JP6185041B2 - Glycerol production promoter derived from Staphylococcus epidermidis, antibacterial peptide production promoter derived from skin epidermis keratinocytes, and their use as an external preparation for skin protection - Google Patents

Description

  The present invention is based on one or a series of promoting functions of glycerol production from Staphylococcus epidermidis, which is a skin resident bacterium, that is, a useful bacterium, and production of antibacterial peptides derived from skin epidermis keratinocytes. The present invention relates to a drug for protecting the skin from external stress and harmful bacteria and an external preparation for skin protection.

  In human skin, symbiotic microorganisms form a population as a skin resident flora. As skin resident bacteria, Staphylococcus epidermidis, Staphylococcus aureus, Corone bacteria and the like live, and the protective function from the outside of the skin is maintained by the balance of these bacteria. However, due to environmental factors such as rapid temperature changes, dryness, strong cleaning agents and chemicals, or psychosomatic factors such as trauma, mental stress, and poor general condition due to illness, these microbiota balances are disrupted and take It is known that the protection function fails (Non-Patent Document 1).

  For example, Staphylococcus epidermidis, known as a useful fungus for skin, decomposes sebum to produce glycerol, and prevents physical growth such as preventing dry skin and controlling the pH of the skin surface to suppress the propagation of various bacteria. It plays a role of protective function. Therefore, when the ratio of Staphylococcus epidermidis decreases, these protective functions are not exerted and may cause skin diseases. In order to prevent such skin diseases, moisturizers, antibacterial agents, bactericidal components, and the like have been widely used in external preparations for skin. However, some of these drug components also suppress the protective function of aerobic bacteria Staphylococcus epidermidis. There is concern that it will end.

  Staphylococcus epidermidis also stimulates skin keratinocytes, promotes the production of antibacterial peptides from skin keratinocytes, and plays a role in strengthening the biological protective function of the living body. As antibacterial peptides produced from skin epidermal keratinocytes, human β-defensin-2 (hBD2: human β-defensin-2) and human β-defensin-3 (hBD3: human β-defensin-3) are known. These antibacterial peptides have been reported to exhibit antibacterial action against Staphylococcus aureus, which is a harmful bacterium of the skin, and serve as a biological barrier to protect the skin from harmful bacteria (Non-patent Document 2).

  Conventionally, carbohydrates such as gluco-oligosaccharides have been used as an assimilation source for the purpose of promoting the growth of Staphylococcus epidermidis (Patent Document 1), but not only useful Staphylococcus epidermidis but other S. aureus, etc. In some cases, the protective function of the skin was not achieved because of its ability to assimilate against harmful bacteria. In addition, when sugar is applied to the skin, it often gives a sticky feeling, which causes another problem that it is difficult to formulate the feel.

  Staphylococcus aureus is known to be present abundantly in the skin of people with dry skin or rough skin, and patients with atopic dermatitis. It is also known that toxins produced by Staphylococcus aureus cause further deterioration of the skin condition. It is thought that it is important in order to keep the skin surface state healthy to effectively sterilize this Staphylococcus aureus and maintain Staphylococcus epidermidis having a skin protecting function.

  The role of Staphylococcus epidermidis in the skin is similar to that of lactic acid bacteria and bifidobacteria, which are useful bacteria of the intestinal tract. These useful bacteria in the intestine are known to adjust the pH in the intestine, suppress the growth of harmful bacteria such as Clostridium perfringens and Escherichia coli, and have a protective function in the intestine.

  In the intestinal tract, useful bacteria such as lactic acid bacteria and bifidobacteria are ingested to improve the intestinal environment, but in general, these useful bacteria reach the intestine and become settled. Enteric type encapsulation was necessary. In addition, in order to increase useful bacteria in the intestines, a method of ingesting food and drink containing various oligosaccharides selectively assimilated by dietary fiber and useful bacteria has been performed. However, these methods may cause excessive intestinal fermentation or diarrhea depending on the individual constitution. As one method for solving such a problem, a method using a heat-treated dead cell of lactic acid cocci was provided (Patent Document 2).

  The heat-treated dead cells of Enterococcus faecalis can promote the growth of bifidobacteria in the intestinal tract, suppress the growth of harmful bacteria, and improve the intestinal protective function by ingesting them as food and drink. Moreover, since it is a dead microbe, the process as food / beverage products is easy, and since unnecessary fermentation does not advance, there is little influence on the storage stability of food / beverage products.

  As the heat-treated dead cells of lactic acid bacteria, microorganisms classified into Enterococcus faecalis can be preferably exemplified, but Enterococcus faecalis EC-12 strain (accession number FERM BP-10284) is particularly preferable, and Enterococcus faecalis is used. The 16S rDNA of the EC-12 strain (Accession No. FERM BP-10284) is registered as “AB15482” in the National Institute of Genetics.

Japanese Patent Laid-Open No. 2005-002087 JP 2004-051530 A

Gallo, R.D. L. Et al., Journal of Investigative Dermatology 131, P1974-1980 (2011). Kisich, K.M. O. Et al., Journal of Investigative Dermatology 127: 2368-2380 (2007).

  In order to keep the skin surface state healthy, means and methods for protecting the skin from external stress such as dryness and harmful bacteria are indispensable. In the present invention, glycerol from Staphylococcus epidermidis and the production of antibacterial peptides against harmful bacteria such as Staphylococcus aureus derived from skin epidermis keratinocytes, through one or a series of promoting functions, from the outside such as drying It is an object of the present invention to provide an agent for protecting the skin from stress and harmful bacteria and an external preparation for skin protection.

  As a result of investigating various lactic acid bacteria and intensive studies, the present inventors have found that Enterococcus faecalis EC-12, which is a lactic acid bacterium, produces glycerol derived from staphylococcus epidermidis in the heat-treated dead cell. It has been found that there is an action of promoting the production of antibacterial peptides derived from skin epidermal keratinocytes. In addition, when applied to the skin, it is very smooth and excellent in touch and imparts a feeling-improving effect to any type of aqueous, oily, emulsion, and powder external preparations.

That is, the present invention includes the following items.
[1] A glycerol production promoter derived from Staphylococcus epidermidis containing an enterococcus lactic acid bacterium as an active ingredient.
[2] The glycerol production promoter derived from Staphylococcus epidermidis according to item 1, wherein the Lactococcus of the genus Enterococcus is Enterococcus faecalis.
[3] The glycerol production promoter derived from Staphylococcus epidermidis according to Item 1 or 2, wherein the Enterococcus lactic acid cocci is Enterococcus faecalis EC-12 (Accession number FERM BP-10284).
[4] The glycerol production promoter derived from Staphylococcus epidermidis according to items 1 to 3, wherein the Enterococcus lactic acid cocci are dead cells.
[5] The glycerol production promoter derived from Staphylococcus epidermidis according to Items 1 to 4, wherein the Enterococcus lactic acid cocci are heat-treated dead cells.
[6] A skin epidermal keratinocyte-derived antibacterial peptide production promoter containing Enterococcus lactic acid cocci as an active ingredient.
[7] The antibacterial peptide production promoter derived from skin epidermis keratinocytes according to Item 6, wherein the Enterococcus lactic acid cocci is Enterococcus faecalis.
[8] The antibacterial peptide production promoter derived from skin epidermal keratinocytes according to Item 6 to 7, wherein the Enterococcus lactic acid cocci is Enterococcus faecalis EC-12 (Accession number FERM BP-10284).
[9] The antibacterial peptide production promoter derived from skin epidermis keratinocytes according to Item 6 to 8, wherein the Enterococcus lactic acid cocci are dead cells.
[10] The antibacterial peptide production promoter derived from skin epidermal keratinocytes according to Item 6 to 9, wherein the Enterococcus lactic acid cocci are heat-treated dead cells.
[11] The antibacterial peptide is human β-defensin-2 (hBD2: human β-defensin-2) or human β-defensin-3 (hBD3: human β-defensin-3). Antibacterial peptide production promoter.
[12] An external preparation for skin protection comprising the glycerol production promoter derived from Staphylococcus epidermidis according to Items 1 to 11 and the antibacterial peptide production promoter derived from skin keratinocytes.

  The agent or the external preparation for skin protection according to the present invention produces human β-defensin-2 and human β-defensin-3 which are antibacterial peptides against glycerol derived from Staphylococcus epidermidis and Staphylococcus aureus derived from skin keratinocytes. Can be promoted. That is, by promoting the production of glycerol, dry skin can be prevented, and the protective function of the skin such as adjusting the pH of the skin surface to suppress the propagation of germs is promoted. On the other hand, the production of antibacterial peptides against Staphylococcus aureus from skin epidermis keratinocytes can be promoted, and the skin can be protected from harmful bacteria on the skin. Therefore, the health of the skin surface state can be maintained through one or a series of promoting functions. Furthermore, when applied to the skin, it is smooth and extremely excellent in touch, and any type of external preparation such as aqueous, oily, emulsion or powder can give a feeling improvement effect.

Heat-treated dead cells of Enterococcus faecalis EC-12 and aloe vera polysaccharide as a plant polysaccharide are added to the culture solution of Staphylococcus epidermidis standard bacterium (ATCC 12228), and the amount of glycerol produced is quantified. Represents the result. The result of quantifying the amount of glycerol produced by adding heat-treated dead cells of Enterococcus faecalis EC-12 into a culture solution of skin resident bacteria collected from the human eyebrows. The result of having quantified the amount of glycerol extract | collected from the human cheek in the human monitor test using the soft lotion containing the heat-processed dead cell of Enterococcus faecalis EC-12 is represented. A human normal keratinocyte culture is cultured in a medium containing heat-treated dead cells of Enterococcus faecalis EC-12, RNA is extracted from the human normal keratinocyte cell line, and human β-defensin-2 (hBD2 ), The expression level of human β-defensin-3 (hBD3) mRNA was measured by real-time PCR.

  The glycerol production promoter derived from staphylococcus epidermidis, the antibacterial peptide production promoter derived from keratinocytes of the skin, or the external preparation for skin protection of the present invention contains Enterococcus lactic acid cocci as an active ingredient. The enterococcus lactic acid cocci are preferably heat-treated dead cells of Enterococcus faecalis, in particular Enterococcus faecalis EC-12.

The drug or the external preparation for skin protection of the present invention has the following characteristics.
1. Promotes glycerol production from Staphylococcus epidermidis.
2. Furthermore, by using in combination with plant polysaccharides, glycerol production derived from Staphylococcus epidermidis is synergistically promoted.
3. Promotes antimicrobial peptide production against harmful bacteria such as Staphylococcus aureus derived from skin epidermis keratinocytes.
4). Via one or a series of these promoting functions, the function of protecting the skin from external stress such as dryness and harmful bacteria is enhanced.
5. When applied to the skin, it is smooth and very excellent in touch, and any type of external preparation such as aqueous, oily, emulsion, and powder can provide a feeling-improving effect.

  The culture method of Enterococcus lactic acid cocci is not particularly limited, including conventionally known lactic acid bacteria culture methods, but the heat-treated dead cells can be produced by the following method. From the culture obtained by culturing Enterococcus lactic acid bacteria according to a conventional method, the cells are collected by, for example, filtration, centrifugation, etc., washed with water, suspended in water, etc., and heat-treated. Depending on the concentration, it can be adjusted by drying. You may carry out using the spray-drying normally used, a freeze dryer, etc. In some cases, enzyme treatment, surfactant treatment, grinding / pulverization treatment can be performed before and after sterilization treatment by heating or the like, or before and after drying treatment, and those obtained by these treatments are also included in the present invention. Of dead cells or processed products thereof.

