JP6984910B2 - Ingredients for skin cosmetics and skin cosmetics containing them - Google Patents

Description

The present invention relates to a raw material for skin cosmetics and a skin cosmetic prepared by blending the raw materials. The raw material for skin cosmetics according to the present invention has an excellent usability, a skin whitening effect, and a strong antioxidant power. Therefore, the skin cosmetic according to the present invention is extremely easy to use, and if it is continued to be used, it is effective in improving the firmness of the skin and suppressing the occurrence of wrinkles and stains on the skin.

Conventionally, some raw materials for cosmetics having a good feeling of use such as smoothness and familiarity when applied to the skin, and some skin cosmetics using the raw materials have been introduced (for example, Patent Document 1). However, conventional cosmetic raw materials with good usability depend on oil-based raw materials for imparting smoothness at the time of application, and oil-based raw materials such as liquid paraffin, squalane, and silicone oil can be used as water-in-oil type or oil-in-water oil. Many of them are emulsified in a mold, so the sticky feeling peculiar to oil content cannot be avoided. Further, even a lotion that does not use an oily raw material tends to contain a polyhydric alcohol as a moisturizer, so that a sticky feeling often remains on the skin after use (for example, Patent Document 2).

Japanese Patent Application Laid-Open No. 2012-207001 Japanese Unexamined Patent Publication No. 5-155479 Japanese Patent Publication No. 2012-528152

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Sept.2011, p.6189-6198 Genetic and Biochemical Map for the Biosynthesis of Occidiofungin, an Antifungal Produced by Burkholderia contaminans Strain MS14 Biochemical and Biophysical Research Communications 380 (2009) 328-332 Biosynthesis of an antifungal oligopeptide in Burkholderia contaminans strain MS14

The present invention is a raw material for cosmetics for skin, which has an excellent feeling of use, a whitening effect on the skin, and a strong antioxidative power, and is extremely easy to use. An object of the present invention is to provide a skin cosmetic that has the effect of improving the skin and suppressing the occurrence of wrinkles and stains on the skin.

In the process of studying Burkholderia contaminans, the present inventor has excellent usability and whitening effect when the culture solution is used as a raw material for skin cosmetics. It was found that it is rich in antioxidant properties, and as a result of further research, the present invention was completed.

Burkholderia contamination is a type of bacterium belonging to the genus Burkholderia, and the cyclic glycopeptide occidiofungin extracted from the culture medium is the growth of fungi such as mold and yeast. It is known to have antifungal properties that inhibit burkholderia (Patent Document 3 and Non-Patent Document 1).

The problem of the present invention can be solved by the following raw materials for skin cosmetics and skin cosmetics.

(1) A raw material for skin cosmetics, which comprises or contains a culture solution of Burkholderia contaminans containing occidiofungin.

(2) A raw material for skin cosmetics used to give an excellent feeling of use to skin cosmetics, which is a substance produced by barkholderia contaminance containing ocidiofundin or a substance thereof. A raw material for skin cosmetics, which is characterized by containing.

(3) It is a raw material for skin cosmetics used to give an excellent feeling of use to skin cosmetics , and is characterized by being ossidiofundin or containing ossidiofundin. Ingredients for skin cosmetics.

(4) A skin cosmetic containing the raw material for skin cosmetic according to (1) above.

(5) The skin cosmetic characterized by comprising a skin cosmetic material according as main raw material in (1).

(6) A skin cosmetic containing the raw material for skin cosmetic according to the above (2) or (3).

(7) A skin cosmetic containing the raw material for skin cosmetics according to (2) or (3) above as a main raw material.

The raw material for skin cosmetics according to (1) above is made of or blended with a culture solution of barkholderia contaminance containing ossidiofundin, and is easily adapted to the skin. Immediately after it has become familiar with the skin, the skin feels moisturized, the skin becomes smooth and squeaky, and the skin becomes soft .
Further, when the raw material for skin cosmetics is applied to the skin, the water content of the stratum corneum increases, so that the skin is moisturized, the evaporation amount of the transepidermal water content is suppressed, and the barrier function of the skin is enhanced. Therefore, since the viscoelasticity of the skin is improved, the skin becomes soft and the firmness of the skin is improved. That is, the softness, smoothness, and moistness of the skin are improved.

