KR102463782B1 - A whitening composition and its manufacturing method by bio-renovation of Ligustrum Janponicum Extract - Google Patents

Abstract

The present invention relates to a whitening composition by biorenovation of an extract of Gwangju and a method for preparing the same, and inhibits the activation of B16F10 melanoma cells (including apoptosis and aggression relief) by using an active ingredient of a natural product, a natural product, the enzymatic function of a microorganism. It aims to help skin whitening while increasing the stability of the skin and reducing side effects by generating a reactant.
The composition for whitening by biorenovation of the extract of Gwangju according to the present invention consists of a mixed reaction of the microorganism, Bacillus amyloliquefaciens , and the extract of Gwangju.
According to the present invention, a method for preparing a whitening composition by biorenovation of a Kwangnam tree extract comprises: a first step of preparing the Kwangno tree extract; A second step of selecting and culturing Bacillus amyloliquefaciens as a microorganism and then centrifuging to obtain a microbial pellet; and a third step of mixing and reacting the microbial pellets obtained in the second step with the Kwangnam tree extract prepared in the first step, wherein the first step is extracting the flowers and leaves of Kwangnam tree with ethanol and then concentration under reduced pressure. Extract the sagebrush extract.

Description

A whitening composition and its manufacturing method by bio-renovation of Ligustrum Janponicum Extract

The present invention relates to a whitening composition using Gwang tree, and more particularly, a whitening composition by biorenovation of Gwang tree extract, which is effective for skin whitening from the active ingredient of Gwang tree, which is a natural product, while increasing skin stability and reducing side effects, and a method for manufacturing the same is about

This section provides background information related to the subject matter of the present application and is not necessarily prior art.

Human skin color is changed by various factors such as aging, UV rays, and stress, and it is determined by the concentration and distribution of melanin inside the skin. is also affected by

Melanocytes are phenolic biopolymers that determine hair color and skin color. They synthesize and secrete melanin, a complex of black protein, to prevent skin damage caused by UV rays, as well as to absorb toxic drugs. perform an important role.

Melanin is absorbed into the skin in various forms and processes and is present in the basal layer of the skin, and serves to protect the skin from UV rays and external damage. In particular, UV rays can trigger an inflammatory response through many mechanisms, including direct activation of keratinocytes and other cells that activate components of the skin immune system to release inflammatory mediators. UV-induced DNA damage activates the binding of p53 to the promoter of the POMC (pro-opiomelanocortin) gene to generate α-MSH (melanocyte-stimulating hormones), and the produced α-MSH serves to send a signal to melanocytes. Stimulates cAMP (cyclic adenosine monophosphosphate) production by binding to MC1R (melanocortin 1 receptor) phosphorylates CREB is a transcription factor that regulates the expression of various genes, including the expression of microphthalmia-associated transcription factor (MITF). MITF regulates the expression of melanin-producing enzymes, tyrosinase-related protein (TRP)-1, 2, and tyrosinase. Among them, melanin production starting from tyrosinase enzyme is converted into dopaquinone (DOPA-quinone) through tyrosinase enzyme through dopa (3,4-dihydroxy phenylalanine), which is DOPA through enzymatic reaction and auto-oxidation reaction Melanin is formed through -chrome.

When the synthesis of melanin occurs excessively in the skin, the skin tone is darkened, and spots, freckles, and the like occur. Therefore, by inhibiting the synthesis of melanin in the skin, it is possible not only to realize skin whitening by brightening the skin tone, but also to improve skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones, and genetic causes. have. Therefore, in the prior art, a substance having inhibitory activity against tyrosinase, such as hydroquinone, ascorbic acid, kojic acid, and glutathione, is mixed with cosmetics such as ointments and essences to the skin. Whitening was realized or skin hyperpigmentation such as spots and freckles was improved. However, hydroquinone exhibits a certain whitening effect, but there is a problem that the blending amount must be limited to a very small amount due to severe skin irritation. There is this. In addition, thiol-based reactants such as glutathione and cysteine not only have a characteristic unpleasant odor, but also have a problem in transdermal absorption, and their glycosides and derivatives are also highly polar, so it is difficult to use them as ingredients in cosmetics.

