KR100698533B1 - A Method for Preparing Compositions for Preventing and Treating Antimicrobial Infenction and Antitumor Action using Opuntia humifusa Fermented Liquids - Google Patents

Abstract

The present invention relates to a method for preparing a Winter Fermented Cactus ( Ountia humifusa ) fermentation broth that has anticancer activity against cervical cancer cell lines, and a composition for preventing and treating antibacterial infection and anticancer activity using the same. Yeast is added to provide a composition having excellent anticancer activity and an antioxidant function.

Fermentation broth provided in the present invention is excellent in anti-cancer activity and can provide a health functional composition excellent in antioxidant capacity by the addition of oligomeric proanthocamide (OPC).

Winter Cactus, Health Drink, Anticancer Activity    

Description

A method for preparing compositions for preventing and treating antimicrobial infenction and antitumor action using opuntia humifusa fermented liquids

1 is a block diagram showing the manufacturing process of the winter cactus fermentation broth of the present invention,

2 is a graph showing the element content ratio of each part in the frozen cactus fermentation broth freeze-dried of the present invention,

Figure 3 is a graph showing the anticancer activity effect by the winter cactus cactus crushed liquid obtained by the present invention, a is stem methanol extract, b is stem ethanol extract, c is stem water extract, d is fruit methanol extract, e is fruit Ethanol extract, f represents the leaf extract, respectively

Figure 4 is a graph showing the anticancer activity of the winter cactus fermentation broth obtained by the present invention, a is a winter cactus cactus fermentation broth, b is a winter cactus cactus fermentation broth.

The present invention relates to a method for preparing a wintering cactus ( Opuntia humifusa ) fermentation broth that is excellent in anti-cancer and anti-oxidant activity and useful for the prevention and treatment of diseases, and a composition for preventing and treating antimicrobial infections and anti-cancer effects.

There are more than 4,000 kinds of cactus on the planet. Among them, cactus with fruit is called palm cactus and has been used for food and food substitute since ancient times. According to the herbaceous tree, palm cactus is known to be effective in bronchial, asthma, cough, lung disease, gastritis, constipation, enteritis, nephritis, hypertension, diabetes, heart disease, neuralgia, arthritis. Drinking these palm cactus juice has a diuretic effect, activation of intestinal motility and appetite-boosting effects, and has recently been used in skin diseases, rheumatism and burn treatment. Palm cactus is being re-examined as a functional plant as it is known for its improvement in digestive function, wound healing effect, and effect on rheumatoid arthritis. As a study on the physiological activity of cactus, Paik recently confirmed the antioxidant function as well as the anti-ulcer activity by cactus fruit powder.

Opuntia ficus-indica var.saboten is a tropical plant belonging to the Cactaceae family, and it is a naturalized plant native to Jeju Island in Korea. On the other hand, palm cactus is developed and marketed as a variety of favorite foods, and according to the component analysis, it contains a large amount of dietary fiber, vitamins and flavonoids. As such, various studies on the efficacy and composition of Jeju palm cactus have been reported. In particular, studies on the activation of immune system cells in rats, antimicrobial effect and antioxidant activity against six food poisoning microorganisms such as E. coli Effects have been announced.

However, Jeju baeknyeoncho is not known to survive in the cold of minus 20 ^o C, so it is impossible to live in the middle of Korea. Currently, Opuntia humifusa, a type of palm cactus, is known to overwinter at minus 20 ^o C. Its characteristics and composition are also known to be different from those of the centennial plant growing on Jeju Island. Woldong cactus is a perennial plant that can be cultivated for years to decades and can be grown in Yeongnam, Honam, and Gangwon-do. As bioactive substances have been concentrated for many years, they have excellent functionality and various physiological activities such as antioxidant, antibacterial and anti-ulcer effects are known, but their effects on anticancer activity are not known.