Further, if the raw material for skin cosmetics described in (1) above is continuously used, the tyrosinase activity in the cells forming the skin is lowered and the production of melanin is suppressed, so that a whitening effect is produced on the skin. Further, since the raw material for skin cosmetics has antioxidant properties, it is possible to suppress the occurrence of wrinkles and stains on the skin.

In addition, since the culture solution of Burkholderia contamination containing ossidiofundin has antifungal properties, by blending the raw material for skin cosmetics described in (1) above, the skin cosmetics are preservative. Gender can be imparted.
As described above, the raw material for skin cosmetics according to (1) above is extremely useful as a raw material for skin cosmetics.

The term "skin" as used herein refers to all tissues covering the surface of the human body.

In addition, "skin cosmetics" include all cosmetics that are used in contact with the skin, and are not only cosmetics that are applied to the skin and used, but also cleaning cosmetics for cleaning the skin. It also includes bath cosmetics that are used by dissolving in hot water for bathing.

The Burkholderia contamination culture medium is a so-called Burkholderia contamination culture medium in which all or most of the Burkholderia contamination cells have been removed by centrifugation or a filter. It is a supernatant and contains osidiofundin.

The raw material for skin cosmetics according to (2) above is a substance produced by barkholderia contaminance containing osidiofundin, or contains the substance, so that it easily adapts to the skin. Immediately after it has become familiar with the skin, the skin feels moisturized, the skin becomes smooth and squeaky, and the skin becomes soft. Therefore, it is extremely useful as a raw material for skin cosmetics used to impart an excellent usability to skin cosmetics.

Since the raw material for skin cosmetics described in (3) above is osidiofundin or contains ossidiofundin, it easily adapts to the skin and moisturizes the skin immediately after it has been applied to the skin. It feels smooth, the skin is smooth and squeaky is not felt, and the skin is softened. Therefore, it is extremely useful as a raw material for skin cosmetics used to impart an excellent usability to skin cosmetics.

Said skin cosmetic composition according to (4), according to the above (1) from those containing skin cosmetic material, said the same raw material for skin cosmetic according to (1) Show the effect.
That is, the skin cosmetic according to (4) above is extremely easy to use, and if it is continued to be used, the intracellular tyrosinase activity of the skin is reduced and the production of melanin is suppressed, so that a whitening effect on the skin is achieved. Occurs. In addition, since such skin cosmetics have antioxidant properties, it is possible to suppress the occurrence of wrinkles and stains on the skin. As described above, the skin cosmetic according to (4) above is extremely useful for improving the skin and preventing photoaging.
Further, since the culture solution and the produced substance of Burkholderia contamination contain ossidiofundin and have antifungal properties, the skin cosmetics described in (4) above have antiseptic properties. Will be.

Since the skin cosmetics described in (5) above are mainly made of the raw materials for skin cosmetics described in (1) above, are they the same as the skin cosmetics described in (4) above? Shows more effect.
That is, the skin cosmetic according to (5) above is extremely easy to use, and if it is continued to be used, the intracellular tyrosinase activity of the skin is reduced and the production of melanin is suppressed, so that a whitening effect on the skin is achieved. Occurs. In addition, since such skin cosmetics have antioxidant properties, it is possible to suppress the occurrence of wrinkles and stains on the skin. As described above, the skin cosmetic according to (5) above is extremely useful for improving the skin and preventing photoaging.
Further, since the culture solution and the produced substance of Burkholderia contamination contain ossidiofundin and have antifungal properties, the skin cosmetics according to (5) above have antiseptic properties. It becomes a thing.

Since the skin cosmetic according to the above (6) contains the raw material for the skin cosmetic according to the above (2) or (3), the skin according to the above (2) or (3). It has the same effect as raw materials for cosmetics.
That is, the skin cosmetic according to (6) above is easy to be applied to the skin, and the skin is moisturized immediately after being applied to the skin, and the skin becomes smooth and smooth without squeaks. It has an excellent feeling of use such as softening of the skin.
Further, since the culture solution and the produced substance of Burkholderia contamination contain ossidiofundin and have antifungal properties, the skin cosmetics described in (6) above have antiseptic properties. Will be.

