KR20000039570A - Cosmetics composition containing fermentation broth of lactobacillus - Google Patents

Cosmetics composition containing fermentation broth of lactobacillus Download PDF

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KR20000039570A
KR20000039570A KR1019980054944A KR19980054944A KR20000039570A KR 20000039570 A KR20000039570 A KR 20000039570A KR 1019980054944 A KR1019980054944 A KR 1019980054944A KR 19980054944 A KR19980054944 A KR 19980054944A KR 20000039570 A KR20000039570 A KR 20000039570A
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lactic acid
lactobacillus
fermentation broth
kfcc
acid bacteria
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KR1019980054944A
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KR100302504B1 (en
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이창학
이용두
김기환
엄지홍
백영진
허철성
한지섭
김현미
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이세복
나드리화장품 주식회사
이은선
주식회사 한국야쿠르트
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Dermatology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Birds (AREA)
  • Epidemiology (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Cosmetics (AREA)

Abstract

PURPOSE: Cosmetics composition containing fermentation broth of lactobacillus such as Bifidobacterium longum (KFCC 10870), Lactobacillus casei (KFCC 10813), Lactobacillus acidophilus (KCTC 3170) and Streptococcus thermophilus (ATCC 19987) is provided which has excellent effects of skin softening, whitening and moisturizing. CONSTITUTION: Fermentation broth obtained from culturing of at least one microorganism from Bifidobacterium longum (KFCC 10870), Lactobacillus casei (KFCC 10813), Lactobacillus acidophilus (KCTC 3170) and Streptococcus thermophilus (ATCC 19987) is added in cosmetics. Bifidobacterium longum (KFCC 10870) isolated from intestine of Korean produces lactic acid and vitamine B. Lactobacillus casei (KFCC 10813) is used for the production of dairy products and Lactobacillus acidophilus (KCTC 3170) has an excellent ability of inhibiting growth of other microorganisms. Streptococcus thermophilus (ATCC 19987) has a very good ability of producing lactic acid. At least one of lactobacillus is inoculated in the culture medium which comprises skim milk or whey and cultured at 36-37°C and pH 7.0-7.4. After the culture broth is centrifuged to remove cell mass and high molecular weight precipitate. The amount of culture broth is 0.0001-30 weight % of cosmetics.

Description

유산균 발효액 및 이를 함유하는 화장료 조성물Lactic acid bacteria fermentation broth and cosmetic composition containing same

본 발명은 피부 개선에 효과가 있는 고기능성의 유산균 발효액 및 이를 함유하는 화장료 조성물에 관한 것이다.The present invention relates to a highly functional lactic acid bacteria fermentation broth effective for skin improvement and a cosmetic composition containing the same.

효모나 곰팡이 박테리아 같은 미생물은 발효주, 발효유와 같은 식품이나 의약품 제조에 유용하게 사용되어 왔으며, 근래에는 화장품 산업에서 사용 예가 증가되고 있다. 예를 들면, 미생물을 사용하여 각종 효소, 히아루론산(hyaluronic acid), 리놀레닉산(linolenic acid), 락토오스(lactose), 락토퍼옥시다아제(lactoperoxidase)와 기타 다른 생리 활성 물질을 만들고 있다.Microorganisms, such as yeast and fungal bacteria, have been useful for the production of foods and pharmaceuticals, such as fermented liquor, fermented milk, and in recent years, their use in the cosmetic industry is increasing. For example, microorganisms are used to make various enzymes, hyaluronic acid, linolenic acid, lactose, lactoperoxidase and other bioactive substances.

유산균은 인체 내에서 유익한 역할을 하는 세균으로서 탄수화물 같은 당을 이용하여 유산(젖산)을 생성하는 균이다. 지금까지 400여종이 발견되었으며, 그 중 상품성이 있는 것은 18종으로 알려져 있다.Lactobacillus is a bacterium that plays a beneficial role in the human body and generates lactic acid (lactic acid) using sugars such as carbohydrates. Over 400 species have been discovered so far, 18 of which are known for their commercial value.

화장품 원료 제조에 사용되는 유산균으로는, 스트렙토코커스 이퀴(streptococcus equi), 같은 속의 쥬에피데미쿠스(streptococcus zooepidermicus: 대한민국 특허공고 제94-998호)와 락토바실러스 불가리쿠스(lactobacillus bulgaricus: 대한민국 특허공개 제80-146호), 락토코커스(lactococcus), 스트렙토코커스 써모필러스(streptococcus thermophilus: 대한민국 특허공개 제92-2112호), 효모(Yeast: FR-2718752) 등이 있다.Lactic acid bacteria used in the manufacture of cosmetic raw materials include Streptococcus equi, Streptococcus zooepidermicus (Korean Patent Publication No. 94-998) and Lactobacillus Bulgaricus (Korean Patent Publication) 80-146), lactococcus, streptococcus thermophilus (Korean Patent Publication No. 92-2112), yeast (Yeast: FR-2718752), and the like.

이러한 유산균들을 적절한 배양액 중에서 배양하여 얻어지는 발효액들은, 미백, 보습, 각질층의 턴오버(turn over) 속도 증가에 의한 피부의 소프트닝(skin softning), 피부 주름살 완화등에 효과가 있는 것으로 보고되고 있으나, 실제 이들 발효액을 함유하는 화장품의 이러한 기능성 효과는 발효액 중의 유효성분에 의하기 보다는 화장품에 첨가된 다른 원료를 통하여 나타내는 경우가 많았다.Fermentation broths obtained by culturing these lactic acid bacteria in an appropriate culture medium are reported to be effective in skin softening, skin wrinkle reduction by skin whitening, moisturizing, and increasing turnover rate of the stratum corneum. Such functional effects of cosmetics containing fermentation broth were often expressed through other raw materials added to cosmetics rather than by active ingredients in fermentation broth.

