KR20190016253A - Fermentation method of lactic acid bacteria using berry and its use - Google Patents
Fermentation method of lactic acid bacteria using berry and its use Download PDFInfo
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- KR20190016253A KR20190016253A KR1020170100235A KR20170100235A KR20190016253A KR 20190016253 A KR20190016253 A KR 20190016253A KR 1020170100235 A KR1020170100235 A KR 1020170100235A KR 20170100235 A KR20170100235 A KR 20170100235A KR 20190016253 A KR20190016253 A KR 20190016253A
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- berry
- skin
- cbg
- berries
- fermented product
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
Description
본 발명은 베리를 이용한 유산균 발효 방법 및 이의 용도에 관한 것이다. The present invention relates to a fermentation method of lactic acid bacteria using berry and its use.
피부는 외부를 덮고 있는 기관으로 바깥쪽에서부터 표피, 진피 및 피하지방층의 독특한 세 개의 층으로 구성되어 있다. 표피는 중층편평상피(stratified squamous epithelium)의 각질형성세포가 대부분을 차지하고 있으며, 콜라겐 섬유와 탄력 섬유와 같은 기질 단백질로 이루어진 진피는 표피 아래에 위치하며, 여기에 혈관, 신경, 땀샘 등이 있다. 피하지방층은 지방세포로 구성되어 있다. 상기 구조로 이루어진 피부는 체온 조절과 다양한 외부 자극으로부터 몸을 보호함과 동시에 내측의 수분을 막는 장벽 기능이 있다. 이를 위하여, 피부의 표피층에서는 분화되는 과정을 통하여 수분을 함유하면서 얇은 각질층(stratum corneum)으로 발달하게 된다.The skin is an organs covering the outside, consisting of three distinct layers of skin from the outside, the epidermis and the subcutaneous fat layer. The epidermis is mainly composed of stratified squamous epithelium keratinocytes. The dermis, composed of matrix proteins such as collagen fibers and elastic fibers, is located under the epidermis, and there are blood vessels, nerves, and sweat glands. The subcutaneous fat layer is composed of adipocytes. The skin composed of the above-mentioned structure has a barrier function to protect the body from body temperature control and various external stimuli while blocking the inner moisture. To this end, the epidermis of the skin develops into a thin stratum corneum containing water through differentiation.
따라서 피부 보습의 정도는 화장품, 피부 과학 및 의약품 등의 분야에서 중요한 요소로 인식되고 있다. 즉, 각질층의 보습상태는 피부의 외형(피부의 유연성 및 거친 정도)과 직접적으로 연관되어 있으며, 건강상태의 척도가 된다. 또한, 각질층의 피부 수분 함량은 피부의 컨디션 및 투과성과 관련되는 피부 막 항상성에 영향을 미치므로, 화장품 또는 바르는 의약품의 흡습도가 달라질 수 있어 피부 보습이 중요한 요소로 작용하고 있다.Therefore, the degree of skin moisturization is recognized as an important factor in the field of cosmetics, skin science, and medicine. That is, the moisturizing condition of the stratum corneum is directly related to the appearance of the skin (flexibility and roughness of the skin) and is a measure of health condition. In addition, since the skin moisture content of the stratum corneum influences the skin membrane homeostasis related to the condition and permeability of the skin, the moisture absorption of the cosmetic or the medicine to be applied can be changed, thus moisturizing the skin is an important factor.
최근 들어, 환경오염 등의 이유로 강한 자외선에 노출되는 경우가 많아졌고, 이에 의한 피부 보습량의 손실이 증가하면서 피부 노화가 가속화되고, 피부의 손상에 의한 관련 질병의 발병률이 증가하고 있는 추세이다. 광노화에 의해 피부가 보습 및 탄력을 잃지 않도록 피부의 보습력을 유지해주고, 또한 그 효과를 오랫동안 지속시켜주는 화장료 조성물 또는 건강기능 식품에 대한 요구는 점점 증가하는 추세이다. In recent years, there have been a lot of cases of exposure to strong ultraviolet rays due to environmental pollution and the like, and as a result, the loss of skin moisturizing amount due to the increase in skin moisture has accelerated the aging of the skin and the incidence of the related diseases caused by skin damage is increasing. There is an increasing demand for a cosmetic composition or a health functional food which maintains the moisturizing power of the skin so that the skin does not lose its moisturizing and elasticity due to photoaging, and that the effect is maintained for a long time.
한편, 일반적으로 베리류는 딸기류의 관목 등의 작은 열매로 안토시아닌을 포함하고 있는 것으로 블루베리, 아사이베리, 라즈베리, 스트로베리, 블랙베리, 오디(mulberry), 커런트(currant), 크랜베리, 체리, 구스베리, 아세롤라, 엘더베리 등이 있다. 이러한 베리류의 열매는 수분, 탄수화물, 단백질, 그 밖의 다량의 식이 섬유소와 칼륨, 망간, 철분으로 이루어져 있으며, 달고 신맛이 강하며 붉은빛의 갈색을 띠고, 베리류 등이 갖고 있는 특유한 색의 붉은색과 푸른빛을 띠는 페놀 안토시아닌, 기타 플라보노이드와 같은 식물성 영양소 성분은 항암, 항염증, 항산화 작용을 한다고 보고되어 있다. 베리류 중에서 블루베리는 미국 타임지 선정 암예방 10대 수퍼푸드에 속할 정도로 항산화 능력이 우수하고 그 기능이 다양한 것으로 알려져 있다. 특히, 안토시아닌이 많이 함유되어 있어 항산화 능력이 우수하며 시력개선, 암과 심장 질환 및 요로 감염증 예방에서부터 기억력 감퇴 및 운동 능력 감소와 같은 노화 지연 등 다양한 건강 기능적 효능이 알려져 있다.On the other hand, berries generally contain anthocyanin as a small fruit such as a berry of a strawberry, such as blueberry, assai berry, raspberry, strawberry, blackberry, mulberry, currant, cranberry, cherry, gooseberry, Acerola, and elderberry. These berries are composed of water, carbohydrates, proteins, and other large amounts of dietary fiber, potassium, manganese, and iron. They are sweet and sour, reddish brown, and have a distinctive red color Plant nutrients such as blue phenolic anthocyanins and other flavonoids have been reported to have anti-cancer, anti-inflammatory and antioxidant properties. Among the berries, blueberries are known to have excellent antioxidant ability and various functions to belong to the top ten super foods of the United States Time magazine to prevent cancer. Especially, it has excellent antioxidant ability because it contains a lot of anthocyanins, and various health functional effects such as improvement of eyesight, prevention of cancer and heart disease and urinary tract infection, decline of memory and decrease of exercise capacity are known.
