KR20180124424A - Novel strain of Fusidium coccineum spp., and composition for improving skin beauty comprising a culture solution of the strain - Google Patents
Novel strain of Fusidium coccineum spp., and composition for improving skin beauty comprising a culture solution of the strain Download PDFInfo
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- KR20180124424A KR20180124424A KR1020170058901A KR20170058901A KR20180124424A KR 20180124424 A KR20180124424 A KR 20180124424A KR 1020170058901 A KR1020170058901 A KR 1020170058901A KR 20170058901 A KR20170058901 A KR 20170058901A KR 20180124424 A KR20180124424 A KR 20180124424A
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- fusidium
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Abstract
Description
신규 후시디움 코씨네움 균주 및 그 균주 배양액을 포함하는 피부미용 개선용 조성물에 관한 것이다.A novel foshium coccinium strain and a culture solution of the strain.
후시디움 속 (Fusidium sp.) 미생물은 곰팡이이며, 스테로이드계 항생물질인 후시딘산 (fusidic acid)은 후시디움 코씨네움 (Fusidium coccineum) 균주로부터 얻을 수 있다. 후시딘산은 그람양성균, 특히 포도상구균감염증에 유효하며 내복 및 외용약으로도 사용되고 있다. 약물에서 후시딘산은 소듐 후시데이트 (sodium fusidate)의 형태로 사용되고 있다.Fushimi Stadium in (Fusidium sp.) Fungi and microorganisms, the steroidal husidin San antibiotics (fusidic acid) are co-Fushimi Stadium Stadium Cine (Fusidium coccineum strains. Fucidin is effective against Gram-positive bacteria, especially staphylococcal infections, and is also used as an underwear and external medicine. Fusidic acid in the drug is used in the form of sodium fusidate.
중국 공개특허 101812498호는 후시딘산의 발효 생산 방법에 관한 것으로, 후시디움 코씨네움 (F. coccineum) 균주를 이용하여 발효 공정을 통해 후시딘산을 생산하는 방법을 개시하고 있다. Chinese Patent Publication No. 101812498 relates to a method for fermenting fucidin acid, and discloses a method for producing fucindinic acid through a fermentation process using F. coccineum strain.
그러나, 후시디움 코씨네움 (F. coccineum) 균주 자체에 대한 연구는 많이 진행되지 않았으며, 특히 후시디움 코씨네움 균주를 화장품 등의 피부 미용 개선 용도로 사용한 연구는 전무하다. However, much research has not been conducted on the F. coccineum strain itself, and there have been no studies using Fushidium coccinum strain for skin cosmetic improvement.
일 양상은 후시디움 코씨네움(Fusidium coccineum) 균주 (수탁번호: KCCM12013P)를 제공한다.One aspect is Fusidium coccineum strain (accession number: KCCM12013P).
다른 양상은 후시디움 코씨네움(Fusidium coccineum) 균주 배양액을 유효성분으로 포함하는 피부미용 개선용 조성물을 제공한다.Another aspect is Fusidium < RTI ID = 0.0 > coccineum ) culture medium as an active ingredient.
다른 양상은 후시디움 코씨네움 균주 배양액을 제조하는 방법을 제공한다.Another aspect provides a method of producing a fusidium coccinium culture.
일 양상은 후시디움 코씨네움(Fusidium coccineum) 균주 (수탁번호: KCCM12013P)를 제공한다.One aspect is Fushidium cochineum (Fusidium coccineum) Strain (Accession No .: KCCM12013P).
상기 "후시디움 코씨네움(Fusidium coccineum)"은 곰팡이의 일종으로서, 식물로부터 분리한 것일 수 있다. 상기 식물은 그 종류를 한정하지 않으나, 예를 들어 은행나무 잎일 수 있다. 상기 균주는 식물 샘플을 감자한천배지에 접종하여 배양된 콜로니를 순수 분리 배양하는 방법에 의해 분리된 균주일 수 있다. 상기 균주는 미황색 집락을 형성하며 생장하는 것일 수 있다. 상기 균주는 감자한천배지에서 생장하는 것일 수 있다.The " Fusidium " coccineum " is a kind of fungus which may be isolated from a plant. The plant is not limited in its kind but may be, for example, a ginkgo leaf. The strain is obtained by inoculating a plant sample with potato agar medium The strain may be a strain isolated by a method of purely isolating and culturing colonies. The strain may be one which grows to form a pale yellow colonies. The strain may be grown in a potato agar medium.
다른 양상은 후시디움 코씨네움(Fusidium coccineum) 균주 배양액을 유효성분으로 포함하는 피부미용 개선용 조성물을 제공한다.Another aspect is Fusidium < RTI ID = 0.0 > coccineum ) culture medium as an active ingredient.
상기 피부미용 개선은 피부 장벽 강화, 피부 보습, 피부 세포 재생, 피부 활력 증가, 항산화, 항염증, 또는 항아토피일 수 있다. 후시디움 코씨네움 균주 배양액은 줄기세포 배양액에 비해 피부 장벽 강화 효과, 피부 보습 효과, 피부 세포 재생 효과, 피부 활력 증가 효과, 항산화 효과, 항염증 효과, 또는 항아토피 효과가 우수하므로, 상기 후시디움 코씨네움 균주 배양액을 포함하는 조성물은 피부 장벽 강화, 피부 보습, 피부 세포 재생, 피부 활력 증가, 항산화, 항염증, 또는 항아토피용 조성물로 사용될 수 있다.Such skin aesthetic enhancement may be skin barrier enhancement, skin moisturization, skin cell regeneration, increased skin vitality, antioxidant, anti-inflammatory, or anti-atopic. The culture medium of the Fusidium coccinum strain is superior to the stem cell culture solution in that it has a skin barrier strengthening effect, a skin moisturizing effect, a skin cell regeneration effect, a skin vitality increasing effect, an antioxidative effect, an antiinflammatory effect or an antiatopic effect, A composition comprising a culture of a nociceptic strain may be used as a composition for skin barrier strengthening, skin moisturizing, skin cell regeneration, skin vitality enhancement, antioxidant, anti-inflammatory, or anti-atopic composition.
용어, "피부 장벽 강화"란 피부 가장 바깥쪽에 위치하여 수분과 영양 손실을 막아주는 피부 장벽의 기능이 증진되는 모든 작용을 의미할 수 있다.The term " skin barrier enhancement " may refer to any action that is located at the outermost position of the skin, thereby enhancing the function of the skin barrier to prevent moisture and nutritional loss.
용어, "피부 보습"이란 피부 수분을 유지하거나 수분 손실을 방지하는 모든 작용을 의미할 수 있다.The term " skin moisturizing " may mean any action that maintains skin moisture or prevents moisture loss.
용어, "피부 세포 재생"이란 피부의 일부분이 소실되었을 경우 그 부분을 보충하거나 피부 세포의 증식을 증진시키는 모든 작용을 의미할 수 있다.The term " skin cell regeneration " may mean any action that, when a portion of the skin is lost, replenishes that part or promotes proliferation of the skin cells.
용어, "피부 활력 증가"란 피부의 세포 내 미토콘드리아의 활성이 증가하여 피부의 생기 또는 활력을 증가시키는 모든 작용을 의미할 수 있다.The term " increased skin vitality " may refer to any action that increases the activity of mitochondria in the cells of the skin, thereby increasing the vitality or vitality of the skin.
용어, "항산화"란 산화를 억제하는 모든 작용을 의미할 수 있다. 상기 항산화 작용에 의해 피부 노화가 예방, 개선, 또는 억제될 수 있다.The term " antioxidant " may mean any action that inhibits oxidation. The antioxidative action can prevent, ameliorate, or inhibit skin aging.
용어, "항염증"이란"염증 억제 또는 개선"과 혼용될 수 있으며, 면역 반응이 완화되어 NO 생성이 억제되는 모든 작용을 의미할 수 있다.The term " anti-inflammatory " can be used interchangeably with " inflammation inhibition or improvement ", and may mean all actions in which the immune response is alleviated and NO production is suppressed.
용어, "항아토피"란 "아토피 억제, 개선 또는 완화"과 혼용될 수 있으며, 아토피 피부염(atopic dermatitis)의 증상인 가려움증(소양증), 피부건조증, 습진 등을 억제, 개선 또는 완화시키는 모든 작용을 의미할 수 있다.The term " anti-atopy " can be used interchangeably with " inhibiting, improving or alleviating atopy ", and includes all actions that inhibit, ameliorate, or alleviate the symptoms of atopic dermatitis (pruritus) It can mean.
용어, "개선"이란 상태의 완화 또는 치료와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미할 수 있다.The term " improvement " may mean any action that at least reduces the degree of symptomatology, such as a condition associated with relief or treatment of a condition.
상기 균주는 식품, 토양, 바다 등에서 분리한 것일 수 있다.The strain may be isolated from food, soil, sea or the like.
일 구체예에 따른 조성물에서, 상기 균주는 후시디움 코씨네움(Fusidium coccineum) 균주(수탁번호: KCCM12013P)일 수 있다. In a composition according to one embodiment, the strain may be a Fusidium coccineum strain (accession number: KCCM12013P).