발효 배양액 중 유효 물질의 양이 불충분한 점을 보완하기 위한 방법으로, 얻어진 배양액 속의 고분자를 단백질 분해 효소를 이용하여 저분자로 분해시키거나(대한민국 특허공고 92-5642호, 대한민국 특허공고 80-1462호), 가수분해 효소를 이용하여 저분자화하는 방법(대한민국 특허공고 제94-000998호)이 공지되어 있다. 이런 방법으로 발효액 속에 유효 물질의 양이 증가되기는 하지만, 발효, 효소분해의 2단계 공정을 거쳐야 하므로, 제조 비용과 시간이 증가하고, 작업 효율이 떨어지게 된다.In order to compensate for the insufficient amount of the active substance in the fermentation broth, the polymer in the obtained broth is decomposed into low molecules using proteolytic enzymes (Korean Patent Publication No. 92-5642, Korean Patent Publication No. 80-1462). And a method of low molecular weight using a hydrolase (Korean Patent Publication No. 94-000998) are known. In this way, the amount of the active substance is increased in the fermentation broth, but it must go through a two-step process of fermentation and enzymatic degradation, which increases manufacturing cost and time, and decreases working efficiency.

본 발명에는 종래 기술이 가지는 이 같은 문제점을 해결하여, 보다 효율적인 방법으로 다량의 유효성분을 포함하는 유산균 발효액 및 이를 함유하는 화장료 조성물을 제공하고자 한다.The present invention is to solve such problems with the prior art, to provide a lactic acid bacteria fermentation broth containing a large amount of active ingredients in a more efficient manner and a cosmetic composition containing the same.

본 발명자들은 이러한 점을 감안하여 우수한 효능, 효과를 나타내는 발효액을 생성하는 미생물을 검색한 결과, 한국형 유산균 비피도 박테리아 론검(Bifidobacteria longum: KFCC 10870), 락토바실러스 카제이(Lactobacillus casei: KFCC 10813), 락토바실러스 애시도필러스(Lactobacillus acidophilus: KCTC 3170), 스트렙토코커스 써모필러스(Streptococcus thermophilus: ATCC 19987)의 4종의 미생물이 특히 유용함을 확인하였으며, 특히 한국형 유산균 비피도 박테리아 론검을 포함한 2종 이상의 균을 복합 배양하면 시너지(synergy) 효과를 일으켜 더 우수한 효과를 나타내는 유산균 발효액을 얻을 수 있다는 사실을 확인하여 본 발명을 완성하였다.In view of the above, the present inventors have searched for microorganisms that produce fermentation broths having excellent efficacy and effectiveness, and as a result, Korean lactic acid bacteria Bifidobacteria longum (KFCC 10870), Lactobacillus casei (KFCC 10813), Lactobacillus acidophilus (KCTC 3170) and Streptococcus thermophilus (ATCC 19987) were found to be particularly useful, especially two or more species including Lactobacillus nasal bacteria Completion of the present invention was confirmed by the fact that the combined culture of the bacteria to produce a synergistic effect (synergy) to obtain a fermentation broth that shows a better effect.

도 1은 본 발명의 실시예와 비교예의 유산균 발효액을 피부에 도포한 90분 후에 증가된 보습도를 일렉트로 임피던스로 나타낸 그래프이며,Figure 1 is a graph showing the moisture retention increased by 90 minutes after applying the lactic acid bacteria fermentation broth of the Examples and Comparative Examples of the present invention on the skin,

도 2는 본 발명의 실시예와 비교예의 유산균 발효액의 티로시나제 억제율을 나타내는 그래프이다.2 is a graph showing the tyrosinase inhibition rate of the lactic acid bacteria fermentation broth of the Examples and Comparative Examples of the present invention.

따라서, 본 발명에 의하면, 유산균을 배양액에 접종하고 배양하여 얻어지는 유산균 발효액에 있어서, 상기 유산균으로 한국형 유산균 비피도 박테리아 론검(Bifidobacteria longum: KFCC 10870)과, 락토바실러스 카제이(Lactobacillus casei: KFCC 10813), 락토바실러스 애시도필러스(Lactobacillus acidophilus: KCTC 3170) 및 스트렙토코커스 써모필러스(Streptococcus thermophilus: ATCC 19987)로 이루어진 군에서 선택된 1종 이상의 복합 균주를 사용하여 얻어지는 것을 특징으로 하는 유산균 발효액이 제공된다.Therefore, according to the present invention, in the lactic acid bacteria fermentation broth obtained by inoculating and incubating the culture with lactic acid bacteria, Korean lactic acid bacteria Bifidobacteria longum (KFCC 10870) and Lactobacillus casei (KFCC 10813) are used as the lactic acid bacteria. , Lactobacillus ashophilus (KCTC 3170) and Streptococcus thermophilus (Streptococcus thermophilus: ATCC 19987) is provided a lactic acid bacteria fermentation broth characterized in that obtained using at least one complex strain selected from the group consisting of. .

본 발명에 사용되는 한국형 비피도 박테리아 론검(KFCC 10870)은 한국인의 장에서 분리해 낸 균으로 유산 생성 능력과 비타민 B군 생산 능력이 탁월하여, 요구르트 등의 식품 생산에 이용되지만, 고기능성 화장료 생산과 관련한 용도가 검토된 바는 없다.Korean Bifidobacteria Rongum (KFCC 10870) used in the present invention is a bacterium isolated from Korean intestines and has excellent lactic acid production ability and vitamin B group production ability, but is used in food production such as yogurt, but produces high functional cosmetics. No use has been reviewed.

락토바실러스 카제이는 유제품 생산에 주로 사용되어 왔으며 역가가 가장 높은 것으로 알려져 있다.Lactobacillus casei has been used mainly in dairy production and is known to have the highest titer.