As a patent document for skin whitening, Patent Registration No. 10-1326690 discloses skin containing ginsenoside F1 (20-O-β-D-glucopyranosyl-20(S)-protophanaxatriol) as an active ingredient. It is a cosmetic composition for whitening, and Patent Registration No. 10-2010235 discloses black orchid (Liparis nervosa (Thunb.) Lindl.), pheasant fern (Plagiogyria japonica Nakai), Burmannia cryptopetala Makino and Utricularia yakusimensis Masam. It is a cosmetic composition having a skin moisturizing and skin whitening function containing a mixed extract extracted from a mixture of tyermanni Moore) extract as an active ingredient.

Registered Patent No. 10-1326690 Registered Patent No. 10-2010235 Registered Patent No. 10-0894714

The present invention is to solve the above-mentioned problems, and inhibits the activation of B16F10 melanoma cells from the active ingredient of the natural product, Gwangnae, through the reaction of microorganisms (including apoptosis, alleviation of aggression) and produces a compound that inhibits melanin synthesis An object of the present invention is to provide a whitening composition by biorenovation of Kwangnam extract that increases skin stability and reduces side effects, and a method for preparing the same.

The whitening composition by biorenovation of the Kwangnam tree extract according to the present invention is characterized in that it consists of a mixed reaction of the microorganism Bacillus amyloliquefaciens and the Kwangno tree extract.

According to the present invention, a method for preparing a whitening composition by biorenovation of a Kwangnam tree extract comprises: a first step of preparing the Kwangno tree extract; A second step of selecting and culturing Bacillus amyloliquefaciens as a microorganism and then centrifuging to obtain a microbial pellet; A third step of bioconversion to inhibit the activation of melanoma cells through the reaction of the microbial pellet obtained in the second step and the Kwangnamu extract prepared in the first step through the active ingredient of the Kwangnae extract, In the third step, the microbial pellet is washed with PG buffer, suspended in a PG buffer suspension, and then reacted with the Kwangnam tree extract.

According to the whitening composition by biorenovation of the Gwang tree extract according to the present invention and the method for preparing the same, the active ingredient of Gwangmyeon, which was not used for whitening, inhibits the activation of B16F10 melanoma cells through the reaction with microorganisms (death, alleviation of aggression) ) and produces a compound that inhibits melanin synthesis, thereby helping skin whitening, and as a natural product, it has the effect of increasing the stability of the skin and reducing side effects.

Therefore, it can be used without skin trouble by being added to skin whitening compositions such as cosmetics such as skin, lotion, cream, essence, pack, foam cleansing, etc. or medicines such as ointment for external use. .

1 is a process diagram of a method for preparing a whitening composition by biorenovation of an extract of Gwangju tree according to the present invention.
Figure 2 is a graph of the HPLC chromatogram of the general Kwangnae and the renovation reaction of the Kwangnae extract according to the present invention and Bacillus amyloliquefaciens.
3 to 9 show the test results of the whitening composition by biorenovation of Gwangnam extract according to an embodiment of the present invention,
3 is a graph showing the B16F10 cell viability,
Figure 4 is a graph showing the amount of melanin (Melanin) content,
Figure 5 is a graph showing the tyrosinase (tyrosinase) activity test results,
6 to 9 are images showing the results of Western blot analysis of the whitening composition by biorenovation of the Gwangnam tree extract of the present invention.

In the following description of the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description thereof will be omitted. In addition, the terms described below are terms defined in consideration of functions in the present invention, which may vary according to intentions or customs of users and operators. Therefore, the definition should be made based on the content throughout this specification.

The whitening composition by biorenovation of the Gwangnam extract according to the present invention, the activation of B16F10 melanoma cells from the active ingredient of Gwangju through the biorenovation bioconversion technique by the microorganism Bacillus amyloliquefaciens It is produced as a biorenovation reactant with excellent antioxidant properties that can inhibit

1. Guangxin extract.

Gwangnam is an evergreen broad-leaved arboreous tree distributed in Jeollanam-do, Gyeongsangnam-do coast, Jeju, island regions, Japan and Taiwan as a plant belonging to the oleaceae family. In addition, it is known that the leaves contain syringin, and components such as ursolic acid, oleanolic acid, and acetyl-oleanolic acid are contained in the peel and flesh. It has been reported that the leaves of Gwangnam are smooth in nature and non-toxic, improve eyesight, and are effective in relieving pain, stomatitis, periodontitis, acute bacterial dysentery, malignant tumors, and burns. Until now, research on the whitening effect of Gwangnam is not yet known, and it is an object to obtain a composition capable of whitening from the active ingredient (syringin) of Gwangmyeon through the present invention.

2. Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ).

As a microorganism for microbial innovation, Bacillus amyloliquefaciens was selected.

Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) was searched for microorganisms existing in Jeju traditional salted fish, for example, and among them, Bacillus amyloliquefaciens isolated from Jeju soybean soy sauce was selected.

That is, the present invention can help to promote the economy and promote the local government by utilizing the local government's special products in that the present invention uses Gwangnam, which is cultivated in large quantities in Jeju Island, and Jeju's traditional salted fish.

The overall process of the method for producing a whitening composition by biorenovation of a mulberry extract according to the present invention includes preparation of a mulberry extract - selection and culture of microorganisms - biorenovation - centrifugation / filtration - freeze-drying, and each process is as follows (Fig. see 1).

1. Manufacture of extracts from mulberry.

Gwangnam tree extract is manufactured through alcohol extraction, and the process is alcohol extraction - filtration - concentration under reduced pressure - freeze-drying The specific process is as follows.

go. spirit extraction.

Extract alcohol using 0.9-1.1 L of ethanol (preferably 70% Ethanol) with respect to 300 g of leaves (can be pulverized into small sizes).

Conditions for ethanol extraction are 57-63°C, 2 hours 40 minutes to 3 hours 20 minutes, preferably 60°C, 3 hours.

me. percolation.

The alcohol-extracted extract contains a solid, and the solid is filtered with, for example, a filter paper (paper filter).

All. Concentration (vacuum concentration).

In order to increase the concentration of the active ingredient of the extract that has undergone the filtration process, the extract is concentrated, preferably concentrated under reduced pressure (it can be freely carried out under normal reduced pressure concentration conditions, so the conditions for concentration under reduced pressure are not limited to specific values), and ethanol is removed in this process do.

la. freeze-dried.

The concentrated solution subjected to reduced pressure concentration may be stored by freeze-drying at, for example, minus 110° C., and the freeze-drying process is selected as necessary.

The above alcohol extraction process is the optimal process for extraction without destroying the active ingredients of the mulberry tree.

2. Microbial selection and culture.

Bacillus amyloliquefaciens selected as a microorganism was cultured in Nutrient broth (Beef extract 3.0 g/L, Peptone 5.0 g/L) for 18 hours to 22 hours, preferably for 20 hours according to optimal growth conditions. After culturing for 20 hours, the microorganisms are centrifuged at a speed of 4,800 to 5,200 rpm, 3 to 5° C., 8 minutes to 12 minutes, preferably 5,000 rpm, 4° C., for 10 minutes to obtain a microbial pellet.

3. Biorenovation (biotransformation reaction).

go. reaction synthesis.

The obtained microbial pellet was washed with PG buffer (50 mM phosphate pH 7.2 and 2% glycerol), and suspended in 5 ml of PG buffer (the reason for suspending is to remove foreign substances contained in the medium for culturing microorganisms in a clean state) In order to induce a reaction, and to accurately determine the concentration of glycerol in the composition). Thereafter, the suspension and the K. serrata extract are mixed {Gwangnae extract: microbial pellet: PG buffer = 1: 9-11: a weight ratio of 45-55, preferably 1: 10: 50 } and 120-180 rpm at 27-33 ° C. The reaction solution is synthesized by culturing for 70 hours to 74 hours at a rate of , preferably 72 hours at 30° C. at a rate of 150 rpm. The mixing ratio is an optimal ratio for recovering the antioxidant active ingredient of the Gwangnam extract.

me. Check whether biorenovation reactants are generated.

It may further include a step of confirming whether the biorenovation reactant (reactant) reacts. After the reaction is completed, centrifugation is performed to collect the supernatant as a biorenovation reactant, and concentrated under reduced pressure in DMSO (dimethyl sulfoxide, dimethyl sulfoxide). After dissolution, the bioconversion reaction product is checked by HPLC (high performance liquid chromatography).

All. separation sympathy.

Based on the HPLC result, a peak for the biorenovation reaction product generated using Prep-HPLC (preparative high performance liquid chromatography) was separated. After confirming the purity of the separated sample through HPLC, it is identified through LC-Mass (Liquid chromatography-mass spectrometry) and NMR (Nuclear magnetic resonance) Since they are the same, a detailed description will be omitted).

4. Centrifugation and filtration.

The biorenovation reaction is centrifuged at a speed of 4,800-5,200 rpm, 3-5° C., 8-12 minutes, preferably 5,000 rpm, 4° C., 10 minutes, and filtered through filter paper.