일 구체예에 따른 조성물에서, 상기 배양액은 균주를 배양하여 얻은 균체 배양액에서 균체를 제거한 것일 수 있다. In the composition according to one embodiment, the culture may be one wherein the cells are removed from the culture medium obtained by culturing the strain.
일 구체예에 따른 조성물에서, 상기 배양액은 후시딘산(fusidic acid)을 포함하는 것일 수 있으나, 이에 한정되지 않는다.In a composition according to one embodiment, the culture may be, but is not limited to, include fusidic acid.
상기 배양액은 조성물 총 중량에 대하여 0.001 중량% 내지 80 중량%, 예를 들면, 0.01 중량% 내지 60 중량%, 0.01 중량% 내지 40 중량%, 0.01 중량% 내지 30 중량%, 0.01 중량% 내지 20 중량%, 0.01 중량% 내지 10 중량%, 0.01 중량% 내지 5 중량%, 0.05 중량% 내지 60 중량%, 0.05 중량% 내지 40 중량%, 0.05 중량% 내지 30 중량%, 0.05 중량% 내지 20 중량%, 0.05 중량% 내지 10 중량%, 0.05 중량% 내지 5 중량%, 0.1 중량% 내지 60 중량%, 0.1 중량% 내지 40 중량%, 0.1 중량% 내지 30 중량%, 0.1 중량% 내지 20 중량%, 0.1 중량% 내지 10 중량%, 0.1 중량% 내지 5 중량%, 0.1 중량% 내지 3 중량%, 0.1 중량% 내지 2중량%로 포함될 수 있다.The culture solution may be used in an amount of 0.001 to 80 wt%, for example, 0.01 to 60 wt%, 0.01 to 40 wt%, 0.01 to 30 wt%, 0.01 to 20 wt% %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20% From 0.05% to 10%, from 0.05% to 5%, from 0.1% to 60%, from 0.1% to 40%, from 0.1% to 30%, from 0.1% to 20% % To 10 wt%, 0.1 wt% to 5 wt%, 0.1 wt% to 3 wt%, and 0.1 wt% to 2 wt%.
일 구체예에 따른 조성물은 필라그린 (Filaggrin)의 발현을 증가시키는 것일 수 있고, 구체적으로 필라그린의 발현을 증가시켜 피부 장벽 강화 효과를 나타내는 것일 수 있다.The composition according to one embodiment may be one that increases the expression of filaggrin, and more specifically, the expression of pilar green to increase the skin barrier.
일 구체예에 따른 조성물은 세라미드 합성효소 3 (Ceramide synthase 3), b-글루코세레브로시다제 (b-Glucocerebrosidase), 또는 AQP3 (Aquaporin 3)의 발현을 증가시키는 것일 수 있고, 구체적으로 세라미드 합성효소 3, b-글루코세레브로시다제 , 또는 AQP3의 발현을 증가시켜 피부 보습 효과를 나타내는 것일 수 있다.The composition according to one embodiment may increase the expression of
일 구체예에 따른 조성물은 세포 내 미토콘드리아 활성을 증가시키는 것일 수 있고, 구체적으로 섬유아세포 내 미토콘드리아 활성을 증가시켜 피부 세포 재생 또는 피부 활력 증가 효과를 나타내는 것일 수 있다.The composition according to one embodiment may be one that increases intracellular mitochondrial activity, specifically, increases mitochondrial activity in fibroblasts, thereby exhibiting skin cell regeneration or skin vitality enhancement.
일 구체예에 따른 조성물은 IL-1a, IL-6, 또는 TSLP의 발현을 억제하는 것일 수 있고, 구체적으로 IL-1a, IL-6, 또는 TSLP의 발현을 억제하여 항염증 또는 항아토피 효과를 나타내는 것일 수 있다.The composition according to one embodiment may inhibit the expression of IL-1a, IL-6, or TSLP, and specifically inhibits the expression of IL-1a, IL-6, or TSLP to produce anti- .
일 구체예에 따른 조성물은 상기 배양액을 유효한 양, 또는 유효 성분으로서 포함할 수 있다. 상기 유효한 양은 개체에 따라 적절하게 선택할 수 있다. 질환 내지 상태의 중증도, 개체의 연령, 체중, 건강, 성별, 개체의 추출물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 투여 기간, 상기 조성물과 배합 또는 동시 사용되는 다른 조성물을 포함한 요소 및 기타 생리 내지 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다.The composition according to one embodiment may contain the above culture medium as an effective amount or as an active ingredient. The effective amount may be appropriately selected depending on the individual. The severity of the disease or condition, the age, weight, health, sex, sensitivity of the individual to the extract, time of administration, route and rate of excretion, duration of administration, factors including other compositions combined or co- May be determined according to factors well known in the art, physiology or medical field.
일 구체예에 따른 피부미용 개선용 조성물은 화장품학적으로, 식품적으로, 또는 약학적으로 허용가능한 부형제 또는 담체를 더 포함할 수 있다. 상기 담체는 부형제, 붕해제, 결합제, 활택제, 또는 그 조합일 수 있다. 상기 부형제는 미결정 셀룰로오즈, 유당, 저치환도 히드록시셀룰로오즈, 또는 그 조합일 수 있다. 상기 붕해제는 전분글리콜산 나트륨, 무수인산일수소 칼슘, 또는 그 조합일 수 있다. 상기 결합제는 폴리비닐피롤리돈, 저치환도 히드록시프로필셀룰로오즈, 히드록시프로필셀룰로오즈, 또는 그 조합일 수 있다. 상기 활택제는 스테아린산 마그네슘, 이산화규소, 탈크, 또는 그 조합일 수 있다.The composition for improving skin care according to one embodiment may further comprise a cosmetically, pharmaceutically or pharmaceutically acceptable excipient or carrier. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low substituted hydroxy cellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, calcium monohydrogen phosphate anhydrous, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or combinations thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.
상기 조성물은 비경구 투여 제형으로 제형화될 수 있다. 비경구 투여 제형은 주사제, 또는 피부외용제일 수 있다. 피부외용제는 크림, 겔, 연고, 피부 유화제, 피부 현탁액, 경피전달성 패치, 약물 함유 붕대, 로션, 또는 그 조합일 수 있다. The composition may be formulated into a parenteral dosage form. The parenteral dosage form may be an injection or an external preparation for skin. The external skin preparation may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, a lotion, or a combination thereof.
상기 피부외용제는 통상 화장품이나 의약품 등의 피부외용제에 사용되는 성분, 예를 들면 수성성분, 유성성분, 분말성분, 알코올류, 보습제, 증점제, 자외선흡수제, 미백제, 방부제, 산화방지제, 계면활성제, 향료, 색제, 각종 피부 영양제등을 필요에 따라서 적절하게 배합할 수 있다.The external preparation for skin is usually used as a component used in external skin preparations such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, an antiseptic, , Coloring agents, various skin nutrients, and the like can be appropriately blended as needed.
상기 피부외용제는, 에데트산이나트륨, 에데트산삼나트륨, 시트르산나트륨, 폴리인산나트륨, 메타인산나트륨, 글루콘산 등의 금속봉쇄제, 카페인, 탄닌, 벨라파밀, 감초추출물, 글라블리딘, 칼린의 과실의 열수추출물, 각종생약, 아세트산토코페롤, 글리틸리틴산, 트라넥삼산 및 그 유도체 또는 그 염등의 약제, 비타민 C, 아스코르브산인산마그네슘, 아스코르브산글루코시드, 알부틴, 코지산, 글루코스, 프룩토스, 트레할로스 등의 당류등도 적절하게 배합할 수 있다.The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, fructose, fructose and other herbal medicines, various herbal medicines, tocopherol acetate, glycyrrhizic acid, Sugars such as trehalose and the like can also be appropriately compounded.