Since the skin cosmetic according to the above (7) is mainly made of the raw material for the skin cosmetic according to the above (2) or (3), the skin cosmetic according to the above (6). Shows the same or better effect as the fee.
That is, the skin cosmetic according to (7) above is easy to be applied to the skin, and immediately after it is applied to the skin, the skin feels moisturized, and the skin becomes smooth and smooth without squeaks. It has an excellent feeling of use such as softening of the skin.
Further, since the culture solution and the produced substance of Burkholderia contamination contain ossidiofundin and have antifungal properties, the skin cosmetics described in (7) above have antiseptic properties. Will be.

It is a chromatogram of the component of the production substance extracted from the culture medium of Burkholderia contamination. It is a graph which shows that the raw material for skin cosmetics which concerns on this invention reduces the tyrosinase activity in the B16 melanoma cell. It is a graph which shows that the raw material for skin cosmetics which concerns on this invention has a free radical scavenging action. It is a chromatogram of the component of the concentrate of the washing liquid which mixed water and acetonitrile in a ratio of 9: 1 containing the substance which produces Burkholderia contamination.

In the present invention, a culture solution of Burkholderia contamination and a production substance are used. Therefore, in the following, [1] a medium for culturing Berkholderia contamination, [2] a culture medium obtained by culturing Berkholderia contamination, and [3] a Berkholderia obtained from such a culture medium. The substance produced by contamination and [4] ossidiofundin obtained from such a substance will be described in this order.

In the present invention, "%" means "mass%" unless otherwise specified. Further, the numerical range represented by using "~" in the present invention means a range including the numerical values before and after "~" as the lower limit value and the upper limit value.

[1. Culture medium〕
As the medium for culturing Burkholderia contamination, a general medium may be used, and a liquid medium is preferable from the viewpoint of ease of handling.
For example, when using an SCD medium (Soybean-Casein Digest Broth) product manufactured by Nippon Pharmaceutical Co., Ltd., the products include casein peptone (17 g / L), soybean peptone (3 g / L), and dipotassium monohydrogen phosphate (dipotassium monohydrogen phosphate). Since it is composed of 2.5 g / L), glucose (2.5 g / L), sodium chloride (5 g / L), etc., 1 L of purified water is added to this product to dissolve it, and then a high-pressure steam sterilizer is used. It can be sterilized at 121 ° C. for about 15 minutes to prepare a liquid SCD medium.

[2. Culture solution]
Burkholderia culture Delia Kontaminansu, for example, were inoculated with Burkholderia Kontaminansu (10 ³ cfu / mL) in SCD medium of the liquid (250 mL), allowed to stand 3 days at 30 ° C. in a constant temperature environment It may be cultured. The culture solution (supernatant) according to the present invention can be obtained by separating and removing the cells from the liquid medium containing the cells of Berkholderia contamination thus obtained by centrifugation or filtration. ..

For example, the liquid medium containing the cells was centrifuged at a relative centrifugal acceleration of 10,000 G for 30 minutes while maintaining the product temperature at 4 ° C., and separated into a supernatant and a precipitate, and the obtained supernatant was subjected to membrane. By filtering with a filter (pore size 0.22 μm), the culture medium of Berkholderia contamination according to the present invention (hereinafter referred to as “culture solution A”) can be prepared.

Since the culture solution A of Burkholderia contamination obtained by the above operation exhibits antifungal properties, it is considered that ocidiofundin is present at a concentration of 1 μg / mL or more.

[3. Produced substance]
An example of the method of extracting the produced substance from the above-mentioned culture solution A will be described.
An appropriate amount (for example, 50 g) of the culture solution A is weighed, ammonium sulfate (15 g) is added thereto, and the mixture is stirred and mixed for about 2 hours while cooling to about 0 ° C. The obtained mixture is centrifuged at a relative centrifugal acceleration of 10,000 G for 20 minutes by adjusting the product temperature to about 5 ° C., and separated into a supernatant liquid and a precipitate. The supernatant is discarded, and acetonitrile (about 1.5 mL) having a concentration of 35% is added to the precipitate and kneaded to dissolve the precipitate. The obtained solution is filtered using a membrane filter having a pore size of 0.22 μm to obtain an acetonitrile solution (about 8 mL) containing a Burkholderia contamination-producing substance. Next, a substance produced by Burkholderia contamination (hereinafter referred to as “produced substance B”) may be separated from the above-mentioned acetonitrile solution using a high performance liquid chromatograph device.

