Composition for inhibiting lipofuscin accumulation or removing lipofuscin comprising culture or its extract of bacillus ginsengisoli

Abstract

The present invention discloses a composition for inhibiting lipofuscin accumulation or removing lipofuscin. The composition contains a strain of the genus Bacillus, a lysate thereof, a culture thereof, or an extract of said lysate or culture. The composition provides an anti-aging or whitening effect by inhibiting lipofuscin accumulation or removing lipofuscin. Furthermore, the composition provides an effect of preventing, improving, or treating diseases caused by lipofuscin accumulation, such as skin hyperpigmentation, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), myocardial infarction, and age-related macular degeneration.

Description

Technology Field

[0001] The present invention discloses a composition for inhibiting lipofuscin accumulation or removing lipofuscin, and a strain of the genus Bacillus having activity for inhibiting lipofuscin accumulation or removing lipofuscin. Background Technology

[0002] Lipofuscin is a type of metabolic waste produced when various metabolites are digested or processed within intracellular liposomes; it is a yellowish-brown pigment granule that is physiologically present in the heart muscle. As lipofuscin accumulates in cells, cell function deteriorates and the rate of aging accelerates.

[0003] The hearts of long-lived elderly individuals take on a brownish tint and shrink due to an increase in lipofuscin pigment; this phenomenon is known as cardiac brown atrophy. Similar pigmentation can occur in the liver, known as hepatic brown atrophy. Furthermore, cases of lipofuscin pigment deposition in the heart, liver, and kidneys have been reported in cancer patients or those suffering from wasting diseases such as severe pulmonary tuberculosis for a long time. Meanwhile, the most prominent phenomenon associated with skin aging is pigmentation, such as melasma and age spots, which results from the accumulation of lipofuscin. Consequently, lipofuscin is also referred to as "age spots inside the body." Therefore, inhibiting the production and / or accumulation of lipofuscin to prevent it from accumulating in skin cells is crucial for preventing skin aging.

[0004] As mentioned above, lipofuscin, which naturally occurs during cellular metabolic processes, accumulates in larger quantities within cells not only with aging but also when cellular function declines or there is repeated exposure to external toxins. Furthermore, accumulated lipofuscin interferes with normal cellular function and accelerates the aging process. Lipofuscin is found not only in skin cells but also in the liver, kidneys, heart muscle, retina, adrenal glands, nerve cells, and ganglion cells; it is known to cause various degenerative diseases when it accumulates in vital organs. For example, if lipofuscin accumulates in heart muscle, it can worsen the condition and lead to myocardial infarction, while accumulation in the retina or nerve cells can cause macular degeneration or Alzheimer's disease. Additionally, if it accumulates in hormone-secreting organs such as the adrenal glands, liver, and pituitary gland, it can cause abnormalities in the metabolic function of bodily hormones, including growth hormone and sex hormones.

[0005] Conventional techniques for removing lipofuscin have primarily involved dermatological laser procedures. Additionally, substances for preventing skin aging have been used indirectly, and there are virtually no known substances for directly removing lipofuscin. To date, there has also been no known information regarding the inhibition or removal of lipofuscin accumulation using Bacillus strains. Prior art literature

[0006] (Patent Document 0001) KR 10-2016-0049435 A The problem to be solved

[0007] In one aspect, the present disclosure aims to provide a composition for inhibiting lipofuscin accumulation or removing lipofuscin.

[0008] In another aspect, the present disclosure relates to Bacillus ginsengisolyi Heritage GR22, with the novel accession number KCCM12963P ( Bacillus ginsengisoli The purpose is to provide the Heritage GR22 strain. means of solving the problem

[0009] In one aspect, the present disclosure relates to Bacillus ( Bacillus A composition for inhibiting lipofuscin accumulation or removing lipofuscin is provided, comprising as an active ingredient a strain of the genus ) a lysate, a lysate thereof, a culture thereof, or an extract of said lysate or culture.

