KR101694500B1 - Cosmetic composition comprising fermented rice root extract and method for preparing thereof - Google Patents

Abstract

The present invention provides a cosmetic composition for skin whitening comprising a fermented extract of rice (Oryza sativa L.) roots as an active ingredient, and a process for producing a fermented extract of rice roots.

Description

[0001] The present invention relates to a cosmetic composition comprising a rice root-root fermentation extract and a method for producing the same,

The present invention relates to a cosmetic composition comprising a rice-root-fermented extract and a method for producing the same.

The skin color of a person is determined by pigmentation, distribution and concentration of melanin such as spots and freckles, and the more melanin, the darker the skin color. Melanin is the last product in the metabolism of tyrosine, a kind of amino acid. It is a pigment molecule present in most organisms. It is a polymer of phenolic compounds and exists mainly as a melanin protein which binds strongly to globulin. It does not dissolve in water but dissolves in alkaline solution or concentrated sulfuric acid. Melanin has the ability to block more than a certain amount of ultraviolet rays, which keeps skin temperature and protects skin from ultraviolet rays. Cells that make melanin are called melanocytes, and melanin is made from cell organelles called melanocytes in melanocytes. Melanocytes extend melanocyte-bearing protrusions to deliver melanin to other cells around the keratinocyte. Melanin is present in cells in many different ways, and changes in skin color occur when melanocytes regulate the diffusion and aggregation of melanocytes.

Melanin is produced by an enzyme called tyrosinase, which is a complex and sequential chemical reaction. When melanosomes are filled with melanin, they migrate to the epidermis through keratinocytes and develop color. Melanogenesis in melanocytes is the first step in the formation of tyrosinase, and melanosomes should be formed, melanosomes appear in the form of small carriers and germinate from the Golgi apparatus. Man has two kinds of melanomas, one is Eumelanosome, egg-shaped and about 0.5 ~ 2㎛ in size. Another is the Pheomelanosome, which is spherical and 0.5 μm in size. In this melanoma, brown or black pigment contains sulfur. It is a dark colored eumelanin and yellow or reddish brown pigment, which contains a deep concentration of sulfur and forms a brightly colored peromelanin. Hair, and color of the eyes.

Waste designated by the Ministry of Environment means a substance that is not required for human life or business activities by burning materials, sludge, waste oil, etc. Agricultural wastes are substances that are deemed to be worthless after agricultural production activities are completed. Typical examples are rice roots after harvest, paprika leaves after harvest, starch foil after starch extraction from potatoes and potatoes, and residue left over from oil palm oil.

Rice roots are always submerged in water, and the agricultural waste is abandoned due to its low quality and useful value. It is very old history that started to use agricultural waste in Korea, and mud bricks made of rice straw have been used as building materials as a representative example. So far, agricultural waste has been widely used as a substitute for building materials. In recent years, plants such as poplar, willow, acacia, sugar cane and sweet potato have been used as biofuels in the development of "alternative raw materials" have.

The utilization of industrial wastes promotes resource circulation, thereby exceeding the limit of limited resources and positively contributing to value creation.
A related prior art is Korean Patent Laid-Open Publication No. 1315638.

The object of the present invention is to provide a cosmetic composition which exhibits whitening effect together with skin stability.

Another object to be solved by the present invention is to provide a cosmetic composition showing inhibition of tyrosinase activity and inhibition of melanin formation.

Another object of the present invention is to provide a rice root-roasted fermented extract exhibiting excellent tyrosinase activity inhibition and melanin production inhibitory effect.

The cosmetic composition for skin whitening of the present invention contains a fermented extract of rice ( Oryza sativa L. ) roots as an active ingredient.

The method for producing the rice-root-fermented extract of the present invention comprises preparing a medium containing the root extract of rice, inoculating the medium with the K. pneumoniae strain KCCM11570P, and culturing the medium inoculated with the strain Culture and fermentation.

The present invention can provide a cosmetic composition showing whitening effect with skin safety.

The present invention can provide a cosmetic composition that exhibits inhibition of tyrosinase activity and inhibition of melanin formation.

The present invention can provide a rice root-roasted fermented extract exhibiting excellent tyrosinase activity inhibition and melanin production inhibitory effect.