일 구체예에 따른 조성물은 화장료 조성물일 수 있다. 이 경우, 배양액은 화장수(스킨로션), 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양 로션, 마사지크림, 영양 크림, 모이스쳐 크림, 핸드크림, 파운데이션, 에센스, 영양 에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션, 바디클렌저, 현탁액, 겔, 분말, 페이스트, 마스크팩 또는 시트 또는 에어로졸 조성물을 포함하는 제형으로 제조될 수 있다. 이러한 제형의 조성물은 당해 분야에서 통상적인 방법에 따라 제조될 수 있다. 상기 화장료 조성물은 보존제, 안정화제, 계면활성제, 용해제, 보습제, 에몰리언트제, 자외선 흡수제, 방부제, 살균제, 산화 방지제, pH 조정제, 유기 및 무기 안료, 향료, 냉감제 또는 제한제 등을 더 포함할 수 있다. 상기 보습제 등의 추가 성분의 배합량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 당업자가 용이하게 선정 가능하며, 그 배합량은 조성물 전체 중량을 기준으로 0.001 내지 5 중량%, 구체적으로 0.01 내지 3 중량%일 수 있다.The composition according to one embodiment may be a cosmetic composition. In this case, the culture solution can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizing cream, a hand cream, a foundation, , A soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, a suspension, a gel, a powder, a paste, a mask pack or a sheet or an aerosol composition. Compositions of such formulations may be prepared according to methods conventional in the art. The cosmetic composition may further contain preservatives, stabilizers, surfactants, solubilizers, moisturizers, emollients, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH regulators, organic and inorganic pigments, fragrances, have. The amount of the additional component such as the above-mentioned moisturizing agent and the like can be easily selected by those skilled in the art within a range not to impair the purpose and effect of the present invention. The amount of the additional component is 0.001 to 5% by weight, % ≪ / RTI >
일 구체예에 따른 조성물은 식품 조성물일 수 있다. 이 경우, 당해 기술분야에 공지되어 있는 통상적인 건강기능식품의 제형으로 제제화될 수 있다. 상기 식품 조성물은 예를 들어 산제, 과립제, 정제, 환제, 캅셀제, 현탁액, 유제, 시럽제, 침제, 액제, 엑스제 등의 일반적인 제형으로 제조될 수도 있고, 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 젤리, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등의 임의의 건강식품 형태로 제조될 수도 있다. 상기 건강식품의 제제화를 위해 식품학적으로 허용 가능한 담체 또는 첨가제를 사용할 수 있으며, 제조하고자 하는 제형의 제조에 당해 기술분야에서 사용 가능한 것으로 공지되어 있는 임의의 담체 또는 첨가제가 이용될 수 있다. 상기 첨가제로서 각종 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 이외에도 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 첨가제 성분은 독립적으로 또는 조합하여 사용할 수 있으며, 첨가제의 비율은 조성물 총 중량을 기준으로 0.001 내지 5 중량%, 구체적으로 0.01 내지 3 중량%일 수 있다.The composition according to one embodiment may be a food composition. In this case, it can be formulated into a conventional health functional food formulation known in the art. The food composition may be prepared by conventional formulations such as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, And may be manufactured in the form of any health food such as confectionery, pizza, ramen, other noodles, gums, jellies, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes. A pharmaceutically acceptable carrier or excipient may be used for the formulation of the health food, and any carrier or additive known to be usable in the art for the preparation of the formulation to be produced may be used. As the above-mentioned additives, there can be used various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Carbonating agents, and the like. In addition, it may contain natural fruit juice, fruit juice beverage and flesh for the production of vegetable beverages. These additive components may be used independently or in combination, and the proportion of the additive may be 0.001 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition.
상기 식품 조성물 중의 상기 배양액의 함량은 사용 목적(예방 또는 개선)에 따라 적합하게 결정될 수 있다. 일반적으로, 전체 식품 중량의 0.01 내지 15 중량%로 포함할 수 있으며, 음료로서 제조될 경우 100 mL를 기준으로 0.02 내지 10 g, 구체적으로 0.3 내지 1 g의 비율로 함유할 수 있다. The content of the culture medium in the food composition may be suitably determined according to the intended use (prevention or improvement). Generally, it may contain 0.01 to 15% by weight of the total food weight, and when it is prepared as a beverage, it may contain 0.02 to 10 g, specifically 0.3 to 1 g, based on 100 mL.
상기 음료는 상기 추출물 이외의 다른 성분을 더 포함할 수 있으며, 통상적으로 음료에 사용되는 다양한 향미제 또는 천연 탄수화물 등을 더 함유할 수 있다. 상기 천연 탄수화물로는 단당류(예: 포도당, 과당 등), 이당류(예: 말토즈, 수크로즈 등), 다당류(예: 덱스트린, 시클로덱스트린 등)와 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이 함유될 수 있다. 또한, 향미제로서 천연 향미제(예: 타우마틴, 스테비아 추출물 등) 및 합성 향미제(예: 사카린, 아스파탐 등)를 함유할 수 있다. 상기 천연 탄수화물의 비율은 음료 100 mL 당 일반적으로 약 1 내지 20g, 구체적으로 약 5 내지 12g으로 함유될 수 있다.The beverage may further contain ingredients other than the above extract, and may further contain various flavors or natural carbohydrates commonly used in beverages. Examples of the natural carbohydrate include conventional sugars such as monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugars such as xylitol, sorbitol, erythritol, Of sugar alcohols may be contained. In addition, as a flavoring agent, a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.) may be contained. The ratio of the natural carbohydrate may be generally about 1 to 20 g, specifically about 5 to 12 g per 100 mL of beverage.
일 구체예에 따른 조성물은 약학 조성물일 수 있다. 구체적으로, 상기 약학 조성물은 피부 질환을 예방 또는 치료하기 위한 약학 조성물일 수 있다. 상기 피부 질환은 피부 장벽 기능 손상에 의한 질환, 피부 노화, 피부 상처, 피부 흉터, 또는 피부 염증, 아토피 피부염일 수 있다. 용어, “예방”은 질병의 발생을 억제하는 것을 포함한다. 용어, “치료”는 질병의 발전의 억제, 경감, 또는 제거를 포함한다.The composition according to one embodiment may be a pharmaceutical composition. Specifically, the pharmaceutical composition may be a pharmaceutical composition for preventing or treating a skin disease. The skin disease may be a disease caused by skin barrier function impairment, skin aging, skin wounds, skin scarring or skin inflammation, or atopic dermatitis. The term " prevention " includes inhibiting the occurrence of a disease. The term " treatment " includes inhibiting, alleviating, or eliminating the development of the disease.
상기 피부 장벽 기능 손상은 피부 장벽의 기능이 저하되거나 손상되어 피부에 나타나는 모든 변화를 의미할 수 있다. 예를 들어, 피부 주름 증가, 건조, 피부염, 아토피 피부염, 알레르기성 피부염, 여드름 등을 포함할 수 있으나, 이에 한정되지 않는다. 일 구체예에 따른 약학 조성물은 필라그린의 발현을 증가시켜 피부 장벽 기능을 강화하여 피부 장벽 기능 손상을 예방 또는 치료할 수 있다.Damage to the skin barrier function may mean all the changes that appear on the skin due to the degradation or damage of the skin barrier function. For example, it may include, but is not limited to, increased wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne and the like. The pharmaceutical composition according to one embodiment of the present invention can enhance the expression of pillar green to enhance the skin barrier function, thereby preventing or treating damage to the skin barrier function.
상기 피부 노화는 나이가 들어가면서 피부에 나타나게 되는 유형과 무형상의 모든 변화를 의미할 수 있다. 예를 들어, 피부 주름 증가, 건조, 상처 치유 능력의 저하, 피부 면역 기능의 저하 등을 포함할 수 있으나, 이에 한정되지 않는다. 일 구체예에 따른 약학 조성물은 피부 장벽 강화, 피부 수분 유지, 피부 수분 손실 방지, 피부 세포 재생, 항산화 등의 효과에 의해 피부 노화를 예방 또는 치료할 수 있다.The aging of the skin can mean all types and intangible changes that appear on the skin as the age goes on. For example, it may include, but is not limited to, increased skin wrinkles, drying, decreased wound healing ability, decreased skin immune function, and the like. The pharmaceutical composition according to one embodiment can prevent or treat skin aging by the effects of strengthening skin barrier, maintaining skin moisture, preventing loss of skin moisture, regenerating skin cells, and antioxidation.
상기 피부 상처 또는 피부 흉터는 피부 세포가 재생, 분화, 증식되어 잃어버렸던 연속성을 다시 유지하거나, 반흔의 교원질 합성을 방해하고 교원질 분해를 촉진하여 개선 또는 치유될 수 있는 모든 질환을 의미할 수 있다. 예를 들어, 타박상, 찰과상, 열상 등의 상처, 비후성 반흔, 켈로이드 반흔 등의 흉터를 포함할 수 있으나, 이에 한정되지 않는다. 일 구체예에 따른 약학 조성물은 피부 수분 유지, 피부 수분 손실 방지, 피부 세포 재생, 항염증 등의 효과에 의해 피부 상처 또는 피부 흉터를 예방 또는 치료할 수 있다.The skin wound or skin scar may mean any disease that can be improved or remedied by maintaining the continuity of skin cells regenerated, differentiated, proliferating, interfering with the synthesis of scarring collagen and promoting collagen degradation. But are not limited to, scars such as bruises, abrasions, lacerations, scars such as hypertrophic scars, keloid scars, and the like. The pharmaceutical composition according to one embodiment can prevent or treat skin wounds or skin scars due to the effects of skin moisture retention, prevention of skin moisture loss, skin cell regeneration, and anti-inflammation.