The components of the product B obtained by the above operation were measured by a high performance liquid chromatograph mass spectrometer (LC-MS). The operating conditions at that time are as follows a to d.
a) HPLC: LC-2030C 3D, LCMS-2020 manufactured by Shimadzu Corporation
b) Column: Discovery® HS F5-5, 5 μm, 20 mm × 2.1 mm
c) Flow velocity: 0.25 mL / min
d) Solvent: acetonitrile / 0.1% formic acid water 0min: acetonitrile 0%
5min: 25% acetonitrile
11min: 35% acetonitrile
15min: 95% acetonitrile

As a result, as shown in FIG. 1, the position and molecular weight of the peak that appears largely in the chromatogram of the product B are described on page 331 of Non-Patent Document 2 (Biochemical and Biophysical Research Communications 380 (2009) 328-332). It was confirmed that the peak positions and molecular weights (1200.6, 1216.6) of ossidiofundin described in "Isolation of antifungal compound production by strain MS14" and "Fig. 4" in the right column match.
Therefore, it was found that the above-mentioned Burkholderia contamination-producing substance contained a high concentration of ocidiofundin.

[4. Osidi Ofunjin]
An example of a method for separating ocidiofundin from the above-mentioned product B will be described.
Product B (190 mL) is concentrated on an evaporator to 1.5 mL, loaded onto the SPE cartridge head and washed with a wash solution (10 mL) containing water and acetonitrile in a 9: 1 ratio. Next, the washing liquid (10 mL) containing the product B after washing is concentrated to 1.5 mL with an evaporator to obtain a concentrated washing liquid.

Osidiofundin can be separated from the concentrate of this cleaning liquid by using a high performance liquid chromatograph (HPLC). The operating conditions at that time are as follows a to d.
a) HPLC: LC-2030C manufactured by Shimadzu Corporation
b) Column: InertSustain® C18, 3 μm, 2.1 mm × 150 mm
c) Flow velocity: 0.2 mL / min
d) Solvent: 0.1% Acetnitonyl formate / 0.1% Water formic acid 0min: 0.1% Acetnitonyl formate 10%
1min: 0.1% Acetnitonyl formate 10%
15min: 0.1% Acetnitonyl formate 90%
30min: 0.1% Acetnitonyl formate 90%

As shown in FIG. 4, the peak at the position of 12.13 minutes in the chromatogram of the concentrate of the washing liquid is due to ossidiofundin (molecular weight 1216.6). Such ossidiofundin can be obtained as solid ossidiofundin (4.8 mg) by elution from the column using a washing solution in which water and acetonitrile are mixed in an appropriate ratio, and then evaporating the liquid with an evaporator. can.

[Test example]
The present inventor conducted the following test on the culture solution A of Burkholderia contamination prepared in the above-mentioned preparation example, and was able to confirm the effect. Therefore, this culture solution A can be used as it is as a raw material for skin cosmetics. It can also be blended with a moisturizer, oil, etc. as one of the raw materials constituting the raw material for skin cosmetics. Further, the culture solution A can also be used as a main raw material for skin cosmetics.

<< Test Example 1 >> Use test of culture solution A (a) Test method For four skilled sensory test technicians, apply culture solution A to almost the entire skin of each face, let it pass for 5 minutes, and before and after that. Each person's feelings were evaluated on a scale of 5 points (1 to 5 points), and the average value was calculated. The criteria for evaluation were "1 point: not felt, 2 points: neither, 3 points: slightly felt, 4 points: felt, 5 points: very felt".
For control, a liquid SCD medium not inoculated with Burkholderia contamination was used.

(B) Test results The test results are shown in Table 1.

(B) Findings From the above test results, the culture solution A easily blends into the skin, the skin feels moisturized after use, the skin becomes smooth and squeaky, and the skin becomes soft. Can be understood.

<< Test Example 2 >> Skin function measurement test to which culture solution A was applied (a) Test method A female in her 30s was used as a subject, and the entire forearm was washed with soap to remove sweat and dirt, and then the temperature was immediately constant. After entering a constant humidity chamber (room temperature 21.0 ± 0.2 ° C., humidity 45.0 ± 0.5%) and acclimatizing for 20 minutes, the first skin function measurement was performed.
Then, the culture solution A was applied to the inside of the forearm of the subject, and the change in skin function was measured over time.
For skin function measurement, "corneal layer water content" is used to evaluate the degree of dryness of the skin, and Tewameter (manufactured by Khazaka) is used to evaluate the barrier function of the skin. ) Is used for "percutaneous moisture evaporation", and a cute meter (Cutometer: manufactured by Khazaka) is used for "skin viscous elasticity" (skin softness and skin firmness) to evaluate the elasticity of the skin. The measurement was performed three times each, and the average value was calculated.
Further, a relative value with the passage of time when the first measured value (before applying the culture solution A) was set to 100 was calculated for each elapsed time.
For control, a liquid SCD medium not inoculated with Burkholderia contamination was used.