FIG. 1 is a view showing cytotoxicity of rice root extract. FIG.
FIG. 2 is a view showing the cytotoxicity of the rice root-roots extract of the present invention obtained by fermenting the rice seed root extract with the use of a strain of Kupiabidus vulcanis of Deposit No. KCCM11570P.
FIG. 3 is a chart for confirming the whitening effect of the rice root extract before fermentation. FIG.
FIG. 4 is a chart showing the whitening effect of the rice root-roots fermentation extract. FIG.

Hereinafter, specific embodiments of the present application will be described in more detail. However, the techniques disclosed in this application are not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention.

In order to solve the above problems, the inventors of the present invention continued research to produce a natural material showing excellent whitening effect without skin toxicity, and found that the rice root was used as a fermentation strain and a specific strain of Cupriavidus basilensis And the fermented extract fermented showed a remarkable whitening effect, skin stability and moisturizing effect. The fermentation strain was identified and named as JHskin3-6, and it was entrusted to Korea Microorganism Conservation Center on September 19, 2014 and received KCCM11570P.

The cosmetic composition according to one embodiment of the present invention may contain a fermented extract of rice ( Oryza sativa L. ) roots as an active ingredient.

The rice (Oryza sativa L.) is an annual herbaceous plant of the alopecia plantus. The East Indian is planted in rice fields and fields with edible crops from the Wonsan. Its height is about 1m. Leaves are thin and long. When it matures, the end of the stem comes out and the flower grows around the end of July to August. Rice is called "rice", and about 40% of the world's population is made up of rice. More than 20 species are known to belong to the rice plant, but most of them are cultivated rice (O. sativa). The crops used in rice are hawthorn and rice straw, and the root of rice, which is the underground part of rice, is mainly abandoned as agricultural waste.

In the present invention, the rice root means the root of rice ( Oryza sativa L. ), and may specifically include a ground portion of rice, more specifically, a ground portion of rice paddy. After the rice is harvested, it is possible to create industrial value by utilizing the underground portion of rice, which is classified as ordinary waste.

The fermented extract of the rice seed roots can be prepared by fermenting the rice root extract. The above-mentioned rice root extract can be obtained by a conventional plant extraction method. For example, rice roots can be dried by sunlight or hot air and then extracted with an extraction solvent.

As the extraction method, an extract containing the active ingredient can be obtained by extraction treatment by an arbitrary method such as, for example, hot water extraction, reflux cooling extraction, ultrasonic extraction, cold water extraction, percolation and the like. Concretely, it can be extracted once to five times by the extraction method of hot water extraction.

When the mixture is extracted by cold extraction, it can be extracted by heating in an extractor equipped with a reflux condenser at 50 to 100 ° C for 4 to 20 hours or by immersing at 5 to 37 ° C for 1 to 15 days. In the case of cold beating and percolation, it takes about 1 to 200 hours. In the case of extracting hot water, it can be performed for 0.5 to 72 hours at a temperature close to the reflux temperature of the solvent, depending on the type and temperature of the solvent used. In particular, tinctures or fluid extracts or extracts leached into functional alcohols can be used.

The extraction solvent may be water, an organic solvent or a mixture thereof, and the organic solvent may be an anhydrous or hydrated lower alcohol of C1-4. For example, as the extraction solvent, polar solvents such as water, ethanol, propanol, methanol, n-butanol, acetone, and 1,3-butylene glycol; Non-polar solvents such as ether, hexane, benzene, chloroform, and ethyl acetate; A mixed solvent of the non-polar solvent and the polar solvent; Solvents such as alkaline water, vegetable oils such as soybean oil and sesame oil; Or mixtures thereof. Specifically, water and a mixed solvent of at least one of ethanol, methanol, n-butanol, and propanol, or 1,3-butyleneglycol can be used.

The extraction solvent may be 1 to 50 times, 1.5 to 20 times, and more preferably 2 to 6 times the extraction volume of the dried rice roots.

More specifically, the methanol extract of dried rice roots is extracted with 1 to 3 times the volume of methanol at 20 to 30 ° C for 1 to 72 hours, and the solvent is evaporated to obtain a first extract. The first extract After adding n-butanol 1 to 3 times its volume and extracting with hot water at 20 to 30 ° C for 1 to 72 hours, the solvent is evaporated and concentrated and lyophilized to obtain a rice root extract powder.