상기 피부 염증 질환은 면역 자극에 의해 유발되는 모든 질환을 의미할 수 있다. 예를 들어, 아토피성 피부염, 습진, 지루성 피부염, 건선, 여드름 등을 포함할 수 있으나, 이에 한정되지 않는다. 일 구체예에 따른 약학 조성물은 피부 장벽 강화, 피부 수분 유지, 피부 수분 손실 방지, 항염증, 항아토피 등의 효과에 의해 피부 염증 질환을 예방 또는 치료할 수 있다.The skin inflammatory disease may mean all diseases caused by immune stimulation. But are not limited to, for example, atopic dermatitis, eczema, seborrheic dermatitis, psoriasis, acne, and the like. The pharmaceutical composition according to one embodiment of the present invention can prevent or treat skin inflammation diseases by enhancing skin barrier, maintaining skin moisture, preventing skin moisture loss, anti-inflammation, and anti-atopy.
다른 양상은 상기 조성물을 개체에게 투여하는 단계를 포함하는 개체의 피부미용을 개선시키는 방법을 제공한다. 상기 조성물에 대해서는 상술한 바와 동일하다. Another aspect provides a method of improving skin beauty of an individual comprising administering the composition to a subject. The composition is the same as described above.
상기 방법은 구체적으로, 개체의 피부 장벽 기능을 강화시키는 방법, 개체의 피부 수분을 유지 또는 수분 손실을 효율적으로 방지하는 방법, 개체의 피부 세포를 재생하는 방법, 개체의 피부 활력을 증가시키는 방법, 개체 내 항산화 작용을 증진시키는 방법, 개체의 염증을 개선 또는 치료하는 방법, 개체의 아토피 피부염을 개선, 치료 또는 예방하는 방법일 수 있다.Specifically, the method may include a method of enhancing a skin barrier function of an individual, a method of efficiently maintaining skin moisture or preventing moisture loss of an individual, a method of regenerating skin cells of an individual, a method of increasing skin vitality of an individual, A method of improving the antioxidant activity in an individual, a method of improving or treating inflammation of an individual, a method of improving, treating or preventing atopic dermatitis of an individual.
용어, "투여하는", "도입하는" 및 "이식하는"은 상호교환적으로 사용되고 일 구체예에 따른 조성물의 원하는 부위로의 적어도 부분적 국소화를 초래하는 방법 또는 경로에 의한 개체 내로의 일 구체예에 따른 조성물의 배치를 의미할 수 있다. 일 구체예에 따른 조성물의 배양액 또는 배양액 성분의 적어도 일부를 생존하는 개체 내에서 원하는 위치로 전달하는 임의의 적절한 경로에 의해 투여될 수 있다.The terms " administering ", " introducing " and " transplanting " are used interchangeably and refer to a method of delivering a composition according to one embodiment ≪ / RTI > may refer to the arrangement of the composition according to the invention. May be administered by any suitable route that delivers at least a portion of the culture broth or culture medium components of the composition according to one embodiment to the desired location within the living organism.
투여는 당업계에 알려진 방법에 의하여 투여될 수 있다. 투여는 예를 들면, 정맥내, 근육내, 경구, 경피 (transdermal), 점막, 코안 (intranasal), 기관내 (intratracheal) 또는 피하 투여와 같은 경로로, 임의의 수단에 의하여 개체로 직접적으로 투여될 수 있다. 상기 투여는 전신적으로 또는 국부적으로 투여될 수 있다.Administration can be by any method known in the art. Administration may be by any means directly administered to a subject, such as by intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration, . The administration can be systemically or locally administered.
상기 개체는 포유동물, 예를 들면, 사람, 소, 말, 돼지, 개, 양, 염소, 또는 고양이일 수 있다. 상기 개체는 피부미용 개선, 예를 들어 피부 장벽 강화, 피부 보습, 피부 세포 재생, 피부 활력 증가, 항산화, 항염증, 또는 항아토피 효과를 필요로 하는 개체일 수 있다.The subject may be a mammal, such as a person, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat. The subject may be an individual in need of skin aesthetic improvement, such as skin barrier enhancement, skin moisturization, skin cell regeneration, increased skin vitality, antioxidant, anti-inflammatory, or anti-atopic effect.
상기 투여는 후시디움 코시네움 균주 배양액을 개체당 일당 0.1 mg 내지 1,000 mg, 예를 들면, 0.1 mg 내지 500 mg, 0.1 mg 내지 100 mg, 0.1 mg 내지 50 mg, 0.1 mg 내지 25 mg, 1 mg 내지 1,000 mg, 1 mg 내지 500 mg, 1 mg 내지 100 mg, 1 mg 내지 50 mg, 1 mg 내지 25 mg, 5mg 내지 1,000 mg, 5 mg 내지 500 mg, 5 mg 내지 100 mg, 5 mg 내지 50 mg, 5 mg 내지 25 mg, 10mg 내지 1,000 mg, 10 mg 내지 500 mg, 10 mg 내지 100 mg, 10 mg 내지 50 mg, 또는 10 mg 내지 25 mg을 투여하는 것일 수 있다. 다만, 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성별, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있고, 당업자라면 이러한 요인들을 고려하여 투여량을 적절히 조절할 수 있다. 투여 횟수는 1일 1회 또는 임상적으로 용인가능한 부작용의 범위 내에서 2회 이상이 가능하고, 투여 부위에 대해서도 1개소 또는 2개소 이상에 투여할 수 있으며, 매일 또는 2 내지 5일 간격으로 총 투여 일수는 한번 치료 시 1일에서 30일까지 투여될 수 있다. 필요한 경우, 적정 시기 이후에 동일한 치료를 반복할 수 있다. 인간 이외의 동물에 대해서도, kg당 인간과 동일한 투여량으로 하거나, 또는 예를 들면 목적의 동물과 인간과의 기관(심장 등)의 용적비(예를 들면, 평균값) 등으로 상기의 투여량을 환산한 양을 투여할 수 있다.The administration may be carried out in a culture medium of the fusidium oxycinum strain in an amount of 0.1 mg to 1,000 mg, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg From 1 mg to 500 mg, from 1 mg to 100 mg, from 1 mg to 50 mg, from 1 mg to 25 mg, from 5 mg to 1000 mg, from 5 mg to 500 mg, from 5 mg to 100 mg, from 5 mg to 50 mg , 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dose may be variously prescribed depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and responsiveness of the patient, These factors can be taken into account to appropriately adjust the dosage. The number of doses may be at least once per day or within the range of clinically acceptable side effects and may be administered at one or two or more doses at the site of administration and may be administered daily or every two to five days The number of days of administration can be administered from one day to 30 days at one time of treatment. If necessary, the same treatment may be repeated after the appropriate time. For an animal other than a human, the dosage may be the same as that of a human per kg, or the dosage may be converted into a dose in terms of a volume ratio (for example, an average value) One dose may be administered.
다른 양상은 후시디움 코씨네움(Fusidium coccineum) 균주를 배양하는 단계; 배양 후 얻어진 균체 배양액을 원심분리하여 균체를 제거하고 상등액을 회수하는 단계; 및 회수한 상등액을 여과하여 잔존 균체를 제거하는 단계;를 포함하는 후시디움 코씨네움 균주 배양액을 제조하는 방법을 제공한다.Another aspect is Fusidium < RTI ID = 0.0 > coccineum ); Centrifuging the cell culture solution obtained after culturing to remove the cells and recovering the supernatant; And filtering the recovered supernatant to remove the remaining microbial cells. The present invention also provides a method for producing a fusidium coccinium culture.
상기 후시디움 코씨네움(F. coccineum) 균주에 대해서는 상술한 바와 동일하다.The above-mentioned F. coccineum strain is the same as described above.
일 구체예에 따른 방법에서, 상기 균주는 후시디움 코씨네움(Fusidium coccineum) 균주(수탁번호: KCCM12013P)일 수 있다.In a method according to one embodiment, the strain may be a strain of Fusidium coccineum (accession number: KCCM12013P).
상기 배양은 20 내지 35℃, 25 내지 30℃에서 12 내지 120시간, 24 내지 100시간, 48 내지 80시간 동안에서 수행될 수 있다.The culturing can be carried out at 20 to 35 DEG C, 25 to 30 DEG C for 12 to 120 hours, 24 to 100 hours, 48 to 80 hours.
상기 방법에 의해 제조된 후시디움 코씨네움 균주 배양액은 필라그린의 발현을 증가시켜 피부 장벽을 강화할 수 있고, 세라미드 합성효소 3, b-글루코세레브로시다제 또는 AQP3의 발현을 증가시켜 피부 보습 효과를 나타낼 수 있고, 세포 내 미토콘드리아 활성을 증가시켜 피부 세포 재생 또는 피부 활력 증가 효과를 나타낼 수 있고, 항산화 효과를 나타낼 수 있고, IL-1a, IL-6 또는 TSLP의 발현을 억제하여 항염증 또는 항아토피 효과를 나타낼 수 있다. 이러한 후시디움 코씨네움 균주 배양액의 피부 장벽 강화, 피부 보습, 피부 세포 재생, 피부 활력 증가, 항산화, 항염증 또는 항아토피 효과는 줄기세포 배양액에 비해 매우 우수하다. 따라서, 상기 방법에 의해 제조된 후시디움 코씨네움 균주 배양액은 화장품, 식품, 약학 조성물 등 다양한 용도로 이용될 수 있다.The fusidium coccinum culture prepared by the above method can enhance the expression of pillar green to enhance the skin barrier and increase the expression of
일 양상에 따른 신규한 후시디움 코씨네움(F. coccineum) 균주에 의하면, 균주의 다양한 활성을 응용하여 상기 균주를 피부 미용 개선 용도로 사용할 수 있다.According to a novel aspect of the present invention, a novel F. coccineum strain can be used for improving skin beauty by applying various activities of the strain.