Currently, it is a product that uses palm cactus, and functional drinks that are extracted and sold along with herbal medicines such as stems, leaves, or roots are sold on the market. In addition, as the antioxidant and antibacterial activity is known, it is marketed as a functional soap, cosmetic raw materials and packs. However, as the manufacturing method of the functional beverage is extracted at high pressure and high temperature, a lot of vitamins weak in heat are destroyed, and it is difficult to recognize the efficacy of the effect. Therefore, the invention using the new palm cactus fermentation broth has begun.

Furthermore, fermentation broth manufacturing method of Baeknyeoncho in Jeju, which is similar to wintering cactus, is prepared by mixing sugar or vinegar with food or food additives such as sugar, vinegar, etc. in Korean Patent Application No. 1994-0039828 A method for producing a composition using cactus fruits, such as lychees, pickles, beverages, and liquor of cactus fruits, prepared by extractive purification, is disclosed.

In addition, Korean Patent Application Publication No. -0013602 (Method for preparing functional food using palm cactus) discloses a method of manufacturing syrup, jelly, extract, etc. using the fruit, stem, or leaf of palm cactus. However, such a manufacturing method is not possible to prepare a beverage capable of physiological activity.

Therefore, in the present invention, by applying pressure to the wintering cactus itself, and then adding yeast, or lactic acid bacteria, the anti-cancer activity of cervical cancer, which has not been known until now, is reinforced, and at the same time, oligomeric proanthamide (OPC) in the fermentation broth prepared The present invention was completed by focusing on the addition of) to prepare novel functional raw materials that maximize taste and antioxidant activity.

That is, it is an object of the present invention to provide a method for producing a winter cactus fermentation broth that enhances antioxidant activity without increasing vitamins and increases anticancer activity by fermentation microorganisms.

Another object of the present invention is to provide a composition for the prevention and treatment of antimicrobial infections and anti-cancer effects using the winter cactus palm cactus fermentation broth excellent in antioxidant activity.

According to one aspect of the present invention, in order to achieve the above object,

Adding 4-6% (w / v) brown sugar to the fruit, stem or root ground liquid of the washed Cactus Opuntia humifusa ;

Inoculating the brown sugar addition liquid simultaneously or sequentially with lactic acid bacteria and yeast within a total amount of 5-15% (v / v); And

 Provided is a method for producing a palm cactus fermentation broth comprising the step of fermenting the inoculum at 35 to 45 ° C. for 2 to 10 days.

According to another aspect of the present invention,

Prevention of antimicrobial infections and anti-cancer effects, which comprises the pulverized liquid and oligomeric proanthocamide (OPC) in a weight ratio of 1 to 10: 0.1 to 5: 0.01 to 5 in the palm cactus fermentation broth prepared as described above. A therapeutic composition is provided.

Hereinafter, a method for preparing a winter cactus fermentation broth of the present invention will be described with reference to FIG. 1.

First, a pulverized liquid is prepared by grinding the washed winter cactus fruit or stem or root or a mixture thereof. Existing palm cactus could use only fruit or stem, but the winter cactus used in the present invention is characterized by high efficiency because it can be used not only fruit, stem but also root.

Brown sugar is added to the obtained winter cactus crushed liquid. In this case, brown sugar is added to supplement sugar for the purpose of effectively performing fermentation, and its content is usually about 4-6% (w / v).

Then, as a microorganism for fermentation, lactic acid bacteria or yeast is added. At this time, lactic acid bacteria or yeast can be inoculated simultaneously or sequentially in the total amount of the strain solution grown to 10 ⁹ cells / ml within the range of 5-15% (v / v). If the total amount of inoculated bacteria is less than 5%, the inoculated strain is killed and no fermentation is performed. If the inoculated strain is 15% or more, the anti-cancer activity of the fermentation broth is reduced due to the shortening of the fermentation period. to be.

The type of beneficial microorganisms to be used is not particularly limited, yeast include Saccharomyces process three Levy jiae (Saccharoces cerevisae) a, and the lactic acid bacteria is Lactobacillus Planta room (Lactobacillus plantarum, ATCC8014) or Lactobacillus casei (Lactobacillus Use of casei ATCC7469) is preferred in that it is commonly available.

In addition, the number of yeasts and lactic acid bacteria added for fermentation is not limited to this, but it is preferable that they are usually available about 10 ⁹ cells / ml.