락토바실러스 애시도필러스는 식·음료 및 일부 화장료 제조에 이용되었으며, 다른 미생물의 생장을 억제하는 능력이 뛰어난 것으로 알려져 있다.Lactobacillus ashdophyllus has been used in the manufacture of food, beverages and some cosmetics, and is known for its ability to inhibit the growth of other microorganisms.

스트렙토코커스 써모필러스는 고온성 균으로 유산 생산 능력이 뛰어나 식·음료 뿐만 아니라 영양 보습의 화장료 조성물 등에도 이용되어 왔다.Streptococcus thermophilus is a thermophilic bacterium and has excellent lactic acid production ability, and has been used in cosmetic compositions for nutritional moisturizing as well as food and beverage.

본 발명에서는 한국형 비피도 박테리아 론검과, 락토바실러스 카제이, 락토바실러스 애시도필러스 및 스트렙토코커스 써모필러스로 이루어진 군에서 선택된 1종 이상의 균을, 통상적으로 사용되는 유산균 배양액, 예를 들어 스킴밀크 함유 배지나 유장(whey)함유 배지에 접종하여 배양할 수 있다. 배양액의 pH는 7.0 - 7.4로 유지하고, 배양시 온도 36 - 37℃, 통기량 1VVM의 조건을 유지하는 것이 바람직하다. 얻어진 발효액을 원심분리하여 고분자 침전물과 균체를 제거하면, 고보습, 미백, 주름 억제 또는 완화 효과가 우수한 본 발명의 유산균 발효액을 얻을 수 있다.In the present invention, one or more bacteria selected from the group consisting of Korean Bifido bacteria Rongum, Lactobacillus casei, Lactobacillus ashidophyllus and Streptococcus thermophilus are commonly used lactic acid bacteria culture medium, for example, containing skim milk. The culture may be inoculated in a medium or whey-containing medium. The pH of the culture solution is preferably maintained at 7.0-7.4, and maintained at a temperature of 36-37 ° C and aeration rate of 1 VVM at the time of culture. When the obtained fermentation broth is centrifuged to remove the polymer precipitate and the cells, the lactic acid bacterium fermentation broth of the present invention excellent in high moisturizing, whitening, anti-wrinkle or alleviating effect can be obtained.

본 발명의 방법에 따라 유산균 복합 배양으로 나타나는 발효액의 뛰어난 효과는 유산균의 대표적 대사 산물인 유산(젖산), 히아루론산 뿐 아니라 각 균주가 가지는 소수의 특정 대사 산물이 나타내는 시너지 효과라고 할 수 있다. 유산균 발효액의 유효 성분으로는 히아루론산과 젖산을 포함하여 40여종의 효소 단백질, 지질, 유해균 사멸 기능의 길항성 물질, 비타민 A, B1, B2, B6, B12, E, 아스코빈산, 무기질 등의 물질이 있는 것으로 분석되고 있다.The excellent effect of the fermentation broth in the lactic acid bacteria complex culture according to the method of the present invention can be said to be a synergistic effect of a few specific metabolites of each strain as well as lactic acid (lactic acid), hyaluronic acid which is a representative metabolite of lactic acid bacteria. The active ingredients of lactic acid bacteria fermentation broth include over 40 kinds of enzyme proteins, including hyaluronic acid and lactic acid, lipids, antagonists of killing harmful bacteria, vitamins A, B1, B2, B6, B12, E, ascorbic acid, and minerals. It is analyzed that there is.

본 발명에 의한 유산균 발효액은 화장수, 크림, 로션, 에센스 등의 화장료 조성물에 첨가할 수 있으며, 그 양은 전체 화장료 조성물 중량에 대하여, 중량으로 0.0001 내지 30%가 되도록 하는 것이 바람직하다. 중량 0.0001% 미만에서는 함량이 미미하여 원하는 고기능 효과를 나타내지 못하며, 30% 이상인 경우에는 함량 증가에 따른 효과의 증가가 거의 없어 경제성이 떨어진다.The lactic acid bacteria fermentation broth according to the present invention can be added to cosmetic compositions such as lotion, cream, lotion and essence, and the amount thereof is preferably 0.0001 to 30% by weight based on the total weight of the cosmetic composition. If the weight is less than 0.0001%, the content is insignificant and does not exhibit the desired high-functional effect. If the content is 30% or more, the effect of increasing the content is hardly increased and economic efficiency is low.

이하 본 발명을 실시예 및 비교예에 의하여 설명한다. 하기 실시예는 본 발명의 이해를 용이하게 하기 위해 제공하는 것일 뿐, 본 발명이 이 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described by way of examples and comparative examples. The following examples are provided only to facilitate understanding of the present invention, but the present invention is not limited to these examples.