다른 양상에 따른 피부미용 개선용 조성물에 의하면, 상기 후시디움 코씨네움(F. coccineum) 균주 배양액을 피부 장벽 강화, 피부 보습, 피부 세포 재생, 피부 활력 증가, 항산화, 항염증 또는 항아토피 용도로 사용할 수 있다.According to another aspect of the present invention, there is provided a composition for improving skin beauty, comprising the F. coccineum culture broth as a skin barrier enhancement, skin moisturizing, skin cell regeneration, skin vitality enhancement, antioxidant, anti- Can be used.
다른 양상에 따른 후시디움 코씨네움 균주 배양액을 제조하는 방법에 의하면, 피부미용 개선 효과가 있는 후시디움 코씨네움 균주 배양액을 제조할 수 있으므로, 제조된 배양액을 화장품, 식품, 약학 조성물 등 다양한 분야에 응용할 수 있다.According to another aspect of the present invention, there is provided a method for producing a fusidium coccinium culture, which comprises culturing the cultured Fusidium coccinium strain in a cosmetic, food, pharmaceutical composition, .
도 1은 필라그린(Filaggrin)의 mRNA 상대수준(relative level)을 나타낸 그래프이다. (-): 음성대조군인 무처리군. ** p<0.01 vs (-) control, * p<0.05 vs (-) control.
도 2는 bGC 및 CerS3의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.
도 3은 보습인자인 AQP3의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.
도 4는 항염증 효과를 확인한 결과로서, 사이토카인인 IL-1a의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.
도 5는 항염증 효과를 확인한 결과로서, 사이토카인인 IL-6의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.
도 6은 항아토피 효과를 확인한 결과로서, TSLP의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군. (+): 자극유도 실시군.
도 7은 세포 재생 및 활력 효과를 확인한 결과로서, 상대 세포 생존력 (무처리군대비%)을 나타낸 그래프이다. Control: 음성대조군인 무처리군.
도 8은 항산화력을 나타낸 그래프이다. (-): 음성대조군인 무처리군; AA: 양성대조군인 AA처리군; 후시디움배양액: 후시디움 코씨네움 배양액 1%(v/v) 처리군; 줄기세포배양액: 줄기세포 배양액 1%(v/v) 처리군. Figure 1 is a graph showing mRNA relative levels of filaggrin. (-): Negative control group. ** p <0.01 vs (-) control, * p <0.05 vs (-) control.
Figure 2 is a graph showing mRNA relative levels of bGC and CerS3. (-): Negative control group.
FIG. 3 is a graph showing the mRNA relative level of the moisturizing factor AQP3. (-): Negative control group.
FIG. 4 is a graph showing mRNA relative levels of cytokine IL-1 a as a result of confirming anti-inflammatory effects. (-): Negative control group.
FIG. 5 is a graph showing the relative level of mRNA of IL-6, a cytokine, as a result of confirming the anti-inflammatory effect. (-): Negative control group.
FIG. 6 is a graph showing mRNA relative levels of TSLP as a result of confirming the anti-atopic effect. (-): Negative control group. (+): Stimulation induction group.
FIG. 7 is a graph showing relative cell viability (% of untreated group) as a result of confirming cell regeneration and vitality effect. Control: Negative control group.
8 is a graph showing antioxidant activity. (-): negative control group, negative control group; AA: positive control group AA treatment group; Fusidium culture medium: treated with 1% (v / v) of fusidium cochineum culture; Stem cell culture:
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나, 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
실시예Example 1: One: 후시디움Fushidium 코씨네움Coincidence (( F. F. coccineumcoccineum )) 균주의 분리 및 동정Isolation and Identification of Strain
충북, 제천에서 수집한 건조된 은행잎을 멸균증류수 100ml에 넣고 1시간 동안 교반하여 미생물의 분리를 실시하였다. 교반이 완료된 샘플을 취하여 감자한천배지(제조사: Beckton Dickinson, 미국)에 접종하여 배양된 균사 콜로니 30종을 선별하였다. 선별된 균주는 ㈜마크로젠에 위탁하여 Fungi ITS 부분을 선택적으로 분석하여 NCBI Blast를 통해 기준 균주(Type strain)들과 상동성을 분석하였다. 분석결과 CXHB-8로 분리된 흰색 균사의 곰팡이가 후시디움 코씨네움(Fusidium coccineum)과 상동성이 98% 이였으며, 기존에 알려진 기준 균주와 표현 형질의 차이가 있는 것으로 확인되었다. 구체적으로, 기존에 알려진 후시디움 코씨네움 표준균주는 백색의 집락을 형성하며 생장하지만, 상기 분리한 균주는 미황색 집락을 형성하였다. 또한, 기존의 후시디움 코씨네움 표준균주는 영양조건이 까다롭지 않은 반면, 상기 분리한 균주는 감자한천배지 외에서는 성장하지 않거나 성장이 어려운 것을 확인할 수 있었다. 따라서, 분리된 균주를 후시디움 코씨네움 CX-3FU로 명명하였으며, 이를 한국 미생물 보존센터에 2017년 4월 14일자로 기탁하고, 기탁번호 KCCM12013P를 부여받았다. Dried ginkgo leaves collected from Chungbuk and Jecheon were added to 100 ml of sterilized distilled water and stirred for 1 hour to isolate the microorganisms. The stirred sample was taken and 30 mycelial colonies cultured in potato agar medium (manufactured by Beckton Dickinson, USA) were selected. The selected strains were consigned to Macrogen Co., Ltd., and the Fungi ITS part was selectively analyzed and the homology with the reference strains was analyzed through NCBI Blast. As a result, it was confirmed that the fungus of white mycelia separated by CXHB-8 had a homology of 98% with Fusidium coccineum . Specifically, the previously known Fushidium cochineum standard strain grows to form a white colonies, but the isolated strain forms a pale yellow colon. In addition, while the conventional Fushidium cochinum strain is not suited to nutritional conditions, it can be confirmed that the isolated strain does not grow or grow outside the potato agar medium. Thus, the isolated strain was named Fushidium coccumenum CX-3FU, deposited at the Korean Microorganism Conservation Center on Apr. 14, 2017, and assigned the deposit number KCCM12013P.
실시예Example 2: 2: 후시디움Fushidium 코씨네움Coincidence (( F. F. coccineumcoccineum ) 균주 배양액의 제조) Preparation of culture broth
상기 실시예 1에서 분리된 후시디움 코씨네움(F. coccineum)의 배양액을 제조하였다.A culture solution of F. coccineum isolated in Example 1 was prepared.
구체적으로, 분리된 후시디움 코시네움 CX-3FU는 감자한천배지에서 최적의 생육환경을 보이는 것으로 확인되었다. 감자한천배지는 5g을 멸균증류수 100ml과 혼합하여 121℃ 15분간 멸균하여 사용하였다. 멸균 후 25℃로 냉각하고, 분리된 CX-3FU 균체 1g을 접종하였다. 접종된 배지는 28℃ 항온기에서 100rpm으로 교반하여 배양을 실시하였고, 배양기간은 7일이었으며, 7일 이후 0.2μm 필터를 통과시켜 필터멸균하여 후시디움 코씨네움의 배양액으로 사용하였다. Specifically, it was confirmed that the isolated fusidium cochineum CX-3FU showed optimal growth environment in the potato agar medium. 5 g of potato agar medium was mixed with 100 ml of sterilized distilled water and sterilized at 121 ° C for 15 minutes. After sterilization, the mixture was cooled to 25 DEG C and 1 g of the separated CX-3FU cells was inoculated. The inoculated medium was incubated at 28 ° C with stirring at 100 rpm and cultured for 7 days. After 7 days, the cells were sterilized by passing through a 0.2 μm filter and used as a culture medium of fusidium coccinium.
비교예Comparative Example 1: 줄기세포 배양액의 제조 1: Preparation of stem cell culture liquid
인간 유래 줄기세포 배양액을 제조하였다. 구체적으로, 인간 유래 줄기세포를 ATCC(Rockville, MD, USA)에서 구매하였으며, 10%의 소태아혈청(FBS, ATCC, Rockville, MD, USA)과 1%의 항균/항진균 용액(Antibiotic/Antimycotic: AA, ATCC, Rockville, MD, USA)을 첨가한 DMEM (HyClone, Logan, UT, USA)을 이용하여 37, 5%, CO2 조건의 인큐베이터에서 배양하였다. 배양액은 원심분리를 실시하여 세포를 제거 후 상층 배양액만 사용하였다.Human stem cell culture was prepared. Human-derived stem cells were purchased from ATCC (Rockville, MD, USA) and cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS, ATCC, Rockville, MD, USA) and 1% Antibiotic / Antimycotic solution using the AA, ATCC, Rockville, MD, USA), a DMEM (HyClone, Logan, UT, USA was added) 37, 5%, and cultured in an incubator of CO 2 conditions. The culture was centrifuged to remove the cells and only the upper layer culture was used.