In order to ferment the useful microorganism input liquid obtained in this way, fermentation is carried out at 35 to 45 ° C. for 2 to 10 days to effectively prepare the wintering cactus fermentation broth of the present invention while preventing the destruction of vitamins under high temperature and high pressure. At this time, the fermentation step may be repeated depending on the presence or absence of fermentation.

At this time, the presence or absence of fermentation is determined by checking the pH. That is, if the pH is around 4.0 is completed, if it does not meet the additional fermentation to determine whether the pH 4.0 and then complete the fermentation step. After fermentation, the total number of lactic acid bacteria is 10 ⁷ ~ 10 ⁸ cells / ml and the total number of yeast is 10 ⁸ ~ 10 ⁹ cells / ml. However, it may increase slightly as the storage period elapses.

The composition obtained by containing the crushed liquid and oligomeric proanthocymidine (OPC) in the weight ratio of 1-10: 0.1-5: 0.01-5 in the wintering cactus fermentation broth prepared in this way was prevented against antimicrobial infection and anticancer activity. The therapeutic effect can be exerted. At this time, out of the above range is not preferable because the harmony of anti-cancer activity, antioxidant capacity is broken.

At this time, the grinding liquid is added as an excipient, and is more preferable in terms of toxicity or anticancer action than adding general chemicals.

In addition, OPC is usually a substance having an antioxidant action, in the present invention serves to enhance the antioxidant action in the fermentation broth expressing the antioxidant action.

Furthermore, in the present invention, by providing the winter cactus fermentation broth in a liquid type and a powder type, it can be provided as a cosmetic raw material excellent in health functional beverages and anti-cancer effect and antioxidant activity, respectively.

EMBODIMENT OF THE INVENTION Hereinafter, the form for implementing this invention is demonstrated in detail, taking an example. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Preparation Example 1 Preparation of Fermentation Broth of Winter Cactus Cactus

Washed palm cactus fruit, stems, roots (total 100 kg) was ground to a size of less than 2.O × 2.O mm using a grinder to prepare a palm cactus grinding liquid. 5% (w / v) of brown sugar was added to the obtained pulverized liquid.

Then, 10% (v / v) of Saccharoces cerevisae (10 ⁹ cells / ml) culture medium was added to the wintering cactus crushed liquid, and then Lactobacillus plantarum (ATCC8014) as lactic acid bacteria. (10 ⁹ cells / ml) The culture was inoculated with 10% (V / V).

The inoculum was fermented at 40 ° C. for about 10 days to prepare a palm cactus fermentation broth of the present invention. The pH of the prepared fermentation broth was confirmed to be near 4.O and the step was completed. After fermentation, the total number of lactic acid bacteria was 10 ⁷ cells / ml and the total number of yeasts was about 10 ⁸ cells / ml.

By using the prepared palm cactus fermentation broth, the antioxidant activity of the palm cactus fermentation broth of the present invention was reinforced by 1-10% by weight ratio to increase the antioxidant activity, but not in the pulverized liquid before fermentation, but the antibacterial and anticancer activity was increased after fermentation. Was observed.

Example 1 Analysis of General Components of Winter Cactus Fermentation Broth

The general components of the fermentation broth of overwintering cactus prepared in Preparation Example 1 were analyzed by A.O.A.C.

The crude fibers of the samples were measured with a crude fiber analysis device (Fibertec system, Sweden) using acid and alkali decomposition methods. Moisture was measured three times with an infrared moisture meter (Kett, FD-240, Japan), and the crude protein content was determined by using the Kjeltec-1035 (Auto sampler system, Sweden) to determine the total nitrogen content using a Kjeltec method. Calculated by multiplying. Crude fat and crude ash were measured according to the A.O.A.C method. The dietary fiber was added to α-amylase in a dry powder and heated for 15 minutes in boiling water, followed by 0.2 N hydrochloric acid (pH 1.5) and hydrolyzed at 40 ° C. for 6 hours using pepsin.