(B) Test results a) Changes in the water content of the stratum corneum The water content of the stratum corneum is 132 (control is 127) 15 minutes after the application and 132 minutes after the application, when the water content of the stratum corneum is 100 before the application of the culture solution A, and 30 minutes after the application. 129 (control is 126) was shown.
It can be said that the larger the value of the water content of the stratum corneum, the more moisturized the skin.

b) Changes in the amount of transepidermal water loss
The transepidermal water loss amount was 92 (control: 108) 60 minutes after the application, where 100 was before the application of the culture solution A.
As for the amount of percutaneous water evaporation, it can be said that the smaller the value, the smaller the amount of water evaporation from the skin surface, and the higher the barrier function of the skin.

c) Changes in skin viscoelasticity (skin softness)
The skin viscoelasticity (softness of the skin) was 118 (control 101) 5 minutes after the application, when the value before the application of the culture solution A was 100.
As for skin viscoelasticity (skin softness), it can be said that the larger the value, the softer the skin.

d) Changes in skin viscoelasticity (skin firmness)
The skin viscoelasticity (skin firmness) shows 107 (control remains 100) 15 minutes after application and 112 (control 101) 30 minutes after application, where 100 is before the application of the culture solution A. Indicated.
It can be said that the larger the value of skin viscoelasticity (skin elasticity), the stronger the elasticity of the skin.

(C) Findings:
From the above test results, it was confirmed that when the culture solution A was applied to the skin, the skin was moisturized, the barrier function of the skin was improved, the skin was softened, and the firmness of the skin was strengthened.

Generally, melanin is formed by the action of the enzyme tyrosinase, which is biosynthesized in pigment cells. Therefore, in order to prevent and improve dark skin and skin spots and freckles, it is effective to inhibit the activity of tyrosinase involved in the production of melanin.
The present inventor conducted a test to confirm the presence or absence of an action of inhibiting the activity of tyrosinase in the culture solution A.

<< Test Example 3 >> Test for confirming intracellular tyrosinase activity inhibitory action (a) Preparation of test substance a) Culture solution A was used as the test substance. Also, as a control, a liquid SCD medium not inoculated with Burkholderia contamination was used.
b) Phosphate buffer saline (PBS) was used as the solvent for the test substance.
c) The test substance was continuously diluted at a concentration of 100% to 2 times the common ratio to a total concentration of 8 (0.78%, 1.56%, 3.13%, 6.25%, 12.5%). 8 types of 25.0%, 50.0% and 100.0%) were prepared.

(B) Test method a) were seeded B16 melanoma cells 9.9 × 10 ³ cells / 99μL 1 per well in 96-well plates and cultured for 24 hours in a C0 ₂ incubator. Two plates were prepared, one for measuring cell viability and the other for measuring intracellular tyrosinase activity.
b) After 24 hours of culturing, 1 μL of the above test substance was added to each well to adjust the final concentrations to 0.0078%, 0.0156%, 0.0313%, 0.0625% and 0.125%, respectively. .. 0.25%, 0.5%, 1.0%, and cultured for 72 hours in a C0 ₂ incubator. In addition, 3 wells were used for each test substance, and PBS was used for the blank.
c) cultured for 72 hours after, the Cell Counting Kit-8 to the plate for measuring cell viability was added 10 [mu] L, were incubated for 2 hours in a C0 ₂ incubator.
d) After 2 hours of culturing, the 96-well plate was shaken at 270 rpm for 5 seconds and the absorbance at 450 nm (OD ₄₅₀ ) was measured using a microplate reader.
e) The medium of the plate for measuring intracellular tyrosinase activity was removed, and the cells were washed once with 200 μL PBS.
f) 90 μL of PBS containing 0.5% Triton X-100 was added to each well of the 96-well plate, and 10 μL of 10 mM L-DOPA solution was added.
g) Immediately after the addition (0 minutes later), the 96-well plate was shaken at 270 rpm for 5 seconds, and the absorbance at 475 nm (OD ₄₇₅ ) was measured using a microplate reader.
h) After a 96-well plate and allowed to stand for 60 minutes in a C0 ₂ incubator, it was measured _{OD 475} again (after 60 minutes).
i) The cell viability of the test substance was determined from _{OD 450.} The intracellular tyrosinase activity was determined from the difference between 0 minutes and 60 minutes after _{OD 475 (ΔOD 475} ). In addition, the intracellular tyrosinase activity per cell was determined from _{ΔOD 475} / OD _450.

