KR101883825B1 - Manufacturing method of fermented cynanchum wilfordii and fermented cynanchum wilfordii manufactured by same method - Google Patents

Abstract

The present invention relates to a method for producing fermented Bacillus subtilis using a Rythopus oryzae strain and a fermented Bacillus subtilis prepared by the method. Fermented white water produced by the method of the present invention using the Rissofus orientalis can be usefully used as a composition for antioxidant and anti-inflammation by enhancing the antioxidative and anti-inflammatory effects.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a method for producing fermented Baechuo, and a fermented Baechu-fermented Baechu-based fermented Baechu-

The present invention relates to a process for producing fermented fermented milk and a fermented fermented milk produced by the process.

Hassuo is a perennial herbaceous perennial native to China, native to China. It is possible to cultivate in the whole country because it is wild, cold tolerance, insecticide, and resistance to weather in the mountainside of each country. There are two kinds of red and white seaweeds. White seaweeds have the efficacy of nourishing tonic, blood, blood, echinacea, and leukemia, and are useful for the treatment of diseases such as debilitating anemia, anemia, early white hair, nervous breakdown, ). The enemy Suseo is a kind of aesthetics such as interpolation, exorcise, blood, breeze, 寧心安 神, 爱 精 精, 골 骨, It has an efficacy that it is a year-old unhealthy, and it has the efficacy that it is a year-round fire. In the Korean Pharmacopoeia, the standard of herbal medicines is defined by dividing the raw seaweed into seaweed and the white seaweed into white seaweed.

Baeksoo is a member of the Asclepiadaceae family, and is sometimes referred to by other names such as mockery, dwarf, sunshine, new mockery, and root. In one room, the herbaceous plant root (root of root) was dried. In addition, the roots of white roots are one-component materials, and have an effect of darkening gray hair in particular. Unripe radish is effective in constipation by promoting intestinal motility.

The roots of Bacillus subtilis include active ingredients such as cynanchol, cynandione A, cardiac glycosides, amino acids and sugars. Double synandione A exhibits anti-inflammatory activity by inhibiting the phosphorylation of IkB-a and the migration of NF-kB into the nucleus.

In recent years, researches are being conducted to utilize the roots of papaya or papaya oyster roots or their extract components as materials for medicines, health functional foods, and cosmetics. For example, Korean Patent No. 10-1041437 discloses a composition for treating vascular disease in a composition containing an extract of Bacillus subtilis as an active ingredient. In addition, there has been reported a use of a combination extract of Bacillus subtilis mixed with other herbal materials for the prevention of hair loss and promotion of hair growth, and for the treatment of sleep disorders of Bacillus subtilis extract.

Accordingly, the inventors of the present invention have completed the present invention by confirming that the fermented Bacillus anthracnose produced by using Rizufus orientalis, a kind of Bacillus subtilis, has an antioxidative and anti-inflammatory effect.

It is an object of the present invention to provide a process for producing fermented Bacillus subtilis having an antioxidative and antiinflammatory activity enhanced by using a Rythopus oryzae strain.

It is another object of the present invention to provide a fermented soybean curd prepared by the above method.

It is still another object of the present invention to provide a cosmetic composition or a health functional food containing the fermented white ginseng extract or the extract thereof produced by the above method as an active ingredient.

In order to achieve the above object, the present invention provides a method for producing fermented Bacillus subtilis having enhanced antioxidant and anti-inflammatory activity, comprising culturing the Bacillus subtilis strain inoculated with the Rispuss oryzae strain.

In addition, the present invention provides a fermented soybean curd prepared by the above method.

The present invention also provides an anti-inflammatory cosmetic composition containing fermented white ginseng or an extract thereof produced by the above method as an active ingredient.

In addition, the present invention provides an anti-inflammatory health functional food comprising fermented white ginseng or an extract thereof produced by the above method as an active ingredient.

The present invention also provides an antioxidant cosmetic composition comprising fermented white ginseng or an extract thereof produced by the above method as an active ingredient.

In addition, the present invention provides an antioxidant health functional food containing fermented white ginseng or its extract prepared by the above method as an active ingredient.