After the enzyme treatment, sodium hydroxide was neutralized, and then pancreatin was added thereto, followed by shaking at 40 ° C for 1 hour. 95% ethanol was added to the digested reaction solution and left to stand for 1 hour.

Insoluble precipitate produced during standing was centrifuged at 3,000 rpm for 20 minutes to separate total fiber. The precipitate was dissolved in distilled water and centrifuged at 3,000 rpm to dry the precipitate at 100 ° C. to form an insoluble dietary fiber, and 95% ethanol was added to the supernatant and allowed to stand for 1 hour. After separation, it was dried and made into soluble dietary fiber, and the total amount of insoluble and soluble dietary fiber was used as the total dietary fiber amount.

To analyze crude fat, sample was dried 1-2 hours at 90 ℃ in advance, ether was extracted with ether, and Soxhlet fat extractor was used to continuously extract lipids from food for 16-32 hours, and then ether was removed from the extract. The Soxhlet extraction method using the residue obtained by drying at 100 degreeC as a crude fat was used.

The general component analysis results of the fermentation broth of the obtained winter cactus are summarized in Table 1 below.

Energy calorie (kcal / 100g) Crude protein (%) Crude lipid (%) Crude carbohydrate (%) stem  86 0.92 0.12 20 Root 121 2.4 0.12 28 Fruit  65 1.04 0.08 15

As shown in the table above, the water content of stem was 88.4%, crude protein was 8.1%, crude fat was 1.1%, crude ash was 12.4%. Fruit moisture content was 89.1%, crude protein was 4.6%, crude fat was 1.5. % And crude ash were 7.8%, indicating no significant difference in moisture and crude fat contents of stems and fruits. However, the contents of stalks contained roughly twice as much crude protein and crude ash as fruits.

When comparing the energy calories, the stem and root showed 1.3 and 1.9 times higher calories than the fruit. Carbohydrates were similar in stem and root, but fruit was as low as 15%.

Example 2 Elemental and Mineral Analysis of Winter Cactus Fermentation Broth

Lyophilized fermentation broth was pulverized with a grinder to compare elemental analysis ratios for winter cactus fermentation broth, and was supported by Elemental Analyzer (FISONS, EA 1110) from Kyungpook National University. H), and nitrogen (N) was compared by quantitative analysis.

Analysis of Inorganic Components

For comparison and analysis of inorganic components, I received analysis and support from Busan Branch of Korea Institute of Basic Science.

Assay method is dry method, and the pretreatment process is to make fermentation broth into dry powder, add 0.2-0.3 g of each sample to Teflon vessel, add 5 ml of HNO ₃ (high purity, GR grade by Merck), and then open the stopper. Heated at 100-150 ° C. Through this process, the acid was completely blown out, and again, 3 ml of HNO ₃ and 0.5 ml of HCl were added thereto, and the stopper was closed and heated under pressure at about 100 ° C. for about a day.

After the container was completely cooled, the stopper was opened to confirm that the sample was completely dissolved in acid, and then heated again until about 1 ml of acid remained while the stopper was opened. Dilute with distilled water (× 100) and use ICP analyzer (Jobin-Yvon, JY 38 Plus, France) with ICP-AES to calculate calcium (Ca), magnesium (Mg), zinc (Zn), iron (Fe) and phosphorus (P). ), Respectively.

In addition, winter cactus was fermented by site and then lyophilized to compare the composition ratio of the element content and the results are shown in FIG. As shown in FIG. 2, when looking at the site-specific element content, no significant differences were found, such as 38.4-39.6% carbon, 43.3-44.4% oxygen, 5.6-6.2% hydrogen, and 0.9-1.8% nitrogen. Sulfur was not measured in all site-specific fermentations.

When comparing elemental analysis and general composition for the fermentation broth by site, no special difference was recognized. Although the composition of each part may be the same, the inorganic ingredient may be an important variable when developing various foods.

In this regard, the inorganic components for each part were analyzed by ICP and the results are shown in Table 2 below.