  The water used for washing and suspending includes pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, etc., and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Accordingly, the water that can be used as the extraction solvent in the present invention includes purified water, heat-treated water, ion-exchanged water, physiological saline, various buffer solutions, and the like.

  Enterococcus lactic acid cocci obtained as described above are glycerol production promoters derived from Staphylococcus epidermidis, antibacterial peptide production promoters against harmful bacteria such as Staphylococcus aureus derived from skin epidermis keratinocytes, It can be used as an external preparation for skin protection that maintains the health of the skin surface state through a series of promoting functions.

In the glycerol production promoter derived from Staphylococcus epidermidis and the antibacterial peptide production promoter derived from skin epidermis keratinocytes according to the present invention, 5 × 10 ¹¹ or more, particularly preferably 5 × 10 ¹² , of the above-mentioned heat-treated dead cells. It is preferable that the suspension is adjusted so as to contain at least one. Moreover, in the external preparation for skin protection used on a daily basis, it is preferable that the formulation is adjusted so that one third of the same amount is applied per use.

(Form of external preparation for skin protection)
The external preparation for skin protection according to the present invention is suitable for use in ampules, capsules, powders, granules, liquids, gels, bubbles, emulsions, sheets, mists, sprays, etc. 1) pharmaceuticals, 2) quasi drugs 3) topical or systemic skin external preparations (for example, skin lotions, emulsions, creams, ointments, lotions, oils, basic cosmetics such as packs, facial soaps such as solid soap, liquid soap, hand wash, etc. Makeup such as skin cleanser, massage agent, cleansing agent, hair remover, hair remover, shaving treatment, after shave lotion, pre-show lotion, shaving cream, foundation, lipstick, blusher, eye shadow, eyeliner, mascara, etc. Up cosmetics, perfumes, nail enamels, nail enamels, nail enamel removers, poultices, plasters, tapes, 4) Medicinal or / and cosmetic preparations applied to the scalp and hair (eg shampoos, rinses, hair treatments, pre-hair treatments, permanent solutions, Hair dyes, hair styling, hair styling agents, hair growth and hair nourishing agents, poultices, plaster agents, tape agents, sheet agents, aerosol agents, etc.) 5) Bathing agents used in bath water, 6) Others, odor prevention Agents, deodorants, antiperspirants, sanitary products, sanitary cotton, wet tissue and the like.

(Constituents of external preparation for skin protection)
In addition, such external preparations for skin protection can be produced by optionally selecting and using the components and additives exemplified below as needed, and the blending amount in these prescription systems is particularly Although not specified, it is generally considered that 0.0001 to 50% is preferable.

(1) Various fats and oils Avocado oil, almond oil, fennel oil, sesame oil, olive oil, orange oil, orange rafa oil, sesame oil, cocoa butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, cucuin nut oil, safflower oil , Shea fat, liquid shea fat, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, owl, palm Oil, beef tallow, pork tallow, squalene, squalane, pristane or hydrogenated products of these oils (hardened oil, etc.).

(2) Waxes Beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax and the like.

(3) Mineral oils Liquid paraffin, petrolatum, paraffin, ozokelide, ceresin, microcrystalline wax, etc.

(4) Fatty acids Lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, lanolin fatty acid Synthetic fatty acids such as natural fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid and isopentanoic acid.

(5) Alcohols Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, phenoxyethanol, and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol .

(6) Polyhydric alcohols Ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol and the like.

(7) Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyl decyl dimethyloctanoate, cetyl lactate, myristyl lactate, Diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, and the like.

(8) Metal soaps Aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, zinc undecylenate and the like.

(9) gum, sugar or water-soluble polymer compound gum arabic, benzoin rubber, dammar gum, guaiac oil, Irish moss, karaya gum, tragacanth gum, carob gum, quinseed, agar, casein, lactose, fructose, sucrose or ester thereof, Trehalose or its derivatives, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C ₂ -C ₄ ) chitin or chitosan to which alkylene (C ₂ -C ₄ ) oxide such as ethylene oxide is added Low molecular chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or salt thereof, hyaluronic acid or salt thereof, chondroitin sulfate or salt thereof, heparin, ethylcellulose, methylcellulose, cal Polymethyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylate, polyethylene oxide, polypropylene oxide, etc. Alkylene oxide or its crosslinked polymer, carboxyvinyl polymer, polyethyleneimine and the like.

(10) Surfactant Anionic surfactant (alkyl carboxylate, alkyl sulfonate, alkyl sulfate ester salt, alkyl phosphate ester salt), cationic surfactant (alkyl amine salt, alkyl quaternary ammonium salt), amphoteric Surfactant: Carboxylic acid type amphoteric surfactant (amino type, betaine type), sulfate ester type amphoteric surfactant, sulfonic acid type amphoteric surfactant, phosphate ester type amphoteric surfactant, nonionic surfactant ( Ether type nonionic surfactant, ether ester type nonionic surfactant, ester type nonionic surfactant, block polymer type nonionic surfactant, nitrogen-containing type nonionic surfactant), other surfactants ( Natural surfactants, protein hydrolyzate derivatives, polymer surfactants, surfactants containing titanium and silicon, fluorocarbons System surfactant), and the like.

(11) Various vitamins Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), Riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acids, pantothenic acids, biotins, choline, inositols, vitamin C group: vitamin C acid or derivatives thereof, vitamin D group: Ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotaxosterol, vitamin E group: vitamin E or derivatives thereof, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), me Dione (vitamin K3), menadiol (vitamin K4), other essential fatty acids (vitamin F), carnitine, ferulic acid, .gamma.-oryzanol, orotic acid, vitamin P such (rutin, eriocitrin, hesperidin), vitamin U and the like.

(12) Various amino acids Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine , Arginine, ornithine, histidine and the like, and their amino acid derivatives such as sulfate, phosphate, nitrate, citrate or pyrrolidone carboxylic acid.

(13) Various additives derived from plant or animal raw materials These are processed in a conventional manner according to the type and form of the product to be added (for example, grinding, milling, washing, hydrolysis, fermentation, purification) , Compression, extraction, fractionation, filtration, drying, pulverization, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc., arbitrarily selected and combined), arbitrarily selected from various materials You can use it.

  The solvent used for extraction may be selected in consideration of the intended purpose and type of the product to be provided or the processing to be performed later. Usually, water, methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol 1 type selected from various organic solvents such as lower alcohols such as isobutanol or hydrous lower alcohols, polyhydric alcohols such as propylene glycol, 1,3-butylene glycol and glycerin or hydrous polyhydric alcohols, acetone and ethyl acetate Alternatively, it is desirable to use a mixture of two or more. However, when it is not preferable to contain an organic solvent depending on the application, it is sufficient to use only water or ethanol that can be easily removed after extraction, and it may be used alone or in any mixture with water. The thing extracted and extracted may be sufficient.

  In addition, when using additives derived from plant or animal-based ingredients for systemic or topical external preparations and cosmetics, it protects skin and hair, moisturizes, improves touch and texture, imparts flexibility, and stimulates Relaxation, stress relief by fragrance, cell activation (cell aging prevention), suppression of inflammation, improvement of skin and hair quality, prevention of rough skin and its improvement, hair growth, hair growth, hair loss prevention, imparting gloss, cleansing effect, In addition to cosmetic effects such as fatigue relief, blood flow promotion, and warm bath effects, effects such as flavoring, deodorization, thickening, antiseptic, and buffering can also be expected.

  In addition to this, various cosmetic and pharmaceutical effects of each of the raw materials known so far are expected, and by combining these, the intended effect of the present invention can be enhanced, It is also possible to make a product that expects a functional effect.