(C) Test results The test results are as shown in FIG.
That is, from FIG. 2, it was confirmed that the tyrosinase activity in the B16 melanoma cells was significantly reduced in the range of the concentration of the culture solution A in the range of 0.25% to 1%.

(2) Findings From the above test results, it is understood that the culture solution A having a concentration of 0.25% to 1% significantly reduces the tyrosinase activity in the B16 melanoma cells and thus acts to suppress the production of melanin. can.

In general, it is known that free radicals such as active oxygen having strong oxidizing power cause damage to living tissues and cells and cause skin aging. Therefore, the cosmetic raw material preferably has an excellent antioxidant power for suppressing / eliminating the generation and action of free radicals.
The present inventors have obtained DPPH (2,2-Diphenyl-1-picrylhydrazyl) for the culture solution A.
Was used to perform a test to confirm the presence or absence of free radical scavenging action.
DPPH is a compound having a maximum absorption of 517 nm in a radical state, and the DPPH scavenging rate is the rate of decrease in the absorbance of DPPH at 517 nm when an antioxidant is added to DPPH and reduced. The antioxidant capacity of antioxidants can be evaluated. That is, the larger the DPPH elimination rate, the higher the antioxidant capacity.

<< Test Example 4 >> Free radical scavenging action confirmation test (a) Preparation of test substance a) Culture solution A was used as the test substance. Also, as a control, a liquid SCD medium not inoculated with Burkholderia contamination was used.
b) The test substance was continuously diluted 10-fold from 100% concentration to a total of 6 concentrations (0.001%, 0.01%, 0.1%, 1.0%, 10.0%, 6 types of 100.0%) were prepared.

(B) Preparation of reagent a) A 3.1 mg DPPH solution is dissolved in 50 mL of ethanol to prepare a 0.16 mM DPPH solution, which is further diluted 20-fold with ethanol to a 0.008 mM DPPH solution. Was adjusted to and used in the test.
b) 1.44 mL of acetic acid was dissolved in purified water and the volume was increased to 100 mL. Further, 2.05 g of sodium acetate was dissolved in purified water, and the volume was increased to 100 mL. This acetic acid solution and sodium acetate solution were mixed at a ratio of 1: 8 to prepare 0.25 M acetic acid Buffer, and the pH was confirmed to be 5.5 by a pH meter.

(C) Test method a) 20 μL of the test substance was added to a 96-well plate. Two groups of test substance wells were prepared, one of which was DPPH (+) and the other of which was DPPH (-).
b) 60 μL of ethanol was added to the DPPH (+) well. In addition, 100 μL of ethanol was added to the DPPH (−) well.
c) 80 μL of 0.25 M acetate Buffer was added to all wells, a plate seal was attached to a 96-well plate, and the mixture was incubated at 30 ° C. for 5 minutes.
d) After 5 minutes, 40 μL of the DPPH solution was added to the DPPH (+) well, and the mixture was stirred for 30 seconds using a plate mixer. After stirring, the reaction was carried out at 30 ° C. for 20 minutes.
e) After 20 minutes, the absorbance at ₅₁₇ nm (OD 517) was measured with a microplate reader.
f) Using the obtained OD ₅₁₇ , the DPPH elimination rate of the culture solution A with respect to the control was calculated.

(D) Test results The test results are as shown in FIG. The vertical axis of the graph (DPPH elimination) is a relative value (%) of the elimination rate of the test substance when the elimination rate of DPPH by the control (SCD medium) is 0%.
From FIG. 3, it was confirmed that the DPPH scavenging ability was high within the range of the concentration of the culture solution A from 0.001% to 100.0%.

(E) Findings From the above test results, it can be recognized that the culture solution A having a concentration of 0.001% to 100.0% has antioxidant properties, and acts to suppress the occurrence of wrinkles and stains on the skin. Can be understood.

