Effect of Temperature and pH on Inflow of Germanium Ion into Yeast Cells pH Germanium content (ppm) Temperature (℃) Germanium content (ppm) 4.0 1775 20 2552 5.0 3168 30 3845 6.0 4638 35 5344 6.5 5836 40 4789 7.0 3556 45 2321 8.0 2567 50 1842

Example 5: Comparison of germanium content in yeast cells by incubation time

As shown in Table 4, the germanium solution mixed with the yeast cells at pH 6.5, the temperature of 35 ℃, which is the optimal condition for the culture, was added to the reactor to compare the germanium content introduced into the yeast cells according to the culture time. Incubation for 15 to 25 hours was shown to induce maximum germanium ions into the cells.

Comparison of Germanium Content in Cells According to Incubation Time Response time (hr) Germanium content in cells (ppm) 5 2389 10 3913 15 4680 20 5541 25 5060 30 3681

As described and demonstrated in detail above, the present invention provides a high concentration of biogermanium and a method for producing the same in large quantities. According to the method for producing high content biogermanium of the present invention, after culturing the yeast strain to isolate only the cells, the germanium solution is added to the cells and cultured the cells and germanium under constant conditions to introduce germanium ions into the cells, high yield Biogerium yeast can be produced, and the recovery of germanium is possible, such as the limitation of yeast cell production when producing germanium by adding germanium during yeast culture and difficulty in recovering germanium ions remaining in the medium after the culture is completed It is very effective in solving the problem.

Claims (1)

i) Yeast is incubated in a medium consisting of 0.3 to 0.5% peptone, 0.1 to 0.3% yeast extract, 3 to 5% glucose, 0.1 to 0.3% malt extract and water (rest) at 25 to 35 ° C for 24 to 48 hours. To produce the cells; And, ii) The yeast cells produced in the electrical process are separated and mixed in a germanium solution having a germanium concentration of 3000 to 5000 rpm in a ratio of 1: 0.5 to 2 (volume ratio), and mixed at a pH of 5 to 7, and a temperature of 30 to 40 ° C., 15 to 15 Method for producing a high content of biogermanium yeast comprising the step of incubating for 25 hours to induce the introduction of germanium ions into the cells.