[0010] In an exemplary embodiment, the composition may include a culture of a strain of the genus Bacillus or an extract thereof as an active ingredient.

[0011] In an exemplary embodiment, the strain is Bacillus ginsengisoli ( Bacillus ginsengisoli ) and Bacillus thuringiensis ( Bacillus thuringiensis It may be one or more of ).

[0012] In an exemplary embodiment, the strain is Bacillus ginsengisoli ( Bacillus ginsengisoli It could be ).

[0013] In an exemplary embodiment, the strain is Bacillus ginsengisolyi Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli It could be Heritage GR22)

[0014] In an exemplary embodiment, the composition may be an anti-aging composition.

[0015] In an exemplary embodiment, the composition may be a whitening composition.

[0016] In an exemplary embodiment, the composition may prevent, improve, or treat a disease caused by lipofuscin accumulation.

[0017] In an exemplary embodiment, the disease caused by lipofuscin accumulation may include one or more selected from the group consisting of skin hyperpigmentation, sarcopenia, progeria, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), myocardial infarction, age-related macular degeneration, neuronal ceroid lipofuscinoses (NCL), acromegaly, denervation atrophy, and chronic obstructive pulmonary disease (COPD).

[0018] In an exemplary embodiment, the skin hyperpigmentation may be one or more selected from the group consisting of melasma, liver spot, freckle, lentigo, nevus, pigmentation caused by drugs and pigmentation caused by inflammation.

[0019] In an exemplary embodiment, the composition may be a cosmetic composition, a food composition, or a pharmaceutical composition.

[0020] In an exemplary embodiment, the composition may contain an active ingredient in an amount of 0.01 to 99.99 weight%.

[0021] In another aspect, the present disclosure relates to Bacillus ginsengisolyi Heritage GR22 (accession number KCCM12963P) Bacillus ginsengisoli Provides the Heritage GR22 strain.

[0022] In an exemplary embodiment, the strain may have lipofuscin accumulation inhibitory or lipofuscin removal activity. Effects of the invention

[0023] In one aspect, the technology disclosed in the present disclosure has the effect of providing a composition for inhibiting lipofuscin accumulation or removing lipofuscin.

[0024] In another aspect, the technology disclosed in this disclosure is Bacillus ginsengisoli Heritage GR22, with novel accession number KCCM12963P ( Bacillus ginsengisoli It has the effect of providing Heritage GR22 strains. Brief explanation of the drawing

[0025] Figure 1 shows the 16S rDNA partial sequence of the Bacillus ginsengisolyi Heritage GR22 strain according to one embodiment. Figure 2 shows a graph confirming the lipofuscin removal efficacy of a Bacillus genus strain culture in fibroblasts according to one experimental example. Specific details for implementing the invention

[0026] The present disclosure will be described in detail below.

[0027] In one aspect, the present disclosure relates to Bacillus ( Bacillus A composition for inhibiting lipofuscin accumulation or removing lipofuscin is provided, comprising as an active ingredient a strain of the genus ) a lysate, a lysate thereof, a culture thereof, or an extract of said lysate or culture.

[0028] In this document, "active ingredient" refers to a component that exhibits the intended activity alone or can exhibit the intended activity in combination with a carrier, etc., which is inactive in itself.

[0029] The above 'crushed material' refers to a product obtained by crushing the microorganisms themselves by chemical or physical force.

[0030] The term "culture" above refers to a substance comprising some or all of the substances contained within a culture medium in which microorganisms are cultured, regardless of the form of the culture. For example, the culture may include metabolites or secretions that are byproducts of microbial culture. Additionally, the microorganisms themselves may also be contained within the culture.

[0031] In an exemplary embodiment, the culture may be the culture itself obtained by culturing a microorganism according to the present disclosure in a suitable liquid medium, a filtrate (filtrate or supernatant after centrifugation) from which the microorganism has been removed by filtering or centrifuging the culture, a lysate obtained by ultrasonically treating the culture or treating the culture with a lysozyme, or a concentrated powder obtained by concentrating and freeze-drying the culture.

