JPH04235905A - Cosmetics - Google Patents

Abstract

PURPOSE:To obtain a melanin inhibitor comprising an extract of cells of bacterium belonging to the genus Streptococcus or Lactococcus as an active ingredient. CONSTITUTION:A cosmetic comprising an extract of cells of bacterium such as Streptococcus thermophilus YIT 2,001 strain (FERM P-11,891) or Lactococcus lactis subsp. cremoris (FERM P-11,790) extracted with water or an alcohol or a mixture thereof as an active ingredient. The extract has excellent tyrosinase inhibitory activity, is safe and a cosmetic blended with the extract has excellent beautifying effects and good feeling in use.

Description

[Detailed description of the invention]

0001

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a melanin production inhibitor, and more particularly to a melanin production inhibitor having an excellent tyrosinase activity inhibitory effect and a cosmetic containing the same.

0002

2. Description of the Related Art The dark discoloration of human skin caused by sunburn and the like is said to be caused by the synthesis of melanin in pigment cells present in the basal layer of the epidermis. Melanin is synthesized by oxidative polymerization of tyrosine, a type of amino acid, by tyrosinase, an oxidizing enzyme.

Conventionally, vitamin C, cysteine, arbutin, kojic acid, glutathione, hydroquinone, and the like have been known as substances that inhibit melanin production by inhibiting tyrosinase activity. Furthermore, as melanin production-inhibiting substances derived from natural products, extracts of plants belonging to the Umbelliferae family, extracts of placenta, and the like are known.

0004

[Problems to be Solved by the Invention] However, these conventional melanin production inhibitors generally have poor stability, may not be sufficiently effective, or may have safety issues, and are not always sufficient. It wasn't satisfying. Therefore, it has been desired to develop a melanin inhibitor that is stable, highly safe, has a strong melanin production inhibiting effect, and has good incorporation into cosmetics, and a cosmetic containing the melanin inhibitor. .

[0005]

[Means for Solving the Problems] Under these circumstances, the present inventors have made extensive studies and have found that a bacterial cell extract of a specific microorganism belonging to the genus Streptococcus or Lactococcus, which is used as a starter for fermented milk and cheese. The present invention was completed based on the discovery that the compound has an extremely excellent tyrosinase activity inhibiting effect, is highly safe, and can be widely incorporated into cosmetics.

That is, the present invention provides a melanin production inhibitor whose active ingredient is an extract extracted from cells of the genus Streptococcus or Lactococcus with water or alcohol, and cosmetics containing the same.

The melanin production inhibitor of the present invention is produced by extracting cells of the genus Streptococcus or Lactococcus with water or alcohol.

The microorganism used in the present invention is not particularly limited as long as it belongs to the genus Streptococcus or Lactococcus, but Streptococcus thermophilus strain YIT2001 (Streptococcus thermophilus)
Thermophilus YIT2001 strain (also referred to as Streptococcus thermophilus ST-1 strain) or Lactococcus lactis subsp. cremoris YIT2058 strain is particularly preferred.

Streptococcus thermophilus YI
The mycological properties of the T2001 strain and Lactococcus lactis subspecies cremoris YIT2058 strain are as follows.

(1) Streptococcus thermophilus YIT2001 strain morphology
Growth of Gram-positive cocci anaerobically
+ catalase
-hemolytic
−
Gas production from glucose -10
Growth at °C
Growth at -45℃
+6.5% salt tolerance
-Growth at pH 9.6
-40%bil
Growth in e - Coagulability of skim milk
+methylene blue
-NH3 production
-Hydrolysis of aesculin
-Sugar fermentability *1: Arabinose
-xylose
−Rhamnose
-mannose
-galactose
+Sucrose
+Cellobiose
-lactose
+Trehalose -Melibiose -Raffinose -
meletitose
-Mannitol
-Sorbitol
-glucose
+ - is negative + is positive *1: The basal medium for sugar fermentation property test is CPR as an indicator.
An ILS medium containing (chlorophenol red) was used.

(2) Lactococcus lactis subsp. cremoris YIT2058 strain morphology
Gram-positive cocci catalase
- gas production from glucose
Growth at -10℃
Growth at +45°C - Coagulation of skimmed milk
- Gas production from gluconic acid
-Nitrate reduction
-Sugar fermentability *2: Arabinose
-xylose
−Rhamnose
-ribose
−Mannose +
fructose
+galactose
−Sucrose
-maltose
−Cellobiose
+Lactose -
trehalose
−Melibiose
−Raffinose
-Meletitose
-Mannitol
-Sorbitol -Aesculin
- Salicin
-Amygdalin
-Sorbose
-Inositol -Inulin
−Starch
-α-methyl-D-glucoside
-glucose
+dextrin
-Glycogen
-- is negative + is positive *2: The basal medium for sugar fermentation property test is CPR as an indicator.
A medium based on ILS containing .