The fermentation of the above-mentioned rice root extract can be obtained by fermenting the root extract of rice with the use of Cupriavidus basilensis sp. Strain of KCCM11570P . By fermenting the root extract of rice with the above strain, inhibition of tyrosinase activity and inhibition of melanin synthesis are remarkably enhanced, and a natural whitening extract without toxicity can be provided.

 The rice roots fermentation extract may be contained in an amount of 0.001 to 30% by weight, more preferably 0.02 to 10% by weight, more preferably 0.05 to 5.0% by weight based on the total weight of the composition. And there is no problem in skin safety.

The composition of the present invention can be used as a whitening cosmetic composition, which can provide an excellent whitening effect by inhibiting tyrosinase activity and inhibiting the production of melanin.

The composition according to the invention may be formulated containing a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example as a solution, a gel, a solid, a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic form (liposome) In the form of ionic fibrin dispersions, or in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions may be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening agents, gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, Such as fillers, emulsifiers, emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics May contain adjuvants commonly used in the field of cosmetics or dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

In addition, the composition of the present invention may contain a skin absorption promoting substance to increase the skin improving effect.

In addition, the composition of the present invention may contain, in addition to the rice roots fermentation extract, other ingredients capable of synergizing with the skin protection effect, within the range not impairing the anti-aging, moisturizing and whitening effect of the present invention can do.

A method for preparing a rice-root fermented extract according to an embodiment of the present invention comprises preparing a culture medium containing the rice root extract, inoculating the culture medium with the Kugelavus vulcanis strain of Deposit No. KCCM11570P, Culturing and fermenting the inoculated medium.

The rice root extract can be prepared as described above with respect to the cosmetic composition according to one embodiment of the present invention. The prepared rice root extract can be used as a powder state by evaporating a solvent. More specifically, a rice root extract powder which is freeze-dried after concentration and evaporation of a solvent can be used.

The medium may comprise the rice root extract powder, for example, by sterilizing and heating a mixture containing the rice root extract powder and distilled water. The mixture may be from 1:99 to 99: 1, in particular from 99: 1 to 1: 1, more specifically from 99: 1 to 4: 1, by weight of the roots root extract powder: distilled water. Within this range, fermentation by Kupirabidus vulcanisis is effective.

The culture medium can be cultured at 24 to 32 ° C, specifically at 26 to 30 ° C, for 24 to 128 hours, more specifically, for 36 to 96 hours, inoculated with the strain Kupirabidus vulcanis of Accession No. KCCM11570P.

After the cultivation, the cells are removed from the culture medium and filtered to produce a rice roots fermented extract.

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to examples, comparative examples and test examples. However, these examples, comparative examples and test examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples, comparative examples and test examples.

[Test Example 1] Isolation and identification of fermentation strains

The face of three 25 year old female skin was wiped with sterile gauze soaked with sterilized water to collect a sample, and the supernatant of the sample was taken by shaking in sterilized distilled water. 0.1 ml of the sample diluted to 10 ^-4 times (volume) was added to the R2A agar medium (0.5 g / L of yeast extract, 0.5 g / L of Proteose Peptone No. 3, 0.5 g / L of casamino acid, L of silver starch, 0.5 g / L of soluble starch, 0.3 g / L of sodium pyruvate, 0.3 g / L of dipotassium phosphate, 0.05 g / L of magnesium sulfate and 15 g / L of agar) Respectively.

A total of 17 microorganisms were isolated by visual observation according to the phenotype of the cultured colonies. Using the microorganisms, the rice root extract was fermented in the same manner as in Example 1 to obtain a remarkably excellent whitening effect And the strains were isolated.

Genomic DNA of the isolated strain was isolated and PCR was performed to amplify 16s rDNA. Specifically, 25 μl of 2X Tag DNA Polymerase Master Mix (MgCl _2, dNTP, reaction buffers addition / Promega, USA) was added to 1 μl of DNA, and the forward primer (27F: 5'-AGAGTTTGATCCTGGCTCAG-3 ' 1 μl of reverse primer (No. 1492R: 5'-TACGGYTACCTTGTACGACTT-3 ') of No. 2 was added to each well, and distilled water was added to make 50 μl of the final reaction solution. The denaturation at 95 ° C for 5 minutes was followed by denaturation at 95 ° C for 30 seconds, primer binding at 52 ° C for 45 seconds, and DNA synthesis at 72 ° C for 45 seconds, followed by 30 cycles of DNA synthesis at 72 ° C for 8 minutes Lt; RTI ID = 0.0 > 4 C. < / RTI >

Search the nucleotide sequence of the amplified fragment obtained from the genetic bank database (Genbank database) results, Ku-free Avi Bar Douce siren system to identify as corresponding to the (Cupriavidus basilensis) to paste the named person JHskin3-6 separated, and in 2014 09 On the 19th day, it was entrusted to the Korea Microorganism Conservation Center and received the accession number KCCM11570P.