Fermented white water produced by the method of the present invention using the Rissofus orientalis can be usefully used as a composition for antioxidant and anti-inflammation by enhancing the antioxidative and anti-inflammatory effects.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing a method for producing a fermented white bean.
Fig. 2 is a photograph of a white water fermented by using Rizoffus oryzae.
FIG. 3 is a graph showing cytotoxicity of the fermented Bacillus subtilis extract. FIG. The Y axis represents the cell survival rate as compared to the control group, and the X axis represents the concentration of the fermented white water extract (CW-RO) treated.
4 is a graph showing the inhibitory effect of nitrogen monoxide production on the fermented white water extract. The Y axis represents the nitrite concentration, and the X axis represents the concentration of the fermented white water extract (CW-RO) treated and the LPS treatment.
FIG. 5 is a graph showing the inhibitory effect on the tumor necrosis factor (TNF-α) expression of the fermented Bacillus subtilis extract. The Y-axis represents the concentration of TNF-α, and the X-axis represents the concentration of the fermented white water extract (CW-RO) and LPS treatment.

Hereinafter, the present invention will be described in detail.

The present invention provides a method for producing fermented Bacillus subtilis with enhanced antioxidant and anti-inflammatory effects, which comprises culturing a Bacillus subtilis strain with inoculation.

The strain may be used at a concentration of 1 x 10 ³ condia / ml to 1 x 10 ⁹ condia / ml, specifically 1 x 10 ⁴ condia / ml to 1 x 10 ⁸ condia / ml, more specifically 1 x 10 ⁵ condia / 1 × 10 may be inoculated in a concentration of ⁷ condia / ㎖. In one embodiment of the invention, the strain may be inoculated at a concentration of 1x10 < ⁶ > condia / ml.

The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. If the incubation temperature is out of the above range, the activity of the Ricorpops oryzae strain is lowered and fermentation may not occur. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days. If the incubation period is too short, the fermentation does not occur sufficiently. If the incubation period is too long, the antioxidant ability and antiinflammatory activity of the fermented white rot can be lowered.

In a specific embodiment of the present invention, the present inventors have produced fermented Bacillus subtilis using a Rythopus oryzae strain.

In addition, the present invention provides a fermented soybean curd prepared by the method of the present invention.

The above fermented feed water has the above-described characteristics. For example, the production method of the present invention can be carried out by inoculating a strain of Bacillus subtilis oryzae. The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days.

In a specific embodiment of the present invention, the fermented white wool produced by the method of the present invention has an increased anti-inflammatory component, cinanideone content, as well as a radical scavenging ability. On the other hand, suppression of nitric oxide production or tumor necrosis factor expression was observed (see Tables 1 to 2 and 3 to 5).

The present invention also provides an anti-inflammatory cosmetic composition containing fermented white ginseng or an extract thereof produced by the method of the present invention as an active ingredient.

The above fermented feed water has the above-described characteristics. For example, the production method of the present invention can be carried out by inoculating a strain of Bacillus subtilis oryzae. The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days.

The fermented white water produced by the production method of the present invention may further comprise drying and extracting. The drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying. In one embodiment of the present invention, the drying may be freeze-drying.

The extraction can be carried out using water, alcohol or a mixture thereof as a solvent. The alcohol may specifically be a C ₁ to C ₄ lower alcohol, and more specifically may be ethanol. In one embodiment of the invention, the solvent may be water. As the extraction method, any conventional method known in the art can be used, such as extraction under reduced pressure, hot water extraction, filtration, hot water extraction, immersion extraction, reflux extraction, cold extraction, steam extraction, room temperature extraction or ultrasonic extraction. In one embodiment of the invention, the extraction method may be hot water extraction.

The extraction may be performed for 10 minutes to 4 hours, specifically 30 minutes to 3 hours, more specifically 50 minutes to 2 hours. In one embodiment of the invention, the extraction can be carried out for one hour. In addition, the number of times of extraction can be performed once to three times, specifically once or twice. In one embodiment of the present invention, the extraction may be performed twice.