한편, 한국등록특허 제1706848호는 비타민 C를 이용한 베리 발효물의 제조방법을 개시하고 있으며, 한국등록특허 제1008954호는 앗싸이 베리의 발효물을 포함하는 화장료 조성물, 약학조성물 및 앗싸이 베리의 발효방법을 개시하고 있다. 하지만, 본 발명의 베리를 이용한 유산균 발효 방법 및 이의 용도에 대해 아직까지 개시된 바가 없다.Korean Patent No. 1706848 discloses a method for producing a fermented berry using vitamin C, and Korean Patent No. 1008954 discloses a cosmetic composition, a pharmaceutical composition, and a method for fermenting a berry, which comprises a fermented product of a berry, Lt; / RTI > However, the method of fermenting lactic acid bacteria using the berry of the present invention and its use have not yet been disclosed.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명에서는 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21, KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(Lactobacillus rhamnosus CBG-C14, KACC91981P) 균주를 각각 접종하여 발효물을 제조하였을 때, 발효 전보다 총 폴리페놀 함량 및 항산화 활성이 증가하였으며, 베리 열매 추출물보다 피부 보습효과가 우수할 뿐만 아니라, 다른 유산균 균주인 락토바실러스 플랜타룸(KCTC21004) 및 락토바실러스 람노서스(KCTC3237) 균주를 각각 접종하여 제조한 베리 열매 발효물에 비해 경피수분손실량(TEWL: transepidermal water loss) 및 각질층 두께가 현저히 감소하는 것을 확인함으로써, 본 발명을 완성하였다. SUMMARY OF THE INVENTION The present invention has been made in view of the above-described needs, and it is an object of the present invention to provide a method of producing Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) in berry fruit extracts of five kinds (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) C21, KACC92083P) and Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14, KACC91981P) were inoculated respectively, and the total polyphenol content and antioxidant activity of the fermented product were higher than those before fermentation, (TEWL: transepidermal water (TEWL)) compared with the fermented product of Berry fruit prepared by inoculating Lactobacillus plantarum (KCTC21004) and Lactobacillus lambosus (KCTC3237) strain, loss and the thickness of the stratum corneum decreased remarkably, thereby completing the present invention.
상기 과제를 해결하기 위하여, 본 발명은 1) 선별된 베리 열매를 세척하는 단계; 2) 상기 단계 1)에서 세척한 베리 열매에 물을 첨가하고 분쇄한 후 추출하는 단계; 3) 상기 단계 2)의 베리 열매 추출물에 유산균을 접종하여 발효시키는 단계; 및 4) 상기 단계 3)의 베리 열매 발효물을 동결건조하여 분말화하는 단계;를 포함하는 피부 보습용 베리 열매 발효물의 제조 방법을 제공한다.In order to solve the above-mentioned problems, the present invention provides a process for producing a berry fruit, comprising: 1) washing selected berry fruit; 2) adding water to the berry fruit washed in step 1), pulverizing and extracting the berry fruit; 3) fermenting the berry fruit extract of step 2) by inoculating the lactic acid bacteria; And 4) lyophilizing the fermented product of berry fruit in step 3) and pulverizing the berry fruit fermented product.
또한, 본 발명은 상기 제조 방법으로 제조된 피부 보습용 베리 열매 발효물을 제공한다.In addition, the present invention provides a fermented product of berry fruit for skin moisturizing, which is produced by the above-mentioned method.
또한, 본 발명은 상기 피부 보습용 베리 열매 발효물을 유효성분으로 함유하는 피부 보습용 화장료 조성물을 제공한다.The present invention also provides a cosmetic composition for moisturizing the skin, comprising the fermented product of berries for moisturizing the skin as an active ingredient.
또한, 본 발명은 상기 피부 보습용 베리 열매 발효물을 유효성분으로 함유하는 피부 보습용 건강기능식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for skin moisturizing comprising the above fermented product of berries for moisturizing skin as an active ingredient.
본 발명의 베리 열매 발효물은 자외선 조사에 의해 유발된 피부 수분의 손실을 억제하는 피부 보습 효과가 우수하므로, 이를 유효성분으로 포함하는 피부 보습용 화장료 또는 건강기능식품에 유용하게 이용될 수 있다.The berry fruit fermented product of the present invention is excellent in skin moisturizing effect for suppressing the loss of skin moisture caused by irradiation with ultraviolet rays, and thus can be usefully used as a skin moisturizing cosmetic or health functional food containing the active ingredient.
도 1은 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21, KACC92083P) 균주(A) 및 락토바실러스 람노서스 CBG-C14(Lactobacillus rhamnosus CBG-C14, KACC91981P) 균주(B)를 각각 접종한 후, 발효시간에 따른 발효물의 pH 변화를 확인한 결과이다.
도 2는 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주(A) 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주(B)를 각각 접종한 후, 발효시간에 따른 상기 2종의 유산균 균주의 생균수 변화를 확인한 결과이다.
도 3은 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주(A) 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주(B)를 각각 접종하여 발효 전과 후의 총 폴리페놀의 함량변화를 확인한 결과이다.
도 4는 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주(A) 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주(B)를 각각 접종하여 발효 전과 후의 항산화 활성 변화를 확인한 결과이다.
도 5는 동물모델 마우스에 자외선(UV)을 처리한 후, 본 발명의 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주를 이용하여 제조한 블랙베리 열매 발효물(UV-LPFB)과 비교대조군인 블랙베리 열매 물 추출물(UV-WEB) 및 블랙베리 열매 에탄올 추출물(UV-EEB)을 각각 식이하였을 때, 마우스 피부의 경피수분손실량(TEWL: transepidermal water loss)을 비교한 결과이다. Cont.는 아무것도 처리하지 않은 정상군이며, UV는 자외선 처리군이다.
도 6은 동물모델 마우스에 자외선(UV)을 처리한 후, 유산균 균주에 따라 제조한 블랙베리 열매 발효물을 식이하였을 때, 마우스 피부의 경피수분손실량(TEWL: transepidermal water loss)을 비교한 결과이다. CBG-C21 FB는 본 발명의 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주를 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며; CBG-C14 FB는 본 발명의 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며, 비교대조군으로 사용한 KCTC21004 FB 및 KCTC3237 FB는 락토바실러스 플랜타룸(KCTC21004) 균주 및 락토바실러스 람노서스(KCTC3237) 균주를 각각 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며, Cont.는 아무것도 처리하지 않은 정상군이며, UV는 자외선 처리군이다.
도 7은 동물모델 마우스에 자외선(UV)을 처리한 후, 본 발명의 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주를 이용하여 제조한 블랙베리 열매 발효물(UV-LPFB)과 비교대조군인 블랙베리 열매 물 추출물(UV-WEB) 및 블랙베리 열매 에탄올 추출물(UV-EEB)을 각각 식이하였을 때, 마우스 피부의 각질층 두께를 비교한 결과이다. Cont.는 아무것도 처리하지 않은 정상군이며, UV는 자외선 처리군이다.