Specific plants (herbal medicines) as raw materials include, for example, almonds (toen), ai (indigo leaves), aokazura (Keifu Fuji), Aoki (Aoki), Aogiri or Kenashia Ogiri (Ao Tung), Aotsu Samurai (wooden candy) , Aotenma, acacia, red ginseng, red spruce `` fruit '', red clover, red grapes, red ginseng (logwood), red wrinkles (red buds), red crested or red crested or reddish (root), red crested or reddish (ground yellow), Agi (Aki), Aki larch, Akinire (crusted skin), Aguay Guassou, Aguahe (Omiten coconut palm), Akebi (Kitsu), Asa (Asakojin), Maruba morning glory or morning glory (Chicken), Hydrangea (hydrangea), Ashitaba (Tomorrow), Azuki (Red Azuki), Asunaro, Acerola, Asenyaku (Asenyaku), Athira (Shokuyokanna), Anise, Abiyu, A Biurana, oilseed rape, Abemaki `` fruit '', avocado, ama, achacha (Amacha), Achacharu, Amadokoro (Jade bamboo), Amana (a light-hearted), amaranth (Hyuyu, Hagatetou, Himogeito, Senninkoku, Sugimorigateto, Hosoaogeito, Hasoagoito Aozzuran, Amaranthus Hippochondriax), Amigasayuri or Baimo (shellfish mother), Aristolomeria (Yurizuisen),
Arcana (Arcanna), Argarobo (Cabbage), Altea, Arnica, Alpinia or Katsumadai (Suzukuku), Aloe (Amber), Aloe Vera, Angelica, Apricot or Honanzu (Anzujin), Ansokkou (benzoic incense), Yellow support, Igakozorina (Gold) Head), Ikarisou or Yachima Taikoshi (Yinokukaku), Rabbits (Ginseng Grass), Japanese Knotweed (Tiger root), Yew (1st place), Strawberry, Fig (No flower `` fruit, leaf ''), Ichihatsu (first), Ichibibi (winter) Yuzu), Ichiyakusou (one herb), Ginkgo (Ginkgo `` Seeds, Leaves ''), Itohimehagi (Toshishi), Carob, Taizuna (Talexiki), Inubiyu (Honagainubiyu), Rice `` Seeds, Seed coat '', Inondo `` Seeds '', Ibuki Muskweed, nettle, ylang-ylang, iwa tobacco (Iwabuchi), iwahiba or iwamatsu (wind mushroom), common bean, oolong , Fennel (Wolf), Wheat (Chibusanoki), Ukiyagara (Triple), Uguisukagura `` Fruit '', Himewyogaku, Turmeric (Uranium), Duckweed (Floating), Usbasaishin or Keirinsaishin or Oshusaishin (spicy) , Usbunia mushrooms, Usbunichai mallow, phalaenopsis, crabs (summer hay), Udo or Shishido (Clamming live, single lived, Tang Dynasty), Unyadegato (Unkalia, Cats claw), Uba tea, Ume (Plum plum `` seed, pulp ''), Vulgaris, Rumors (Circulatum), Eel mandarin (Chen), Eunbok, Estragon, Ezoukogi (Saigoka), Suzuran, Echinashi (Eurasian pteris), Enishitake (Ezotake), Ebisugusa or Cassia Tora (Keriko), El Campre Berry "fruit", Elemi, Elingi or Prerotor Suerinji, Enju (Spirit, Satsuma Rice), Pea, Ogi or Kibanaogi (Yellow), Narco Yuri or Kagura-mabanaru Yuri (Yosei), Ohisano Yuki or Sasanoyuki, Ouren (Yo Ren), Ogata Hakken, Okarasuuri (Carocon), Ooguruma (Civil engineering fragrance), Otsu ヅ rafuji (Preventive self), Psyllium (car front child, car front grass), Greater thistle (Luozhou leakage, Bald eagle),
Toucan (palm), toucan cactus, toucan or kerla (white birch), tossed beetle (banaba), tossed fruit (fruit), tossed grass (cereal essence), toad catechu, toad or hawthorn (toilet) Barley), Oka, Okazeri (snake bed), Okra `` fruit '', Oguruma (overturn), Oguri higotai (Zhengzhou leakage, indigo head), Panax ginseng or Tochibaninjin (Ginseng), Hypericum pertussis or Pterotus pertussis or Hypericum perforatum (Brother cut grass), Odorikosou (continuous), Onamomi (Circle ear), Onigomi, Onikoro or Tokoro or Nagadokoro (Hikai), Oninoyara (Tempa), Onyuri or Sasayuri or Hakatayuri (Yuri), Onoelan, Ononis, Halibut楡), Ominae (defeated soy sauce), Yuuko, Dutch mustard (Creso) ), Dutch jellies, Dutch ville, Dutch honey bees, olives `` fruits, seeds, leaves '', oregano, orange `` fruits, fruit peels '', silkworms (ripening yellow), cacao `` fruits, peels, seeds '', oysters (柿蒂 `` leaves '' ``), Kagikazura (Katsufuji Rin), Kakidooshi or Kantorisou (Lotus), Kakiran, Gaku Hydrangea, Cassia, Casinoki (fruit berries), Gadju (莪 朮), Kashiwa (Shinoki, Kashiwa), Cascarilla , Cascara grada, gypsophila, gypsophila, cynomolgus (Kinshato), kaniwa, carnation, valerian (valerian), hippopotamus, birch or fern birch (white birch), gerbera, pumpkin, kapoknoki `` seed '', kahoku sansho (potato), Catfish (carp yellow), chamomile or roman chamomile, chamomile (weed), cam cam (camo duck), color, chrysanthemum, crow cucumber or cinnacarus Woori (Wang Yuan), Osh bishak (half-summer), Oats, Garana `` Seeds '', Karaho (Koh-Pak), Karaya, Garyu, Karin (Kiso), Garcinia, Kawamidori, Kawara-Psycho (Koryocho, Shirakaba), Kawajisha , Kawaratake, Kawaranadeshiko (stone bamboo) or Ezokawara Nadeshiko (buckwheat, buckwheat), Kawaraninjin (blue potato), kawara mugwort (bamboo chin), kansui (licorice), licorice (licorice), cantalara, candelilla, kanto, canna, Orchid, raspberry (Ezo strawberry, Dutch strawberry, shrimp strawberry, nava white strawberry, maple strawberry, European raspberry), kiwi `` fruit, leaf '', kikarasuuri (guro-root), kyo-kyo (kikyo, bellflower root), chrysanthemum (chrysanthemum,
(Shimakangi, Kakusen nogi), Kikutaniku, Kisame (Cruciferous), Gigiri (Sheep's Root), Pheasant (Cultivation), Kizuta, Kidachi Aloe, Kidachihakka, Kina, Kinanoki (Shinkona, Akakinoki), Quinua (Quinoa), Yellowfin Kimameme, Gymnema sylvestre, Kimenkaku, cabbage, cabbage `` Immature fruit '', caraway, cucumber, gyoryu (Nishikawa willow, Taiyanagi), Kiraja saponaria, kiraya, Kiranso (golden acne grass), kumquat `` fruit '', kinkiju, Ginsekai, Kimbusen, Kimma, Kimihiki (Sentsukusa), Kinran, Ginlan, Kinryohen, Guapeba Vermela, Guava "fruit", Guayule, Quercus infectria (garlic), kukui nuts, kugenumaran, wolfberry , Earth bone skin) (Winter), Kudzu (Kuzukone), Kusunoki, Gooseberry `` Fruit '', Ginseng (Yellow Flower), Cutty River, Gardenia (Mountain), Kunugi (Mouse Saw), Kupuas, Kubeba, Kumazasa, Kumatsutsura (Horse Grass), Clara (bitter), cranberry `` fruit '', chestnut `` seed, fruit, astringent skin '', curcrigo latifolia `` fruit '', grapefruit `` fruit and leaf '', buckthorn, black moth (rescue), kamebahikokoshi or kurobanahiokikoshi or hikikoshi (life extension) Grass), cloves (chome, clove), gumbainazuna (cough, cough), kei-gai (butterfly, 荊芥 ho), celosia (fowl crown, chicken clover), gekkavidin, bay bay (laurel wreath), Kenashi Sartor Iibara (dawn, mountain return) , Gentian,
Gennoshouko (Oldweed Grass), Kemponashi (Kugushi), Kokisekikoku, Kishumikan (Koji, Tachibana, Obenimikan, Fukurekan, Sagamikouji, Ponkan, Santara (Tachibana)), Kousinbara (Monthly Flower), Koiusihakka, Kozo '', Kocha (tea), Kouhone (river bone), Kohon (Tsubakimoto, Karamotomoto), Koryang, Koryokukyo (Kora Kyo), Cilantro "fruit", Kouren (Huo Ren), Koganebana (Yellow Gon), Cowberry (Etsutachibana) ), Coconut `` fruit '', goshuyu (gow), gosho strawberry (covered bonsai), pepper (pepper), cosmos, copaiba balsam, coffee `` seed, leaves '', kobushi or magnolia (hot pepper), burdock (beef bowl, beef coconut) , Coconut lees, sesame (sesame seeds), sesame seeds (genus ginseng), rubbish (gomicho), saneka or binankazura or pine bush, wheat (wheat), rice or rice Rice bran `` Red cocoon, white birch '', rice oil, cola acminata `` seed '', cola bella `` seed '', colony llya, fenugreek `` fruit '', colombo, kondurango, kombu, konjac, comfrey (sardine grass), sisal ( Sisal Asa), Sai Hairan, Sakaneran (Ezosa Kanelan), Sasima button eel or cinnabar eel or cinnamon (wei Sensen), Sakura (Oshima cherry, Yamazakura, Oyamazakura, Edhigashi, Bean cherry, Miyazakura, Yoshizakura, Takazakura, Takanezakura, Takanezakura, Takane cherry `` Leaf, flowers, fruits, bark (cherry bark) '', cherries, pomegranates, Sasa, Sababaginran, Sasanka, Sagimodaka (Sawamata), sweet potatoes, sugar cane, sugar beet, saffron jujube (Sanbanjin, Safflower) ), Sapodilla, Potea mariyo, pomelo `` fruit '', savonso, salmon berry `` fruit '', salamander ginger (sand hemp), salamander (sand indigo sashimi), salvia (sage), scallops (bamboo rape), sawagurumi (mountain walnuts), sankaku cactus, Sangre de Grado (croton), Sansikuyoso, Sanshichininjin (Ginseng Ginseng), Sanshuyu (Yamamuro), Sansho (Yamamuro), Sandzukon (Yamadone), Shia (Kalite), Cyanoki "fruit", Shiitake ( Shiitake mushrooms), Zion (purple mushrooms), winged bean,
Dikitaris, Sikkunshi (Princess), Perilla or Blue-tailed or Chimmento or Catamenzo (Shiyo leaf, Shiso-ko), Rosewood, Shinanoki, Shinohonoki, Shirarenyou (ream), Shimeji Shirotamogitake, Shimoklen (hot pepper), Shimotsuke, potato, shajin (shasan), jasmine (matsurika), jade beard (barley winter), jujube sage, shukukonka miso, shukushamitsu (sand jelly, shredded sand), juzudama , Shurro `` fruit '', Shunran, Zououyashi, ginger (ginger), Zouzan hydrangea (Tsuneyama), ginger (sardine, root), shozu `` fruit '', shirakashi `` seed '', silane (bikkyu), lantern butterfly `` seed '', White clover (clover), white clover (chicken bone grass) , White squirrels (smoke squirrels), Shirobanatsuta "flowers", Shirobana Lupine, Shirominanten (southern Tenmi), Cinconacylbra, Ginkgo biloba (Mizuka, Suzuka, Senncho), Cinnamon, Cinnamomum cassia (Kaishin), Suika , Honeysuckle (gold and silver flowers, Shinobu winter), sorrel (acid model), swim berry `` fruit '', star apple, stevia, strawberry `` fruit '', succulent (Xu Longan), sugina (question), purslane (horse tooth soap, horse tooth soap Child), plum `` fruit '', sulgaran (oran), Japanese red pine `` cone '', Japanese larch, Japanese cypress, Japanese walnut, Japanese hawthorn, Japanese dandelion, Japanese horse chestnut (Maroniae), Japanese pear `` fruit '', Japanese cherry tree, Elder, American nezu (Juniper, Juniper), Seyo Yarrow (milfoil), rose, pearl butterfly, mistletoe, mint moth or wild mint (Secret, Kibanano Sekkok) (Dendrobium, sarcophagus),
Cedron (Boshushuboku), mallow, Hiroha Senega, Senega, Seri, Celery, Senkyu (river cucumber), Senshinren (Pennial ream), Sendan, Sennin cactus, Senna `` fruit, leaf '', Sennin (Otsuka), Senbu ), Senboku, soka (grass), elephant chew, soshinwabai, buckwheat `` seed '', some potato, broad bean, daiou (daihuang), radish, soybean, daidai (orange peel, fruit), takasagorihigotai (southeast indigo sashimi), Takasaburo (Cirrus lotus), Takatodai (Daikan), Takarabi (Spider spine), Dark sweet cherry `` fruit '', hollyhock, ginger perilla (thyme, perilla), Tachidokoro (hikai), Tachiyanagi, Tamarind `` seed '', onion, Tamshiba (powder), taranoki (fruit, leaves, root bark), tarayou (large leaf winter blue, one leaf tea, bitter tea, bitter tea), dahlia (ten Kubotan), Tarwi, Tankimi Bean, Tanshichiwa, Tanjin (Tanjin), Dandelion (Kangei) or Shirobana Dandelion or Mouko Dandelion, Danmaru, Cherry `` Fruit '', Tigaya `` Fruit, Root, Bud '', Chikutsutsuninjin (Bamboo Ginseng), Chicory , Chancapiedra, Tulip, Chocolate-no-stay, Korean thistle (Artichoke), Korean dragonfly (Dahi), Korean senile (Amber), Korean mugwort (Bamboo leaf), Choreimaitake (Amber), Chorogi, Apricot , Tsukushi Kaneran, Tsukuritake (mushroom), Tsuchitakebi, camellia, camellia, tsumekusa (lacquered weed), Tsuyukusa (duck eel), Tsuruazuki (red bean), Tsurutsuchiakebi, Tsurudokudami (what neck rice cake), Tsuruna (Apricot apricot), Tsurunjin (four leaves) ), Tsurume, Tsuwabuki, Deiko, Teuchiguru , Dubery `` fruit '', tengusa, tencha (bamboo tea), tendaiyaku (glaze), red pepper (winter coconut), red pepper, red pepper (crown), red snapper (marigold), red snapper (marigold), red pepper, tounabena (River cut), corn or corn hair (southern cocoon), red potato mochi (woman Sadako), turkey or chorus lindow (dragon gall), toksa (wood bandits), dokudami (ten medicines), tokoshi (gush roots), toshishi or mamedaoshi Or prunus, tomato (bark, “bark, leaf, root”), ash (bark), tomato, tragacanth, triacanthus “seed”, turkey, tormentilla, dronoki, troloaoi, tongamame, nigerian berry “fruit”, Chinese yam or yam (Yakuyaku), Nagi Ikada (Butcher Bloom), Naginata Koju, Nazuna, Rapeseed, Natsume Tatehaki (sword beans), Natsumikan, Natsume (Otsuka), Naniwaibara (Golden Cherry), Navena (Continued), Nameko, Peanut (Peanut), Nanten (Nami Tenami), Namba Karamushi (Hamama), Niosummille, Nigaki ( (Nakigi), Nigamugi (Anguish),
Nikuzuku, Kay or Nikkei or Ceylon Nikkei or Yab Nikkei (Cinnamon) or Keishi (Katsura), Flying Plover, Leek (Chicken), Elderberry (bone, `` fruit, flower, stem, leaf ''), garlic (Otsuka), Nurde ( Quintuplet), leek, nemno or nebu or nebi or nemlinoki or zigokubana (community), nozami (big eagle), nobara (fruit), nosen kazura (Lingwei flower), nogate tou, yarrow, nodake (maehu), nobara, noomo, pineapple `` Fruit '', hibiscus (Busouge, Furiinbusouge, Roselle), Hailindo, Hauchiwa, Pau Dorse, Hakama Labosi (bone fracture), Hakukayumatou, Hakusanchidori, Hakusen (white tinea), Hakulberry `` fruit '', Hakobe (breeding), Hazel (lion), basin (roth root), basil, lotus (lotus, lotus, lotus), parsley (holland jelly) Hadakamugi, Batata, Hachiku or Madake (bamboo shoots), patchouli, mint (thin load, light load leaf), pepper beans, pearl barley (yokuinin), hanasuge (wisdom mother), banana, hana mint, hanabishi (shitsurishi, shishitsuri), vanilla bins, papaya, Papaver, Pahalobobo, hub `` whole plant, stem, leaf '', paprika, scallop, clam or honey beetle (Vulgaris), Hamasge (Katsutsuki), Hamabishi (Shitsuko), Hermanus (Maika flower) , Hamaboufu (beach windbreak), Hamelis, Palm, Rose (rose), Balata, Balata rubber tree, Agaric (Agaric, White-faced agaric, Usukimorinokasa),
Haran, Pariurus (Parus edulis), Parietalia, Harkanran, Balsamina (Turureishi, Nigauri), Harnile (Crustacea, Birch skin, Bamboo leaf), Barbasco, Harlindow, Handaikai (Popular Ocean), Panoki, Hiogi (Gardenfish), Chicknut carrot (participant), pecan nuts, ganbanana (stone wall, vine pearl flower), Higurmadaria, Higotai, Hishi (Hishimi), pistachio, beet, Hitotsuba (stone wall), Hitsubei Nishida, daisies (Daisy), Hinata Wild boar, cypress, cypress, cypress, hiba, castor, sunflower, green pepper, himeuzu (tengu), himematake (Karimaritake, hiromatsutake), hime-myo-ran, pimenta `` fruit '', beak, cypress `` Fruits '', chickpeas, lentils, bellows oysters, Hirohakinakusa (white翁), loquat "fruits, leaves, stems", betel wax (large clothes skin, coconut), puer tea (common ear tea), Japanese quail (southern rattan), Japanese pufferfish, fukidanpopo (sect winter flower, winter winter leaf), Fujibakama (Orchid grass) ), Fuji bean (Tomodo), Fujimodoki (Chrysanthemum cherry, spikelet), Grapes `` Fruits, pericarp, seeds, leaves '', Beech, Fuyumushinatsusatake (Cycium cordyceps), Brazilian licorice, Brazilian carrot, Brazilian palm, Brazilian elephant palm, Black currant `` fruit '', blackberry, branyen, plum `` fruit '', full ceria, blueberry (Prunus japonica), prunes, floral blanka, blond psyllium, bundo (mung bean), hazelnut, loofah, hetsukaran, safflower (safflower), Henken, Belladonna, Berry "fruit", Persair, Pervianburg, Pereschia grandif Rear, bean diatom or nettle (scenic), boysenberry `` fruit '', broom or sardine or nendo or hahagigi kokia (silk), spinach, spinach (daffodil, acute pupa, pelagic grass), pouteria sapota, pouteria・ Luma, Hono, Bofu (windproof), spinach, physalis (Torune), honoki (Japanese welfare, magnolia), bokeh (wooden moss), Hosoba Aragiku (Kayoyoku), Hosobaoguruma, Hosobanaokera (蒼朮), Hosowanakirinso ( Kageten 37), Hosobabarengiku, Bodaiju (bamboo tree), buttons (peony `` flowers, leaves, stems, bark ''), button pi (peony skin), hops, jojoba, borage (Lulichisha), Bordeaux, white berry `` fruit ”, Hononiku (meat jugo, Taigei), Honsekkok (iron crust stone, sparrow mountain stone agate), Mizul tennan show (Tennan star), Maitake (maiko), Maou (hemp) Yellow), maca, macadamia nuts, margaret (mokushungiku), makuri (marine grass), mugwa (mulberry white bark `` bark, leaf ''), magnolia sprengeri, mazaran uba, masalanzuba de ceara, masheira de boi, Mashua (Tamanowenhalen, Kyukonkinrenka), Matatabi (wood tengu), Matsukasa, pine `` leaves, bark, root '', Matta Orillo, Matsuhodo (茯苓), Matico (Cordon sillo), Mayoram (Hanahakka), Malvano jardin (Bitter), Mulberry `` Fruit '', Quince, Malene (Billow Muzu squid), Mango, Mangosteen, Manzanya (Andesca chamomile),
Manshugurumi, Mandarin `` fruit '', Mannentake (reishi), pheasant (fruit berry `` fruit), Mishima Saiko (Shibako), Mizuohobako or Mizuasagao (Dragon tongue grass), Mizokakushi (half-sided lotus), Misohagi , Mitsagi willow or elder willow (Hongshu), Mitsuwashi, Mitsuba, Midori Zapote, Midori Hakka, Mimosa, Miyoga, Miracle fruit `` fruit '', Myrrh, Milobaran, Mugiwaragiku, Mukuge (Kiso), Mukunoki, Mukuroji (Life extension skin), Munya (Purple root), purple saplings or murasakikibu (purple pearls), purple corn, purple samurai fuji (Kunming chicken blood wisteria), megakansa opuntica, red-bellied herb, mebougi, melaroika, melissa, merilot, melon `` fruit '', mouko mugi , Mokkirin, moje (pepper), ilex, mok Kou (wood incense), Japanese maple peach, peach (peach `` leaves, seeds, flowers, fruit ''), coconut palm, Morero cherry `` fruit '', Morohaya (burlap), Yakawa Murasakiimo, Yakuchi (Michichi), cornflower ( Cornflower), cornflower mint, yacon, yashabushi (arrow wheel) or Himeyashabushi or Obayayashabushi `` fruit, pericarp, fruit ear '', willow willow, starfish (octagonal gold plate), mistletoe (willow parasitism), willow (kawayanagi, tachiyanagi, weeping willow, red willow, Pussy willow, Incolyana, Kinuyanagi, Koryanagi, Eunryu willow, Miyama willow, palm, willow, Obayanagi, Tairikuki willow, Fox willow, Doronoki, Yanagidae `` leaves, stems '', cigarettes, gold beetle Spirit), pokeweed (commercial land), Japanese alder (Yamatake), Japanese yam (Plum) ),
Porcupine, Eucalyptus, Yukinoshita (Tiger ear grass), Yucca or Flavifolia, Yuzu `` fruit '', Lily, Yorigusa, Mugwort (Leaf), Ligan (Raimaru), Lime `` Fruit '', Rye, Lacanca `` Fruit '', Raspberry `` Leaf '' , Fruit '', latania (chlamelia), lacquer or shallot (palm white), lavender, longan (long-eyed meat), agave (Aonu agave, fuchlin agave), ryocha (green tea), apple `` fruit, seeds, leaves, root '', gentian, Lubus or Suabisimus (Suzuryu), Ruritama Thistle (Vulgaria chintai, Xinjiang indigo sashimi), Reishi (Toothpick, toothpick core), Lettuce (Chisha), Red currant `` Fruit '', Red Pitaya, Lemon `` Fruit '', Lemongrass, Forsythia or Chinese Forsythia (Region), Forsythia, Wintersweet, Wacoal, Lokon Ashine), loganberry "fruit", rosemary (Rosmarinus officinalis), rose hip (Novara), wasabi, Watafujiutsugi (dense Komuhana), burnet (Chinire), and the like.