5. Concentrate under reduced pressure.

In order to increase the concentration of the active ingredient of the biorenovation reactant that has undergone the filtration process, the biorenovation reactant is concentrated under reduced pressure (it can be freely carried out under normal reduced pressure concentration conditions, so the conditions for concentration under reduced pressure are not limited to specific values).

6. Freeze drying.

The concentrate that has been concentrated under reduced pressure can be stored by freeze-drying, and the freeze-drying process is selected as needed.

The whitening composition according to the present invention is added so as to satisfy a content of 0.01 to 25% by weight in cosmetics, external drugs, and the like.

Preferred examples of the whitening composition according to the present invention are as follows.

<Example>

1. Manufacturing.

go. Gwangnam extract: 300 g of leaves of Gwangnam (raw material collected in Hallim-eup, Jeju-si, Jeju-si, Jeju-si, Jeju Special Self-Governing Province, provided by the Jeju Resources Plant Research Institute) are heated with 1 L of 70% ethanol at 60°C for 3 hours to extract alcohol. An extract was prepared.

me. Microorganism: Bacillus amyloliquefaciens (KCTC 43033) strain acquired from the Microbial Center was used for the biorenovation reaction, and Nutrient Broth (Beef extract 3.0 g/L, Peptone 5.0 g/L) medium required for the growth of the strain was used. and incubated for 72 hours at 30° C. at a speed of 150 rpm. After culturing this strain for 20 hours, centrifugation was performed at 5,000 rpm, 4° C., for 10 minutes to obtain a microbial pellet. The obtained microbial pellet was washed with PG buffer (50 mM phosphate pH 7.2 and 2% glycerol), and suspended in 5 ml of PG buffer.

All. biorenovation.

After mixing the extract and microorganisms at 30 ° C. for 72 hours at a speed of 150 rpm, a sample of the biorenovation reaction product is generated through separation and identification, and the supernatant is concentrated under reduced pressure through centrifugation and freeze-dried at -110 ° C. did.

2. Test.

The test of this example is to confirm the protein expression levels of TRP-1, TRP-2, MITF, and tyrosinase involved in cell viability, melanin synthesis inhibition, tyrosinase activity inhibition, and melanin synthesis using B16F10 melanoma cells.

go. cell culture.

α-Melanocyte-stimulating hormone (α-MSH, α-melanocyte-stimulating hormone) and L-3,4-dihydroxyphenylalanin (L-DOPA) used in this test were obtained from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA) was purchased and used. B16F10 melanoma cells were purchased from ATCC (American Type Cell Culture, USA), and Dulbecco's modified Eagle's containing 10% Fetal Bovine Serum (FBS, Gibco BRL Co., Grand Island, NY, USA) and 1% penicillin/streptomycin. medium (DMEM, Gibco BRL Co. Grand Island, NY, USA) was cultured in an incubator at 37 °C, 5% CO ₂ and subcultured once every 3 days.

me. HPLC Analysis of Biorenovation Bioconversion

HPLC comparative analysis of the sample was performed using Shimadzu SpectroMonitor 3200 digital UV/Vis detector (Phenomenex 4 μm Hydro-RP 80 Å, 250×4.6 mm), and purified water (0.1% TFA (Trifluoroacetic acid)) and Acetonitrile were used as mobile phases. was used. The composition of the mobile phase is A: H ₂ O 99.9: TFA (Trifluoroacetic acid) 0.1, B: Acetonitrile, starting with A: 90%, B: 10%, and analyzed under the conditions of A: 0%, B: 100% at 30 min. did. In addition, analysis was performed under UV 254 nm, which is the maximum absorption wavelength, 10 μl of a sample dissolved in ethanol, a solvent flow rate of 1.0 mL/min, a column temperature of 40° C., and gradient conditions.

Figure 2 is a graph of HPLC chromatograms of B. amyloliquefaciens (KCTC 43033) extract (BA), which is a microorganism used for biorenovation (Biorenovation), bioconverted L. japonica extract (LJ), and bioconverted L. japonica extract (LJBR). As a result, it was confirmed that a new peak that does not exist in LJ was generated at LJBR 14-17.5min. This suggests that the unknown reactant present in the LJ may have been converted into a new reactant through metabolism with microorganisms.

All. Cell viability.

Cytotoxicity of B16F10 melanoma cells (LJ), bioconverted L. amygdala extract (LJBR), and B. amyloliquefaciens extract (BA) used for biorenovation was measured by MTT assay. B16F10 melanoma cells were seeded in a 24-well plate at 1.0 × 10⁴ cells/well and cultured for 24 hours at 37°C, 5% CO₂ incubator. Then, α-MSH (200 nM) and 25, 50, and 100 μg/mL was treated with a concentration of and cultured for 72 hours.