[0032] In an exemplary embodiment, the culture may be a culture medium.

[0033] In an exemplary embodiment, the culture may be a culture obtained by culturing a strain in a liquid medium and then removing the strain.

[0034] In an exemplary embodiment, the culture may be a concentrate obtained by concentrating the culture medium.

[0035] The term "extract" above refers to a product obtained by extracting the crushed material or culture, regardless of the extraction method, extraction solvent, extracted component, or form of the extract. The term "extract" is a broad concept that includes all substances that can be obtained by processing or treating them through other methods, such as concentration, drying, or fractionation, after extraction.

[0036] In an exemplary embodiment, the extract may be extracted with one or more extraction solvents selected from the group consisting of water, anhydrous or hydrated alcohols having 1 to 6 carbon atoms (e.g., methanol, ethanol, propanol, or butanol), propylene glycol, butylene glycol, dipropylene glycol, glycerin, acetone, ethyl acetate, chloroform, methylene chloride, butyl acetate, diethyl ether, dichloromethane, hexane, and mixtures thereof.

[0037] In an exemplary embodiment, the extract may be extracted with a solvent selected from water, alcohol, or a mixture thereof, preferably C1 to C4 lower alcohols or a mixture thereof, and more specifically, with an aqueous solution of methanol or ethanol.

[0038] In an exemplary embodiment, the composition may include a culture of a strain of the genus Bacillus or an extract thereof as an active ingredient.

[0039] In an exemplary embodiment, the composition may include a culture solution of a Bacillus genus strain as an active ingredient.

[0040] In an exemplary embodiment, the strain may be isolated from ginseng.

[0041] In an exemplary embodiment, the strain may be isolated from ginseng and soil of the ginseng rhizosphere.

[0042] In an exemplary embodiment, the strain is Bacillus ginsengisoli ( Bacillus ginsengisoli ) and Bacillus thuringiensis ( Bacillus thuringiensis It may be one or more of ).

[0043] In an exemplary embodiment, the strain is Bacillus ginsengisoli ( Bacillus ginsengisoli It may be desirable in terms of lipofuscin removal activity to be )

[0044] In an exemplary embodiment, the strain is Bacillus ginsengisolyi Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli It could be Heritage GR22)

[0045] In an exemplary embodiment, the Bacillus ginsengisolyi heritage GR22 strain may have the 16S rDNA partial sequence of SEQ ID NO. 1.

[0046] In an exemplary embodiment, the composition may be an anti-aging composition.

[0047] In this document, "for anti-aging" refers to a use for preventing, delaying, and / or improving aging phenomena caused by internal factors, including genetic factors, and external factors, including ultraviolet rays. The above composition provides the effect of preventing, improving, or treating diseases or symptoms related to cellular aging by reducing the level of cellular aging.

[0048] In an exemplary embodiment, the composition may be a skin anti-aging composition.

[0049] In an exemplary embodiment, the composition may be a whitening composition.

[0050] In an exemplary embodiment, the composition may prevent, improve, or treat a disease caused by lipofuscin accumulation.

[0051] In an exemplary embodiment, the disease caused by lipofuscin accumulation may include one or more selected from the group consisting of skin hyperpigmentation, sarcopenia, progeria, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), myocardial infarction, age-related macular degeneration, neuronal ceroid lipofuscinoses (NCL), acromegaly, denervation atrophy, and chronic obstructive pulmonary disease (COPD).

[0052] In an exemplary embodiment, the skin hyperpigmentation may be one or more selected from the group consisting of melasma, liver spot, freckle, lentigo, nevus, pigmentation caused by drugs and pigmentation caused by inflammation.

[0053] In an exemplary embodiment, the composition may help whiten and / or improve skin tone by inhibiting skin hyperpigmentation.

[0054] In an exemplary embodiment, the composition may be a cosmetic composition, a food composition, or a pharmaceutical composition.

[0055] In an exemplary embodiment, the cosmetic composition may additionally include functional additives and ingredients included in general cosmetic compositions. The functional additives may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extracts. Other ingredients included in the formulation may include oil components, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc.