  As seaweeds, seaweed [Green algae: Chlorella vulgaris, Chlorella pyrenoidosa, Chlorella ellipsoidia, Aonori (Usba aonori, Sugiaonori, Hiraaoori, Boaonori, Hosoeda Aonori), Anaaoosa (Aosa)] (Macombu, Resilibu, Hosocome, Mitsuishi Kombu), Wakame, Hirome, Aowakame, Giant Kelp (Macrocystis Integrifolia, Neocitus Ruetkeana), Hijiki, Hibamata, Umiuchiwa, Usubaumiuchiwa Umiuchiwa, Akaba Umiuchiwa, Konami Uchiwa, Okinawa Uchiwa, Usuki Uchiwa, Etsumi Uchiwa], seaweed [Red algae: Hijirimen, Maca (Tengusa), Hiraksa, Syria, Oaxa Caterpillar, Yatabegusa, Yuukiri, Shimatengusa, Tosakanori, Togekirinsai, Amakusakirinsai, Giraffe, Juniper ginkgo rhinoceros, Tsunomata, Totsuka (Yahazutsu notamata), Ezotsunotamata, Togetu no Takita , Yaleus banori, Kagius banori, Susius banori, Haius banori, Akamo mizunori] and the like.

  In addition, other algae such as green algae (Chlamydomonas: Chlamydomonas, Akayukimo, Dunaliella: Dunaliella, Chlorococcus: Chlorococcus, Kuwanomimo: Kuwanomimo, Volboxa: Ougemawari, Volbox, Palmera, Yotsumeo, Aomido: Chrysanthemum, Aoumidro, Tirguimidro, Hibidoro: Hibidoro, Aosa: Anaaaosa, Amiaosa, Nagaaosa, Kawanori: Kawanori, Friccella, Shiogusa Magama genus: Magamata, Iwagata: Fusaikawa, Surichogata, Heraiwata, Kurokita, Hanemo, Genus: Mill, Chromel, Sakibutimil, Nagamil, Hira Lemon, Casanori genus: Casanori, Jusmo genus: Jujusmo, Tamajusmo, Mizojuzumo, Mika ヅ kimo, Korekaete, Tsutsumimo genus: Kyukougusa, Hitogususa genus: Humanegusa, Hirohatohigususa, Umushihimoe, Sakikimoe Sumiremo: Sumiremo, Hoshimamidro, Fushinashimidoro etc.).

  Cyanobacteria (Suizenjinori: Suizinori, Aoko, Nenju: Kawatake, Ishijelly, Hassai, Yuremo, Rasenmo (Spirulina): Spirulina, Trichodesmium (Iaciasio), etc.