실시예 1Example 1

한국형 유산균 비피도 박테리아 론검(Bifidobacteria longum KFCC 10870), 락토바실러스 카제이(Lactobacillus casei KFCC 10813), 락토바실러스 애시도필러스(Lactobacillus acidophilus KCTC 3170), 스트렙토코커스 써모필러스(Streptococcus thermophilus ATCC 19987)의 4종의 미생물을 같은 농도(106cells/ml)로 박토트립톤 1.0%(W/V), 박토펩톤 0.5%(W/V), 스킴 밀크 10%(W/V), 트리스 완충액으로 조성된 3ℓ 배양액에 30ml 접종(각 7.5ml씩)하여, 수소 이온 농도(pH) 7.0∼7.4, 온도 36∼37℃, 통기량 1VVM을 유지하면서, 6시간 동안 배양하였다. 6시간째에 미리 준비한 60%의 포도당 용액과 10%의 펩톤 용액을 배양액 내의 포도당 농도가 6%, 프로테오스 농도가 1%로 유지되도록 공급하였다. 24시간동안 유가 배양한 후 배양을 종결하여 발효액을 얻었다. 위의 발효액을 6,000g에서 30분간 원심 분리하여 고분자 침전물과 유산균체가 제거된 유산균 발효액(1,500g)을 얻었다.Korean Lactobacillus Bifidobacteria longum KFCC 10870, Lactobacillus casei KFCC 10813, Lactobacillus acidophilus KCTC 3170, and 4 Streptococcus thermophilus of Streptococcus thermophilus Microorganisms of the species were composed of bactotriptone 1.0% (W / V), bactopeptone 0.5% (W / V), skim milk 10% (W / V) and Tris buffer at the same concentration (10 6 cells / ml). 30 ml inoculation (7.5 ml each) was inoculated in 3 L culture medium, and cultured for 6 hours while maintaining a hydrogen ion concentration (pH) of 7.0 to 7.4, a temperature of 36 to 37 ° C, and an aeration of 1 VVM. At 6 hours, 60% glucose solution and 10% peptone solution prepared in advance were supplied to maintain the glucose concentration in the culture solution at 6% and the proteose concentration at 1%. After incubation for 24 hours, the culture was terminated to obtain a fermentation broth. The fermentation broth was centrifuged at 6,000 g for 30 minutes to obtain lactic acid bacteria fermentation broth (1,500 g) from which polymer precipitates and lactic acid bacteria were removed.

실시예 2Example 2

한국형 비피도 박테리아 론검과 스트렙토코커스 써모필러스를 106cells/ml의 농도로 각 15ml씩 총 30ml로 하여 실시예 1과 같은 조건으로 배양하고 처리하여, 유산균 발효액(1,500g)을 얻었다.Korean Bifidobacteria Rongum and Streptococcus thermophilus were incubated and treated under the same conditions as in Example 1 with 30 ml of 15 ml each at a concentration of 10 6 cells / ml to obtain a lactic acid bacteria fermentation broth (1,500 g).

비교예 1Comparative Example 1

실시예 1에서 한국형 비피도 박테리아 론검 대신에 락토바실러스 불가리쿠스(lactobacillus bulgaricus: ATCC 12278)를 사용한 것 이외에는 동일한 방법으로 실시하여, 유산균 발효액(1,500g)을 얻었다.The lactic acid bacteria fermentation broth (1,500 g) was obtained in the same manner as in Example 1, except that Lactobacillus bulgaricus (ATCC 12278) was used instead of the Korean Bifido bacteria rongum.

비교예 2Comparative Example 2

실시예 2에서, 한국형 비피도 박테리아 론검 대신에 락토바실러스 애시도필러스를 사용한 것 이외에는 동일한 방법으로 실시하여, 유산균 발효액(1,500g)을 얻었다.In Example 2, lactic acid bacteria fermentation broth (1,500 g) was obtained in the same manner except that Lactobacillus ashidophilus was used in place of the Korean Bifido bacteria rongum.

비교예 3Comparative Example 3

실시예 2에서, 한국형 비피도 박테리아 론검 대신에 락토바실러스 카제이를 사용한 것 이외에는 동일한 방법으로 실시하여, 유산균 발효액(1,500g)을 얻었다.In Example 2, lactic acid bacteria fermentation broth (1,500 g) was obtained in the same manner except that Lactobacillus cascai was used instead of the Korean Bifido bacteria Rongum.

비교예 4Comparative Example 4

실시예 1에서, 4종의 균주를 사용하는 대신 한국형 유산균 비피도 박테리아 론검만을 106cells/ml의 농도로 30ml 사용한 것 이외에는 동일한 방법으로 실시하여, 유산균 발효액(1,500g)을 얻었다.In Example 1, lactic acid bacteria fermentation broth (1,500 g) was obtained in the same manner, except that only 30 ml of Korean lactic acid bacteria Bidodo bacteria Rongum was used at a concentration of 10 6 cells / ml instead of four strains.

비교예 5Comparative Example 5

실시예 1에서, 4종의 균주를 사용하는 대신 스트렙토코커스 써모필러스만을 106cells/ml의 농도로 30ml 사용한 것 이외에는 동일한 방법으로 실시하여, 유산균 발효액(1,500g)을 얻었다.In Example 1, lactic acid bacteria fermentation broth (1,500 g) was obtained in the same manner except that only 30 ml of Streptococcus thermophilus was used at a concentration of 10 6 cells / ml instead of four strains.

제조예 1Preparation Example 1

표 1의 조성물을 순서대로 계량하고 균일하게 혼합한 후 정제수로 잔량을 채워, 실시예 및 비교예에 따라 제조한 유산균 발효액을 함유하는 유연화장수(스킨)를 제조하였다.The composition of Table 1 was measured in sequence and uniformly mixed, followed by filling the remaining amount with purified water, thereby preparing a flexible softener (skin) containing a lactic acid bacteria fermentation broth prepared according to Examples and Comparative Examples.

유연화장수Flexible Cosmetics 중량(%)weight(%) 정제수Purified water 잔량Remaining amount 글리세린glycerin 2.002.00 1,3 부틸렌 글리콜1,3 butylene glycol 2.002.00 구연산Citric acid 0.010.01 에탄올ethanol 10.0010.00 방부제antiseptic 미량a very small amount 색소Pigment 미량a very small amount 향료Spices 미량a very small amount 유산균 발효액Lactic acid bacteria fermentation broth 10.0010.00

제조예 2Preparation Example 2

하기 표 2의 제 1 상을 75℃로 균일하게 혼합한 후 75℃로 미리 혼합한 제 2 상을 제 1상에 가하여 균일화 시킨 후 45℃까지 냉각하여 제 3 상을 가하고 25℃까지 냉각하여, 실시예 및 비교예에 따라 제조한 유산균 발효액을 함유하는 로션을 제조하였다.After uniformly mixing the first phase of Table 2 below at 75 ° C., the second phase pre-mixed at 75 ° C. was added to homogenize the first phase, cooled to 45 ° C., added to the third phase, cooled to 25 ° C., Lotions containing lactic acid bacteria fermentation broths prepared according to Examples and Comparative Examples were prepared.