실험 방법 1: Experimental Method 1: mRNAmRNA 발현 조사(실시간 RT- Expression studies (RT- PCRPCR ))
사람 각질형성세포주인 HaCaT를 6-웰 플레이트에 5×105 세포/웰 접종하여 10%의 FBS와 1%의 AA가 첨가된 배지를 이용하여 24시간 배양하였다. 그 후 후시디움 코씨네움 배양액 또는 줄기세포 배양액, Poly I:C, 및 IL-4를 FBS가 첨가되지 않은 배지에 24 시간 동안 추가 배양하였다. 전체 RNA를 추출하기 위해 각 웰에 RNA iso (Takara Bio Inc., Otsu, Japan) 1 mL을 첨가하여 세포를 용해시키고 클로로포름(Sigma-Aldrich, St. Louis, MO) 200 μL를 첨가하여 14,000 rpm에서 10분 동안 원심분리 하였다. 상층액을 취하여 새로운 튜브에 옮기고 같은 양의 이소프로판올 (Sigma-Aldrich, St. Louis, MO)을 첨가한 후 14,000 rpm에서 10분 원심 분리하여 RNA를 분리하였다. 99% 에탄올 (Sigma-Aldrich, St. Louis, MO)을 이용하여 7,500 rpm에서 원심 분리하여 2회 세척하고 건조시킨 후 디에틸피로카보네이트(DEPC)-water (Ambion, Austin, TX, USA)에 녹였다. Nanodrop 2000 (Thermo Fisher Scientific, Waltham, MA, USA)를 이용해 RNA를 정량 하였고, 전체 RNA 2 μg을 DEPC와 함께 70℃에서 5분 동안 가열시킨 후, 역전사 프리믹스(ELPIS-Biotech, Daejeon, Korea)에 넣고 최종 부피가 20 μL가 되도록 하였다. 42℃에서 55분, 70℃에서 15분 동안 반응시켜 cDNA를 합성하여 중합효소연쇄반응(PCR)에 사용하였다. 얻어진 cDNA로부터 유전자들을 증폭시키기 위해 5배 희석시킨 cDNA 2 μL를 프라이머 1 μL, DEPC 7 μL, SYBR 그린 마스터 믹스(Life Technologies, Grand Island, NY, USA) 10 μL와 함께 StepOne Plus RT-PCR 시스템(Applied Biosystems, Foster City, CA, USA)을 이용하여 PCR을 실시하였다. 반응 조건은 50℃ 2분, 95℃ 10분에서 반응 후 95℃ 10초와 60℃ 1분을 40회 반복하여 증폭시켰다. 표 1에 증폭하고자 하는 유전자들의 프라이머 염기서열을 기재하였다.HaCaT, a human keratinocyte cell line, was inoculated to a 6-well plate at 5 × 10 5 cells / well and cultured for 24 hours using a medium supplemented with 10% FBS and 1% AA. Then, Fosmidium cochineum culture or stem cell culture solution, Poly I: C, and IL-4 were further cultured in a medium to which no FBS was added for 24 hours. To extract total RNA, 1 mL of RNA iso (Takara Bio Inc., Otsu, Japan) was added to each well to dissolve the cells and 200 μL of chloroform (Sigma-Aldrich, St. Louis, MO) And centrifuged for 10 minutes. The supernatant was transferred to a new tube and the same amount of isopropanol (Sigma-Aldrich, St. Louis, MO) was added and the RNA was isolated by centrifugation at 14,000 rpm for 10 minutes. Centrifuged at 7,500 rpm using 99% ethanol (Sigma-Aldrich, St. Louis, MO), washed twice, dried and dissolved in diethyl pyrocarbonate (DEPC) -water (Ambion, Austin, TX, USA) . RNA was quantitated using Nanodrop 2000 (Thermo Fisher Scientific, Waltham, Mass., USA) and 2 μg of total RNA was heated with DEPC at 70 ° C for 5 minutes, then transferred to a reverse transcription primer (ELPIS-Biotech, Daejeon, Korea) To a final volume of 20 μL. The reaction was carried out at 42 ° C for 55 minutes and at 70 ° C for 15 minutes, and cDNA was synthesized and used for PCR. To amplify genes from the resulting cDNA, add 2 μL of 5-fold diluted cDNA to the StepOne Plus RT-PCR system (10 μL) with 1 μL of primer, 7 μL of DEPC, and 10 μL of SYBR Green Master Mix (Life Technologies, Grand Island, Applied Biosystems, Foster City, CA, USA). The reaction was carried out at 50 ° C for 2 minutes and at 95 ° C for 10 minutes, followed by 40 cycles of 95 ° C for 10 seconds and 60 ° C for 1 minute. Table 1 shows the primer sequences of the genes to be amplified.
번호number
β-Actin
실험 방법 3: 자료분석 및 통계처리Experiment method 3: Data analysis and statistical processing
세포 분석에 대한 그룹간 통계는 스튜던트티이검정(student's t-test)을 사용하였다. 두군 간의 유의적 차이에 의한 통계적 검정 후 (-) 대비 *P < 0.05, **P < 0.01, (+)대조군 대비 #P < 0.05, ##P < 0.01 로 표현하였다.Group-to-group statistics for cell analysis were Student's t-test. (P <0.05), ** P <0.01, and (+) # P <0.05 and P <0.01 compared to the control group, respectively.
실험 방법 4: 세포증식 Experimental Method 4: Cell proliferation 촉진능Acceleration ability 평가 evaluation
실시예와 비교예의 세포증식 촉진능 비교를 위해 인간 섬유아세포주인 Hs68 섬유아세포에서 MTT(3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) 시약을 이용한 측정법으로 실험을 수행하였다.In order to compare the cell proliferation promoting activities of the examples and the comparative examples, experiments were carried out with Hs68 fibroblasts, a human fibroblast, using MTT (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyltetrazolium bromide) reagent .
인간 섬유아세포주인 Hs68 섬유아세포를 2×104 세포/웰 농도로 96-웰 플레이트에 분주한 후, 37℃, 5% CO2 조건의 배양기에서 24시간 동안 배양하였다. 이후 실시예와 비교예의 샘플을 각각 10㎍/m의 농도로 첨가하는 배지로 세포배양액을 교체해 준 다음 24시간 동안 추가로 배양하였다. 배양 후 각 웰의 배지를 제거하고 DPBS 세척 과정을 1회 거친 다음 0.5mg/mL 농도의 MTT 시약을 첨가하여 37℃에서 4시간 동안 반응시켰다. 반응과정에서 형성된 포르마잔 크리스탈(formazan crystal)을 MTT 시약을 제거한 다음 DMSO(dimethyl sulfoxide)로 용해시킨 다음 분광광도계를 이용하여 590nm에서 흡광도를 측정하였다. 세포 생존율은 샘플 없이 배지만 첨가한 대조군의 평균 흡광도 값에 대한 백분율로 나타내었다.Human fibroblast Hs68 fibroblasts were divided into 96-well plates at a concentration of 2 × 10 4 cells / well and cultured in an incubator at 37 ° C. and 5% CO 2 for 24 hours. Thereafter, the cell culture medium was replaced with a medium containing 10 μg /
실험예Experimental Example 1: 피부 장벽 강화 효과 확인 1: Confirm skin barrier strengthening effect
후시디움 코씨네움(F. coccineum) 배양액의 피부 장벽 강화 효과를 확인하기 위한 실험을 하였다.Experiments were conducted to confirm the skin barrier strengthening effect of F. coccineum culture.
구체적으로, 상기 실험 방법 1에 따라, 사람 각질형성세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, 필라그린의 mRNA 상대수준을 확인하였다.Specifically, in accordance with
도 1은 필라그린(Filaggrin)의 mRNA 상대수준(relative level)을 나타낸 그래프이다. (-): 음성대조군인 무처리군. ** p<0.01 vs (-) control, * p<0.05 vs (-) control.Figure 1 is a graph showing mRNA relative levels of filaggrin. (-): Negative control group. ** p <0.01 vs (-) control, * p <0.05 vs (-) control.