[0056] In an exemplary embodiment, the cosmetic composition is not particularly limited in its formulation and can be appropriately selected according to the purpose. For example, it may be prepared in one or more formulations selected from the group consisting of skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, but is not limited thereto.

[0057] In an exemplary embodiment, when the formulation of the composition is a paste, cream, or gel, animal fiber, plant fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0058] In an exemplary embodiment, when the formulation of the composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and particularly in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0059] In an exemplary embodiment, when the formulation of the composition is a solution or an emulsion, a solvent, a solvating agent, or an emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0060] In an exemplary embodiment, when the formulation of the composition is a suspension, a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used as a carrier component.

[0061] In an exemplary embodiment, when the formulation of the composition is a surfactant-containing cleansing agent, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyl taurate, a sarcosinate, a fatty acid amide ether sulfate, an alkylamidobetaine, an aliphatic alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a linolenic acid derivative, or an ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0062] In an exemplary embodiment, the food composition may be in a liquid or solid form, such as various foods, beverages, chewing gum, tea, vitamin complexes, health supplements, etc., and may be used in the form of powder, granules, tablets, capsules, or beverages. For each food composition of a specific form, ingredients commonly used in the field can be appropriately selected and combined by a person skilled in the art without difficulty according to the form or purpose of use, and a synergistic effect may occur when applied simultaneously with other raw materials.

[0063] In an exemplary embodiment, the liquid formulation may include various flavoring agents or natural carbohydrates as additional ingredients, such as in conventional beverages. Examples of the natural carbohydrates include disaccharides such as monosaccharides, glucose, and fructose; polysaccharides such as maltose and sucrose; conventional sugars such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. As for the flavoring agents, natural flavoring agents (taumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) may be advantageously used. The proportion of the natural carbohydrate may generally be about 1 to 20 g, and in one aspect, about 5 to 12 g per 100 ml of the composition.

[0064] In an exemplary embodiment, the food composition may, in one aspect, include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In another aspect, it may include fruit pulp for the production of natural fruit juices and vegetable beverages. The above ingredients may be used independently or in combination. The proportion of the additive may vary, but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition.

[0065] In an exemplary embodiment, the pharmaceutical composition may further contain pharmaceutical adjuvants such as preservatives, stabilizers, hydrating agents or emulsification promoters, salts and / or buffers for osmotic pressure regulation, and other therapeutically useful substances, and may be formulated into various oral or parenteral administration forms according to conventional methods.

[0066] In an exemplary embodiment, the oral dosage form is, for example, a tablet, pill, hard and soft capsule, liquid, suspension, emulsifier, syrup, powder, granule, granule, pellet, etc., and these formulations may contain a surfactant, a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine), and a lubricant (e.g., silica, talc, stearic acid and its magnesium or calcium salt, and polyethylene glycol). The tablet may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidine, and, in some cases, may contain pharmaceutical additives such as disintegrants, absorbents, coloring agents, flavoring agents, and sweeteners, such as starch, agar, alginic acid, or its sodium salt. The tablet may be manufactured by conventional mixing, granulation, or coating methods.

[0067] In an exemplary embodiment, the parenteral administration form may be a transdermal administration form, such as an injection, drops, ointment, lotion, gel, cream, spray, suspension, emulsion, or patch, but is not limited thereto.

[0068] In an exemplary embodiment, the determination of the dosage of the active ingredient is within the level of a person skilled in the art, and the daily dosage of the drug varies depending on various factors such as the stage of progression, onset time, age, health condition, and complications of the subject to be administered, but based on adults, the composition may be administered in divided doses of 1 μg / kg to 200 mg / kg in one aspect and 50 μg / kg to 50 mg / kg in another aspect, divided into 1 to 3 doses per day.

[0069] In an exemplary embodiment, the pharmaceutical composition may be a topical skin preparation, and the topical skin preparation may include any of the formulations of medicines applied externally to the skin.