<< Test Example 5 >> Use test of ossidiofundin Dissolve ossidiofundin obtained by the method of separating ossidiofundin from the above-mentioned product B in water to prepare a 0.002% aqueous solution. , A use test was conducted in which this was applied to the skin.

(B) Test method For four skilled sensory test technicians, apply an aqueous solution of ossidiofundin to almost the entire facial skin, let it pass for 5 minutes, and give each person's sensations before and after that a perfect score of 5 (5 points). We asked them to evaluate with 1 to 5 points) and calculated the average value. The criteria for evaluation were "1 point: not felt, 2 points: slightly not felt, 3 points: neither, 4 points: slightly felt, 5 points: felt".
Water was used for control.

(B) Test results The test results are shown in Table 2.

(B) Findings From the above test results, the aqueous solution of Osidiofundin is easy to adapt to the skin, the skin feels moisturized after use, the skin becomes smooth and squeaky, and the skin becomes smooth. It can be understood that it becomes soft.
Therefore, the aqueous solution of ocidiofundin can be used as it is as a raw material for skin cosmetics. It can also be blended with a moisturizer, oil, etc. as one of the substances constituting the raw material for skin cosmetics. Further, an aqueous solution of ossidiofundin can be used as a main raw material for skin cosmetics.

<< Example 1 >>
A raw material for skin cosmetics containing a culture solution of Burkholderia contamination containing the above-mentioned ossidiofundin (culture solution A) was produced.
Purified water and culture solution A in the blending amounts shown in Table 3 were stirred and mixed, and 1,3 butylene glycol was further added and mixed by stirring to obtain a raw material for skin cosmetics.

The obtained raw material for cosmetics for skin was easy to adapt to the skin, smoothed the skin after application, and improved the softness, smoothness, and moistness of the skin.

Further, since the obtained raw material for cosmetics for skin contains 50% of the culture solution A, the result of Test Example 3 above (the culture solution A reduces the intracellular tyrosinase activity at a concentration of 0.25% to 1%). As is clear from the above, it inhibits the intracellular tyrosinase activity and exerts a skin whitening effect.
Further, as is clear from the results of Test Example 4 above (the culture solution A has a concentration of 0.001% to 100% and a high DPPH scavenging ability), the raw material for cosmetics for skin has antioxidant properties. , It has the effect of suppressing the occurrence of wrinkles and stains on the skin.

Further, the obtained raw material for skin cosmetics was enclosed in a glass container and stored at room temperature for 1 month, but the quality was not deteriorated without spoilage.

<< Example 2 >>
A skin cosmetic containing the raw material for skin cosmetic obtained by the method of Example 1 above was produced.
Polyoxyethylene (POE = 60) hardened castor oil was added to the purified water in the amount shown in Table 4 and dissolved by stirring, then citric acid and trisodium citrate were added and dissolved by stirring, and then other raw materials shown in Table 3 were added. Was stirred and dissolved to obtain a skin cosmetic.

The obtained skin cosmetics were easy to adapt to the skin, smoothed the skin after application, and improved the softness, smoothness, and moistness of the skin.

Further, since the obtained skin cosmetics contain 1% of the raw materials for skin cosmetics obtained in Example 1 and therefore contain 0.5% of the culture solution A, the above test. As is clear from the results of Example 3 (culture solution A reduces the intracellular tyrosinase activity at a concentration of 0.25% to 1%), it inhibits the intracellular tyrosinase activity and exerts a skin whitening effect.
Further, as is clear from the results of Test Example 4 above (the culture solution A has a high DPPH scavenging ability at a concentration of 0.001% to 100%), the skin cosmetic has antioxidative properties, and therefore the skin. It has the effect of suppressing the occurrence of wrinkles and stains.

Further, the obtained skin cosmetics were enclosed in a glass container and stored at room temperature for 1 month, but they did not spoil and no deterioration in quality was observed.

The raw material for skin cosmetics and the skin cosmetic according to the present invention are mainly used in the beauty field because they can improve the firmness of the skin and suppress the occurrence of wrinkles and stains on the skin. ..

Claims (3)

A raw material for skin cosmetics, which comprises or contains a culture solution of Burkholderia contaminans containing occidiofungin. A skin cosmetic containing the raw material for skin cosmetic according to claim 1. A skin cosmetic containing the raw material for skin cosmetics according to claim 1 as a main raw material.

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