도 1에 나타낸 바와 같이, 피부장벽 구성에 중요한 인자인 필라그린의 발현율을 RT-PCR로 분석한 결과, 후시디움 코씨네움 배양액을 처리하면 무처리군 대비 약 8배(후시디움 코씨네움 배양액 1% 처리), 약 10배(후시디움 코씨네움 배양액 5% 처리) 증가하는 것으로 확인되었다. 또한, 비교예의 줄기세포 배양액보다 8배 이상 발현율이 차이가 남을 확인할 수 있었다. 따라서, 후시디움 코씨네움 배양액은 각질형성세포 분화를 일으켜 피부 장벽 개선을 도와주는 필라그린의 발현을 증가시키므로, 피부 장벽 강화 효과가 매우 우수함을 알 수 있었다. As shown in FIG. 1, when the expression ratio of filaggrin, which is an important factor in constituting the skin barrier, was analyzed by RT-PCR, it was found that when the Fosmidomocinum culture medium was treated, it was about 8 times that of the
실험예Experimental Example 2: 피부 보습 효과 확인 2: Confirm skin moisturizing effect
후시디움 코씨네움(F. coccineum) 배양액의 피부 보습 효과를 확인하기 위한 실험을 하였다.Experiments were conducted to confirm skin moisturizing effect of F. coccineum culture.
2-1. 2-1. 보습인자Moisturizing factor CerS3CerS3 및 And bGC의bGC's 발현 증가 효과 확인 Confirmation of increased expression
후시디움 코씨네움 배양액에 의해 보습인자인 세라미드 합성효소 3 (Ceramide synthase 3: CerS3) 및 b-글루코세레브로시다제 (b-Glucocerebrosidase: bGC) 의 발현이 증가하는지 확인하기 위한 실험을 하였다.(Ceramide synthase 3: CerS3) and b-glucocerebrosidase (bGC) were increased by the culture medium of Fosmidium cocinum.
구체적으로, 상기 실험 방법 1에 따라, 사람 각질형성세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, bGC 및 CerS3의 mRNA 상대수준을 확인하였다.Specifically, in accordance with
도 2는 bGC 및 CerS3의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.Figure 2 is a graph showing mRNA relative levels of bGC and CerS3. (-): Negative control group.
도 2에 나타낸 바와 같이, 피부 보습에 중요한 인자인 bGC, CerS3 유전자의 발현율을 측정하기 위해 RT-PCR로 분석한 결과, 후시디움 코씨네움 배양액을 처리하면 무처리군 대비 약 2배(후시디움 코씨네움 배양액 1% 처리) 증가하는 것으로 확인 되었다. 그러나 처리농도가 증가할 수록 보습인자의 발현율이 다소 감소되는 경향을 보이는 것으로 확인되었다. 또한, 비교예의 줄기세포 배양액 대비 발현율이 2배 가량 좋은 것으로 확인되었다.As shown in Fig. 2, when RT-PCR was performed to measure the expression ratios of bGC and CerS3 genes, which are important factors for skin moisturization, it was found that when Fusiodium kosineum culture medium was treated, And 1% treatment of coccinium cultures). However, as the concentration of the treatment increased, the expression rate of the moisturizing factor tended to decrease. In addition, it was confirmed that the expression ratio of the stem cell culture fluid of the comparative example was about twice as good.
2-2. 2-2. 보습인자Moisturizing factor AQP3의AQP3 발현 증가 효과 확인 Confirmation of increased expression
후시디움 코시네움 배양액에 의해 보습인자인 아쿠아포린 3 (Aquaporin 3: AQP3)의 발현이 증가하는지 확인하기 위한 실험을 하였다.Experiments were conducted to determine if the expression of Aquaporin 3 (AQP3), which is a moisturizing factor, is increased by the fucoidiumcocineum culture.
구체적으로, 상기 실험 방법 1에 따라, 사람 각질형성세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, AQP3의 mRNA 상대수준을 확인하였다. 양성대조군으로 레티놀 1μM을 사용하였다.Specifically, in accordance with
도 3은 보습인자인 AQP3의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.FIG. 3 is a graph showing the mRNA relative level of the moisturizing factor AQP3. (-): Negative control group.
도 3에 나타낸 바와 같이, 피부 보습에 중요한 인자인 AQP3 유전자의 발현율을 측정하기 위해 RT-PCR로 분석한 결과, 후시디움 코씨네움 배양액을 처리하면 무처리군 대비 약 1.5배(후시디움 코씨네움 배양액 1% 처리) 증가하는 것으로 확인 되었다. AQP3 의 경우 고농도로 처리되었을 때 발현율이 감소되었으므로, 최적 발현 농도는 후시디움 코씨네움 배양액 1%로 확인되었다. As shown in FIG. 3, in order to measure the expression ratio of AQP3 gene, which is an important factor in skin moisturization, by RT-PCR, it was found that when Fusidium coccinium cultures were treated, about 1.5
실험예Experimental Example 3: 항염증 및 3: Anti-inflammatory and 항아토피Anti-atopic 효과 확인 Check the effect
후시디움 코씨네움(F. coccineum) 배양액의 항염증 및 항아토피 효과를 확인하기 위한 실험을 하였다.Experiments were carried out to confirm the anti-inflammatory and anti-atopic effects of F. coccineum culture.
3-1. 염증 인자 IL-1a 및 IL-6의 억제 효과 확인3-1. Identification of inhibitory effects of inflammatory factors IL-1a and IL-6
후시디움 코시네움 배양액에 의해 염증 인자로서 중요한 사이토카인으로 알려진 IL-1a와 IL-6 유전자의 발현이 억제되는지 확인하기 위한 실험을 하였다.Experiments were performed to confirm the inhibition of the expression of IL-1α and IL-6 genes, which are known to be important cytokines as inflammatory factors, by the culture medium of fosmidium coccineum.
구체적으로, 상기 실험 방법 1에 따라, 사람 각질형성세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, IL-1a 및 IL-6의 mRNA 상대수준을 확인하였다. 양성대조군으로 덱사메타손 (Dexamethasone: Dex) 1μM을 사용하였다.Specifically, in accordance with
도 4는 항염증 효과를 확인한 결과로서, 사이토카인인 IL-1a의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.FIG. 4 is a graph showing mRNA relative levels of cytokine IL-1 a as a result of confirming anti-inflammatory effects. (-): Negative control group.
도 5는 항염증 효과를 확인한 결과로서, 사이토카인인 IL-6의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군.FIG. 5 is a graph showing the relative level of mRNA of IL-6, a cytokine, as a result of confirming the anti-inflammatory effect. (-): Negative control group.
도 4 및 도 5에 나타낸 바와 같이, 후시디움 코씨네움 배양액은 양성대조군인 덱사메타손 보다 IL-1a 및 IL-6 억제율이 약 2배 가량 높은 것으로 확인되었다. 한편, IL-6의 경우 후시디움 코씨네움 배양액 5% 농도보다, 1% 농도에서 억제율이 뛰어난 것으로 확인되었다. 따라서, 후시디움 코씨네움 배양액은 항염증 효과가 우수함을 알 수 있었다.As shown in FIG. 4 and FIG. 5, it was confirmed that the fusidium cochineal culture supernatant was about twice as high as that of the positive control group, dexamethasone, for the inhibition of IL-1 a and IL-6. On the other hand, it was confirmed that IL-6 had a higher inhibition rate at a concentration of 1% than the concentration of 5% concentration of Fosmidium cochineum. Therefore, it was found that the fusidium cochineum culture medium had an excellent anti-inflammatory effect.
3-2. 가려움증 인자 3-2. Itching factor TSLP의TSLP's 억제 효과 확인 Confirmation of inhibition
아토피의 발생 시 소양증이 발생되는데, 이때 TSLP(Thymic Stromal Lymphopoietin) 인자가 중요한 인자로 작용한다. 후시디움 코시네움 배양액에 의한 TSLP의 억제율을 측정하기 위해 RT-PCR로 확인하였다.Atopic dermatitis occurs when pruritus occurs, in which TSLP (Thymic Stromal Lymphopoietin) factor plays an important role. RT-PCR was performed to determine the inhibition rate of TSLP by the fusidium oxycinum culture.
구체적으로, 상기 실험 방법 1에 따라, 사람 각질형성세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, TSLP의 mRNA 상대수준을 확인하였다. 양성대조군으로 덱사메타손 (Dexamethasone: Dex) 1μM을 사용하였다.Specifically, in accordance with
도 6은 항아토피 효과를 확인한 결과로서, TSLP의 mRNA 상대수준을 나타낸 그래프이다. (-): 음성대조군인 무처리군. (+): 자극유도 실시군.FIG. 6 is a graph showing mRNA relative levels of TSLP as a result of confirming the anti-atopic effect. (-): Negative control group. (+): Stimulation induction group.
도 6에 나타낸 바와 같이, 후시디움 코씨네움 배양액의 경우 TSLP의 발현 억제 효과가 양성대조군인 덱사메타손과 유사한 수준으로 확인되었다. 따라서, 후시디움 코씨네움 배양액은 항아토피 효과가 우수함을 알 수 있었다.As shown in FIG. 6, in the case of Fosmidomocinum culture solution, the inhibitory effect of TSLP expression was confirmed to be similar to that of the positive control dexamethasone. Therefore, it was found that the fusidium cochineum culture medium had an excellent anti-atopic effect.
실험예Experimental Example 4: 세포 증식 효과 확인 4: Confirm cell proliferation effect
후시디움 코씨네움 배양액에 의한 세포의 재생과 활력을 측정하기 위한 실험을 하였다.Experiments were carried out to measure the regeneration and vitality of the cells by the Fosmidium cochineum culture.