[0070] In an exemplary embodiment, the composition may contain an active ingredient in an amount of 0.01 to 99.99 weight%.

[0071] In another exemplary embodiment, the composition may contain an active ingredient in an amount of 0.01 wt% or more, 0.1 wt% or more, 0.5 wt% or more, or 1 wt% or more, and 99.99 wt% or less, 90 wt% or less, 80 wt% or less, 70 wt% or less, 60 wt% or less, 50 wt% or less, 40 wt% or less, 30 wt% or less, 20 wt% or less, 10 wt% or less, 5 wt% or less, 3 wt% or less, or 1 wt% or less.

[0072] In another aspect, the present disclosure relates to Bacillus ginsengisolyi Heritage GR22 (accession number KCCM12963P) Bacillus ginsengisoli Provides the Heritage GR22 strain.

[0073] In an exemplary embodiment, the strain may have lipofuscin accumulation inhibitory or lipofuscin removal activity.

[0074] The present disclosure will be described in more detail below through examples. These examples are solely for illustrating the present disclosure, and it will be obvious to those skilled in the art that the scope of the present disclosure is not to be interpreted as being limited by these examples.

[0075] Example 1. Isolation and Identification of Microorganisms

[0076] 1 g of ginseng and soil from the ginseng rhizosphere cultivated in Anseong-si, Gyeonggi-do, South Korea was added to 50 mL of PBS buffer (phosphate buffered saline) and stirred for 30 minutes, after which it was filtered using filter paper (Whatman filter paper, 2 μm). The filtrate was plated onto solid medium M17 Agar and incubated statically at 30 ℃ for 2 days. Subsequently, strains showing microbial colonies were subcultured at least 2 times and up to 4 times to finally isolate a single strain.

[0077] After culturing the above-mentioned isolated microorganism in a liquid medium, genomic DNA was extracted from the microorganism and amplified by the PCR (polymerase chain reaction) method using 27F (5'-AGAGTTTGATCMTGGCTCAG-3', SEQ ID NO. 2) and 1492R (5'-TACGGYTACCTTGTTACGACTT-3', SEQ ID NO. 3) primers, which are commonly used for bacterial identification, and the sequence was analyzed. As a result of searching the analyzed nucleotide sequence in the Gene Bank, the isolated strain was identified as Bacillus ginsengisolii ( Bacillus ginsengisoli It was finally confirmed that it has 97% homology with the DCY53 (KCTC 13945) strain, and thus Bacillus ginsengisolyi Heritage GR22 ( Bacillus ginsengisoli The strain was named Heritage GR22, and on March 5, 2021, the strain was deposited with the Korean Culture Collection of Microorganisms, the depositary institution, and assigned deposit number KCCM12963P. The 16S rDNA partial sequence of the Bacillus ginsengisolyi Heritage GR22 strain of sequence number 1 is shown in Figure 1.

[0078] In the manner described above, strains exhibiting microbial colonies were subcultured a minimum of 2 times and a maximum of 4 times to finally isolate single strains. After culturing in liquid media, genomic DNA was extracted from the microorganisms and amplified by PCR using 27F (5'-AGAGTTTGATCMTGGCTCAG-3', SEQ ID NO. 2) and 1492R (5'-TACGGYTACCTTGTTACGACTT-3', SEQ ID NO. 3), primers commonly used for bacterial identification, and the sequences were analyzed. As a result of searching the analyzed nucleotide sequences in the Gene Bank, the isolated strain was identified as Bacillus thuringiensis ( Bacillus thuringiensis It was confirmed that it belongs to ). The isolated strain is Bacillus thuringiensis ( Bacillus thuringiensis It had 98% homology with strain KU4.

[0079] Examples 2. Preparation of microbial culture solution

[0080] The Bacillus ginsengisoli and Bacillus thuringiensis strains identified in Example 1 were each inoculated into M17 medium containing 0.5% glucose and cultured at 30°C for one day. After microbial culture, the cells were removed by centrifugation at 12,000 rpm, and the microbial culture solution was recovered. The recovered solution was concentrated by ultrafiltration (10 KDa cutoff) and washed with distilled water. Subsequently, the concentrate was dried using a freeze-dryer to produce a powder.