Then, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide, Sigma-Aldrich, St. Louis, MO, USA) solution was added and incubator at 37 °C, 5% CO₂ After reacting for 4 hours, the supernatant was removed, and the formed pellet was dissolved by adding DMSO (dimethyl sulfoxide), placed in a 96 well plate, and absorbance was measured using a spectrophotometer (Thermo Fisher Scientific, Vantaa, Finland) at a wavelength of 570 nm. .

As can be seen from FIG. 3 , when BA, LJ, and LJBR were treated at concentrations of 25, 50, and 100 μg/mL, BA, LJ, and LJBR showed cell viability of about 98% or more at the measured concentrations. Therefore, subsequent experiments were carried out at concentrations of 25, 50, and 100 μg/mL that do not show cytotoxicity.

la. Evaluation of melanin production inhibitory activity.

To confirm the melanin synthesis inhibitory efficacy of LJ and LJBR, B16F10 melanoma cells were seeded in a 6-well plate at 4.0 × 10⁴ cells/well and cultured for 24 hours in an incubator at 37°C, 5% CO₂, α-MSH (200 nM) and LJ and LJBR were treated together at a non-cytotoxic concentration and cultured for 72 hours. After that, the cells were washed with PBS (phosphate buffered saline, Sigma-Aldrich, St. Louis, MO, USA) and treated with trypsin-EDTA (Gibco, Grand Island, NY, USA) to recover the cells and use 1% protease inhibitor ( Cells were lysed by adding RIPA buffer (Sigma-Aldrich, St. Louis, MO, USA) containing Sigma-Aldrich, St. Louis, MO, USA), followed by centrifugation at 13,000 rpm for 40 minutes to separate the pellet and the supernatant. did. 1 N NaOH (Sigma-Aldrich, St. Louis, MO, USA) was added to the separated pellet, heated at 90 °C for 1 hour, and absorbance was measured at a wavelength of 405 nm using a spectrophotometer.

As a result of confirming the effect on melanogenesis during LJ and LJBR treatment, as shown in FIG. 4, compared to the α-MSH alone treatment group, the LJ treatment group showed slight inhibition of melanin synthesis by about 18%, 18%, and 19%, whereas The LJBR treatment group showed inhibition of melanin synthesis by 21%, 25%, and 29%. This result indicates that LJBR bioconverted by biorenovation technique has a superior inhibitory effect on melanin synthesis than conventional LJ.

mind. Tyrosinase (Tyrosinase) production inhibitory activity evaluation.

Tyrosinase is an enzyme involved in melanogenesis. In order to measure the activity of this tyrosinase, B16F10 melanoma cells were dispensed at 4.0 × 10⁴ cells/well in a 6-well plate and cultured at 37°C, 5% CO₂ for 24 hours in an incubator. MSH (200 nM) and samples were treated with each concentration (25, 50, 100 μg/mL) and incubated for 72 hours. Thereafter, cells were recovered by treatment with trypsin-EDTA for 3 minutes, lysed with RIPA buffer (1% protease inhibitor), and centrifuged at 13,000 rpm for 40 minutes to separate the pellet and the supernatant. The separated supernatant was quantified using a BCA protein assay kit (Thermo Scientific, USA), and 0.1 M sodiumphosphate buffer (Samchum Chemical Co., Seoul, Korea) and L-DOPA (Sigma-Aldrich, St. Louis, MO, USA) ) at 2 mg/mL, added to a 96-well plate together with the quantified protein, and reacted at 37°C for 2 hours. After the reaction, tyrosinase activity was measured at 490 nm using a spectrophotometer.

As a result of confirming the effect of LJ and LJBR on tyrosinase acting in the initial stage of melanin synthesis, as shown in FIG. 5, compared with the α-MSH (200 nM) alone treatment group, the LJ treatment group had the highest concentration of 100 μg/mL. While tyrosinase activity was inhibited by 22%, in the LJBR-treated group, 14%, 49%, and 58% of the activity was inhibited at concentrations of 25, 50, and 100 μg/mL. This result shows the possibility that a reactant directly or indirectly involved in the tyrosinase enzyme was generated after bioconversion, thereby inhibiting the tyrosinase enzyme.

bar. Western Blot.