로 션Lotion 중량(%)weight(%) 제 1 상First prize 정 제 수Purified water 잔량Remaining amount 글리세린glycerin 5.005.00 트리에탄올아민Triethanolamine 0.500.50 카르복시 비닐 폴리머Carboxy Vinyl Polymer 0.200.20 방 부 제Preservative 미량a very small amount 제 2 상Second phase 세토 스테아릴 알콜Cetostearyl alcohol 2.002.00 스테아린산Stearic acid 1.001.00 자기 유화형 모노 스테아린산 글리세린Self-Emulsifying Monoglycerol Glycerin 2.002.00 친유형 모노 스테아린산 글리세린Lipophilic Monostearate Glycerin 1.001.00 바 셀 린Petrolatum 1.001.00 모노 스테아린산 폴리옥시에틸렌소르비탄(20 EO)Monostearic acid polyoxyethylene sorbitan (20 EO) 1.501.50 세트퀴올레인산 소르비탄Setquioleic acid sorbitan 1.001.00 유동 파라핀Floating paraffin 5.005.00 스쿠알란Squalane 5.005.00 제 3 상Third phase 향 료Spices 미량a very small amount 유산균 발효액Lactic acid bacteria fermentation broth 10.0010.00

제조예 3Preparation Example 3

실시예 및 비교예에 따라 제조한 유산균 발효액을 함유하고 표 3의 조성을 가진 크림을 제조예 2와 동일한 방법으로 제조하였다.A cream containing a lactic acid bacteria fermentation broth prepared according to Examples and Comparative Examples and having the composition of Table 3 was prepared in the same manner as in Preparation Example 2.

크 림cream 중량(%)weight(%) 제 1 상First prize 정 제 수Purified water 잔량Remaining amount 글 리 세 린Glycerine 5.005.00 1,3-부틸렌 글리콜1,3-butylene glycol 5.005.00 방 부 제Preservative 미량a very small amount 제 2 상Second phase 세토 스테아릴 알콜Cetostearyl alcohol 1.001.00 자기 유화형 모노 스테아린산 글리세린Self-Emulsifying Monoglycerol Glycerin 1.001.00 친유형 모노 스테아린산 글리세린Lipophilic Monostearate Glycerin 2.502.50 마이크로 크리스탈린산 납Micro Crystalline Lead 2.002.00 밀 납Wax 2.002.00 모노 스테아린산 폴리에칠렌 글리콜Monostearic Acid Polyethylene Glycol 2.002.00 모노 스테아린산 소르비탄Mono stearate sorbitan 1.001.00 트리 글리세린Triglycerine 5.005.00 액상 라놀린Liquid lanolin 5.005.00 스쿠알란Squalane 8.008.00 제 3 상Third phase 향 료Spices 미량a very small amount 유산균 발효액Lactic acid bacteria fermentation broth 10.0010.00

제조예 4Preparation Example 4

실시예 및 비교예에 따라 제조한 유산균 발효액을 함유하고 표 4의 조성을 가진 에센스를 제조예 2와 동일한 방법으로 제조하였다.Essences containing the lactic acid bacteria fermentation broth prepared according to Examples and Comparative Examples and having the composition of Table 4 were prepared in the same manner as in Preparation Example 2.

에 센 스essence 중량(%)weight(%) 제 1 상First prize 정 제 수Purified water 잔량Remaining amount 카르복시 비닐 폴리머Carboxy Vinyl Polymer 0.150.15 제 2 상Second phase 방 부 제Preservative 미량a very small amount 글 리 세 린Glycerine 15.0015.00 1,3 부틸렌 글리콜1,3 butylene glycol 5.005.00 에 탄 올ethanol 7.007.00 트리에탄올아민Triethanolamine 0.150.15 폴리옥시 에틸렌 경화 파마자유Polyoxyethylene Cured Perm Oil 1.001.00 초산 토코페롤Tocopherol Acetate 0.500.50 제 3 상Third phase 향 료Spices 미량a very small amount 유산균 발효액Lactic acid bacteria fermentation broth 10.0010.00

비교 제조예 1Comparative Production Example 1

제조예 1에서 유산균 발효액을 첨가하지 않은 것을 제외하고는 동일하게 처리하여 유연화장수를 제조하였다.Except for the addition of lactic acid bacteria fermentation broth in Preparation Example 1 was prepared in the same manner to produce a flexible longevity.

비교 제조예 2Comparative Production Example 2

제조예 2에서 유산균 발효액을 첨가하지 않은 것을 제외하고는 동일하게 처리하여 로션을 제조하였다.A lotion was prepared in the same manner as in Example 2 except that no lactic acid bacteria fermentation broth was added.

비교 제조예 3Comparative Production Example 3

제조예 3에서 유산균 발효액을 첨가하지 않은 것을 제외하고는 동일하게 처리하여 크림을 제조하였다.Except that the lactic acid bacteria fermentation broth was not added in Preparation Example 3 to prepare a cream.

비교 제조예 4Comparative Production Example 4

제조예 4에서 유산균 발효액을 첨가하지 않은 것을 제외하고는 동일하게 처리하여 에센스를 제조하였다.Except not adding the lactic acid bacteria fermentation broth in Preparation Example 4 was prepared in the same manner to prepare an essence.