[0081] Experimental Example 1. Lipofuscin Evaluation of removal activity

[0082] In this experimental example, Bacillus ginsengisoli and Bacillus thuringiensis strains were used as Bacillus strains, and the lipofuscin removal activity of the Bacillus strains was evaluated as follows.

[0083] Normal human dermal fibroblasts (NHDF; Lonza, Switzerland) were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovin serum. All cultures were performed in a 37°C, 5% CO2 incubator. Fibroblasts with induced lipofuscin production were prepared by treating cells with oxidation-inducing substances (40 μM leupeptin, 45 μM FeCl3, and 10 μM H2O2) for 10 days when the cells had grown to 90% confluent. The medium was changed every two days. For the experimental group treated with Bacillus strains, 10 ppm of powder prepared from the Bacillus ginsengisoly culture medium and Bacillus thuringiensis culture medium prepared in Example 2 were applied for 7 days, respectively, 10 days after treatment with the oxidation-inducing substance. Subsequently, the lipofuscin content of each experimental group was measured by the autofluorescence of the lipofuscin pigment using a fluorescence plate reader (Synergy H1 hybrid multi-mode microplate reader, BioTeck). The measurement wavelengths were set to 340–410 nm for excitation and 440–480 nm for emission.

[0084] When fibroblasts were treated with 40 μM leupeptin, 45 μM FeCl3, and 10 μM H2O2, it was confirmed that lipofuscin levels increased to a high concentration of approximately 80% compared to the control group of fibroblasts not treated with oxidation-inducing substances. Furthermore, it was confirmed that the lipofuscin generated in the experimental group of lipofuscin-producing fibroblasts was significantly reduced when treated with culture medium of Bacillus ginsengisoli and Bacillus thuringiensis. In particular, the experimental group treated with Bacillus ginsengisoli culture medium exhibited excellent lipofuscin removal activity (see Fig. 2).

[0085] Examples of formulations of a composition according to one aspect of the present disclosure are described below, but they may be applied to various other formulations, and this is intended only to provide a specific description and not to limit the present disclosure.

[0086] Formulation Example 1. Softening lotion

[0087] A softening lotion was prepared according to a conventional manufacturing method by mixing 0.01 wt% of the Bacillus ginsengisoly culture of Example 2, 3 wt% of glycerin, 2 wt% of butylene glycol, 2 wt% of propylene glycol, 0.1 wt% of carboxyvinyl polymer, 10 wt% of ethanol, 0.1 wt% of triethanolamine, a trace amount of preservative, a trace amount of colorant, a trace amount of fragrance, and the remainder of purified water.

[0088] Formulation Example 2. Nourishing lotion

[0089] A nourishing cosmetic was prepared according to a conventional manufacturing method by mixing 0.01 wt% of the Bacillus ginsengisoli culture of Example 2, 4 wt% of beeswax, 1.5 wt% of polysorbate 60, 0.5 wt% of sorbitan sesquioleate, 5 wt% of liquid paraffin, 5 wt% of squalane, 5 wt% of caprylic / capric triglyceride, 3 wt% of glycerin, 3 wt% of butylene glycol, 3 wt% of propylene glycol, 0.1 wt% of carboxyvinyl polymer, 0.2 wt% of triethanolamine, trace amounts of preservatives, trace amounts of colorants, trace amounts of fragrances, and the remainder of purified water.

[0090] Formulation Example 3. Nourishing Cream

[0091] A nourishing cream was prepared according to a conventional manufacturing method by mixing 0.01 wt% of the Bacillus ginsengisoli culture of Example 2, 10 wt% of beeswax, 1.5 wt% of polysorbate 60, 0.5 wt% of sorbitan sesquioleate, 10 wt% of liquid paraffin, 5 wt% of squalane, 5 wt% of caprylic / capric triglyceride, 5 wt% of glycerin, 3 wt% of butylene glycol, 3 wt% of propylene glycol, 0.2 wt% of triethanolamine, a trace amount of preservative, a trace amount of colorant, a trace amount of fragrance, and the remainder of purified water.