도 8은 동물모델 마우스에 자외선(UV)을 처리한 후, 유산균 균주에 따라 제조한 블랙베리 열매 발효물을 식이하였을 때, 마우스 피부의 각질층 두께를 비교한 결과이다. CBG-C21 FB는 본 발명의 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주를 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며; CBG-C14 FB는 본 발명의 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며, 비교대조군으로 사용한 KCTC21004 FB 및 KCTC3237 FB는 락토바실러스 플랜타룸(KCTC21004) 균주 및 락토바실러스 람노서스(KCTC3237) 균주를 접종하여 제조한 블랙베리 열매 발효물을 처리한 군이며, Cont.는 아무것도 처리하지 않은 정상군이며, UV는 자외선 처리군이다. FIG. 1 is a graph showing the results obtained by comparing the berry fruit extracts of five species (Mackberry, Blueberry, Blackberry, Asaiberry and Raspberry) with Lactobacillus plantarum CBG-C21, KACC92083P strain (A) and Lactobacillus lambosus (B) of CBG-C14 ( Lactobacillus rhamnosus CBG-C14, KACC91981P) were inoculated, respectively.
FIG. 2 is a graph showing the results of a comparison between Lactobacillus plantarum CBG-C21 (KACC92083P) strain (A) and Lactobacillus lambosus CBG-C14 (KACC91981P) in Berry fruit extracts of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) (B) were inoculated, respectively, and the change in viable counts of the two lactic acid bacterial strains was confirmed according to fermentation time.
FIG. 3 is a graph showing the results of a comparison between Lactobacillus plantarum CBG-C21 (KACC92083P) strain (A) and Lactobacillus lambosus CBG-C14 (KACC91981P) in Berry fruit extracts of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) And strain (B) were inoculated, respectively, to determine the change in total polyphenol content before and after fermentation.
FIG. 4 is a graph showing the results of a comparison between the Lactobacillus plantarum CBG-C21 (KACC92083P) strain (A) and the Lactobacillus lambosus CBG-C14 (KACC91981P) in Berry fruit extracts of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) And strain (B) were inoculated, respectively, to examine changes in antioxidant activity before and after fermentation.
FIG. 5 is a graph showing the results of a blackberry fermented product (UV-LPFB) prepared by using Lactobacillus plantarum CBG-C21 (KACC92083P) strain of the present invention after treating an animal model mouse with ultraviolet rays (UV) (TEWL: transepidermal water loss) of mouse skin when diets of UV-WEB and blackberry fruit extract (UV-EEB) were fed, respectively. Cont. Is a normal group that has not treated anything, and UV is an ultraviolet light treatment group.
6 is a result of comparing the transepidermal water loss (TEWL) of mouse skin when blackberry fruit fermented according to a lactic acid bacterium strain was treated after treatment with an ultraviolet ray (UV) to an animal model mouse . CBG-C21 FB is a group treated with a fermented blackberry fruit prepared by inoculating Lactobacillus plantarum CBG-C21 (KACC92083P) strain of the present invention; CBG-C14 FB was treated with the blackberry fermented product prepared by inoculating Lactobacillus lambosus CBG-C14 (KACC91981P) strain of the present invention, and KCTC21004 FB and KCTC3237 FB used as comparative controls were Lactobacillus plantarum KCTC21004) and Lactobacillus lambosus (KCTC3237), respectively. Cont. Is a normal group without any treatment, and UV is a group treated with ultraviolet light.
FIG. 7 is a graph showing the results of a blackberry fermented product (UV-LPFB) prepared using an Lactobacillus plantarum CBG-C21 (KACC92083P) strain of the present invention after treatment with an ultraviolet (UV) (UV-WEB) and blackberry fruit ethanol extract (UV-EEB), respectively. Cont. Is a normal group that has not treated anything, and UV is an ultraviolet light treatment group.
8 shows the results of comparing the stratum corneum thickness of mouse skin when blackberry fruit fermented according to the lactic acid bacteria strain was treated after treating animal model mice with ultraviolet rays (UV). CBG-C21 FB is a group treated with a fermented blackberry fruit prepared by inoculating Lactobacillus plantarum CBG-C21 (KACC92083P) strain of the present invention; CBG-C14 FB was treated with the blackberry fermented product prepared by inoculating Lactobacillus lambosus CBG-C14 (KACC91981P) strain of the present invention, and KCTC21004 FB and KCTC3237 FB used as comparative controls were Lactobacillus plantarum KCTC21004) and Lactobacillus lambosus (KCTC3237), and Cont. Is a normal group without treatment and UV is a group treated with ultraviolet light.
본 발명의 목적을 달성하기 위하여, 본 발명은 In order to achieve the object of the present invention,
1) 선별된 베리 열매를 세척하는 단계;1) washing selected berries;
2) 상기 단계 1)에서 세척한 베리 열매에 물을 첨가하고 분쇄한 후 추출하는 단계; 2) adding water to the berry fruit washed in step 1), pulverizing and extracting the berry fruit;
3) 상기 단계 2)의 베리 열매 추출물에 유산균을 접종하여 발효시키는 단계; 및3) fermenting the berry fruit extract of step 2) by inoculating the lactic acid bacteria; And
4) 상기 단계 3)의 베리 열매 발효물을 동결건조하여 분말화하는 단계;를 포함하는 피부 보습용 베리 열매 발효물의 제조 방법을 제공한다.4) lyophilizing the fermented product of berry in step 3) and pulverizing the berry fruit fermented product to prepare a fermented product of berry fruit for skin moisturization.
본 발명의 피부 보습용 베리 열매 발효물의 제조 방법에서, 상기 베리 추출물은 용매 추출, 물리적 추출, 효소적 추출 등의 다양한 추출법을 이용하여 추출할 수 있으나, 바람직하게는 물리적 추출 방법을 이용하여 추출할 수 있으나, 이에 제한되지 않는다. 상기 추출방법은 유효성분이 파괴되지 않고, 베리 열매로부터 활성이 높은 베리 열매 발효물을 얻을 수 있다. In the method for producing the fermented product of berries for moisturizing skin, the berry extract may be extracted by various extraction methods such as solvent extraction, physical extraction, and enzymatic extraction, but is preferably extracted using a physical extraction method But is not limited thereto. In the above extraction method, a berry fruit fermented product having high activity from berry fruit can be obtained without breaking the effective ingredient.
또한, 상기 베리 추출물은 추출처리에 의해 얻어지는 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 또는 조정제물 또는 정제물 중 어느 하나를 포함하는 것으로 한다.The berry extract may include any one of the extract obtained by the extraction treatment, the diluted or concentrated liquid of the extract, the dried product obtained by drying the extract, or the adjusted product or the purified product.
또한, 상기 베리는 마키베리(Maqui berry), 블루베리(blueberry), 블랙베리(Blackberry), 아사이베리(Acai Berry) 및 라스베리(Raspberry)로 구성된 군으로부터 선택되는 하나 이상일 수 있으나, 이에 제한되지 않는다. In addition, the berry may be at least one selected from the group consisting of Maqui berry, blueberry, Blackberry, Acai berry and Raspberry, but is not limited thereto .