실험예Experimental Example

보습 능력 실험Moisturizing ability experiment

실시예 및 비교예에서 얻어진 유산균 발효액의 보습정도를 측정하기 위해 in vivo 실험으로 사람의 팔뚝 안쪽 피부를 사용하였으며, 1cm2당 1mg의 발효액을 도포하여 30분 간격으로 90분까지의 피부 수분 함유량 정도를 고주파 임피던스 측정장치(SKICON- 200)로 측정하였다.Examples and Comparison was used for upper arm inside the skin of the person in in vivo experiments to measure the moisture degree of the lactic acid fermentation liquor obtained in the example, around the skin moisture content of up to 90 minutes, and 30 minutes by applying the fermentation broth of 1cm 2 per 1mg Was measured with a high frequency impedance measuring apparatus (SKICON-200).

시간(분)Minutes 대조군Control 제조예 1Preparation Example 1 제조예 2Preparation Example 2 비교예 1Comparative Example 1 비교예 2Comparative Example 2 비교예 3Comparative Example 3 비교예 4Comparative Example 4 비교예 5Comparative Example 5 균주(주) Strain Co., Ltd. HCATHCAT HTHT BACTBACT ATAT CTCT HH TT 실험전Before experiment 10.210.2 10.110.1 10.210.2 10.210.2 10.110.1 10.510.5 10.310.3 9.89.8 00 78.678.6 79.279.2 80.680.6 78.778.7 79.279.2 78.478.4 77.577.5 81.481.4 3030 18.318.3 47.547.5 48.248.2 40.240.2 46.346.3 44.444.4 47.347.3 39.439.4 6060 12.212.2 43.043.0 43.643.6 36.436.4 38.438.4 36.836.8 45.245.2 35.735.7 9090 14.214.2 44.244.2 42.642.6 32.732.7 31.731.7 30.630.6 38.838.8 32.432.4 (주)H는 한국형 유산균 비피도 박테리아 론검(Bifidobacteria longum KFCC 10870),C는 락토바실러스 카제이(Lactobacillus casei KFCC 10813),A는 락토바실러스 애시도필러스(Lactobacillus acidophilus KCTC 3170),T는 스트렙토코커스 써모필러스(Streptococcus thermophilus ATCC 19987),B는 락토바실러스 불가리쿠스(Lactobacillus bulgaricus ATCC 12278)* 단위 : μΩ (Note) H is Korean Lactobacillus bifidobacteria bacteria rongeom (Bifidobacteria longum KFCC 10870), C is a Lactobacillus casei (Lactobacillus casei KFCC 10813), A is Lactobacillus ash FIG pillar's (Lactobacillus acidophilus KCTC 3170), T is Streptococcus Thermophilus (Streptococcus thermophilus ATCC 19987), B is Lactobacillus bulgaricus ATCC 12278 * Unit: μΩ

미백 실험Whitening experiment

실시예 및 비교예로 얻어진 유산균 발효액의 미백 효과를 알아보기 위하여 티로시나아제 효소의 억제능력을 확인하였다. 티로시나아제는 생체 내에서 티로신이라는 물질이 산화되는 과정을 촉진하여 멜라닌이 생성되게 도와주는 효소이다. 이 실험에서는 티로시니제(시그마 사제)의 기능을 억제하여 티로신이 산화되고 멜라닌이라는 흑색의 고분자 형성이 억제되는 정도를 보는 방법(pomerantz법)으로 미백 효과를 판정하였다.In order to determine the whitening effect of the lactic acid bacteria fermentation broth obtained in Examples and Comparative Examples, the inhibitory ability of tyrosinase enzyme was confirmed. Tyrosinase is an enzyme that accelerates the process of oxidizing a substance called tyrosine in vivo to help produce melanin. In this experiment, the whitening effect was determined by the method of inhibiting the function of tyrosinase (manufactured by Sigma Co., Ltd.) and oxidizing tyrosine and inhibiting the formation of a black polymer called melanin (pomerantz method).

기질인 3.0mg/ml L-티로신 용액 2ml, pH 6.5 맥베인(Mcllvaine) 완충 용액 2.0ml, 유산균 발효액 2.0ml를 섞은 후, 37.5℃의 항온조에서 15분간 반응시켰다. 형광분석기(Shimadzu)를 이용하여 475nm에서 흡광도를 측정하였다. 억제율은 대조군의 흡광도를 100으로 보았을 때 제조예 1 - 5의 흡광도를 백분율로 구해 100에서 뺀 값을 억제율로 보았다. 대조군은 정제수를 사용하였으며, 결과는 표 6 및 도 2에 나타내었다.2 ml of 3.0 mg / ml L-tyrosine solution, 2.0 ml of pH 6.5 McVeline buffer solution, and 2.0 ml of lactic acid bacteria fermentation broth were mixed and reacted for 15 minutes in an incubator at 37.5 ° C. Absorbance was measured at 475 nm using a fluorescence analyzer (Shimadzu). The inhibition rate was obtained by subtracting 100 from the absorbance of Preparation Examples 1-5 as a percentage when the absorbance of the control group was 100 and viewed as the inhibition rate. As a control, purified water was used, and the results are shown in Table 6 and FIG. 2.

(단위: %)(unit: %) 대조군Control 제조예 1Preparation Example 1 제조예 2Preparation Example 2 비교예 1Comparative Example 1 비교예 2Comparative Example 2 비교예 3Comparative Example 3 비교예 4Comparative Example 4 비교예 5Comparative Example 5 사용균주Use strain HCATHCAT HTHT BCATBCAT ATAT CTCT HH TT 티로시나아제 억제율Tyrosinase inhibition rate 00 67.267.2 58.258.2 49.849.8 44.844.8 42.242.2 48.648.6 39.239.2

in vivo 각질층 턴오버 속도 촉진 효과 실험In vivo stratum corneum turnover speed promoting effect experiment

각질층의 턴오버(turn over) 속도 촉진 효과를 측정하기 위하여, 댄실 클로라이드(dansyl chloride)를 이용하여 제조예 1, 2, 3, 4의 화장료 조성물에 대한 효과를 측정하였다. 댄실 클로라이드는 아미노산의 형광 표시 시약으로 사용되며, 피부의 단백질을 이 염색 시약으로 염색한 후 소실되는 경시 변화를 시각으로 확인하고 대조군과 비교하여 턴오버 속도를 판단하는 데 사용되었다.In order to measure the turn over rate promoting effect of the stratum corneum, the effect on the cosmetic composition of Preparation Examples 1, 2, 3, 4 was measured using dansyl chloride. Dansyl chloride was used as a fluorescence labeling reagent for amino acids, and visually confirmed the change over time after staining the protein of the skin with this staining reagent, and was used to determine the turnover rate compared to the control.