In a specific embodiment of the present invention, the fermented white wool produced by the method of the present invention has an increased anti-inflammatory component, cinanideone content, as well as a radical scavenging ability. On the other hand, suppression of nitric oxide production or tumor necrosis factor expression was observed (see Tables 1 to 2 and 3 to 5).

The cosmetic composition may be added in an amount of 0.1 to 50% by weight, specifically 1 to 10% by weight, based on the total weight of the cosmetic composition. However, the above ratio may vary depending on the form of the cosmetic product, the specific application site and the application capacity.

The cosmetic composition of the present invention includes components commonly used in cosmetic compositions in addition to the above-mentioned fermented white ginseng extract and its extracts, and includes conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, .

The cosmetic composition of the present invention can be prepared in any formulations conventionally produced in the art, and can be prepared, for example, in the form of paste, cream, gel, powder, spray, solution, emulsion, suspension (anhydrous and aqueous) - containing cleansing, anhydrous products (oil and glycol), masks, packs, powders, lotions, soaps, oils, powder foundations, emulsion foundations, wax foundations and the like. Specifically, it can be manufactured in the form of a flexible lotion (skin), a nutritional lotion (milk lotion), a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, Can be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, a chlorofluorohydrocarbon , Propane / butane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent may be used as a carrier component. Examples thereof include fatty acid esters of water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, and the like can be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The present invention also provides an anti-inflammatory health functional food comprising fermented white ginseng or an extract thereof produced by the method of the present invention as an active ingredient.

The above fermented feed water has the above-described characteristics. For example, the production method of the present invention can be carried out by inoculating a strain of Bacillus subtilis oryzae. The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days.

The extract has the above-described characteristics.

In a specific embodiment of the present invention, the fermented white wool produced by the method of the present invention has an increased anti-inflammatory component, cinanideone content, as well as a radical scavenging ability. On the other hand, suppression of nitric oxide production or tumor necrosis factor expression was observed (see Tables 1 to 2 and 3 to 5).

The content of the fermented Bacillus anthracis extract or its extract, which is an active ingredient added to the health functional food, can be determined depending on the purpose. Generally, it may be 0.01 to 90 parts by weight of the total weight of the health functional food.

There is no particular limitation on the form and kind of the health functional food. The form of the health functional food to which the substance can be added may be a tablet, a capsule, a powder, a granule, a liquid or a ring.

The health functional food of the present invention may contain various flavoring agents or natural carbohydrates as an additional ingredient such as a normal health functional food. Such natural carbohydrates may include sugar alcohols such as monosaccharides such as glucose and fructose, polysaccharides such as disaccharides such as maltose and sucrose, dextrin, cyclodextrins, xylitol, sorbitol, erythritol and the like . As the sweetening agent, natural sweetening agents such as tau Martin and stevia extract, synthetic sweetening agents such as saccharine or aspartame, and the like may be used.

In addition to the above, the health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, Alcohol, and the like. These components may be used independently or in combination. The proportion of such additives may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides an antioxidative cosmetic composition comprising fermented white ginseng or an extract thereof produced by the method of the present invention as an active ingredient.

The above fermented feed water has the above-described characteristics. For example, the production method of the present invention can be carried out by inoculating a strain of Bacillus subtilis oryzae. The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days.

The extract has the above-described characteristics.

In a specific embodiment of the present invention, the fermented white wool produced by the method of the present invention has an increased anti-inflammatory component, cinanideone content, as well as a radical scavenging ability. On the other hand, suppression of nitric oxide production or tumor necrosis factor expression was observed (see Tables 1 to 2 and 3 to 5).

The present invention also provides an antioxidant health functional food containing fermented white ginseng or an extract thereof produced by the method of the present invention as an active ingredient.

The above fermented feed water has the above-described characteristics. For example, the production method of the present invention can be carried out by inoculating a strain of Bacillus subtilis oryzae. The cultivation may be carried out at a temperature of 20 ° C to 40 ° C, specifically, 25 ° C to 35 ° C, more specifically, 28 ° C to 32 ° C. In one embodiment of the invention, the incubation temperature may be 30 < 0 > C. The incubation period may be from 2 days to 6 days, more specifically from 3 days to 5 days. In one embodiment of the invention, the incubation period may be four days.