또한, 상기 베리 열매는 85% 이상 숙성된 것이 바람직하지만, 이에 제한되지 않는다.In addition, the berry fruit is preferably aged at least 85%, but is not limited thereto.
또한, 상기 유산균은 락토바실러스 플랜타룸(Lactobacillus plantarum) 균주, 락토바실러스 람노서스(Lactobacillus rhamnosus) 균주 또는 이들의 혼합 균주일 수 있으며, 바람직하게는 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21)(KACC92083P) 균주, 락토바실러스 람노서스 CBG-C14(Lactobacillus rhamnosus CBG-C14)(KACC91981P) 균주(한국등록특허 제1655852호 참고) 또는 이들의 혼합 균주일 수 있으나, 이에 제한되지 않는다. 상기 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21)(KACC92083P) 균주는 농업생명공학연구원에 2015년 7월 10일자로 기탁하였다.The lactic acid bacterium may be a Lactobacillus plantarum strain, a Lactobacillus rhamnosus strain, or a mixed strain thereof. Preferably, the lactic acid bacteria are Lactobacillus plantarum CBG-C21, (KACC92083P), Lactobacillus rhamnosus CBG-C14 (KACC91981P) strain (see Korean Patent No. 1655852), or a mixed strain thereof, but is not limited thereto. The Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) (KACC92083P) was deposited at the Institute of Agricultural Biotechnology on July 10, 2015.
또한, 상기 단계 3)에서 베리 열매 추출물에 1.0×107 내지 1.0×109 CFU/㎖의 유산균을 접종하는 것일 수 있으며, 바람직하게는 1.0×108 CFU/㎖의 유산균을 접종하는 것일 수 있으나, 이에 제한되지 않는다.In step 3), the berry fruit extract may be inoculated with 1.0 × 10 7 to 1.0 × 10 9 CFU / ml of lactic acid bacteria, preferably 1.0 × 10 8 CFU / ml of lactic acid bacteria, , But is not limited thereto.
또한, 상기 단계 3)에서 발효는 32~42℃에서 20~28시간 동안 수행하는 것일 수 있으며, 바람직하게는 37℃에서 24시간 동안 수행하는 것일 수 있으나, 이에 제한되지 않는다.In the step 3), the fermentation may be carried out at 32 to 42 ° C for 20 to 28 hours, preferably at 37 ° C for 24 hours, but is not limited thereto.
또한, 상기 단계 3)의 베리 열매 발효물의 유산균 생균수는 6.0×108 내지 1.0×1010 CFU/㎖일 수 있으며, 바람직하게는 6.0×109 CFU/㎖일 수 있으나, 이에 제한되지 않는다.In addition, the number of lactic acid bacteria in the fermented product of berry fruit in step 3) may be 6.0 × 10 8 to 1.0 × 10 10 CFU / ml, preferably 6.0 × 10 9 CFU / ml, but is not limited thereto.
또한, 본 발명은 상기 제조 방법으로 제조된 피부 보습용 베리 열매 발효물을 제공한다.In addition, the present invention provides a fermented product of berry fruit for skin moisturizing, which is produced by the above-mentioned method.
또한, 본 발명은 상기 피부 보습용 베리 열매 발효물을 유효성분으로 함유하는 피부 보습용 화장료 조성물을 제공한다.The present invention also provides a cosmetic composition for moisturizing the skin, comprising the fermented product of berries for moisturizing the skin as an active ingredient.
본 발명의 피부 보습용 화장료 조성물에는 상기 유효성분 이외에 화장품 조성물에 통상적으로 이용되는 성분들을 포함하며, 예컨대 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온 봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭과 같은 통상적인 보조제, 그리고 담체를 포함한다.The cosmetic composition for moisturizing the skin of the present invention includes components commonly used in cosmetic compositions in addition to the above-mentioned effective components, and examples thereof include fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, A foaming agent, a fragrance, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, Lipophilic active agents, conventional adjuvants such as lipid vesicles, and carriers.
본 발명의 피부 보습용 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 스킨, 스킨 소프트너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐로션, 영양로션, 마사지크림, 영양크림, 아이 크림, 모이스쳐 크림, 핸드크림, 에센스, 영양에센스, 팩, 클렌징폼, 클렌징 워터, 클렌징 로션, 클렌징 크림, 바디로션, 바디클렌져, 비누 및 파우더의 화장품 제형으로 제조될 수 있다.The cosmetic composition for moisturizing the skin of the present invention may be prepared in any form conventionally produced in the art and may be in the form of solution, suspension, emulsion, paste, gel, cream, lotion, powder, oil, powder foundation , An emulsion foundation, a wax foundation, and a spray, but is not limited thereto. More specifically, the present invention relates to a cosmetic composition for skin, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizer cream, hand cream, essence, Foam, cleansing water, cleansing lotion, cleansing cream, body lotion, body cleanser, soap and powder.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide Can be used.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로 히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, Propellants such as carbon, propane / butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester , Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
또한, 본 발명은 상기 피부 보습용 베리 열매 발효물을 유효성분으로 함유하는 피부 보습용 건강기능식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for skin moisturizing comprising the above fermented product of berries for moisturizing skin as an active ingredient.
본 발명의 피부 보습용 건강기능식품 조성물에서, 상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 드링크제, 육류, 소시지, 빵, 비스킷, 떡, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 알코올 음료 및 비타민 복합제, 유제품 및 유가공 제품 등이 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다. 상기 건강기능식품은 정제, 캡슐, 분말, 과립, 액상, 환 등의 형태로 제조, 가공될 수 있으나, 이에 한정되지 않으며 법률에 따라 어떤 형태로든지 제조, 가공될 수 있다.In the health functional food composition for skin moisturizing of the present invention, there is no particular limitation on the kind of the food. Examples of the foods to which the above substances can be added include dairy products including dairy products, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, Beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, all of which include health functional foods in a conventional sense. The health functional food may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles, but is not limited thereto and may be manufactured and processed in any form according to the law.
본 발명의 베리 열매 발효물은 식품에 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합량은 그의 사용 목적에 따라 적합하게 결정될 수 있다. 일반적으로, 건강식품 중의 상기 베리 열매 발효물의 양은 전체 식품 중량의 0.1 내지 90 중량부로 가할 수 있다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.The berry fruit fermented product of the present invention can be added directly to the food or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use. Generally, the amount of the fermented berry fruit in the health food may be 0.1 to 90 parts by weight of the total food. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above-mentioned range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.
본 발명의 건강 기능성 음료 조성물은 지시된 비율로 필수 성분으로서 상기 베리 열매 발효물 이외에 함유하는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. The health functional beverage composition of the present invention is not particularly limited to the other components contained in the fermented berry fruit as an essential ingredient in the indicated ratios and may contain various flavors or natural carbohydrates as additional components such as ordinary beverages have. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .
상기 외에 본 발명의 베리 열매 발효물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 베리 열매 발효물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다.
In addition to the above, the berry fruit fermented product of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the berry fruit fermented product of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
제조예Manufacturing example 1. 베리 추출물 및 이의 1. Berry extract and its 발효물Fermentation product 제조 Produce
85% 이상의 숙성된 5종의 베리(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리) 열매를 선별하여 세척한 후, 세척한 베리 열매 1 중량부에 대비하여 1,000 중량부의 물을 첨가하고 마쇄기로 분쇄하여 베리 열매 추출물을 얻은 다음, 면포를 이용하여 여과하였다. 상기 여과한 베리 열매 추출물은 121℃에서 15분간 멸균한 후, 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21, KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(Lactobacillus rhamnosus CBG-C14, KACC91981P) 균주를 1.0×108 CFU/㎖ 농도로 각각 접종하여 37℃에서 24시간 동안 발효시켜 각각의 베리 열매 발효물(이하 '시료'라 함)을 획득하였다. Five kinds of berries (mackberry, blueberry, blackberry, ascorbyl and raspberry) aged 85% or more were selected and washed, 1,000 parts by weight of water was added to 1 part by weight of the washed berries, Followed by pulverization to obtain berry fruit extract, which was then filtered using cotton balls. The filtered berry fruit extract was sterilized at 121 ° C for 15 minutes and then treated with Lactobacillus plantarum CBG-C21 and KACC92083P and Lactobacillus rhamnosus CBG-C14 and KACC91981P, The strain was inoculated at a concentration of 1.0 × 10 8 CFU / ml and fermented at 37 ° C. for 24 hours to obtain fermented berry fruit (hereinafter referred to as "sample").
비교대조군으로 사용한 베리 열매 물 추출물 및 베리 열매 에탄올 추출물은 상기 동일한 방법으로 베리 열매를 선별 및 세척한 후, 베리 열매 1 중량부에 대비하여 500 중량부의 물과 에탄올을 각각 첨가하여 80℃에서 6시간 동안 추출하여 제조하였다.
The water extract of berry fruit and the extract of berry fruit used as a comparative control were selected and washed with the same method as above, and 500 parts by weight of water and ethanol were added to 1 part by weight of the berry fruit, Lt; / RTI >
실시예Example 1. 베리 열매 발효물 1. Fermented fruit berries 의of pH 및 유산균 수 변화 Changes in pH and number of lactic acid bacteria
본 실시예 1에서는 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 각각 접종한 후, 24시간 동안 발효시키면서 2시간마다 발효물을 채취하여 pH를 측정하고, 3시간마다 발효물의 유산균 생균수를 BCP 첨가 평판측정용 배지(Plate Count Agar with Brom Cresol Purple)를 이용하여 측정하였다. 그 결과, 도 1 및 2에 개시한 바와 같이 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물에 대해 본 발명의 락토바실러스 플랜타룸 CBG-C21 균주 및 락토바실러스 람노서스 CBG-C14 균주를 접종하였을 때, 모두 발효시간에 따라 pH 값이 감소되고, 유산균 생균수가 증가되는 것을 확인하였다. 이를 통해 본 발명에 사용된 유산균 균주 모두 pH 2 내지 4에서 생존 및 발육이 가능하고, 내산성이 우수한 것을 확인할 수 있었다.
In Example 1, Lactobacillus plantarum CBG-C21 (KACC92083P) and Lactobacillus lambosus CBG-C14 (KACC91981P) strain were added to berry fruit extracts of five species (Mackberry, Blueberry, Blackberry, Asaiberry and Raspberry) And the fermented product was collected every 2 hours while being fermented for 24 hours. The pH of the fermented product was measured, and the number of lactic acid bacteria in the fermented product was measured using a plate count agar with bromocresol purple (BCP) Respectively. As a result, as shown in Figs. 1 and 2, berry fruit extracts of five kinds (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) were tested for Lactobacillus plantarum CBG-C21 strain of the present invention and Lactobacillus lambda When sucrose CBG-C14 strain was inoculated, the pH value was decreased and the number of lactic acid bacteria viable cells increased with fermentation time. As a result, it was confirmed that all of the lactic acid bacterial strains used in the present invention can survive and develop at
실시예Example 2. 베리 열매 2. Berry fruit 발효물의Fermented 총 폴리페놀 함량 및 항산화 활성 변화 Changes in total polyphenol content and antioxidant activity
본 실시예 2에서는 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매에 대해 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 접종하여 발효 전과 후의 총 폴리페놀 함량 및 항산화 활성을 측정하여 비교하였다.
In Example 2, the berry fruit of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) was treated with Lactobacillus plantarum CBG-C21 (KACC92083P) and Lactobacillus lambosus CBG-C14 (KACC91981P) And the total polyphenol content and antioxidant activity before and after fermentation were measured and compared.
(1) 총 폴리페놀 함량(1) Total polyphenol content
총 폴리페놀 화합물의 함량은 페놀성 물질이 인몰리브덴산(phosphomolybdate)과 반응하여 청색을 나타내는 현상을 이용한 폴린-데니스(Folin-Denis)법을 약간 변형시켜 측정하였다. The total polyphenol compound content was measured by slightly modifying the Folin-Denis method using the phenomenon that the phenolic substance reacted with phosphomolybdate to show blue color.
5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물과 이에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 각각 접종하여 제조한 베리 열매 발효물을 50㎎/mL로 준비하여 이로부터 20㎕를 취해 2㎖의 튜브에서 1.4㎖의 증류수로 희석하고, 0.1㎖의 폴린-데니스 시약(Folin-Denis reagent)을 첨가하여 진탕하였다. 3분 후에 0.2㎖의 탄산나트륨(sodium carbonate) 포화용액을 첨가하고, 전체를 2㎖로 정용하여 잘 혼합한 후에 실온에서 1시간 동안 방치한 후, 분광광도계(spectrophotometer, DU 650, Beckman Coulter Inc., Miami, FL, USA)를 이용하여 725㎚에서 흡광도를 측정하였다. 이때 총 폴리페놀 화합물은 탄닌산(tannic acid)을 이용하여 작성한 표준곡선으로부터 함량을 구하였다. The berry fruit extract of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry), the Lactobacillus plantarum CBG-C21 (KACC92083P) strain and the Lactobacillus lambosus CBG-C14 (KACC91981P) The fermented berry fruit fermented product was prepared at a concentration of 50 mg / mL, and 20 μL of the fermented product was diluted with 1.4 mL of distilled water in a 2 mL tube. Then, 0.1 mL of Folin-Denis reagent was added thereto, Respectively. After 3 minutes, 0.2 ml of a saturated sodium carbonate solution was added, and the whole was adjusted to 2 ml. The mixture was mixed well, allowed to stand at room temperature for 1 hour, and then subjected to spectrophotometer (DU 650, Beckman Coulter Inc., Miami, FL, USA) was used to measure the absorbance at 725 nm. At this time, the total polyphenol compounds were determined from the standard curves prepared using tannic acid.
