KR101116180B1 - The microbial fermented products of androgen from natural products containing sterol and its production method - Google Patents

Abstract

The present invention relates to an androgen-containing fermentation product obtained by fermenting a sterol-containing natural product using a microorganism and a method for preparing the same, and specifically, inoculating a microorganism having an ability to decompose a side chain of sterol to an animal or vegetable natural product including a sterol By fermentation to produce a fermentation product containing androgen to provide a fermentation product containing androgens capable of inhibiting or treating abnormalities in health caused by menopausal disorders and androgen deficiency in men.

Fermentation products containing androgen of the present invention can be usefully used as a health supplement or therapeutic agent in the phenomenon caused by androgen deficiency.

Androgens, animal sterols, vegetable sterols, sterol degrading microorganisms, fermentation

Description

Androgen-containing fermentation products obtained by fermenting a natural product containing sterols using microorganisms and a method for preparing the same {The microbial fermented products of androgen from natural products containing sterol and its production method}

The present invention relates to an androgen-containing fermentation product obtained by fermenting a sterol-containing natural product using a microorganism and a method for preparing the same, and specifically, inoculating a microorganism having an ability to decompose a side chain of sterol to an animal or vegetable natural product including a sterol By fermentation to produce a fermentation product containing androgen, and relates to a fermentation product containing androgens capable of inhibiting or treating abnormal health symptoms caused by menopausal disorders and androgen deficiency in men.

The number of people who complain of menopausal disorders is increasing in proportion to the increase in the elderly population as the life span of people increases. The main cause of these menopausal disorders is the lack of sex hormones or reduced production. As a method of overcoming menopausal disorders, women are receiving hormonal therapy to periodically administer estrogen, a female sex hormone, and men, and hormone therapy to apply androgens, such as testosterone, a male sex hormone. However, such sex hormone replacement therapy is a method of administering synthetic purified female or male sex hormone in a medical institution, and there is a problem that is not readily available to the general public. Therefore, if the fermentation product obtained by fermenting animal or plant natural products can include sex hormones or sex hormone-like substances in the product, it would be possible to suppress or heal the health abnormalities resulting from sex hormone deficiency or imbalance.

In addition, in the case of eggs by reducing or eliminating the cholesterol contained in a large amount through the fermentation process can provide a low-sterol or nosterol egg fermentation products in case of concern about excessive intake of cholesterol in adults.

The present invention inoculates and ferments microorganisms capable of degrading the side chains of sterols into natural products containing sterols such as eggs to produce fermentation compositions containing androgens, thereby causing abnormal health effects caused by menopausal disorders and androgen deficiency in men. It relates to a fermentation product containing androgen that can inhibit or treat and a method for producing the same.

An object of the present invention is to inoculate microorganisms capable of degrading the side chains of sterols into natural products containing a large amount of sterols such as eggs and fermented to produce fermentation compositions containing low and no sterols, and androgen. Provided are compositions comprising androgens capable of inhibiting or treating abnormalities in health caused by androgen deficiency.

In order to achieve the above object, the present invention is inoculated microorganisms, such as Mycobacterium genus, Arthrobacter genus, Norcadia genus, Brevibacterium genus which can decompose the side chains of sterols, preferably Mycobacterium genus microorganisms in natural products containing a large amount of sterols such as eggs To fermentation composition having a final sterol content of 50% by weight or less of the initial sterol content or low sterol or musterol, and androgen.

In the present invention, the culture medium is shaken for 5 days at 30 degrees Celsius in a culture medium containing glucose to use the culture medium as a seed culture.

In the present invention, the seed culture medium of the microorganism during fermentation is used in an amount of 5 to 1000% by weight based on the weight of the natural product including sterol, preferably 10 to 500% by weight.

The sterol-containing natural products usable in the present invention include animal natural products such as eggs, duck eggs, quail eggs, and vegetable natural products such as cacao, soybeans, hemp (fermented herbs, cheonma), nymph, ginseng.

The culture medium was nutrient broth (3g bee extract, 8g peptone, 1000ml water), yeast extract-malt extract broth (yeast extract 4g, malt extract 10g, glucose 4g, water 1000ml), tryptone glucose broth (10g glucose, 1g sodium nitrate, 0.5 g of K ₂ HPO _4, 0.5 g of MgSO _4, 0.5 g of urea, 50 mg of FeCl ₃ , 1 g of tryptic soy broth, and 1000 ml of water) may be used, and the culture medium may vary depending on the microorganism used for fermentation.

The concentration of glucose is preferably 0.1 to 5% by weight of the culture medium weight, more preferably 1.5 to 3.0% by weight is used to add.

In the present invention, the fermentation period is 5 days to 100 days, the fermentation temperature is preferably maintained at 20 to 40 degrees Celsius, more preferably 10 to 40 days, 25 to 35 degrees Celsius.

The present invention is inoculated by fermentation of a microorganism capable of decomposing the side chains of sterols into natural products containing sterols such as eggs and fermented low-sterol or musterol, and by producing a fermentation composition containing androgen to prevent menopausal disorders and androgen deficiency in men By producing a fermentation product containing androgens that can suppress or treat the health problems caused by the health, by using it can easily prevent or treat menopausal disorders resulting from androgen deficiency can be useful for maintaining elderly health in an aging society have.

Hereinafter, the present invention will be described in detail by the following examples.