  Brown alga Sanadagusa: Sanadagusa, Fukurimamiji, Commongusa: Commongusa, Yahazugusa: Ezoyahaz, Yahazugusa, Uraboshiyahaz, Jigamusa: Jigmygusa, Umichiwagusa, Umamisakugakua, Hakusou : Navarimo, Chasomen genus: Motsuki Chasomen, Matsumo genus: Matsumo, Nagamatsumo genus: Nagamatsumo, Okinawa mozuku: Okinawa mozuku, fake Tomozuku: Niseft Mozuku, Futomozuku: Futomozuku, Ishimozuku: Ishimozuku, Kuromo: Kuromo, Nisemozuku: Nisemozuku, Mozuku: Mozuku, Ishige: Ishige, Iroro, Ichimegasa: Ichimegasa Umibosu, Urushigusu: Urushigusa, Keurushigusa, Tobaccogusa, Composita: Composita, Haberodomo: Haberodoki, Havanori: Habanori, Habanori: Habanori, Commonbukuro, Ezobukuro, Ezobukuro Chishima fukuronori genus: Chishima fukuronori, Kagomenori genus: Kagomenori, Muraridori genus: Murachidori, shark genus: shark zusa, genus genus: lobster, genus genus: Yokojimari, Ganoderma genus: cynomolgus, fennel spider: fennel spider, vine genus: turmo, turtle genus: sea turtle, scorpion genus: sujime, misji kumbu genus: misji kumbu, atsuba misji kumbu, kumbu genus: tsutsugara kumbu, oyster kumbu, onikombu, gakonbubu Barracuda, Trollocombu: Trollocombu, Antokume: Antokume, Kajime: Kajime, Tsuruarame, Kurome, Kikuishikombu: Kikuishicombu, Nejikcombu: Nejicombu, Kuroshiome: Kuroshiome, Nekoshikonbu, Aramei Genus: Ainuwakame, Chigaiso, Oniwakame Ezoi Shige: Ezoi Shige, Yabane Moku: Yabane Moku, Rappa Moku: Rappa Moku, Joromoku: Ugano Moku, Jo Lomoku, Hiemoku, Honda Walla: Tamanashimoku, Isomoku, Nagashimamoku, Akamoku, Shidamoku, Honda Walla, Nejimoku, Narasamo, Mametawala, Tatsukuri, Yatsumata Moku, Umitora no Moku, Fushishimoku, Mohmokumoku, Hahamokumoku , Ukimo: Ouokimo, Burukimo: Burukimo, Kayamonori: Kayamonori, etc.).

  Red algae Genus: Sea urchins, Tsukumonori, Anemone crickets, Astragalus, Amaranthus, Amaranthus, Agaricata Genus: Tamaitadaki, Kaikekenori Genus: Kaginori, Kaikekenori, Tengusa: Himetengususa, Hytengususa, Oobusa, Nambugu , Kohira, Yohira, Kinukusa, Hibirodo: Hibirodo, Himebiro, Isomumedo: Isommedo, Aedes: Rhizome, Ryumons: Ryumon, Ranunculaceae, Nisekare Naminohana: Hosobanaminohana, Naminohana, Sangomodoki: Ragaramomodoki, Shiogusagoromo: Shiogusagoromo, Iwanokawa: Etsukiiwanawa, Kinokawa: Kinokawa, Kanonote, Sangori Kyono Shiro, Sakuranori, Nikumukade, Tambanori, Tsurutsuru, Fudarak, Marubahdarak, Isonohana: Nurakusa, Ronulaksa, Oomkadenori, Hirakintoki: Hirakintoki, Matabou: Matabou, Kintoki: Chabokintoki, Kintoki, Matsunori, Rice, Tokamatsu, Hitomatsu, Fushikintoki, Tsunomude, Nagakinito, Crayfish Nagaobane: Naganobane, Funori genus: Hanafonori, Fukurofunori, Mafunori, Karekigusa: Karekigusa, Tosakamodoki: Hosobanatosakamodoki, Hironotosakamodoki, Yatsudegatatosakamodoki, Kurotosakamodokisane, Zotosofomoza Tosakamodoki, Nankaitosakamodoki, Himetosakamodoki, Kinuhada: Kinuhada, Ezotosaka: Ezotosaka, Tsukasanori: Enashikarimenia, Otsukasanori, Hana Tacarimenia, Hoonoo: Honoo, Hikagenoit: Hikagenoit, Usuginu, Nikuhounoo: Nikuhounoo, Venus nago, Venus nago, Susca veni: Suscabeni, Okarugusa: Yamadagusa, Mirin: Mirin, Hosokamiso , Isomocca genus: Isomocca, Yukari genus: Yukari, Hosoyukari, Ibaranori genus: Ibaranori, Saidaibara, Tachiibara, Kagibaranori, Kojinoo: Kizino-o, Isodantsu: Isodantsu, Atsubanori: Atsubanori, Ogonori, Ogonori, Ogori Mizo-okonori, birch, beniogonori, fukureshinori, purple birch, shinkaibanori, spruce birch, kataogonori, ryukyuogono Li, Atlantic scorpion, Ivy genus, Yumigataonori, Kubireogonori, Mosagogonori, Kinubanori, Amanita genus: Hachijotensamodoki, Fushikenori genus: Fushikenori, Namiiwatake, Nymphalanthus Itagususa, Saimi, Hari, Lotus Jigusa: Lotus Jigusa, Sugi nori: Ikanoashi, Hosoibonori, Nobonori, Black Haginansou Genus: gourd, cherries, octopus: octopus, sedge, genus: genus, genus: goat, dals : Dulse, Scorpionus, Anadalus, Weibusa: Uebusca, Benifronori: Benifuronori, Fusunagi: Futsutsunagi, Himefusunagi, Hirohafushitsugi, Otsunagisou, Hirawatagigi, Usuwasugi , Egonori: Egonori, Futogigi, Saeda: Saeda, Chirimomiji: Chirimomiji, Konohori family: Habutaenori, Konohori, Suzuronori, Usubeni: Genus: Kagios banori, Yareus banori, Susius banori, Heius banori, Usubanorimodoki: Usubanorimodoki, Ayanishiki genus: Ayani Genus: Aeginu, Dadia: Enasidadia, Shimadadia: Isohagi, Shimadadia, Dajimodoki: Dajimodoki, Euglena: Morayogusa, Fuyugusa, Makuri: Makri, Yanaginogi, Yuna, Yukinagina , Sasabayanaginori, Sozo genus: Kurosozo, Kobsozo, Hanesozo, Sozonohana, Hanegusa genus: Hananegusa, Hananegusa, Kozanemo genus: Kozanemo, Isomurasaki, Hosozozanemo, Himegoshigusu, Giant genus Isomagusa, Suginasugusa: Suginasusa, Isobashou: Isobasho, Fujimatsumo: Fujimatsumo, Sawweed: Haeshinosakiba, Kawamoku : Kawamoku, Aokawa Mozuku, Himekawa Mozuku, Ideyugogome: Ideyugogome, Okichimozuku: Okimozu, Otosa, Chinomo: Chinomo, Chisujinori, Chisujinori, etc.).

  Axial algae (Shajikumo, Shiratamamo, Hoshitsurimo: Hoshitsurimo, Rikunotumusu, Flaskmo: Himebumo, Chaboflasmo, Tripera, etc.), yellow algae (Hikarimo: Hikarimo, etc.), etc.

  In addition, the materials derived from animal materials include chicken crown extract, cow or pig, human placenta extract, cow or pig stomach, duodenum, intestine, spleen extract or decomposition product thereof, cow or pig brain tissue Extracts, cattle / pig or fish collagen hydrolyzate (acid, alkali, enzyme etc.) degradation products, water-soluble collagen or collagen derivatives such as acylated collagen, cow or pig elastin or elastin hydrolyzate (acid, alkali, Enzymes etc.) or water-soluble elastin derivatives, keratin and its degradation products or their derivatives, silk proteins and their degradation products or their derivatives, porcine or bovine blood cell protein degradation products (globin peptides), cattle or porcine hemoglobin degradation products (Hemin, hematin, heme, protoheme, heme iron, etc.), milk, casein and its degradation products or their derivatives, skim milk powder and their degradation products or their inducements Body, lactoferrin or decomposed product thereof, egg ingredients, fish hydrolyzate, nucleic acid-related substance (ribonucleic acid, deoxyribonucleic acid), or the like.

  In addition, as a material derived from a plant or animal material, any site, cell, tissue, organ, metabolite, etc. derived from a gene recombinant or a cell fusion can be used. Furthermore, for example, an undifferentiated cell group, a cell group in the middle of differentiation, or the like obtained by culturing an arbitrary site, cell, tissue, organ or the like can also be used.

(14) Marine components Seawater such as deep seawater, for example, sea salt, dried seawater, inorganic salt (sodium chloride, magnesium chloride, potassium chloride, etc.), sea mud or mud (from the Dead Sea or Atlantic Ocean) Fango), for example, Italian mud, German fungo, Eifelfango, Freiburg fungo, etc. Sea mud or mud from various locations (containing components: silicon dioxide, titanium dioxide, aluminum oxide, iron oxide, manganese oxide, sodium oxide, potassium oxide, Magnesium oxide, calcium oxide, strontium oxide, sodium, potassium, magnesium, calcium, chromium, iron, copper, nickel, zinc, lead, manganese, arsenic, water), saintoku stone, etc.

(15) Microbial culture metabolite Yeast metabolite, yeast extract, bacterial metabolite, bacterial extract, mold or actinomycete metabolite, mold or actinomycete extract, natto metabolite, natto extract, rice fermentation extract , Fermented rice bran (red rice bran, white birch) extract, Euglena extract or decomposition product thereof or water-soluble derivative thereof, trehalose or derivative thereof, lactic acid fermentation product of raw milk or skim milk powder, fermented lactic acid bacteria of legumes, coconut plant Lactic acid bacteria fermented products.

(16) α-hydroxy acids Glycolic acid, citric acid, malic acid, tartaric acid, lactic acid and the like.

(17) Inorganic pigments Silicic anhydride, magnesium silicate, talc, kaolin, bentonite, mica, titanium mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, Bengala, black iron oxide, gunjo, chromium oxide, chromium hydroxide, carbon black, calamine, etc.

(18) UV absorber / blocker benzophenone derivatives (2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid sodium salt, dihydroxy Dimethoxybenzophenone, dihydroxydimethoxybenzophenone-sodium sulfonate, 2,4-dihydroxybenzophenone, tetrahydroxybenzophenone, etc.), paraaminobenzoic acid derivatives (paraaminobenzoic acid, ethyl paraaminobenzoate, glyceryl paraaminobenzoate, amyl paradimethylaminobenzoate, Octyl paradimethylaminobenzoate), methoxycinnamate derivatives (ethyl paramethoxycinnamate, isopropyl paramethoxycinnamate, octyl paramethoxycinnamate) , 2-methoxyethyl paramethoxycinnamate, sodium paramethoxycinnamate, potassium paramethoxycinnamate, diparamethoxycinnamate mono-2-ethylhexanoate glyceryl, etc.), salicylic acid derivatives (octyl salicylate, phenyl salicylate, homomenthyl salicylate, dipropylene glycol salicylate) , Ethylene glycol salicylate, myristyl salicylate, methyl salicylate, etc.), anthranilic acid derivatives (such as methyl anthranilate), urocanic acid derivatives (urocanic acid, ethyl urocanate etc.), coumarin derivatives, amino acid compounds, benzotriazole derivatives, tetrazole derivatives, Imidazoline derivatives, pyrimidine derivatives, dioxane derivatives, camphor derivatives, furan derivatives, pyrone derivatives, nucleic acid derivatives, allantoin derivatives Conductor, nicotinic acid derivative, vitamin B6 derivative, umbelliferone, esculin, benzyl cinnamate, synoxate, oxybenzone, dioxybenzone, octabenzone, soribenzone, benzoresorcinol, arbutin, guaiazulene, shikonin, baicalin, baicalein, berberine, neoheliopan, esca Rolls, zinc oxide, talc, kaolin, etc.