[0012] Streptococcus thermophilus YIT2001 strain exhibiting the above mycological properties is
No. 1891 (FERM P-11891), Lactococcus lactis subsp.
FERM P-11790) and deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology, Ministry of International Trade and Industry.

[0013] The cells of these microorganisms can be obtained by culturing them under conditions suitable for the growth of the microorganisms and then separating them from the culture supernatant by, for example, centrifugation.

The solvent used for extracting the melanin production inhibitor of the present invention is water or alcohol, and water, methanol, or a mixed solvent thereof is particularly preferred. Extraction is carried out by suspending the bacterial cells in a solvent and allowing them to stand or stir at room temperature to 80°C for 5 minutes to 1 hour. After extraction, insoluble matters are removed and the solvent is distilled off to obtain the melanin production inhibitor of the present invention.

The thus obtained melanin production inhibitor of the present invention has the following physical and chemical properties. (1) pH 4.0-6.0 (2) Various reactions Phenol sulfuric acid method Positive orcinol reaction
Positive BCA protein assay reagent Positive Lowry reaction Positive (3) Ultraviolet absorption (solvent: H 2 O) maximum wavelength 257
~259nm Minimum wavelength 232-234nm (Figure 1)

[0016] By incorporating the melanin production inhibitor of the present invention, a cosmetic having an excellent whitening effect can be obtained. The amount of the melanin production inhibitor added to the cosmetic of the present invention varies depending on the form of the cosmetic, etc., but is usually preferably 0.1 to 10% by weight (in terms of bacterial cell amount).

[0017] The cosmetics of the present invention may be in the form of cream,
Examples include skin cosmetics such as emulsions, lotions, oil-based cosmetics, powder cosmetics, and packs.

In addition to the above-mentioned melanin production inhibitors, the cosmetic of the present invention also contains emulsifiers such as nonionic surfactants, anionic surfactants, and amphoteric surfactants; vegetable oils, animal oils, higher fatty acids, and higher alcohols. , synthetic ester oils, waxes, silicone oils and other oily substances; water, fragrances, preservatives, pigments, skin nutrients, skin activators, moisturizers, ultraviolet inhibitors, p.
An H adjuster and the like can be added.

[0019]

EXAMPLES Next, the present invention will be explained in detail with reference to Examples. Example 1 (1) ILS (Glc) medium (trypticase 1.0
%, yeast extract 0.5%, tryptose 0.3
%, monopotassium phosphate 0.3%, dipotassium phosphate 0.3%, diammonium citrate 0.2%, sodium acetate 0.17%, Salt Sol 0.5%,
Tween 80 0.1%, L-cysteine hydrochloride 0.0
2%, glucose 2.0%) was adjusted to pH 6.8, filled into an Erlenmeyer flask, autoclaved at 121°C for 15 minutes, inoculated with Streptococcus thermophilus YIT2001, and left at 37°C for 24 hours. Culture was performed.

Lactococcus lactis subsp. cremoris YIT2058 is an ILS (Glc)
Using the medium, static culture was performed at 30°C for 20 hours.

After culturing, the bacterial cells and culture supernatant are separated using a centrifuge, and the bacterial cells are suspended in a methanol solution and incubated at 60°C.
After extraction was performed three times for 10 minutes, the bacterial cells were removed and the solvent was further removed under reduced pressure to obtain an extract.

(2) The inhibitory effect on melanin production of the obtained extract was examined using the inhibitory effect on tyrosinase activity, which is an oxidase for melanin production, as an indicator.

Test method Enzyme solution: Tyrosinase (4300 units/mg) 0.1
Dissolve mg in 1 ml of distilled water. Substrate solution: Dissolve L-tyrosine to 10mM. Buffer: 0.4 M phosphate buffer (pH 6.8)
Use.

[0024] In a test tube, add 0.75 ml of buffer solution and 0.0 ml of enzyme solution.
．． After adding 5 ml and 1 ml of the test sample and incubating at 37°C for 10 minutes, 0.75 ml of the substrate solution previously incubated at 37°C was added and allowed to react for 15 minutes. Immediately after the completion of the reaction, 47
Measure the absorbance (B) at 5 nm. In addition, the absorbance (A) in the case where no test sample was added (distilled water added) was measured, and the inhibition rate of tyrosinase activity was calculated from the following formula.