The extract has the above-described characteristics.

In a specific embodiment of the present invention, the fermented white wool produced by the method of the present invention has an increased anti-inflammatory component, cinanideone content, as well as a radical scavenging ability. On the other hand, suppression of nitric oxide production or tumor necrosis factor expression was observed (see Tables 1 to 2 and 3 to 5).

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

However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited thereto.

Risofus Orissa  Fermentation using strain White  Produce

3-year-old white water five (Prince, Korea) and a sterile, 1 × 10 ⁶ condia / ㎖ concentration in the separation tank crispus duck (Rhizopus of here oryzae , RO) (National Academy of Agricultural Science, Korea). This was cultivated at 30 DEG C for 4 days to prepare fermented white pears (FIGS. 1 and 2). At this time, fermented Bacillus subtilis not inoculated with the strain was prepared as a control.

Fermentation Baewooo Oh  Preparation of extract

The hot water extract of fermented Bacillus subtilis and control Bacillus subtilis was prepared by the following method.

Specifically, to prepare a hot-water extract of fermented Bacillus subtilis and control Bacillus subtilis, the fermented white water produced in the above <Example 1> was dried, and 100 g of boiled distilled water was added to 5 g of dried fermented bacilli . The extract was extracted twice using an ultrasonic homogenizer (Power Sonic 420, Hwashin Co., Korea) for 1 hour at a voltage of 50/60 Hz and 700W, and the extract was filtered using Watman No.2 filter paper. The filtrate was concentrated under reduced pressure at a temperature of 50 ° C or below by using a rotary vacuum concentrator (BUCHI, USA), and then freeze-dried to prepare a powder.

Fermentation Baewooo Oh  Toxicity of extracts

The cytotoxicity of fermented Bacillus subtilis extract was confirmed by MTT method.

Specifically, RAW 264.7 cells were inoculated in a 96-well plate at a concentration of 5 × 10 ³ cells / ml per well and cultured for one day. Then, the cells were cultured in the same manner as in Example 2 except that 50 μM, 100 μM, 200 μM or 400 μM Were added to the cells. It was cultured in a 5% CO ₂ incubator at 37 ° C for 3 days, and cytotoxicity was confirmed by the conventional MTT method.

As a result, no cytotoxicity was observed by the fermented white water extract of 400 / / ml concentration (FIG. 3).

< Experimental Example  1> Fermentation White  Active ingredient analysis

The active ingredients of the fermented white water extract prepared in Example 2 were analyzed by UPLC (Ultra Pressure Liquid Chromatography).

Specifically, ACQUITY UPLC BEH C18 (1.7 탆, 2.1 횞 50 ㎜) was used as the column, and solvent A (water containing 0.2% acetic acid) and solvent B (100% acetonitrile) min. &lt; / RTI &gt; The content of cinanideone A was measured at a wavelength of 250 nm using a PDA (Waters, Aquity UPLC) detector. The experiment was expressed as the mean ± standard deviation (mean ± SD) of the values repeated three times.

As a result, as shown in Table 1, the extract of Bacillus subtilis, which is a control group, was 37.85 ± 0.023 mg per 100 g of Bacillus subtilis, but the extract of Bacillus subtilis was 100.77 ± 0.164 mg more than three times (Table 1).

Analysis of active ingredients of fermented Bacillus sample The content of Shinanthion A per 100 g of Bacillus anthracis (mg) CW (Baewo Oh) 37.85 + 0.023 CW / RO (fermented soybeans) 100.77 + 0.164

< Experimental Example  2> Fermentation White  Check antioxidant efficacy

The antioxidant activity of the fermented white water extract prepared in Example 2 was confirmed by analyzing DPPH radical scavenging ability.