그 결과, 도 3에 개시한 바와 같이 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매에 대해 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 이용하여 발효하였을 때, 발효 전보다 총 폴리페놀 함량이 증가된 것을 확인할 수 있었다(표 1 및 표 2). As a result, as shown in FIG. 3, the berry fruit of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) was treated with Lactobacillus plantarum CBG-C21 (KACC92083P) and Lactobacillus lambosus CBG- When C14 (KACC91981P) was fermented, it was confirmed that the total polyphenol content was increased before the fermentation (Table 1 and Table 2).
(2) 항산화 활성 (2) Antioxidant activity
5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매 추출물과 이에 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 각각 접종하여 제조한 베리 열매 발효물을 80% 메탄올을 이용하여 용해하여 이용하였다. 상기 베리 열매 추출물 및 발효물 0.2㎖을 시험관에 첨가한 후 0.2mM의 DPPH 용액(80% 메탄올에 용해) 0.8㎖을 가하여 와류 믹서(vortex mixer)로 10초간 혼합한 후, 실온에서 30분간 방치한 후에 525㎚에서 흡광도를 측정하였다. 하기식을 이용하여 전자공여능(Electron Donating Ability, EDA)을 계산하였으며, 초기 DPPH 농도가 50% 감소될 때까지 필요한 시료의 농도를 EC50(Efficient Concentration)으로 산출하였다. The berry fruit extract of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry), the Lactobacillus plantarum CBG-C21 (KACC92083P) strain and the Lactobacillus lambosus CBG-C14 (KACC91981P) The berry fruit fermented product was dissolved in 80% methanol. 0.2 ml of the berry fruit extract and the fermented product were added to a test tube and 0.8 ml of a 0.2 mM DPPH solution (dissolved in 80% methanol) was added thereto. The mixture was mixed with a vortex mixer for 10 seconds and left at room temperature for 30 minutes The absorbance was measured at 525 nm. Electron donating ability (EDA) was calculated using the following equation. The concentration of the required sample was calculated by EC 50 (Efficient Concentration) until the initial DPPH concentration was reduced by 50%.
EDA(%) = (1 - 시료의 흡광도 / 대조구의 흡광도) × 100EDA (%) = (1 - absorbance of sample / absorbance of control) x 100
그 결과, 도 4에 개시한 바와 같이 5종(마키베리, 블루베리, 블랙베리, 아사이베리 및 라스베리)의 베리 열매에 대해 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 이용하여 각각 발효하였을 때, 발효 전보다 항산화 활성이 증가된 것을 확인할 수 있었다(표 3 및 표 4). As a result, as shown in Fig. 4, the berry fruit of five species (Makiberry, Blueberry, Blackberry, Asaiberry and Raspberry) was treated with Lactobacillus plantarum CBG-C21 (KACC92083P) and Lactobacillus lambosus CBG- C14 (KACC91981P), the antioxidant activity was increased more than before the fermentation (Table 3 and Table 4).
실시예Example 3. 베리 열매 3. Berry fruit 발효물의Fermented 피부 보습효능 평가 Evaluation of skin moisturizing efficacy
상기 실시예 2의 결과를 토대로 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 균주 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 이용하여 발효시켰을 때, 총 폴리페놀 함량이 가장 높고, 항산화 활성이 가장 우수한 블랙베리 열매 발효물을 피부 보습효능 평가에 사용하였으며, 비교대조군으로 블랙베리 열매 물 추출물 및 에탄올 추출물을 사용하였다. 블랙베리 열매 물 추출물, 블랙베리 열매 에탄올 추출물 및 블랙베리 열매 발효물을 분말화하기 위해, 20%의 부형제를 첨가하여 혼합한 후, 동결건조하여 피부 보습효능 평가에 사용하였다.
Based on the results of Example 2 above, when fermentation was carried out using Lactobacillus plantarum CBG-C21 (KACC92083P) and Lactobacillus lambosus CBG-C14 (KACC91981P), the total polyphenol content was the highest and the antioxidant activity was the highest A good fermented blackberry fruit was used to evaluate skin moisturizing efficacy. Blackberry fruit water extract and ethanol extract were used as a comparative control. In order to powder the blackberry fruit water extract, the blackberry fruit ethanol extract and the blackberry fruit fermented product, 20% of excipient was added and mixed, followed by lyophilization to evaluate skin moisturizing efficacy.
(1) 실험동물 및 시료 식이(1) Experimental animal and sample diet
실험동물로는 200~250g의 평균 체중의 9~10주령의 암컷 헤어리스 마우스(HWY/Slc hairless rats)를 SLC사(Shizuoka, Japan)로부터 구입하여 1주 동안 적응시킨 후 사용하였다. 적응기간 중 일반 상태를 관찰하여 건강한 상태의 동물을 시험에 사용하였다. 사육환경은 온도(23±2℃), 습도(55±10%), 명암주기 12시간 (07:00-19:00/조명시간)으로 유지하였다. 시험기간 중 실험동물은 폴리카보네이트(polycarbonate) 케이지(200×320×145mm, Three-shine Co., Daejeon, Korea)에 군당 5마리로 사육하였고 사료는 고형 배합 사료를 급여하는 방법으로 하였으며, 블랙베리 열매 발효물의 처리 농도는 추출물의 식이 공급과 표피 보습 개선에 관한 선행 연구 자료(Choi, Sumin. et al., Journal of Nutrition and Health 2016; 49(5):269~276)를 참고하여 식이 중 1.0%가 포함될 수 있도록 하였다. 블랙베리 열매 발효물을 배합한 고형 배합 사료는 피드랩 코리아(Feedlab Korea, Seoul, Korea)에 위탁하여 제조하였으며, 이는 미세한 입자로 분쇄한 일반 사료와 블랙베리 열매 발효물을 블랙베리 열매 발효물의 농도가 1.0%가 되도록 균일하게 혼합한 후, 실험동물의 먹이로 알맞은 형태로 펠렛화하는 방법으로 제조하였다. 음수는 자외선으로 소독한 상수도수를 자유급이 하였다.Male HWY / Slc hairless rats (9 ~ 10 weeks old) with an average body weight of 200 ~ 250g were purchased from SLC (Shizuoka, Japan) and used for 1 week. Animals in healthy state were used for the test by observing the general condition during the adaptation period. The breeding environment was maintained at temperature (23 ± 2 ℃), humidity (55 ± 10%) and 12 hours of darkness (07: 00-19: 00 / lighting time). During the test period, the animals were housed in a polycarbonate cage (200 × 320 × 145 mm, Three-shine Co., Daejeon, Korea) at a rate of 5 per group. Feeds were fed with solid feed diets, The treatment concentration of fermented fruit was 1.0 (dietary) 1.0 (dietary supplementation) and 1.0 (dietary supplementation) for dietary supplementation and skin moisturization improvement (Choi, Sumin et al., Journal of Nutrition and Health 2016; %. The solid feed formulated with the blackberry fermented product was prepared by commissioning with Feedlab Korea (Seoul, Korea), which was prepared by mixing the general feed and blackberry fermented product pulverized into fine particles with the concentration of blackberry fermented product And 1.0%, respectively, and then pelletized in the form suitable for the feeding of the experimental animals. Negative water was free of tap water which was sterilized by ultraviolet rays.