[Math 1]

Test Results The concentration dependence of the tyrosinase inhibitory activity of the extract is shown in FIG. The sample concentration was expressed as the equivalent amount of bacterial cells contained in 3 ml of the measurement system.

From the above results, the amount of bacterial cells that showed 50% inhibition of tyrosinase activity was 2.9 mg for Streptococcus thermophilus YIT2001, and 2.9 mg for Lactococcus thermophilus YIT2001.
lactis subspecies cremoris YIT205
8 was 4.6 mg.

Example 2 Streptococcus thermophilus YIT2001
were statically cultured at 37° C. for 24 hours using ILS (Glc) medium. In addition, Lactococcus lactis subsp. cremoris YIT2058 is an ILS
(Glc) medium was used for static culture at 30° C. for 24 hours.

After culturing, the bacterial cells and culture supernatant were separated using centrifugation, and the bacterial cells were suspended in distilled water and incubated at 60°C for 10 minutes.
After extraction was performed three times, the solvent was removed under reduced pressure to obtain an extract.

FIG. 3 shows the concentration dependence of the tyrosinase inhibitory activity of the extract obtained by testing in the same manner as in the example. The sample concentration was expressed as the equivalent amount of bacterial cells contained in 3 ml of the measurement system.

From the above results, the amount of bacterial cells that showed 50% inhibition of tyrosinase activity was 3.1 mg for Streptococcus thermophilus YIT2001, and 3.1 mg for Lactococcus thermophilus YIT2001.
lactis subspecies cremoris YIT205
8 was 4.5 mg.

Example 3 Lotion (composition) Streptococcus thermophilus YIT2001
1.0% by weight bacterial cell methanol extract (Example 1) ethanol
10.0% by weight1,
3-butylene glycol
2.0% by weight polyoxyethylene hydrogenated castor oil (50E.O.) 0.05% by weight methyl paraoxybenzoate
0.1% by weight fragrance

0.1% by weight purified water

The total amount of lotion obtained is 100.
It had an excellent whitening effect and a refreshing feeling of use.

Example 4 Emulsion (composition) Streptococcus thermophilus YIT2001
5.0% by weight Bacterial cell methanol extract (Example 1) Stearic acid

2.0% by weight liquid paraffin

6.0 wt% squalane

2.0 wt% sorbitan monostearate
1.
5% by weight polyoxyethylene sorbitan monostearate (20E.O.)
2.0 wt% butyl paraoxybenzoate
0.05% by weight 1,3-butylene glycol
3.0% by weight methyl paraoxybenzoate
0.1% by weight fragrance

0.15% by weight purified water

The emulsion obtained in a total amount of 100 ml had an excellent whitening effect and a moist feeling on use.

Example 5 Cream (composition) Streptococcus thermophilus YIT2001
5.0% by weight Bacterial cell methanol extract (Example 1) Liquid paraffin

23.0% by weight Vaseline

7.0 wt% behenyl alcohol

1.0 wt% stearic acid

2.0 wt% beeswax

2.0 wt% sorbitan monostearate
3.5% by weight polyoxyethylene sorbitan monostearate (20E.O.)
2.5 wt% butyl paraoxybenzoate
0.05% by weight 1,3-butylene glycol
3.0% by weight methyl paraoxybenzoate
0.1% by weight fragrance

0.15% by weight purified water

The cream obtained in a total amount of 100% had an excellent whitening effect and a good feeling of use.

[0034]

[Effects of the Invention] The melanin production inhibitor of the present invention has excellent tyrosinase inhibitory activity and is safe, and cosmetics containing it have an excellent whitening effect and a good feeling of use.

[Brief explanation of the drawing]

FIG. 1 is a drawing showing the ultraviolet absorption spectrum of the melanin production inhibitor of the present invention.

FIG. 2 is a drawing showing the relationship between the tyrosinase activity inhibition rate of the methanol extract of bacterial cells obtained in Example 1 and the amount of bacterial cells extracted with methanol.

FIG. 3 is a drawing showing the relationship between the tyrosinase activity inhibition rate of the hot water extract of bacterial cells obtained in Example 2 and the amount of bacterial cells extracted with hot water.

Claims (2)

[Claims] 1. A melanin production inhibitor containing as an active ingredient an extract obtained from bacterial cells of the genus Streptococcus or Lactococcus with water, alcohol, or a mixture thereof. 2. A cosmetic containing the melanin production inhibitor according to claim 1.