Specifically, 80% ethanol was mixed with DPPH (Sigma) solution to prepare a radical solution having a concentration of 100 μM. The prepared radical solution and each of the extracts prepared in Example 2 were mixed and allowed to stand under light-shielding conditions for 30 minutes. Then, the absorbance of the mixture was measured by a spectrophotometer at a wavelength of 517 nm, and a radical scavenging ability was calculated. As a positive control, ascorbic acid, a typical antioxidant, was used. The experiment was expressed as the mean ± standard deviation (mean ± SD) of the values repeated three times.

As a result, as shown in Table 2, the radical scavenging ability of the fermented white water extract (CW / RO) was 26.30 ± 1.894%, which was about twice that of the control CW extract (Table 2).

Analysis of DPPH Radical Scavenging Ability of Fermented White Water Extracts sample DPPH scavenging (%) CW (Baewo Oh) 11.45 + 0.985 CW / RO (fermented soybeans) 26.30 ± 1.894 Ascorbic acid (10 [mu] M) 20.86 ± 0.293

< Experimental Example  3> Fermentation White  Confirming anti-inflammatory efficacy

&Lt; 3-1 > Confirmation of Nitric Oxide Production Inhibitory Effect

The inhibition of nitrogen monoxide production of the fermented white water extract prepared in Example 2 was analyzed by the following method.

Specifically, RAW 264.7 cells were inoculated to a 96-well plate at a concentration of 2 × 10 ⁴ cells / ml per well and cultured for one day. Then, 50 μM, 100 μM, 200 μM or 400 μM Concentration of fermented Bacillus subtilis extract was added to the cells. This was incubated for 2 hours, and further treated with 1 μg / ml of LPS and incubated for 18 hours. After the culture, the culture solution was mixed with Griess reagent (Promega) at a ratio of 1: 1 and reacted for 10 minutes at room temperature under light shielding conditions. The absorbance of the mixture was then measured with a spectrophotometer at a wavelength of 540 nm.

As a result, the fermented Bacillus subtilis extract inhibited the production of nitrogen monoxide in a concentration-dependent manner (Fig. 4).

&Lt; 3-2 > Confirmation of tumor necrosis factor expression inhibitory effect

The inhibitory effect of the fermented white water extract prepared in Example 2 on tumor necrosis factor expression was analyzed by the following method.

Specifically, RAW 264.7 cells were inoculated to a 96-well plate at a concentration of 2 × 10 ⁴ cells / ml per well and cultured for one day. Then, 50 μM, 100 μM, 200 μM or 400 μM Concentration of fermented Bacillus subtilis extract was added to the cells. The cells were cultured for 2 hours, further treated with 1 μg / ml of LPS and cultured for 24 hours. TNF-α levels were measured by the manufacturer's method using a TNF alpha mouse ELISA kit (Abcam).

As a result, the expression of TNF-α was inhibited in a concentration-dependent manner (FIG. 5) in a statistically significant manner ( ^* p <0.05, ^** p <0.01).

Claims (11)

A method for producing fermented Bacillus subtilis having enhanced antioxidant and anti-inflammatory activity, comprising a fermentation step in which a strain of Rhizopus oryzae is inoculated and cultured.
The method according to claim 1, wherein the strain is inoculated at a concentration of 1 × 10 ³ condia / ml to 1 × 10 ⁹ condia / ml.
The method according to claim 1, wherein the cultivation is carried out at a temperature of 20 ° C to 40 ° C.
The method according to claim 1, wherein the cultivation is performed for 2 days to 6 days.
A fermented beef cattle produced by the method of claim 1.
A fermentation step in which a strain of Rhizopus oryzae is inoculated and cultured; And
And an extraction step of extracting the fermented white water produced by the fermentation step with hot water.
An anti-inflammatory cosmetic composition comprising the fermented white ginseng or the extract of claim 5 as an active ingredient.
The cosmetic composition for anti-inflammation according to claim 7, wherein the extract is extracted using water, a C ₁ to C ₄ lower alcohol or a mixture thereof as a solvent.
An anti-inflammatory health functional food comprising the fermented white ginseng or the extract of claim 5 as an active ingredient.
An antioxidative cosmetic composition comprising the fermented white ginseng or the extract of claim 5 as an active ingredient.
An antioxidant health food containing the fermented white ginseng or the extract of claim 5 as an active ingredient.

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