실험 대상은 나이 20 내지 29세의 건강한 여성 20명이었으며, 10명씩 (가) 군과 (나) 군으로 나누어서 실험을 했다.The test subjects were 20 healthy women aged 20 to 29 years old, and 10 patients were divided into (A) and (B) groups.

먼저, 댄실 클로라이드를 백색 바셀린에 5% 분산시켜 패취 테스트용 반창고(Finn chamber: 직경 1cm)로 피부에 24시간 도포하여 염색하였다. 도포 부위는 실험 대상 20명의 왼팔과 오른팔 안쪽 부위에 각각 8부위였으며, 도포량은 2mg/웰이었다.First, dansyl chloride was dispersed 5% in white petrolatum and applied to the skin for 24 hours in a patch test bandage (Finn chamber: 1 cm in diameter) and dyed. The application sites were 8 sites on the left and right arms of 20 subjects, and the application amount was 2 mg / well.

염색 후, 실험 대상 (가)군 10명의 왼팔 안쪽 각 부위에 제조예 1(유산균 발효액의 종류에 따라 각 7종)과 비교 제조예 1을 도포하게 하였다. (가)군의 오른팔은 제조예 2를, (나)군의 왼팔과 오른팔은 제조예 3과 4를 각각 비교예와 같이 도포하게 하였다. 도포 시간은 하루 2회 아침과 저녁이었으며, 10일 도포 후 아래의 판정 기준에 의해 3명의 판정자가 복수 판정을 했고, 그 결과는 평균값으로 표 7에 나타내었다.After staining, Preparation Example 1 (each of seven species depending on the type of lactic acid bacterium fermentation broth) and Comparative Preparation Example 1 were applied to each part of the left arm of the test subjects (A) group. The right arm of group (A) was prepared in Preparation Example 2, and the left arm and right arm of group (B) were prepared in Preparation Examples 3 and 4 as in Comparative Examples. The application time was morning and evening twice a day. After 10 days of application, three judges made multiple judgments according to the following criteria, and the results are shown in Table 7 as the average values.

판정 기준: 비교예에 비해 소실된 정도Criteria: degree of loss compared to the comparative example

++++ 80% 이상 소실 +++ 60% 이상 소실++++ over 80% lost +++ over 60% lost

++ 40% 이상 소실 + 20% 이상 소실++ More than 40% lost + More than 20% lost

+- 10% 이상 소실 - 10% 미만 소실+-More than 10% lost-less than 10% lost

비교제조예Comparative Production Example 제조예에 함유된 유산균 발효액Lactic Acid Bacteria Fermentation Solution Contained in Preparation Example 실시예 1Example 1 실시예 2Example 2 비교예 1Comparative Example 1 비교예 2Comparative Example 2 비교예 3Comparative Example 3 비교예 4Comparative Example 4 비교예 5Comparative Example 5 제조예 1Preparation Example 1 __ ++++ ++++ ++ ++ ++ ++++ ++ 제조예 2Preparation Example 2 +-+- ++++ ++++ ++ ++ ++ ++++ ++ 제조예 3Preparation Example 3 +-+- ++++++ ++++++ ++++ ++ ++++ ++++ ++ 제조예 4Preparation Example 4 +-+- ++++++++ ++++++ ++++ ++++ ++++ ++++ ++++

in vivo 피부 주름 개선 효과 실험In vivo skin wrinkle improvement effect experiment

유산균 발효액의 피부 주름 개선 효과를 알아보기 위해 표 8와 같은 조건으로 임상 실험을 하였다.In order to examine the skin wrinkle improvement effect of the lactic acid bacteria fermentation broth was carried out under the conditions shown in Table 8.

피험자는 건강한 사람 20명으로, 나이는 30 내지 49세였으며, 피험자를 2 그룹으로 나누어 10명은 실시예 1의 발효액이 들어간 화장료 조성물로 실험하였고, 나머지 10명은 대조군으로 한국형 유산균이 제외된 비교예 1로 실험하였다. 실험 방법은 레플리카(replica)를 안면의 이마 일부분과 왼쪽 눈꼬리에 적용하였다. 0일째 날에 피부를 증류수로 깨끗이 닦은 후, 레플리카(silflo사제) 0.5g에 촉매제 2 내지 3 방울을 섞어 도포하였다. 5분후 조심스럽게 떼어내 이미지 분석기를 이용하여 50배 확대 촬영하였다. 30일이 지난 후와 60일이 지난 후에도 위와 같은 방법으로 피부 레플리카를 촬영한 후 3명의 평가자에 의한 점수기록으로 '매우좋다를 10점', '변화없다를 0점'으로 하여 기록하게 한 다음 3명의 점수를 평균하여 매우좋아졌다-7점 이상, 좋아졌다-5점 이상, 다소 좋아졌다-3점 이상, 변화없다-0점 이상으로 하여 얻어진 결과를 표 9에 나타내었다.The test subjects were 20 healthy people, and the age was 30 to 49 years old. The test subjects were divided into 2 groups, 10 were tested with the cosmetic composition containing the fermentation broth of Example 1, and the remaining 10 were the control group except for Korean lactic acid bacteria. Experiment with Experimental methods were applied to the forehead and left eye of the face. On day 0, the skin was wiped clean with distilled water, and then mixed with 2 to 3 drops of catalyst to 0.5 g of a replica (manufactured by silflo). After 5 minutes, it was carefully removed and 50 times magnified using an image analyzer. After 30 days and 60 days, skin replicas were taken in the same manner as above, and the scores recorded by three evaluators were recorded as 'very good 10 points' and 'no change' 0 points. Table 9 shows the results obtained by averaging three scores, which was very good -7 or more, improved -5 or more, somewhat better -3 or more, and no change -0 or more.