[0092] Formulation Example 4. Pack

[0093] A pack was prepared according to a conventional manufacturing method by mixing 0.01 wt% of the Bacillus ginsengisoli culture of Example 2, 13 wt% of polyvinyl alcohol, 0.2 wt% of sodium carboxymethylcellulose, 0.1 wt% of allantoin, 5 wt% of ethanol, 0.3 wt% of nonylphenyl ether, a trace amount of preservative, a trace amount of colorant, a trace amount of fragrance, and the remainder of purified water.

[0094] Formulation Example 5. Drug for local administration (patch)

[0095] A topical administration agent (patch) was prepared by a conventional method according to the composition listed in Table 1 below.

[0096] Ingredient Name Content (weight%) Bacillus ginsengisoli culture of Example 2 2.0 Beta-1,3-glucan 3.0 diethylamine 0.7 sodium sulfite 0.1 Polyoxyethylene lauryl ether (EO=9) 1.0 Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) 1.0 Viscous paraffin oil 2.5 Caprylic acid ester / Capric acid ester (Cetiol LC) 2.5 Polyethylene glycol 400 3.0 Polyacrylic acid (Carbopol 934P) 1.0 purified water Remaining amount total 100

[0097] Formulation Example 6. Powder

[0098] A powder was prepared by mixing 2g of the Bacillus ginsengisoli culture from Example 2 above and 1g of lactose, and then filling it into an airtight bag.

[0099] Formulation Example 7. Tablet

[0100] 100 mg of the Bacillus ginsengisoli culture from Example 2, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were mixed, and then tablets were prepared by compressing them according to a conventional manufacturing method.

[0101] Formulation Example 8. Capsule

[0102] 100 mg of the Bacillus ginsengisoli culture from Example 2, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were mixed, and then a capsule was prepared by filling it into a gelatin capsule according to a conventional manufacturing method.

[0103] Formulation Example 9. Pill

[0104] 1g of the Bacillus ginsengisoli culture from Example 2 above, 1.5g of lactose, 1g of glycerin, and 0.5g of xylitol were mixed, and then pills were prepared according to a conventional method so that each pill weighed 4g.

[0105] Formulation Example 10. Granules

[0106] 150g of the Bacillus ginsengisoli culture from Example 2, 50mg of soybean extract, 200mg of glucose, and 600mg of starch were mixed, then 100mg of 30% ethanol was added and dried at 60°C to form granules, and then filled into a bag to produce granules.

[0107] Formulation Example 11. Drink

[0108] 50 mg of the Bacillus ginsengisoli culture from Example 2, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide were mixed, and then 300 ml of purified water was added and filled into each bottle with 200 ml. After filling into the bottles, the beverage was prepared by sterilizing at 130°C for 4 to 5 seconds.

[0109] Formulation Example 12. Caramel

[0110] Caramel was prepared by mixing and molding 50 mg of the Bacillus ginsengisoli culture from Example 2 above, 1.8 g of corn syrup, 0.5 g of skim milk, 0.5 g of soy lecithin, 0.6 g of butter, 0.4 g of hydrogenated vegetable oil, 1.4 g of sugar, 0.58 g of margarine, and 20 mg of salt.

[0111] Foregoing, specific parts of the present disclosure have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the present disclosure. Accordingly, the actual scope of the present disclosure is defined by the appended claims and their equivalents.