To measure the expression levels of MITF, TRP-1, TRP-2, and tyrosinase, which are important proteins for melanin synthesis, B16F10 melanoma cells were dispensed in a 6-well plate at 4.0 × 10⁴ cells/well, 37°C, 5% CO ₂ Incubator After incubation for 48 hours, α-MSH (200 nM) and LJ and LJBR were treated together at each concentration (25, 50, 100 μg/mL) and incubated for 48 hours. Then, the cultured cells were washed with PBS, lysed by adding RIPA buffer (1% protease inhibitor), and centrifuged at 13,000 rpm for 40 minutes to separate the pellet and the supernatant. The separated supernatant was quantified using a BCA protein assay kit. After electrophoresis of the quantified supernatant on 10% SDS-PAGE, the protein was transferred to a poly-vinylidene difluoride (PVDF) membrane (Milipore, Burlington, MA, USA). . After that, a solution containing 5% skim milk in 0.05% Tween 20/Tris-buffered saline (0.05% T/TBS) was added to the membrane and blocked at room temperature for 2 hours, and then the primary antibodies MITF, TRP-1, TRP -2, tyrosinase (Santa cruz Biotechnology, USA) was treated and reacted at low temperature (4°C) for 18 hours, washed 4 times with 0.05% T/TBS for 10 minutes, and then Horseradish peroxidase (HRP)-conjugated secondary andtibody Phosphorus anti-mouse IgG (Cell signaling, USA) was diluted with 0.05% T/TBS and 1:10,000 and adhered at room temperature for 2 hours, followed by washing 4 times with 0.05% T/TBS. Proteins were measured using an enhanced chemiluminescence kit (Bio-rad, USA) through an Imaging densitometer (model GS-700, Bio-rad, USA). In addition, MITF, TRP-1, TRP-2, and tyrosinase protein expression levels were expressed in graphs after quantifying the area using the imageJ program (NIH, Bethesda, MD, USA).

As shown in FIGS. 6 to 9, the expression of melanin synthesis-related factors, TRP-1, TRP-2, MITF and tyrosinase, compared to the results of the previous experiment where melanin production was inhibited during LJ treatment for α-MSH-treated cells. did not show concentration-dependent inhibitory activity. On the other hand, the LJBR-treated group showed superior and concentration-dependent whitening activity than the existing extract (LJ). In particular, the expression of TRP-1 and tyrosinase decreased by 89% and 90% at LJBR 100 μg/mL concentration, respectively, making it superior to the untreated group. It showed whitening activity. This is due to the fact that LJ is a natural material, and it is judged that the expression of related factors did not appear consistently due to the multiple components acting in the material. It is presumed to have been shown

Summarizing the above test results, as a result of examining the whitening activity of B16F10 melanoma cells stimulated with α-MSH, the bioconverted reactant (LJBR) using the Gwangnam tree extract (LJ) and the biorenovation technique, the cytotoxicity was determined. As a result of confirming the expression of TRP-1, TRP-2, MITF and tyrosinase involved in melanin production and melanin synthesis within invisible concentrations (25, 50, 100 g/mL), both LJ and LJBR inhibited melanin synthesis, but LJBR showed better activity. In addition, while the expression of factors involved in melanin synthesis was not inhibited in a concentration-dependent manner in LJ, a concentration-dependent inhibitory activity was confirmed in LJBR. did. Therefore, it was confirmed that LJBR has inhibitory activity on melanin synthesis by directly or indirectly affecting the expression of enzymes and proteins related to melanin synthesis. suggest that it can be induced.

Claims (6)

Bacillus amyloliquefaciens , which is a microorganism, a whitening composition by biorenovation of an extract, characterized in that it consists of a mixed reaction of the extract of Gwangju. delete A first step of preparing an extract of Gwangju;
A second step of selecting and culturing Bacillus amyloliquefaciens as a microorganism and then centrifuging to obtain a microbial pellet;
A third step of mixing and reacting the microbial pellets obtained in the second step and the Kwangnam tree extract prepared in the first step,
The first step is a method for producing a whitening composition by biorenovation of a gwang tree extract, characterized in that after extracting the flowers and leaves of gwang tree with ethanol and then extracting the gwang tree extract through concentration under reduced pressure. The method according to claim 3, wherein in the third step, the microbial pellets are washed with PG buffer, suspended in a PG buffer suspension, and then reacted with the Gwangnam tree extract. [Claim 5] The method of claim 4, wherein the method for producing a whitening composition by biorenovation of a Kwangnam tree extract comprises a fourth step of centrifuging the bioconverted renovation reaction product through the third step.








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