시료 식이는 5그룹으로 나누어 정상군(Cont.), UV 처리 대조군(UV), 블랙베리 열매 물 추출물(UV-WEB), 블랙베리 열매 에탄올 추출물(UV-EEB) 및 블랙베리 열매 발효물(UV-LPFB)로 나누어 실시하였다. 전 실험기간 중 일주일에 3회, 2일 간격으로 실험동물의 임상증상과 빈사 및 폐사동물의 유무를 관찰하고 체중을 측정하였으며, 사료 섭취량은 체중 측정과 동일한 시간에 측정하여 군별 평균치를 산출하여 식이 섭취량을 구하였다.
Sample diets were divided into 5 groups and were divided into two groups according to the control group (UV), UV treatment (UV), blackberry fruit extract (UV-WEB), blackberry fruit ethanol extract (UV-EEB) and blackberry fermented fruit -LPFB). The body weight was measured by observing the clinical symptoms, vomiting, and dead animals of the experimental animals three times a week and every two days during the whole experimental period. The feed intake was measured at the same time as the body weight measurement, The intake was determined.
(2) 자외선 조사(2) Ultraviolet irradiation
자외선 조사는 정상군(Cont.)을 제외한 실험군에 8주 동안 주 3회 실시하였고, 자외선은 UV 램프(Mineralight UV Display lamp, UVP, USA)를 사용하였다. 자외선 조사는 1회 최소홍반량(minimal erythemal dose)에 해당하는 97 mJ/cm2의 광량으로 UVB를 이틀 간격으로 주 3회, 8주간 조사하였다. 자외선의 조사는 헤어리스 마우스의 등에서부터 30cm 되는 거리에 광원(Waldmann UV800; 파장: 285-350 nm, 최고파장: 310-315 nm, Waldmann Lichttechnik GmbH, Villingen-Schwenninger, Germany)을 설치하였으며, 정확한 자외선 조사량을 측정하기 위해, 자외선 측정기(Waldmann Lichttechnik GmbH)를 이용하여 광량을 측정하였다.
Ultraviolet irradiation was performed three times a week for 8 weeks except for the normal group (Cont.), And ultraviolet rays were irradiated with a UV lamp (UV light, UVP, USA). Ultraviolet irradiation was performed with UVB at a dose of 97 mJ / cm 2 corresponding to a minimum erythemal dose of 1 time per two days for 3 weeks and 8 weeks. Ultraviolet rays were irradiated using a light source (Waldmann UV800; wavelength: 285-350 nm, maximum wavelength: 310-315 nm, Waldmann Lichttechnik GmbH, Villingen-Schwenninger, Germany) at a distance of 30 cm from the back of the hairless mouse, To measure the dose, the amount of light was measured using an ultraviolet meter (Waldmann Lichttechnik GmbH).
(3) (3) 경피수분손실량Percutaneous water loss (( TEWLTEWL : : transepidermaltransepidermal water loss) 평가 water loss
경피수분손실량 수치는 피부로부터 발산되는 수분량으로, 수치가 높을수록 피부의 보습 기능이 떨어져 있음을 의미하며, 피부 고유의 장벽 기능이 손상되었음을 나타낸다. 경피수분손실량은 항온 및 항습 조건(상대 습도 60±5%, 온도 25.5±0.5℃)에서 실험시작 8주 후에 마지막 자외선 조사 48시간 후, 각 실험군의 헤어리스 마우스의 등 부위 피부에서 델핀 바포미터 와이어리스(Delfin vapometer wireless, SWL 4102, Delfin Technologies Ltd., Finland)를 이용하여 경피수분손실량을 측정하였다. 각 개체별 측정 부위는 동일한 곳으로 하였다. The amount of transdermal water loss is the amount of water emanating from the skin. The higher the value, the lower the moisturizing function of the skin. This indicates that the inherent barrier function of the skin is impaired. The transdermal water loss was measured at 48 hours after the
그 결과, 도 5에 개시한 바와 같이 자외선 처리군(UV)은 자외선조사에 의해 피부 장벽이 손상되어 정상군(Cont., 28.4g/m2/hr)에 비해 33.7g/m2/hr으로, 경피 수분손실량이 증가하였으며, 블랙베리 열매 추출물(UV-WEB 및 UV-EEB)과 블랙베리 열매 발효물을 처리한 경우, 모두 자외선에 의해 증가하였던 피부 수분 손실량이 감소되었으며, 특히 본 발명의 블랙베리 열매 발효물을 처리한 경우, 경피 수분손실량이 23.7g/m2/hr으로 피부 보습 효과가 가장 우수한 것을 확인할 수 있었다.As a result, the UV-treated group (UV), as disclosed in Figure 5 is a 33.7g / m 2 / hr compared to the control group (Cont., 28.4g / m 2 / hr) is the skin barrier is damaged by ultraviolet radiation , The transdermal water loss was increased. In case of treatment with blackberry fruit extract (UV-WEB and UV-EEB) and fermented blackberry fruit, the amount of skin water loss which was increased by ultraviolet rays was decreased, When the fermented product of berries was treated, the transdermal water loss was 23.7 g / m 2 / hr, indicating that the skin moisturizing effect was the most excellent.
또한, 유산균 균주에 따라 블랙 베리 열매 발효물의 경피수분손실량을 비교한 결과, 도 6에 개시한 바와 같이 본 발명의 락토바실러스 플랜타룸 CBG-C21(KACC92083P) 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 이용하여 제조한 블랙베리 열매 발효물의 경우, 비교대조군으로 락토바실러스 플랜타룸(KCTC21004) 및 락토바실러스 람노서스(KCTC3237)를 이용하여 제조한 블랙베리 열매 발효물보다 피부 수분 손실량을 현저하게 감소되는 것을 확인할 수 있었다.
As a result of comparing the amount of percutaneous water loss of the fermented blackberry fruit according to the lactic acid bacteria strain, the lactobacillus plantarum CBG-C21 (KACC92083P) and the lactobacillus lambosus CBG-C14 (KACC91981P) of the present invention, In the case of the fermented product of the blackberry produced by using the strain, the loss of skin water loss was significantly reduced compared with the fermented product of blackberry produced using Lactobacillus plantarum (KCTC21004) and Lactobacillus lambosus (KCTC3237) as comparative control .