Concentration (ppm) Sample AS Ca CD Fe Mg Pb Zn stem  ND 1,800  ND  21  497  ND  710 Root  ND 2,520  ND 12 1,319  ND  610 Fruit  ND  388  ND 10  781  ND 1,627

As shown in the table, 2,520 ppm was detected in the root of Ca, which showed 6.5 times higher calcium content than the fruit. However, Zn showed 2.6 times higher content in fruit.

In conclusion, it could be concluded that it is more advantageous to use the mixed portion in an appropriate ratio than to use only a specific portion.

Example 3. Acute Toxicity Test of Winter Cactus Fermentation Broth

In this example, a safety test for skin irritation of wintering cactus fermentation broth was conducted based on the toxicity test standard of medicines.

This test was conducted in a laboratory set at a temperature of 23 ± 3 ℃, a relative humidity of 50 ± 10%, a ventilation frequency of 13-18 times / hr, a lighting time of 12 hours (7 am to 7 pm), and an illuminance of 150 to 300 Lux. . All the test workers were trained to enter the animal room and wore work clothes, hoods, masks, and gloves.

The temperature and humidity of the animal room were measured every hour by the automatic temperature and humidity meter during the breeding period of the test period, and there was no change that would affect the reliability of the test as a result of environmental measurement.

During the period of acclimation and testing, stainless steel wire cages (220 W × 410 L × 200 H mm) were raised to 5 rats / cage. The box was affixed with an identification card containing the test number, animal number and dose. Feed was freely ingested by irradiation sterilization of solid feed for experimental animals (Cheil Feed Co., Ltd.), and tap water was freely ingested.

The pulverized liquid of palm cactus was used as food, but the palm cactus was administered at a maximum dose of 2,000 mg / kg for the acute toxicity test to ensure the safety of the fermented broth.

Group separation of animals was performed as follows. First, the body weight of all the animals determined to be healthy during the acclimation period was measured, and winter cactus palm cactus fermentation broth was prepared in the same manner as in Preparation Example 1 for each dose immediately before administration to each of five male and female rats.

After fasting overnight before the fermentation broth was administered to animals, a single forced oral administration was performed using an injection tube attached with an oral needle. The dosage amount (ml / kg b.w) was calculated based on the weight immediately before administration.

Then, on the day of dosing, the general condition was observed every hour from 1 hour to 6 hours after administration, and the change of general condition, symptoms of poisoning and the presence of dead animals were observed once daily from the next day to 14 days after administration.

After completion of the observation period, hemorrhage was performed under ether anesthesia to observe the gross abnormalities of internal organs in detail. Gross abnormal organs were stored fixed in 10% neutral formalin solution.

There were no dead animals in male and female rats after palm cactus fermentation, and no clinically specific findings were observed. Autopsy findings showed no change in kidney, liver and lung in both male and female.

In conclusion, palm cactus fermentation broth was found to be nontoxic to rats.

Example 4. Anticancer Effect of Winter Cactus Crushed Liquid and Fermented Liquid

Dulbecos modified Eagle medium (DMEM) and trypsin-ethylenediamine tetraacetic acid (EDTA) for culturing the cervical cancer cell lines KaSki, SiHa and keratinocyte cells HaCaT , Antimycotic-antibiotics were purchased from GIBCO Laboratories (Grand Island, NY), and calf serum (FBS) from Hyclone Laboratories (Logan, UT), 96 well tissue culture plates and tissue culture dishes. Nunc, Inc. (North Aurora Road, IL), and sulforodiamine B (sulphorodiamine B, SRB) were purchased from Sigma.

An ELASA reader was used for absorbance measurement (manufactured by Molecular Device), a cell culture was used for a CO ₂ incubator (manufactured by Vision), and a microscope was used for an inverted microscope (manufactured by Nikon).

The cervical cancer cell lines CasSki, SiHa and keratinocyte cells HaCaT maintained in DMEM / 10% FBS were placed in 96 well plates at 1 × 10 ⁴ / well and incubated in a CO ₂ incubator for 1 day. After 24 hours, each of the extracts were changed to medium and diluted to various concentrations. After 24 hours, the cells were fixed at −20 ° C. with 70% acetone solution and dried to saline the remaining cells in the well using 0.4% SRB solution. After washing and drying, SRB was eluted with 10 mM Trisma base solution, and the absorbance at 562 nm was measured with an ELISA reader.