(19) Whitening agent Paraaminobenzoic acid derivative, salicylic acid derivative, anthranilic acid derivative, coumarin derivative, amino acid compound, benzotriazole derivative, tetrazole derivative, imidazoline derivative, pyrimidine derivative, dioxane derivative, camphor derivative, furan derivative, pyrone derivative, nucleic acid Derivatives, allantoin derivatives, nicotinic acid derivatives, vitamin C or derivatives thereof (vitamin C phosphate magnesium salt, vitamin C glucoside, etc.), vitamin E or derivatives thereof, kojic acid or derivatives thereof, oxybenzone, benzophenone, arbutin, guaiazulene, shikonin , Baicalin, baicalein, berberine, placenta extract, ellagic acid, lucinol, etc.

(20) Tyrosinase activity inhibitor Vitamin C or a derivative thereof (vitamin C phosphate magnesium salt, vitamin C glucoside, etc.), hydroquinone or a derivative thereof (hydroquinone benzyl ether, etc.), kojic acid or a derivative thereof, vitamin E or a derivative thereof, N-acetyltyrosine or derivatives thereof, glutathione, hydrogen peroxide, zinc peroxide, placenta extract, ellagic acid, arbutin, lucinol, silk extract, plant extract (camille, mulberry, gardenia, pearl oyster, clam, clam, mugwort, honeysuckle) , Kihada, dokudami, matsuhod, pearl barley, mussel, hops, hawthorn, eucalyptus, yarrow, altea, keihi, mankaishi, hamamelis, yamagwa, lifelong grass, bellflower, toshishi, zetsugo, gyu, mao, senkyu, dod Tsu, Psycho, storm, littoralis, Scutellaria, peony bark, cranesbill, kudzu root, licorice, Fushi aloe, Cimicifuga, safflower, green tea, black tea, Ali elixir), and the like.

(21) Melanin pigment reduction / decomposition substance Phenylmercury hexachlorophene, mercuric oxide, mercuric chloride, hydrogen peroxide, zinc peroxide, hydroquinone or its derivative (hydroquinone benzyl ether).

(22) Turnover promoting action / cell activator hydroquinone, lactic acid bacteria extract, placenta extract, ganoderma extract, vitamin A, vitamin E, allantoin, spleen extract, thymus extract, yeast extract, fermented milk extract, plant extract (aloe, Ogon, Japanese horsetail, Gentian, Burdock, Shikon, Carrot, Camelis, Hop, Yokuinin, Odori, Semburi, Japanese cypress, Japanese red snapper, Achacha, Hypericum, Cucumber, Necklace, Rosemary, Parsley).

(23) Astringents Succinic acid, allantoin, zinc chloride, zinc sulfate, zinc oxide, calamine, zinc paraphenolsulfonate, potassium aluminum sulfate, resorcin, ferric chloride, tannic acid (including catechin compounds) and the like.

(24) Active oxygen scavenger SOD, catalase, glutathione peroxidase and the like.

(25) Antioxidants Vitamin C or salts thereof, stearic acid ester, vitamin E or derivatives thereof, nordihydrogua cetelenic acid, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), hydroxytyrosol, parahydroxyanisole , Propyl gallate, sesamol, sesamorin, gossypol, propolis, etc.

(26) Lipid peroxide production inhibitor β-carotene, plant extract (sesame culture cell, hamcha, hypericum, hamamelis, clove, melissa, birch, birch, salvia, rosemary, minamitenji, jitsu, ginkgo, green tea).

(27) Anti-inflammatory agent Ictamol, indomethacin, kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d-camphor, dl-camphor, hydrocortisone, guaiazulene, camazulene, chlorpheniramine maleate, glycyrrhizic acid or its Salt, glycyrrhetinic acid or a salt thereof, licorice extract, sicon extract, age extract, propolis, etc.

(28) Antibacterial / bactericidal / disinfectant Acrinol, sulfur, calcium gluconate, chlorhexidine gluconate, sulfamine, mercurochrome, lactoferrin or hydrolyzate thereof, alkyldiaminoethylglycine chloride solution, triclosan, sodium hypochlorite, chloramine T, Sarashi flour, iodine compound, iodoform, sorbic acid or its salt, propionic acid or its salt, salicylic acid, dehydroacetic acid, parahydroxybenzoic acid esters, undecylenic acid, thiamine lauryl sulfate, thiamine lauryl nitrate, phenol, cresol, p- Chlorophenol, p-chloro-m-xylenol, p-chloro-m-cresol, thymol, phenethyl alcohol, O-phenylphenol, Irgasan CH3565, halocarban, hexachlorophene, chlorohexidine, ester Tanol, methanol, isopropyl alcohol, benzyl alcohol, ethylene glycol, propylene glycol, 2-phenoxyethanol, 1,2-pentanediol, zinc pyridione, chlorobutanol, isopropylmethylphenol, nonionic surfactant (polyoxyethylene lauryl ether, Polyoxyethylene nonyl phenyl ether, polyoxyethylene octyl phenyl ether, etc.), amphoteric surfactant, anionic surfactant (sodium lauryl sulfate, lauroyl sarcosine potassium, etc.), cationic surfactant (cetyltrimethylammonium bromide, benzalkco chloride) Nium, benzethonium chloride, methyl rosaniline chloride), formaldehyde, hexamine, brilliant green, malachite green, crystal bio Tsu DOO, Jamaru photosensitive No. 101-containing, photosensitizer No. 201, photosensitizer No. 401, N- long chain acyl basic amino acid derivative and its acid shark salts, zinc oxide, hinokitiol, Sophora root, propolis, and the like.

(29) Moisturizer Glycerin, propylene glycol, 1,3-butylene glycol, hyaluronic acid or its salt, polyethylene glycol, chondroitin sulfate or its salt, water-soluble chitin derivative, water-soluble chitosan derivative, pyrrolidone carboxylic acid or its salt, lactic acid Sodium, mini Sasanishiki extract, natto metabolite, natto extract, loofah water, birch (white birch) or red pine sap

(30) Elastase activity inhibitor diisopropyl fluorophosphate, plant extract (eg, gongoid, hypericum, clara, mulberry leaves, keihi, genokosho, comfrey, salvia, elderberry, bodaiju, button pie), seaweed extract and the like.

(31) Hair preparations Selenium disulfide, alkylisoquinolinium bromide solution, zinc pyrithione, biphenamine, thianthol, castari tincture, pepper tincture, pepper tincture, quinine hydrochloride, strong ammonia water, potassium bromate, sodium bromate, thio Glycolic acid and the like.

(32) Antiandrogens Follicular hormones (estrone, estradiol, ethinyl estradiol, etc.), isoflavones, oxendrone, etc.

(33) Peripheral vascular blood flow promoter Vitamin E or derivatives thereof, assembly extract, garlic extract, carrot extract, aloe extract, gentian extract, toki extract, cephalanthin, carpronium chloride, minoxidil and the like.

(34) Stimulants Pepper tincture, nonyl acid vanillamide, cantalis tincture, ginger tincture, peppermint oil, l-menthol, camphor, benzyl nicotinate and the like.

(35) Metabolic activator Photosensitive element 301, hinokitiol, pantothenic acid or derivatives thereof, allantoin, placenta extract, biotin, pentadecanoic acid glyceride and the like.

(36) Antiseborrheic agent Pyridoxine or a derivative thereof, sulfur, vitamin B6 and the like.

(37) Keratolytic agent Resorcin, salicylic acid, lactic acid, urea and the like.

(38) Oxidizing agent Hydrogen peroxide water, sodium persulfate, ammonium persulfate, sodium perborate, urea peroxide, sodium percarbonate, sodium tripolyphosphate, sodium bromate, potassium bromate, sodium pyrophosphate, Sodium orthophosphate, sodium silicate hydrogen peroxide adduct, sodium sulfate hydrogen peroxide adduct, sodium chloride hydrogen peroxide adduct, β-tyrosinase enzyme solution, mushroom extract, etc.

(39) Hair remover Inorganic reducing agents such as strontium sulfate, sodium sulfide, barium sulfide, calcium sulfide, thioglycolic acid or its salts (calcium thioglycolate, sodium thioglycolate, lithium thioglycolate, magnesium thioglycolate) Strontium thioglycolate) and the like.

(40) Hair swelling agents such as ethanolamine, urea and guanidine.

(41) Dye agent 5-aminoorthocresol, 2-amino-4-nitrophenol, 2-amino-5-nitrophenol, 1-amino-4-methylaminoanthraquinone, 3,3'-iminodiphenol, hydrochloric acid 2 , 4-Diaminophenoxyethanol, 2,4-diaminophenol hydrochloride, toluene-2,5-diamine hydrochloride, nitroparaphenylenediamine hydrochloride, paraphenylenediamine hydrochloride, N-phenylparaphenylenediamine hydrochloride, metaphenylenediamine hydrochloride, orthoaminophenol N-phenylparaphenylenediamine acetate, 1,4-diaminoanthraquinone, 2,6-diaminopyridine, 1,5-dihydroxynaphthalene, toluene-2,5-diamine, toluene-3,4-diamine, nitroparaphenylenediamine , Paraamino Enol, paranitroorthophenylenediamine, paraphenylenediamine, paramethylaminophenol, picramic acid, sodium picramate, N, N'-bis (4-aminophenyl) -2,5-diamino-1,4-quinonediimine, 5 -(2-hydroxyethylamino) -2-methylphenol, N-phenylparaphenylenediamine, metaaminophenol, metaphenylenediamine, 5-aminoorcresol sulfate, 2-amino-5-nitrophenol sulfate, orthoaminophenol sulfate , Orthochloroparaphenylenediamine sulfate, 4,4'-diaminodiphenylamine sulfate, 2,4-diaminophenol sulfate, toluene-2,5-diamine sulfate, nitroparaphenylenediamine sulfate, paraaminophenol sulfate, parasulfate sulfate Nitroorthophenylenediamine, paranitrometaphenylenediamine sulfate, paraphenylenediamine sulfate, paramethylaminophenol sulfate, metaaminophenol sulfate, metaphenylenediamine sulfate, catechol, diphenylamine, α-naphthol, hydroquinone, pyrogallol, phloroglucin, gallic acid, Resorcin, tannic acid, 2-hydroxy-5-nitro-2 ', 4'-diaminoazobenzene-5'-sodium sulfonate, hematein and the like.