[Comparative Example 1] Production of rice root extract

8 kg of rice roots were washed, dried, and then fined. Dried rice roots were extracted with 1.5 times 90% methanol at 25 ° C for 24 hours, and the solvent was evaporated to obtain 284 g of the first extract. To the first extract, 5 kg of n-butanol was added and the mixture was fractionated at 25 캜 for 24 hours, concentrated, and lyophilized to prepare a rice root extract powder.

 [Preparation Example 1] Production of natural root broth root

The rice seed extract powder obtained in Comparative Example 1 was inoculated in a 2 L flask with 10 g of rice root extract powder and 990 g of distilled water, and the mixture was heat-sterilized at 121 캜 for 15 minutes and cooled to 25 캜 to prepare a rice broth natural medium.

[Example 1] Production of rice root roots fermented extract

Cupriavidus basilensis sp. Strain (hereinafter referred to as "strain") of Accession No. KCCM11570P was placed in a flask and incubated at 28 ° C for 2 days at 110 rpm for preincubation.

The pre-cultivated strains were inoculated to the rice seed-roots natural medium prepared in Preparation Example 1 in an amount of 0.01 wt% based on the medium weight, and then cultured at 28 DEG C for 72 hours at 110 rpm. The cells were centrifuged at 9,000 rpm for 30 minutes with a large-capacity centrifuge (2236HR, Zyrogen Co., Ltd.) to remove the cells, followed by filtration through a 0.45 μm filter to produce a roots fermented extract.

[Test Example 2] Cytotoxicity of Rice Root Extract

Human keratinocyte HaCaT (ACTT, CLS 300493, USA) was divided into 3 x 10 ³ cells per well in a 96-well plate with a hemacytometer and cultured at 37 ° C and CO ₂ for 24 hours. The cell culture broth was removed, and the root extract of rice broth prepared in Comparative Example 1 was added to the cells in a concentration of 200 μg / ml, 100 μg / ml, 50 μg / ml, 25 μg / ml, 12.5 μg / ml, 6.25 μg / (Dulbecco's Modified Eagle Medium, Gibco BRL, USA) containing 10% fetal bovine serum (FBS) at a concentration of 1.5625 μg / ml was added to each well in an amount of 100 μl And cultured at 37 ° C and CO ₂ for 24 hours.

In the untreated group, distilled water was treated with the same amount of the extract. In the positive control group, 10% DMSO (dimethyl sulfoxide, Sigma-Aldrich) was treated with the same amount of the extract.

To measure the survival rate of cells, WST-1 (high sensitive Water Soluble Tetrazolium salt) reaction solution was diluted 1/10-fold in DMEM medium, treated with 100 쨉 l per well and allowed to react for 2 hours. Then, ELISA absorption analyzer , Thermo scientific) and the cell activity was calculated according to the following formula 1, and the results are shown in FIG.

<Formula 1>

Cell activity (%) = (absorbance of sample treated group / absorbance of no treated group) * 100

From the results shown in Fig. 1, it can be seen that the rice root extract exhibited cytotoxicity due to a decrease in cell activity in a concentration-dependent manner at a concentration of 25 μg / ml or more.

[Test Example 3] Cytotoxicity of fermented rice extract

The rice seed roots fermented extract prepared in Example 1 was treated at a concentration of 200 μg / ml, 100 μg / ml, 50 μg / ml, 25 μg / ml, 12.5 μg / ml and 6.25 μg / The experiment was carried out in the same manner as in Test Example 1 except for that the cell activity was calculated and the results are shown in FIG.

In the results of FIG. 2, it can be seen that when the rice root roots fermented extract was treated, the cell activity did not show a significant change up to a concentration of 200 μg / ml, and thus the skin stability showed almost no cytotoxicity.