[0112] Name of depositing institution: Korean Culture Collection of Microorganisms Trustee Number: KCCM12963P Date of Deposit: 2021-03-05

Claims

Claim 1 It is a cosmetic composition for anti-aging or whitening, and the composition is Bacillus ( Bacillus ) strains of the genus, lysates thereof, cultures thereof, or extracts of said lysates or cultures are included as active ingredients, and said strains are Bacillus ginsengisoly ( Bacillus ginsengisoli ) and Bacillus thuringiensis ( Bacillus thuringiensis A cosmetic composition for anti-aging or whitening, wherein at least one of ) is present, and the anti-aging or whitening is achieved through inhibition of lipofuscin accumulation or removal of lipofuscin. Claim 2 In claim 1, the strain is Bacillus ginsengisolyi Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli A cosmetic composition for anti-aging or whitening, containing Heritage GR22. Claim 3 A cosmetic composition for anti-aging or whitening according to claim 1, wherein the composition comprises a culture of a strain of the genus Bacillus as an active ingredient, and said culture is obtained by removing the bacterial cells after culturing the strain. Claim 4 A cosmetic composition for anti-aging or whitening according to claim 1, wherein the composition comprises an active ingredient in an amount of 0.01 to 99.99 weight%. Claim 5 A food composition for anti-aging of the skin or improvement of skin hyperpigmentation, and said composition is Bacillus ( Bacillus ) strains of the genus, lysates thereof, cultures thereof, or extracts of said lysates or cultures are included as active ingredients, and said strains are Bacillus ginsengisoly ( Bacillus ginsengisoli ) and Bacillus thuringiensis ( Bacillus thuringiensis A food composition for improving skin anti-aging or skin hyperpigmentation, wherein at least one of ) is present, and the skin anti-aging or skin hyperpigmentation improvement is achieved through inhibition of lipofuscin accumulation or removal of lipofuscin. Claim 6 In claim 5, the strain is Bacillus ginsengisolyi Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli A food composition for improving skin anti-aging or skin hyperpigmentation, based on Heritage GR22. Claim 7 A food composition for improving skin anti-aging or skin hyperpigmentation according to claim 5, wherein the composition comprises a culture of a strain of the genus Bacillus as an active ingredient, and said culture is obtained by removing the bacterial cells after culturing the strain. Claim 8 A food composition for improving skin anti-aging or skin hyperpigmentation, wherein, in claim 5, the composition comprises an active ingredient in an amount of 0.01 to 99.99 weight%. Claim 9 A pharmaceutical composition for the prevention or treatment of diseases caused by lipofuscin accumulation, and said composition is Bacillus ( Bacillus ) strains of the genus, lysates thereof, cultures thereof, or extracts of said lysates or cultures are included as active ingredients, and said strains are Bacillus ginsengisoly ( Bacillus ginsengisoli ) and Bacillus thuringiensis ( Bacillus thuringiensis A pharmaceutical composition for the prevention or treatment of a disease caused by lipofuscin accumulation, wherein the disease comprises one or more of the following: sarcopenia, progeria, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), brown atrophy of the heart, age-related macular degeneration, neuronal ceroid lipofuscinoses (NCL), acromegaly, denervation atrophy, and chronic obstructive pulmonary disease (COPD), and wherein the prevention or treatment of the disease caused by lipofuscin accumulation is achieved through inhibition of lipofuscin accumulation or removal of lipofuscin. Claim 10 In claim 9, the strain is Bacillus ginsengisolyi Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli A pharmaceutical composition for the prevention or treatment of diseases caused by lipofuscin accumulation, Heritage GR22. Claim 11 A pharmaceutical composition for the prevention or treatment of diseases caused by lipofuscin accumulation, wherein the composition comprises a culture of a strain of the genus Bacillus as an active ingredient, and the culture is obtained by removing the bacterial cells after culturing the strain. Claim 12 A pharmaceutical composition for the prevention or treatment of diseases caused by lipofuscin accumulation, wherein the composition comprises 0.01 to 99.99 weight% of an active ingredient. Claim 13 Bacillus ginsengisoli Heritage GR22 with accession number KCCM12963P ( Bacillus ginsengisoli Heritage GR22) strain. Claim 14 In Clause 13, the strain is a strain having lipofuscin accumulation inhibitory or lipofuscin removal activity.

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