(42) Fragrance Natural animal fragrance such as musk, civet, castorium, ambergris, anise essential oil, angelica essential oil, ylang ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guayakwood essential oil, Cumin essential oil, black letter essential oil, cinnamon essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriandel essential oil, perilla essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, gingergrass essential oil, cedar essential oil, spearmint essential oil, western peppermint essential oil, Otsuka perfume essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, winter green essential oil, trout balsam essential oil, buttery essential oil, rose essential oil, palmarosa essential oil, persimmon essential oil, hiba essential oil, sandalwood essential oil, petit gren essential oil, bay essential oil, vetiva essential oil, bergamot Spirit , Peru balsam essential oil, boad rose essential oil, melamine essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary essential oil, Japanese mint essential oil, and other synthetic flavors Fragrance etc.

(43) Dye and Colorant Red Cabbage Dye, Red Rice Dye, Akane Dye, Anato Dye, Ikumi Dye, Turmeric Dye, Enju Dye, Krill Dye, Amber Dye, Caramel, Gold, Silver, Gardenia Dye, Corn Dye, Onion Dye , Tamarind pigment, spirulina pigment, buckwheat whole plant pigment, cherry pigment, laver pigment, hibiscus pigment, grape juice pigment, marigold pigment, purple potato pigment, purple yam pigment, lac pigment, rutin etc.

(44) Others Hormones, sequestering agents, pH adjusters, chelating agents, antiseptic and antibacterial agents, cooling agents, stabilizers, emulsifiers, animal and vegetable proteins and their degradation products, animal and vegetable polysaccharides And its degradation products, animal and plant glycoproteins and their degradation products, blood flow promoters, anti-inflammatory agents, anti-inflammatory agents, anti-allergic agents, wound treatment agents, foam increasing agents, thickeners, enzymes, purified water (electronic Water, small clustering, etc.), oral preparations, deodorants / deodorants, bittering agents, seasonings, etc. are raised, and in combination with these, the effects of the present invention or other cosmetic effects are added Or a synergistic improvement can be expected.

  Examples of Enterococcus lactic acid cocci which are the most remarkable in the objective effect of the present invention are shown in Enterococcus faecalis EC-12, but the present invention is not limited to these Examples.

(Deposit of microorganisms)
Enterococcus faecalis EC-12 is a patent biological deposit center, National Institute of Advanced Industrial Science and Technology (Tsukuba, Ibaraki) on February 25, 2005 (original deposit date). It was deposited at 1-Chome, 1-Chome, Ichi-Chi, 1st 6th. The accession number is FERM BP-10284.

  According to a preferred embodiment of the present invention, the bacterium used as an active ingredient is a bacterium belonging to the genus Enterococcus. More preferably, the bacterium is Enterococcus faecalis.

Examples of Enterococcus faecalis include strains such as Enterococcus faecalis EC-12 (Accession No. FERM BP-10284), ATCC 19433, ATCC 14508, ATCC 23655, IFO 16803, IFO 16804 and the like. Of these, EC-12 is most preferred as the bacterium used as the active ingredient.
Here, the term “mutant strain” means that a specific strain has been mutated by a method well known to those skilled in the art within a range that does not change the properties thereof, or is equivalent to that. The meaning includes things that can be confirmed.

(Obtaining Enterococcus lactobacilli and heat-treated dead cells)
Enterococcus faecalis EC-12 (accession number FERM BP-10284) purchased from Combi Co., Ltd. was used.

(Manufacture of plant polysaccharides)
Aloe vera polysaccharides were produced as plant polysaccharides. After crushing 100 g of aloe vera leaves, squeezing extraction and filtration, ethanol or isopropanol was added to the obtained filtrate and stirred, and then the resulting precipitate was collected, washed again with ethanol or isopropanol, and obtained. The washed product was dried to obtain aloe vera polysaccharide powder.

(Sample adjustment)
Enterococcus lactic acid cocci heat-treated dead cells and aloe vera polysaccharide prepared as described above were mixed to prepare samples 1 to 4 (medium addition final concentration) shown in Table 1.

(Culture of Staphylococcus epidermidis standard bacteria)
As a culture medium, Nutribouillon No. 2 (Nissui Pharmaceutical Co., Ltd.) was used. Samples 1 to 4 (Table value: final concentration% of medium) were added to this medium. Subsequently, Staphylococcus epidermidis standard bacteria (ATCC12228) was inoculated into the medium so as to be 1-5 × 10 ³ CFU / μL, and these were cultured at 37 ° C. for 24 hours. As a control, enterococcus lactic acid cocci heat-treated dead cells were not added (sample 1).

(Method for collecting human skin resident bacteria)
A 3 cm square between the eyebrows was rubbed with a sterile cotton swab 3 times in the vertical direction and 3 times in the horizontal direction, and normal skin bacteria were collected. The swab was then washed in 10 mL of phosphate buffered saline and collected.

(Culture of human skin resident bacteria)
As a culture medium, Nutribouillon No. 2 (Nissui Pharmaceutical Co., Ltd.) was used. Enterococcus Lactococcus heat-treated dead cells were added to this medium so as to have final concentrations of 0.25, 0.5 and 1.0 w / w%. Then, the skin resident bacteria collected by the above method were inoculated into 100 μL medium. These were cultured at 37 ° C. for 24 hours. As a control, heat-treated dead cells were not added (0 w / w%).

(Measurement of glycerol amount)
Each culture solution of the above staphylococcus epidermidis standard bacteria (ATCC12228) and human skin resident bacteria was fractionated. These cultures were vortexed and the amount of glycerol was measured with a glycerol assay kit (Cayman Chemical) according to a conventional method.

(Measurement result of glycerol amount)
The results are shown in FIGS. Enterococcus Lactococcus heat-treated dead cells significantly increased glycerol production from Staphylococcus epidermidis (ATCC 12228) (FIG. 1). An approximately 10-fold glycerol production promoting effect was observed at a final addition concentration of 1.0 w / w% (sample 2) to the medium. In addition, in combination with Aloe vera polysaccharide (0.1 w / w%) (sample 4), glycerol production from S. epidermidis standard bacteria increased synergistically. The addition of Enterococcus lactic acid cocci heat-treated dead cells also increased the amount of glycerol produced in the human skin resident bacterial culture (FIG. 2).

(Human monitor test)
Male and female panelists (subjects: 18 subjects) wash their face every day before going to bed, then enterococcus Lactococcus heat-treated soft skin lotion (skin lotion A) on the right cheek and control on the left cheek. The placebo soft lotion (skin lotion B) to which the lactic acid cocci heat-treated dead cells are not added is used for 7 consecutive days (prescription is the following, soft lotion). After waking up on the 8th day, sterilize 3 cm square on each cheek. Grubbing was performed 3 times in the vertical direction and 3 times in the horizontal direction with a cotton swab to collect glycerol on the skin surface. The swab was then washed in 200 μL of phosphate buffered saline to disperse the metabolites.

(Flexible lotion)
Enterococcus lactic acid cocci heat-treated dead cells 0 or 0.1 w / v%
Ethanol 5.0%
1,3-butylene glycol 8.0%
PEG-60 hydrogenated castor oil 0.2%
Fragrance 0.01%
Methylparaben 0.15%
Citric acid 0.02%
Citric acid 3Na 0.08%
EDTA-2Na 0.03%
The remainder of water 100 was uniformly mixed according to a conventional method to obtain soft lotions (lotion A, lotion B).

(Measurement of glycerol amount)
The dispersion was vortexed and the amount of glycerol was measured with a glycerol assay kit (Cayman Chemical) according to a conventional method.

(Measurement result of glycerol amount)
The results are shown in FIG. When soft lotion containing enterococcus lactobacilli heat-treated dead cells was used, glycerol on the surface of the human monitor skin was significantly increased compared to the soft lotion of the control.

(Sensory evaluation of human monitor test)
For male and female panelists (18 subjects) who conducted a human monitor test, a questionnaire survey was conducted regarding the feeling of use of the soft lotion used in the test and the skin condition before and after the test. The answer results are shown in Table 2.

(Quantitative analysis of human β-defensin-2 (hBD2), -3 (hBD3) mRNA)
Human epidermal keratinocytes (manufactured by Kurabo Industries Co., Ltd.) were cultured to a sub-confluent state using a medium for epidermal cells (Humdia KG2, manufactured by Kurabo Industries Inc.) in a 24-well plate. The medium was further cultured for 3 days in the presence of 5% CO _{2 at} 37 ° C. in a differentiation induction medium in which calcium was added to Humdia KG2, and then replaced with a new medium. Enterococcus Lactococcus heat-treated dead cells were added to this medium so as to have final concentrations of 0.25, 0.5 and 1.0 w / w%. As a positive target, Staphylococcus epidermidis standard bacteria were added so as to be about 1 × 10 ⁴ cells / mL. After 6 hours of culture, total RNA was extracted from human epidermal keratinocytes using RNeasy (Qiagen). Using each extracted RNA as a template, reverse transcription reaction was performed using SuperScriptVILO (Invitrogen) by a conventional method to prepare cDNA.

  PCR was performed using the cDNA. Primers shown in Table 3 were prepared based on the cDNA sequence of human β-defensin-2 (Accession No. NG — 0230301) and the cDNA sequence of human β-defensin-3 (Accession No. NC — 000008). . TAKARA Human Housekeeping Gene Primer Set Human ACTB Primer was used as a primer for β-actin used as an internal standard.

One Step SYBRR PrimeScript ^™ RT-PCR Kit (TAKARA) was used for PCR reaction, and TaKaRa PCR Thermal Cycler Dice was used for real-time PCR analysis by a conventional method. By the real-time PCR analysis, the amounts of human β-defensin-2 and -3 and β-actin mRNA were relatively quantified.

(Evaluation method of human β-defensin-2 and -3 and β-actin mRNA amount)
The amounts of human β-defensin-2 and -3 and β-actin mRNA relative to the amount of β-actin mRNA as an internal standard were evaluated at 100 minutes.
Relative expression level of human β-defensin-2 and -3 mRNA (% of control) = (human β-defensin-2 and -3 mRNA amount) / (β-actin mRNA amount) × 100

(Quantitative analysis results of human β-defensin-2 and -3 mRNA)
The results are shown in FIG. Enterococcus lactic acid cocci heat-treated dead cells also increased β-defensin-2 and -3 mRNA expression levels from human epidermal keratinocytes (FIG. 4).

(Manufacturing examples of various external preparations for skin protection)
1. Sunscreen cream (O / W cream type)
(Oil phase)
2-methoxyhexyl paramethoxycinnamate 5.0%
4-t-butyl-4′-methoxydibenzoylbutane 1.0%
Squalane 10.0%
Vaseline 5.0%
Stearyl alcohol 3.0%
Stearic acid 3.0%
Glyceryl monostearate 3.0%
Polyacrylic acid 1.0%
Antioxidant Appropriate amount Preservative Appropriate perfume Appropriate amount Enterococcus Lactococcus heat-treated dead cells 0.01-1%
(Water phase)
Residual dipropylene glycol 7.0% purified water 100%
Titanium dioxide (alumina surface treatment product) 5.0%
Edetate disodium 0.05%
Triethanolamine 1.0%
(Production method)
The oil phase part and the water phase part are each heated to 70 ° C. to dissolve, the water phase part is sufficiently dispersed in titanium dioxide, the oil phase part is added, and the emulsified particles are homogenized using a homomixer. , Filtered and cooled. As a cooling method, there were a method of gradually cooling with stirring while injecting a refrigerant into the jacket of the production kettle and a method of cooling using a heat exchanger.