[Test Example 4] Inhibitory effect of rice root extract on extracellular melanin synthesis

In order to confirm melanin synthesis inhibitory effect of the rice root extract of Comparative Example 1, experiments were carried out using B16 / F10 melanoma cells (ATCC, CRL-6323) as follows.

B16 / F10 cells were divided into 3 X 10 ⁴ cells per well in a 24-well plate (MTTassay) containing DMEM medium containing 10% FBS and cultured at 37 ° C in a 5% CO ₂ incubator (MCO-20AIC, Sanyo) And cultured for 24 hours. After the culture, the medium was removed, and DMEM medium containing 10% FBS and 50 nM α-MSH (Melanocyte stimulating hormone) was used as a new medium. The rice root extract of Comparative Example 1 was added to the medium at a concentration of 12.5 μg / Mu] g / ml, 3.125 [mu] g / ml and 1.5625 [mu] g / ml, and cultured in a 5% CO ₂ incubator at 37 ° C for 3 days.

In the negative control group, distilled water was treated with the same amount of the extract. As a positive control, α-arbutin, known as a whitening substance, was used at a concentration of 30 μg / ml. The no-treatment group did not contain? -MSH in the medium and did not treat the extract.

100 [mu] l of each of the cultures cultured for 3 days was dispensed in a 96-well plate, and the extracellular melanin secretion amount was calculated by measuring the absorbance at 450 nm using Synergy HT_Multi-reader (BioTek) Is shown in Fig.

<Formula 2>

The amount of extracellular melanin secretion (%) = (absorbance of sample treated group / absorbance of no treated group) * 100

[Test Example 5] Inhibitory effect of fermented rice roots extract on cell melanin synthesis

Except that the rice seed roots fermented extract prepared in Example 1 was treated at a concentration of 25 μg / ml, 50 μg / ml, 100 μg / ml and 200 μg / ml, respectively, instead of the extract of Comparative Example 1 Experiments were carried out in the same manner and the amount of melanin synthesis was calculated. The results are shown in FIG.

3 and 4, it can be confirmed that the rice root-roasted fermented extract inhibits extracellular melanin synthesis in a concentration-dependent manner and exhibits an inhibitory effect on melanin synthesis equivalent to a-arbutin at a concentration of 50 ug / ml.

[Formulation Example 1] Lotion

Using the components listed in Table 1 below, lotions were prepared by conventional methods.

ingredient Content (% by weight) The rice roots fermented extract of Example 1 3.00 Hydroxyethylene cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 5.00 Purified water Balance system 100.0

 [Formulation Example 2] Nutrition lotion

ingredient Content (% by weight) The rice roots fermented extract of Example 1 3.00 Ascorbic acid-2-phosphate magnesium salt 1.00 Water soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.01 Citric acid 0.05 1,3-butylene glycol 3.00 Purified water Balance system 100.0

Korea Microorganism Conservation Center (overseas) KCCM11570P 20140919

<110> International GFC <120> Cosmetic composition comprising fermented rice root extract and          method for preparing <130> AJP14726 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 2 tacggytacc ttgtacgact t 21

Claims (9)

A cosmetic composition for skin whitening comprising a fermented extract of rice ( Oryza sativa L. ) roots as an active ingredient. The method according to claim 1,
Wherein the fermented extract of the rice seed roots is fermented with a rice root extract obtained by extracting at least one of water or an organic solvent as an extraction solvent. The method according to claim 1,
Wherein the fermented extract of rice seeds is obtained by fermenting the rice root extract with a strain of Cupriavidus basilensis sp. Of KCCM11570P . The method according to claim 1,
Wherein the composition inhibits melanin synthesis. The method according to claim 1,
Wherein the composition contains 0.001 to 30% by weight of a fermented extract of the rice seed roots, based on the total weight of the composition. A medium containing rice root extract was prepared,
The culture medium was inoculated with a strain of Kupiabidus vulcanis of the deposit number KCCM11570P, and
And culturing and fermenting the culture medium inoculated with the strain. The method according to claim 6,
Wherein the medium comprises the ratio of the rice root extract to the distilled water at a ratio of 1:99 to 1: 1 by weight. The method according to claim 6,
Wherein the rice seed roots extract is prepared by using at least one of rice roots as an extraction solvent in water or an organic solvent. 9. The method of claim 8,
Wherein the extraction solvent comprises at least one of water and C1-4 anhydrous or hydrated lower alcohol.

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