구체적으로, 상기 실험 방법 4에 따라, 섬유아세포에 실시예의 후시디움 코씨네움 배양액을 1% 또는 5%(v/v) 처리하거나, 비교예의 줄기세포 배양액을 1% 또는 5%(v/v) 처리한 후, 상대 세포 생존력을 확인하였다.Specifically, in accordance with
도 7은 세포 재생 및 활력 효과를 확인한 결과로서, 상대 세포 생존력 (무처리군대비%)을 나타낸 그래프이다. Control: 음성대조군인 무처리군.FIG. 7 is a graph showing relative cell viability (% of untreated group) as a result of confirming cell regeneration and vitality effect. Control: Negative control group.
도 7에 나타낸 바와 같이, 후시디움 코씨네움 배양액은 무처리군 대비 약 20% 이상의 세포증식 효과가 확인되었으며, 그 효과가 농도 의존적으로 증가하는 것을 확인하였다.As shown in FIG. 7, the Fusidium cochineum culture solution showed a cell proliferation effect of about 20% or more as compared with the untreated group, and the effect thereof was confirmed to increase in a concentration-dependent manner.
실험예Experimental Example 5: 항산화 효과 확인 5: Confirmation of antioxidant effect
후시디움 코씨네움 배양액의 항산화 효과를 확인하기 위한 실험을 하였다.Experiments were carried out to confirm the antioxidant effect of Fosmidium cochineum.
구체적으로, 항산화 테스트는 라디칼 소거활성이 있는 항산화제에 의해 정량적으로 탈색되는 DPPH 용액을 이용하여 항산화 활성을 측정하는 방법으로 수행하였다. 먼저, 0.1 mM의 DPPH/80% MeOH를 준비하였다. 다음으로, 0.05 ml의 시료와 아스코르브산(ascorbic acid)을 2.95 ml의 DPPH 용액에 넣었다. 볼텍싱하여 혼합한 후, 실온에서 30분간 암실에서 방치하였다. 517 nm에서 UV를 측정하였다.Specifically, the antioxidant test was carried out by measuring antioxidative activity using a DPPH solution which was quantitatively discolored by an antioxidant having a radical scavenging activity. First, 0.1 mM DPPH / 80% MeOH was prepared. Next, 0.05 ml of sample and ascorbic acid were added to 2.95 ml of DPPH solution. The mixture was vortexed and left in a dark room at room temperature for 30 minutes. UV was measured at 517 nm.
도 8은 항산화력을 나타낸 그래프이다. (-): 음성대조군인 무처리군; AA: 양성대조군인 AA처리군; 후시디움배양액: 후시디움 코씨네움 배양액 1%(v/v) 처리군; 줄기세포배양액: 줄기세포 배양액 1%(v/v) 처리군. 8 is a graph showing antioxidant activity. (-): negative control group, negative control group; AA: positive control group AA treatment group; Fusidium culture medium: treated with 1% (v / v) of fusidium cochineum culture; Stem cell culture:
도 8에 나타낸 바와 같이, 후시디움 코씨네움 배양액은 무처리군 대비 14%의 항산화력을 나타냈고, 줄기세포 배양액은 항산화력이 무처리군 보다 떨어지는 것으로 확인되었다.As shown in FIG. 8, the Fusidium cochineum culture showed an antioxidative power of 14% as compared to the untreated group, and the antioxidative capacity of the stem cell culture solution was found to be lower than that of the untreated group.
<110> Cosmax Co., Ltd. <120> Novel strain of Fusidium coccineum spp., and composition for improving skin beauty comprising a culture solution of the strain <130> PN117190 <160> 26 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Filaggrin-F primer <400> 1 agtgcactca gggggctcac a 21 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Filaggrin-R primer <400> 2 ccggcttggc cgtaatgtgt 20 <210> 3 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Claudin 1-F primer <400> 3 gctctagaat tctcacacgt agtctttccc gct 33 <210> 4 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Claudin 1-R primer <400> 4 gctctagaat tctcacacgt agtctttccc gct 33 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Claudin 4-F primer <400> 5 actttgataa ctgctcctct gac 23 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Claudin 4-R primer <400> 6 ttcgtgtcca gcagagtacc 20 <210> 7 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Alpha-Sphingomyelinase-F primer <400> 7 actttgataa ctgctcctct gac 23 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Alpha-Sphingomyelinase-R primer <400> 8 ttcgtgtcca gcagagtacc 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Ceramide synthase 3-F primer <400> 9 acattccaca aggcaaccat tg 22 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Ceramide synthase 3-R primer <400> 10 ctcttgattc cgccgactcc 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> b-Glucocerebrosidase-F primer <400> 11 acctacactc tctggggacc 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> b-Glucocerebrosidase-R primer <400> 12 ttcagcccac ttcccagacc 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronan synthase 3-F primer <400> 13 cttaagggtt gcttgcttgc 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronan synthase 3-R primer <400> 14 gttcgtggga gatgaaggaa 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin 3-F primer <400> 15 agacagcccc ttcaggattt 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin 3-R primer <400> 16 tcccttgccc tgaatatctg 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-6-F primer <400> 17 tacccccagg agaagattcc 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-6-R primer <400> 18 ttttctgcca gtgcctcttt 20 <210> 19 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha-F primer <400> 19 gctatctggt gcccaggcta t 21 <210> 20 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha-R primer <400> 20 cgacgccaca atccttgtaa t 21 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP-F primer <400> 21 gctatctggt gcccaggcta t 21 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP-R primer <400> 22 cgacgccaca atccttgtaa t 21 <210> 23 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> TARC-F primer <400> 23 cttctctgca gcacatcc 18 <210> 24 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TARC-R primer <400> 24 aagacctctc aaggctttg 19 <210> 25 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Beta-Actin-F primer <400> 25 ggccatctct tgctcgaagt 20 <210> 26 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Beta-Actin-R primer <400> 26 gacaccttca acaccccagc 20 <110> Cosmax Co., Ltd. <120> Novel strain of Fusidium coccineum spp., And composition for improving skin beauty comprising a culture solution of the strain <130> PN117190 <160> 26 <170> KoPatentin 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Filaggrin-F primer <400> 1 agtgcactca gggggctcac a 21 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Filaggrin-R primer <400> 2 ccggcttggc cgtaatgtgt 20 <210> 3 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Claudin 1-F primer <400> 3 gctctagaat tctcacacgt agtctttccc gct 33 <210> 4 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Claudin 1-R primer <400> 4 gctctagaat tctcacacgt agtctttccc gct 33 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Claudin 4-F primer <400> 5 actttgataa ctgctcctct gac 23 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Claudin 4-R primer <400> 6 ttcgtgtcca gcagagtacc 20 <210> 7 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Alpha-Sphingomyelinase-F primer <400> 7 actttgataa ctgctcctct gac 23 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Alpha-Sphingomyelinase-R primer <400> 8 ttcgtgtcca gcagagtacc 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Ceramide synthase 3-F primer <400> 9 acattccaca aggcaaccat tg 22 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Ceramide synthase 3-R primer <400> 10 ctcttgattc cgccgactcc 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> b-Glucocerebrosidase-F primer <400> 11 acctacactc tctggggacc 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> b-Glucocerebrosidase-R primer <400> 12 ttcagcccac ttcccagacc 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronan synthase 3-F primer <400> 13 cttaagggtt gcttgcttgc 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronan synthase 3-R primer <400> 14 gttcgtggga gatgaaggaa 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin 3-F primer <400> 15 agacagcccc ttcaggattt 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin 3-R primer <400> 16 tcccttgccc tgaatatctg 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-6-F primer <400> 17 tacccccagg agaagattcc 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> IL-6-R primer <400> 18 ttttctgcca gtgcctcttt 20 <210> 19 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha-F primer <400> 19 gctatctggt gcccaggcta t 21 <210> 20 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha-R primer <400> 20 cgacgccaca atccttgtaa t 21 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP-F primer <400> 21 gctatctggt gcccaggcta t 21 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP-R primer <400> 22 cgacgccaca atccttgtaa t 21 <210> 23 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> TARC-F primer <400> 23 cttctctgca gcacatcc 18 <210> 24 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TARC-R primer <400> 24 aagacctctc aaggctttg 19 <210> 25 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Beta-Actin-F primer <400> 25 ggccatctct tgctcgaagt 20 <210> 26 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Beta-Actin-R primer <400> 26 gacaccttca acaccccagc 20
Claims (15)
배양 후 얻어진 균체 배양액을 원심분리하여 균체를 제거하고 상등액을 회수하는 단계; 및
회수한 상등액을 여과하여 잔존 균체를 제거하는 단계;를 포함하는 후시디움 코씨네움 균주 배양액을 제조하는 방법. Fusidium coccineum );
Centrifuging the cell culture solution obtained after culturing to remove the cells and recovering the supernatant; And
And filtering the recovered supernatant to remove the remaining microbial cells.
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