2. Sunscreen emulsion (W / O type emulsion)
(Oil phase)
2-methoxyhexyl paramethoxycinnamate 5.0%
Titanium dioxide (hydrophobized product) 5.0%
Zinc oxide (hydrophobized product) 9.0%
Squalane 20.0%
Silicone oil 23.0%
Silicone resin 2.0%
Glycerin diisostearate 2.0%
Organically modified montmorillonite 0.5%
Preservative Appropriate amount of perfume Appropriate amount Enterococcus lactic acid cocci 0.01-1% (appropriate amount)
(Water phase)
Residual 1,3-butylene glycol 5.0% purified water 100%
(Production method)
The oil phase part and the water phase part were each dissolved by heating to 60 ° C., the oil phase part was sufficiently dispersed in titanium dioxide and zinc oxide, the water phase part was added, and the emulsified particles were made uniform using a homomixer. Then, it deaerated, filtered, cooled and accommodated.

3. Sun oil paramethoxycinnamate 2-ethylhexyl 2.0%
Liquid paraffin 56.0%
Isopropyl myristate 10.0%
Silicone oil 30.0%
Silicone resin 2.0%
Antioxidant Appropriate amount of perfume Appropriate amount Enterococcus lactic acid cocci heat-treated dead cells 0.01-1% (appropriate amount)
(Production method)
The silicone resin was added and dissolved in the silicone oil, and then isopropyl myristate was added and mixed with sufficient stirring, and then the remaining components were added and stirred.

4). Powder type foundation (powder)
Zinc myristate treatment talc 20.0%
Sericite 34.6%
Titanium oxide 10.0%
Bengala 1.0%
Yellow iron oxide 4.0%
Black iron oxide 0.4%
Titanium oxide / aluminum oxide Surface treatment mica 15.0%
Nylon powder 5.0%
Enterococcus lactic acid cocci 0.01-1% (appropriate amount)
(Base material)
Squalane 3.0%
Octyldodecyl myristate 3.5%
Vaseline 2.5%
(Other)
Preservative Appropriate amount Perfume Appropriate amount (production method)
The powder component was mixed using a mixer such as a high-speed fluid mixer, and fine particles were formed using a pulverizer. Next, the base material was added and mixed thoroughly using a vertical screw mixer or the like. Furthermore, after pulverizing and granulating using a hammer mill or the like, compression molding was performed.

5. Dual use type foundation (powder)
Silicon-treated titanium oxide 10.0%
Talc 30.0%
Sericite 39.6%
Bengala 1.5%
Yellow iron oxide 5.5%
Black iron oxide 0.4%
Methyl methacrylate powder 3.0%
Enterococcus lactic acid cocci heat-treated dead cells 0.01-1 (appropriate amount)
(Base material)
Squalane 3.0%
Dimethylpolysiloxane 2.0%
Glycerin triisooctanoate 3.0%
Ethyl paraaminobenzoate 2.0%
(Other)
Antiseptic agent

6). Cream (W / O type)
(Phase A)
t-Butylmethoxydibenzoylmethane 3.0%
Diethylaminohydroxybenzoyl hexyl benzoate 2.0%
Dimethicone, caprylyl dimethicone ethoxyglucoside 20.0%
Tri (behenic acid / isostearic acid / eicosanedioic acid) glyceryl 1.0%
Stearyl dimethicone 1.0%
Tocopherol acetate 0.05%
Benzoic acid 0.3%
Diisopropyl sebacate 7.0%
Octocrylene 8.0%
Talc, yellow iron oxide, Mg stearate, titanium dioxide, mica, Al hydroxide, stearic acid 1.4%
(Phase B)
Titanium dioxide, Mg stearate, talc, black iron oxide, Al hydroxide, mica, stearic acid 1.13%
Red and brown iron oxide, Mg stearate, titanium dioxide, mica, water, oxidized Al, stearic acid 0.85%
Titanium dioxide, talc, Mg stearate, Al hydroxide, mica, stearic acid
3.9%
(Phase C)
Residual Mg sulfate as water 100 0.7%
Betaine 1.0%
Hyaluronic acid Na 0.1%
Glycerin 1.0%
(Phase D)
Phenoxyethanol 0.4%
(Phase E)
Titanium dioxide, glycerin, alumina 3.5%
(Phase F)
Dimethylsilylated silica 1.0%
(G phase)
Cyclopentasiloxane, (dimethicone / vinyltrimethylsiloxysilicate) crosspolymer 1.0%
Enterococcus lactic acid cocci heat-treated dead cells 0.01-1 (appropriate amount)
(H phase)
Fragrance 0.3%
(Production method)
The raw materials for phase A were mixed with stirring, the temperature was raised to 70 to 75 ° C., phase B was added to phase A with stirring, and the mixture was stirred until it became uniform. In a separate container, the ingredients of phase C are mixed with stirring, the temperature is raised to 70 to 75 ° C., phase C is added to the AB mixed phase, slowly added with stirring (about 5 minutes), and about 3 minutes after addition of phase C. The mixture was stirred and then naturally cooled. When the temperature became 40 ° C. or lower, D phase was added and stirred for about 1 minute, E phase was added and stirred for about 5 minutes, then F phase was added and stirred for about 1 minute, G phase was added and stirred for about 1 minute. Finally, the H phase was added and stirred for about 2 minutes, and then naturally cooled to room temperature.

7). Moisturizing gel (beauty serum)
(Phase A)
Olive oil fatty acid cetearyl + olive oil fatty acid sorbitan 3.5%
Olive oil unsaponifiable 0.5%
Dicaprylyl carbonate 3.0%
Shea fat 0.5%
Wheat germ oil 0.5%
Cyclomethicone 6.0%
Dimethicone 1.0%
(Phase B)
Residual glycerin as water 100 1.5%
Propylene glycol 1.5%
Polyacrylic acid Na 0.7%
Xanthan gum 0.1%
(Phase C)
Preservatives Appropriate fragrance Appropriate menthol 0.3%
Ethanol 3.0%
Enterococcus lactic acid cocci heat-treated dead cells 0.01-1 (appropriate amount)
(Production method)
A phase and B phase were mixed and prepared. Xanthan gum and polyacrylic acid Na were dispersed by using a batch type disperser, etc., and A phase was added to B phase at 75 ° C., homogenized for several minutes, cooled slowly with stirring, and C phase was added when the temperature reached 40 ° C. (Menthol is dissolved in ethanol).

8). Lip balm (A phase)
Mineral oil 8.0%
Red pigment 1.0%
Yellow pigment 0.3%
Enterococcus lactic acid cocci 0.01-1% (appropriate amount)
(Phase B)
Residual ethylhexyl methoxycinnamate 3.0% with tri (caprylic acid / capric acid) glyceryl 100
Isopropyl myristate 10.0%
(Phase C)
Alkyl (C26-28) methicone 9.0%
Cetanol 1.6%
(Phase D)
PEG-8, tocopherol, ascorbyl palmitate, ascorbic acid, citric acid mixture 0.1%
Propylparaben 0.1%
(Phase E)
Fragrance 0.2%
Mica, titanium oxide mixture 18.4%
Mica, titanium oxide, iron oxide mixture 7.0%
Silica 0.5%
(Production method)
Premix A phase uniformly, add to B phase and mix evenly. Add C phase and heat to 78-80 ° C with slow stirring, then add D and E phases little by little and stir uniformly. Then, it was poured into a mold and naturally cooled to room temperature.

9. Liquid foundation (O / W type)
(Oil phase)
Stearic acid 2.4%
Propylene glycol monostearate 2.0%
Cetostearyl alcohol 0.2%
Liquid lanolin 2.0%
Liquid paraffin 3.0%
Isopropyl myristate 8.5%
Oil-soluble preservative appropriate amount (water phase)
Residual sodium carboxymethyl cellulose with purified water 100 0.2%
Bentonite 0.5%
Dipropylene glycol 4.0%
Triethanolamine 1.1%
Water-soluble preservative appropriate amount (powder)
Titanium oxide 8.0%
Talc 4.0%
Colored pigment appropriate amount Enterococcus lactic acid cocci heat-treated dead cell 0.01-1 (appropriate amount)
(Other)
Perfume appropriate amount (production method)
After the powder was pulverized, an aqueous phase was added and mixed, and an oil phase and a fragrance were added thereto with stirring. After emulsification and dispersion with a homomixer, the mixture was cooled to room temperature.

10. Cream foundation (W / O type)
(Oil phase)
Decamethylcyclopentasiloxane 15.0%
Liquid paraffin 5.0%
Paramethoxycinnamate 3.0%
Polyether-modified dimethylsiloxane 2.0%
(Water phase)
Purified water 100% residual glycerin 5.0%
Water-soluble preservative appropriate amount (powder)
Titanium oxide 10.0%
Talc 3.8%
Coloring pigment appropriate amount Enterococcus lactic acid cocci 0.01-1% (appropriate amount)
(Other)
Perfume

11. Moisturizing cream (O / W type)
(Phase A)
Shea fat 1.5%
Alkyl benzoate (C12-15) 1.3%
Alkyl (C26-28) dimethicone 6.5%
Glyceryl stearate 1.5%
Glyceryl stearate, PEG-100 stearate mixture 3.0%
Hexyl laurate 1.3%
Isododecane 1.3%
Isohexadecane 1.3%
Isopropyl palmitate 1.3%
Tocopherol acetate 0.5%
Enterococcus lactic acid cocci heat-treated dead cells 0.01-1 (appropriate amount)
(Phase B)
Residual carbomer of 100% purified water 0.3%
Glycerin 2.0%
Propylene glycol 2.0%
(Phase C)
Fragrance 0.5%
Phenoxyethanol, methylparaben, ethylparaben, butylparaben, propylparaben, isobutylparaben mixture 0.5%
Tetrahydroxypropylethylenediamine 0.7%
(Production method)
After heating A phase and B phase to 70 degreeC with another container, A phase was added to B phase, it homogenized using a batch type disperser etc., and naturally cooled to 40 degreeC, stirring. Next, Phase C was added to this and further homogenized, and cooled to room temperature.

  The Lactococcus enterococcus according to the present invention promotes the production of glycerol derived from Staphylococcus epidermidis, a skin resident bacterium useful in the skin, and the production of antibacterial peptides derived from skin epidermal keratinocytes. In addition, when applied to the skin, it is smooth and extremely excellent in touch, and imparts good touch to any type of aqueous, oil-based, emulsion, and powder-type external preparations. Therefore, the utilization to the chemical | medical agent which protects the skin which normalizes a skin state or maintains normality, and the external preparation for skin protection is very high.

Claims (3)

A glycerol production promoter derived from Staphylococcus epidermidis containing a dead cell of Enterococcus faecalis EC-12 (Accession Number FERM BP-10284) as an active ingredient. Enterococcus faecalis glycerol production promoter derived from Staphylococcus epidermidis, containing dead cells of Enterococcus faecalis EC-12 (accession number FERM BP-10284) and aloe vera polysaccharide as active ingredients. Enterococcus faecalis Enterococcus faecalis EC-12 (Accession No. FERM BP-10284) containing a dead cell as an active ingredient, an antibacterial peptide production promoter derived from human skin epidermis keratinocytes, wherein the antibacterial peptide is human β The antibacterial peptide production promoter, which is defensin-2 (hBD2: human β-defensin-2) and / or human β-defensin-3 (hBD3: human β-defensin-3).