Cytotoxicity of Each Extract of Cervical Cancer Cell Lines As a result of treating the extracts of wintering cactus to cervical cancer cell lines KaSki, SiHa and keratinocyte cells HaCaT up to 50 ㎍ / ml As can be seen, CaSki and SiHa showed no concentration-dependent cytotoxicity.

On the other hand, the fermentation broth obtained in Preparation Example 1 was lyophilized as it is to see the anticancer effect of the wintering cactus fermentation broth according to the present invention on cervical cancer cell line. In the group (a) treated with the fermentation broth, the growth was markedly reduced due to the concentration-sensitive cytotoxicity in the CaSki cell and SiHa cell lines. Among the cervical cancer cell lines, keratinocyte HaCaT cell lines cancer-cellized by a cause other than HPV, as shown in Figure 4, it was confirmed that no cytotoxicity.

In addition, the group (b) treated with the lyophilized product by diatomaceous earth filtration showed the same tendency as the unfiltered group. This suggests that palm cactus fermentation broth inhibits the growth of CaSki and SiHa cell lines with HPV virus genome.

Example 5 Antibacterial Activity of Winter Cactus Crushed Liquid and Fermented Liquid

In this example, the effect was verified by the diffusion method to confirm the antimicrobial activity using the winter cactus crushed liquid and fermentation broth.

Five types of KCCM40510, KCCM11593, KCCM41294, KCCM40881, and KCCM41291, which are Gram-positive strains that are resistant to Staphylococcus aureus , are distributed by the Korea Microorganism Conservation Center (KCTC), Muller-Hinton Hinton) was inoculated in the medium and pre-cultured at 37 ℃.

The precultured strains were inoculated in Müllerton media to test media and the effects of the first antimicrobial activity on the ingredients shown in Table 3 were verified by the diffusion method using a paper disk.

That is, 60 μl of the component shown in Table 3 was adsorbed onto the paper disc, and then placed on a test medium, incubated overnight at 37 ° C., incubated for 18 hours, and then the ring was measured in mm. Except for repeating the experiment and the results are shown in Table 3 together.

division KCCM40510 KCCM11593 KCCM41294 KCCM40881 KCCM41291 Skin irritation Experimental Example Grinding liquid 18 17 16 17 14 - Fermentation broth 18 15 18 16 11 - Comparative Grinding Liquid 8 8 8 8 8 - Comparative fermentation liquid 12 12 8 8 13 - * Comparative grinding liquid is aloe grinding liquid and fermentation liquid is aloe fermented liquid prepared in the same process

As shown in Table 1, the fermentation broth showed slightly stronger antimicrobial activity than the aloe crushed liquid, but it was not significantly noticeable. It was confirmed that the antibacterial activity was very excellent compared to the liquid.

Preparation Example 2 Preparation of Health Functional Drink

The composition of the health drink using the fermented palm cactus fermentation broth fermented by the above production method is as follows.

   Fermentation broth 99 ml

   OPC ... 1-2 ml

Preparation Example 3 Preparation of Health Functional Powder

The wintering cactus palm cactus fermentation broth having the composition described in 2 above was lyophilized to prepare a total powder formulation of 10 g.

Example 6-Anticancer and Antioxidant Activities of the Compositions Prepared by Grinding Solution, Fermentation Solution, and Preparation Examples 2 and 3

In this sample example, the anticancer and antioxidant activity levels of the pulverized liquid, the fermented broth, and the compositions prepared according to Preparation Examples 2 and 3 were confirmed to verify each activity.

The anticancer activity and antibacterial activity was measured by the method described in the above is dissolved in antioxidant activity Antioxidant activity assay using the DPPH (1,1-diphenyl-2- picrylhydrazyl) samples of various concentrations in 4 mL of methanol 1.5X10 ^- After adding 1 mL of ⁴ M DPPH methanol solution, the solution was left at room temperature for 30 minutes, and the absorbance was measured at 517 nm. The results are shown in Table 4.