Preparation Example: Preparation of Microbial Species Culture Solution

Mycobacterium spp. M3805 strain (KCTC 1829) was inoculated in a nutrient broth medium containing 1% by weight of glucose and shaken at 30 ° C for 5 days, and the culture medium was used as the seed culture solution.

Qualitative analysis by thin layer chromatography

Samples were deposited on a thin layer of silica gel and developed in a mixture of chloroform: ethyl ether (10: 2.5) to dry the solvent, and then checked for the presence of substances such as androgens, which showed ultraviolet absorption under ultraviolet light, and sprayed with 10% sulfuric acid solution at 80 degrees Celsius. -Heat for 1 hour in a 90-degree oven to develop colors and check for materials such as sterols and androgens.

Quantitative Analysis by Gas Chromatography

Using a gas chromatography device with a column for steroid analysis, the substances contained in the sample are separated and detected by a flame detector to check the respective retention time and quantify the amount of the substance from the peak area.

Example 1. Mycobacterium  Bacteria Determination of Glucose Concentrations Suitable for Androgen Production in Fermentation

To determine the glucose concentration suitable for the fermentation of androgens including testosterone from sterols, glucose was added to the broth (nutrient broth; beef extract 3g, peptone 8g, water 1000ml) at 0.75%, 1.5%, 2% and 3% concentrations. Mycobacterium sp. M3805 strain (KCTC 1829) was inoculated in a culture medium containing cholesterol at 0.1% concentration and shaken at 30 ° C. for 20 days. The fermentation broth was extracted with ethyl acetate and analyzed by silica gel thin layer chromatography.

As shown in FIG. 1, the production of testosterone was remarkable in addition to 4-androstenedione at the concentration of 3% glucose. Therefore, it is possible to improve the production of testosterone by adjusting the concentration of glucose to 3% in fermentation broth.

Example 2. Eggs Mycobacterium  Bacteria Confirmation and Changes of Androgen Production in Fermentation

Aseptically add the contents of one egg to 100 mL of the cultivation solution, extract the culture solution every 10 days with shaking culture at 30 degrees Celsius, extract with ethyl acetate solvent, and extract the thin layer chromatography or gas. Chromatographic analysis was performed to confirm the presence of cholesterol and androgen, a degradation product thereof, contained in the fermentation broth.

As shown in FIG. 2, as the fermentation days increase, the amount of sterol decreases and the androgen substance increases, and almost no spot of sterol was observed in the 40-day culture, and the spots of the androgen substances were remarkably confirmed.

In addition, the results of the analysis using gas chromatography to quantify the change over time was as shown in Table 1. Over time, cholesterol was decreased and androgen was accumulated.

 Changes in Sterols and Androgens in Fermentation Broth (per ml) with Fermentation Time Effective date (days) Amount of sterol (μg) Amount of 4-anthrostenedione (μg) Testosterone (μg) 0 1052 - - 10 722 - - 20 526 56 - 30 176 226 - 40 18 354 24

Example 3 Changes in Cholesterol and Androgen According to the Added Egg Volume

The contents of the eggs were mixed, and the mixtures were mixed with 100 ml of microbial culture and 25 ml, 38 ml, 50 ml, and 75 ml of each were mixed and cultured, and the time and amount of androgen for sterol exhaustion and androgen production were examined. .

As shown in FIG. 3, when the amount of the egg contents was changed in the same amount of the seed culture solution, the more the amount added, the longer the fermentation time required to lose the sterol. When 75 ml was added, it could be confirmed that sterol remained in the 50-day fermentation. Table 2 shows the quantitative results of androgens using gas chromatography.

 Changes in Sterols and Androgens According to the Added Egg Volume Egg Content Stock Price 25 ml 38 ml 50 ml 75 ml Number of days spent cultivating cholesterol 25 days 35 days 40 days 60 days or more Production amount of 4-androstendione per ml of fermentation broth (μg) 250 328 366 184 Amount of residual cholesterol per ml of fermentation broth (μg) - - - 194

Experimental Example 4. Effect of Androgen in Fermentation Products

As a result of 3 months of application to the eyebrows in the evening, mixed with 2 g of a fermentation product containing androgen (containing 1440 ㎍ androgen) in a lotion base 45 ml, the color and number of eyebrows were increased as shown in FIG. . These results are physiologically showing that androgens in fermentation products act on the human body and show effects. Therefore, the fermentation product may be usefully applied to alleviate or prevent physiological phenomena occurring in androgen deficiency or degradation by making an appropriate formulation (lotion, patch, capsule, etc.) applicable to the human body.

Figure 1 shows the result of the glucose concentration changes in the fermentation using the bacteria Mycobacterium genus.

Figure 2 shows the androgen production and changes over time in the fermentation of Mycobacterium sp. Bacteria of eggs by TLC.

Figure 3 shows the change in the amount of egg content added by TLC and confirmation of residual cholesterol and androgen production in fermentation broth.

Figure 4 shows the effect on the eyebrow to which the fermentation product of the present invention is applied.

Claims (6)

delete delete delete delete A seed culture solution containing Mycobacterium spp. M3805 strain (KCTC 1829) added to the sterol containing egg at a concentration of 2 to 3% by weight of the culture medium was inoculated in an amount of 5 to 500% by weight, and 20 to 40 degrees Celsius. The final sterol content of the fermentation product, characterized in that incubated for 5 to 100 days at 50 wt% or less of the initial sterol content is a low sterol or sterol-free method for producing a fermentation product containing androgen. delete

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