피험자Subject 10명/(대조군 10명)10 / (10 control) 차수Degree 1차Primary 2차Secondary 3차3rd 4차4th 인원(명)Number of people 10/(10)10 / (10) 10/(10)10 / (10) 10/(10)10 / (10) 10/(10)10 / (10) 사용제품Product used 유연화장수(제조예 1)Soft Cosmetics (Manufacturing Example 1) 로션(제조예 2)Lotion (Manufacturing Example 2) 크림(제조예 3)Cream (production example 3) 에센스(제조예 4)Essence (production example 4) 사용기간Period of use 60일간60 days 60일간60 days 60일간60 days 60일간60 days 사용부위Used part 이마(왼쪽)Forehead (left) 이마(오른쪽)Forehead (right) 눈꼬리(왼쪽)Eye tail (left) 눈꼬리(오른쪽)Eye tail (right)

평가(단위: 명)Evaluation (Unit: person) 제조예 1Preparation Example 1 제조예 2Preparation Example 2 제조예 3Preparation Example 3 제조예 4Preparation Example 4 HCAT실시예 1HCAT Example 1 BCAT비교예 1BCAT Comparative Example 1 HCAT실시예 1HCAT Example 1 BCAT비교예 1BCAT Comparative Example 1 HCAT실시예 1HCAT Example 1 BCAT비교예 1BCAT Comparative Example 1 HCAT실시예 1HCAT Example 1 BCAT비교예 1BCAT Comparative Example 1 매우 좋아졌다Very good 1One 00 22 00 33 00 33 22 좋아졌다Got better 22 1One 44 55 33 33 55 44 다소 좋아졌다Somewhat improved 55 66 33 22 33 66 22 1One 변화없다No change 22 33 1One 33 1One 1One 00 33 계system 1010 1010 1010 1010 1010 1010 1010 1010

표 5, 6, 7의 결과로부터 알 수 있듯이, 한국형 유산균 비피도 박테리아 론검을 포함하는 4종의 복합 균주의 발효액을 사용한 경우에는 사용하지 않은 경우보다 보습력에 있어서 약 35%의 개선 효과와(도 1), 티로시나아제의 억제율에 있어서도 35%의 증가를 확인할 수 있다(도 2). 각질층 턴오버 촉진과 주름 개선에서도 효과가 나타났는데, 특히 각질층 턴오버 촉진 실험에서는 피부 형광 시약인 댄실 클로라이드(dansyl chloride)의 소실속도가 최고 40% 이상 더 빨라짐으로써 각질층의 턴오버 효과가 뛰어난 것으로 판명되었다. 2종의 균을 복합 배양시킬 때에도 한국형 유산균을 포함한 발효액은 포함하지 않은 다른 2종의 유산균 발효액보다 더 나은 효과를 나타내었다.As can be seen from the results of Tables 5, 6, and 7, the fermentation broth of four complex strains containing the Korean lactic acid bacteria Bifidobacterium gum was used and improved about 35% in moisturizing power compared to the case where it was not used. 1), an increase of 35% was also observed in the inhibition rate of tyrosinase (FIG. 2). The effect of promoting stratum corneum turnover and wrinkling has also been shown to be effective. In particular, in the stratum corneum turnover experiment, the loss rate of dansyl chloride, a skin fluorescence reagent, is up to 40% faster, and the turnover effect of the stratum corneum turns out to be excellent. It became. In the case of complex culturing of two bacteria, the fermentation broth containing Korean lactic acid bacteria showed a better effect than the other two lactic acid bacteria fermentation broth that did not contain.

본 발명의 방법에 의하면, 고보습, 미백, 주름 억제 또는 완화 효과가 우수한 유산균 발효액과 이를 함유하는 화장료 조성물을 얻을 수 있다.According to the method of the present invention, a lactic acid bacteria fermentation broth excellent in high moisturizing, whitening, anti-wrinkle or alleviating effect and a cosmetic composition containing the same can be obtained.

Claims (2)

유산균을 배양액에 접종하고 배양하여 얻어지는 유산균 발효액에 있어서, 상기 유산균으로 한국형 유산균 비피도 박테리아 론검(Bifidobacteria longum: KFCC 10870)과, 락토바실러스 카제이(Lactobacillus casei: KFCC 10813), 락토바실러스 애시도필러스(Lactobacillus acidophilus: KCTC 3170) 및 스트렙토코커스 써모필러스(Streptococcus thermophilus: ATCC 19987)로 이루어진 군에서 선택된 1종 이상의 복합 균주를 사용하여 얻어지는 것을 특징으로 하는 유산균 발효액.In the lactic acid bacteria fermentation broth obtained by inoculating the culture with lactic acid bacteria, the Lactobacillus bacterium Bifidobacteria longum (KFCC 10870), Lactobacillus casei (KFCC 10813), Lactobacillus ashdophyllus are used as the lactic acid bacteria. (Lactobacillus acidophilus: KCTC 3170) and Streptococcus thermophilus (Streptococcus thermophilus: ATCC 19987) Lactic acid bacteria fermentation broth characterized in that obtained by using at least one complex strain selected from the group consisting of. 제 1항에 따른 유산균 발효액을 0.0001 내지 30 중량% 함유하는 화장료 조성물.Cosmetic composition containing 0.0001 to 30% by weight of the lactic acid bacteria fermentation broth according to claim 1.
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