(4) 조직학적 관찰 및 표피 두께(epidermal thickness) 측정(4) Histological observation and epidermal thickness measurement
경피수분손실량을 측정한 직후, 각 실험군의 헤어리스 마우스의 등 부위 피부에서 자외선 조사 부위의 피부조직을 생검하여 광학현미경 표본을 제작하였다. 적출해 낸 피부조직을 10% 포르말린에서 고정시킨 후, 수세로서 조직 내에 남아있는 포르말린을 제거하였다. 이후 파라핀 포매 과정을 거쳐 2~4μm의 두께로 절편하여 슬라이드를 제작하고 메이슨 트리크롬(Masson's trichrome) 염색법으로 염색한 후 광학현미경으로 관찰하고 표피 두께를 측정하였다. 각 실험동물의 표피의 두께는 각 표본으로부터 5번의 측정 평균치를 이용하여 평가하였다. Immediately after measuring the amount of transdermal water loss, the skin tissue of the irradiated area of the back skin of the hairless mouse of each experimental group was biopsied and an optical microscope specimen was prepared. The extracted skin tissue was fixed in 10% formalin, and formalin remaining in the tissue was removed by washing with water. The slides were then cut into 2 to 4 μm thickness by a paraffin embedding process, stained with Masson's trichrome staining method, observed with an optical microscope, and the epidermal thickness was measured. The thickness of epidermis of each experimental animal was evaluated using the average of five measurements from each sample.
그 결과, 도 7에 개시한 바와 같이 정상군(Cont.)에 비해 자외선 처리군(UV)은 자외선에 의해 비정상적인 각질 세포 증식으로 인해 각질층 두께가 현저히 증가되었으며, 블랙베리 열매 추출물(UV-WEB 및 UV-EEB)과 블랙베리 열매 발효물을 처리한 경우, 모두 각질층 두께가 감소하였으며, 특히 본 발명의 블랙베리 열매 발효물을 처리한 경우, 각질층 두께가 현저하게 감소한 것을 확인할 수 있었다. As a result, as shown in FIG. 7, the thickness of the stratum corneum was significantly increased due to abnormal keratinocyte proliferation due to ultraviolet rays in the ultraviolet treatment group (UV) compared to the normal group (Cont.), And the blackberry fruit extract UV-EEB) and fermented blackberry fruit decreased the thickness of the stratum corneum. In particular, when the fermented blackberry fruit of the present invention was treated, the thickness of the stratum corneum layer was remarkably reduced.
또한, 유산균 균주에 따라 블랙 베리 열매 발효물의 각질층 두께를 측정하여 비교한 결과, 도 8에 개시한 바와 같이 본 발명의 락토바실러스 플랜타룸 CBG-C21( KACC92083P) 및 락토바실러스 람노서스 CBG-C14(KACC91981P) 균주를 이용하여 제조한 블랙베리 열매 발효물의 경우, 비교대조군으로 락토바실러스 플랜타룸(KCTC21004) 및 락토바실러스 람노서스(KCTC3237)를 이용하여 제조한 블랙베리 열매 발효물보다 각질층 두께가 현저히 감소되는 것을 확인할 수 있었다. 8, the lactobacillus plantarum CBG-C21 (KACC92083P) of the present invention and Lactobacillus lambosus CBG-C14 (KACC91981P (KACC91981P)) of the present invention were measured by comparing the thickness of the stratum corneum of the fermented blackberry fruit according to the lactic acid bacteria strain, ), The thickness of the stratum corneum layer was significantly reduced compared with the blackberry fermented product prepared by using Lactobacillus plantarum (KCTC21004) and Lactobacillus lambosus (KCTC3237) as a comparative control group I could confirm.
Claims (9)
2) 상기 단계 1)에서 세척한 베리 열매에 물을 첨가하고 분쇄한 후 추출하는 단계;
3) 상기 단계 2)의 베리 열매 추출물에 유산균을 접종하여 발효시키는 단계; 및
4) 상기 단계 3)의 베리 열매 발효물을 동결건조하여 분말화하는 단계;를 포함하는 피부 보습용 베리 열매 발효물의 제조 방법.1) washing selected berries;
2) adding water to the berry fruit washed in step 1), pulverizing and extracting the berry fruit;
3) fermenting the berry fruit extract of step 2) by inoculating the lactic acid bacteria; And
4) lyophilizing the fermented product of berry in step 3) and pulverizing the fermented product.
A health functional food composition for skin moisturizing comprising the fermented product of Berry for moisturizing skin according to claim 6 as an active ingredient.
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| KR101535342B1 (en) * | 2013-06-21 | 2015-07-08 | 서원대학교산학협력단 | Cosmetic composition comprising the extract of Aronia melanocarpa and producing method thereof |
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- 2017-08-08 KR KR1020170100235A patent/KR101983041B1/en active IP Right Grant
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101535342B1 (en) * | 2013-06-21 | 2015-07-08 | 서원대학교산학협력단 | Cosmetic composition comprising the extract of Aronia melanocarpa and producing method thereof |
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| KR20210099317A (en) * | 2020-02-04 | 2021-08-12 | (주)케비젠 | Composition for skin regeneration, improvement or treatment damaged by ultraviolet light containing blackberry fermented product as an active ingredient |
| KR20210100802A (en) * | 2020-02-07 | 2021-08-18 | (주)케비젠 | Method for producing fermented spinach and fermented soybean with increased effective component content using lactic acid bacteria |
| KR102283096B1 (en) * | 2020-04-02 | 2021-07-29 | 주식회사 휴먼피부임상시험센터 | Composition for prevention or improvement skin inflammation induced by particlulate matter containing fermented blueberry and black rice extracts |
| KR102290291B1 (en) * | 2020-04-02 | 2021-08-13 | 건국대학교 글로컬산학협력단 | Composition for improving moisturizing skin containing fermented blueberry and black rice extracts |
| CN111388379A (en) * | 2020-04-28 | 2020-07-10 | 西南大学 | Food-grade no-clean hand sanitizer and preparation method thereof |
| KR20220094463A (en) | 2020-12-29 | 2022-07-06 | 원광대학교산학협력단 | Bread and confectionery manufacturing method using Ligularia stenocephala |
| KR102394429B1 (en) * | 2021-03-12 | 2022-05-06 | 주식회사 미그림 | Composition comprising fermented snowberry and its use for improving skin condition |
| KR102630544B1 (en) * | 2023-01-03 | 2024-01-29 | 정현철 | Method for manufacturing praline and praline manufactured therefrom |
| KR102612945B1 (en) * | 2023-07-03 | 2023-12-12 | 주식회사 파인월드 | Cosmetic composition for antioxidation and skin soothing improvement containing acerola extract and houttuynia cordata extract |
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| Publication number | Publication date |
|---|---|
| KR101983041B1 (en) | 2019-05-29 |
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