Classification (1 mg / ml) Anticancer activity (cell growth inhibition rate) (%) Antibacterial Activity (mm) (KCCM40881) Antioxidant activity (%) (DPPH) Skin irritation CaSki SiHa HaCaT Experimental Example Grinding liquid 0 0 0 8 14 - Fermentation broth 60 59 60 16 11 - Preparation Example 2 34 40 48 18 8 - Preparation Example 3 40 37 39 18 13 - Composition of Preparation Example 2 Fermentation broth 99% OPC 1-2% Composition of Preparation Example 3 Fermentation broth 99g OPC 1g

As shown in the table, it was confirmed that the anticancer activity bactericidal activity, antioxidant activity values of the compositions prepared by Preparation Example 2 and Preparation Example 3 was more improved than the fermentation broth itself.

As described above, the wintering cactus fermentation broth provided by the present invention and the composition obtained therefrom are excellent in anticancer activity and the addition of oligomeric proanthocamidine (OPC) can provide a health functional composition excellent in antioxidant activity.

Claims (8)

delete delete delete delete delete A first step of adding 4-6% (w / v) brown sugar to the fruit, stem or root grinding liquid of the washed Cactus Opuntia humifusa ; A second step of inoculating the brown sugar addition liquid simultaneously or sequentially with lactic acid bacteria and yeast within a total amount of 5-15% (v / v); A third step of fermenting the inoculating solution at 35 to 45 ° C. for 2 to 10 days; If the pH of the fermentation broth is less than 4, the second step of inoculating the lactic acid bacteria and yeast in the total amount of 5-15% (v / v) at the same time or sequentially in the second fermentation; And The obtained Winter Cactus fermentation broth, the fruit, stem or root grinding liquid of the Winter Cactus ( Ountia humifusa ) obtained in the first step, and the oligomeric proanthocymidine (OPC) were 1 to 10: 0.1 to 5: 0.1 to 5, respectively. A fifth step of mixing in a weight ratio of; Method for producing a composition for the prevention and treatment of antimicrobial infections, including A first step of adding 4-6% (w / v) brown sugar to the fruit, stem or root grinding liquid of the washed Cactus Opuntia humifusa ; A second step of inoculating the brown sugar addition liquid simultaneously or sequentially with lactic acid bacteria and yeast within a total amount of 5-15% (v / v); A third step of fermenting the inoculum at 35 to 45 ° C. for 2 to 10 days; If the pH of the fermentation broth is less than 4, the second step of inoculating the lactic acid bacteria and yeast in the total amount of 5-15% (v / v) at the same time or sequentially in the second fermentation; And The obtained Winter Cactus fermentation broth, the fruit, stem or root grinding liquid of the Winter Cactus ( Ountia humifusa ) obtained in the first step, and the oligomeric proanthocymidine (OPC) were 1 to 10: 0.1 to 5: 0.1 to 5, respectively. A fifth step of mixing in a weight ratio of; Method for producing a composition for prevention and treatment having anticancer activity comprising a A first step of adding 4-6% (w / v) brown sugar to the fruit, stem or root grinding liquid of the washed Cactus Opuntia humifusa ; A second step of inoculating the brown sugar addition liquid simultaneously or sequentially with lactic acid bacteria and yeast within a total amount of 5-15% (v / v); A third step of fermenting the inoculating solution at 35 to 45 ° C. for 2 to 10 days; If the pH of the fermentation broth is less than 4, the second step of inoculating the lactic acid bacteria and yeast in the total amount of 5-15% (v / v) at the same time or sequentially in the second fermentation; And The obtained Winter Cactus fermentation broth, the fruit, stem or root grinding liquid of the Winter Cactus ( Ountia humifusa ) obtained in the first step, and the oligomeric proanthocymidine (OPC) were 1 to 10: 0.1 to 5: 0.1 to 5, respectively. A fifth step of mixing in a weight ratio of; Method for producing a composition for preventing and treating antioxidant activity, including

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