KR101768921B1 - Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses thereof - Google Patents
Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses thereof Download PDFInfo
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- KR101768921B1 KR101768921B1 KR1020170062461A KR20170062461A KR101768921B1 KR 101768921 B1 KR101768921 B1 KR 101768921B1 KR 1020170062461 A KR1020170062461 A KR 1020170062461A KR 20170062461 A KR20170062461 A KR 20170062461A KR 101768921 B1 KR101768921 B1 KR 101768921B1
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- pediococcus
- fermentation
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Abstract
Description
본 발명은 항균활성이 우수한 발효곡물을 제조하기 위한 스타터용 페디오코커스 애시디락티시 SRCM 102024 균주 및 이의 용도에 관한 것이다.The present invention relates to a strain Pediococcus aceyltracis SRCM 102024 for starter and a use thereof for producing a fermented grain having excellent antibacterial activity.
아름다운 피부에 대한 열망은 나이, 성별을 불문하고 건강한 피부를 원하고 있다. 아름다움에 대한 욕구와 맞물려 건강하고 깨끗한 피부가 유행하고 있다. 이러한 미용동향에 따라 깨끗한 피부유지를 원하지만 현대사회에서는 환경적인 영향으로 여러 가지 고민을 많이 하고 있다. The desire for beautiful skin demands healthy skin regardless of age or gender. Healthy and clean skin is popular with the desire for beauty. In order to maintain clean skin according to these cosmetic trends, many people are worried about environmental problems in modern society.
여드름균(Propionibacterium acnes)은 여드름의 원인균으로 알려져 있고, 피지의 과잉분비와 세포각질의 과다와 축척으로 인해 피지가 모낭속에 남게 되면 모낭의 모루두에 존재하는 피부상주균인 여드름균과 피부포도상구균이 증가하고 이들 균의 지질분해 효소인 리파아제가 피지를 구성하고 있는 지질 중에 트라이글리세라이드를 분해해서 유리지방산으로 변화시킨다. 여드름균은 유리지방산이 모낭벽이나 모공주위의 세포들을 자극해 염증을 일으키고 그 결과 과각화되어 어느 순간 막혀 초기 면포가 생성된다. 유리지방산이 염증을 증폭할 수 있는 효소를 생산하고 이 효소의 작용으로 백혈구가 분해하고 파열된 모낭속의 내용물이 진피 속에 들어가 염증을 일으켜 여드름이 유발된다. Acne bacteria ( Propionibacterium acnes is known to be a causative agent of acne. When sebaceae are left in the hair follicle due to excessive secretion of sebum and excess and scale of cell keratin, acne bacteria and skin staphylococci, which are the skin- Lipase, a lipase of bacteria, breaks down triglyceride into lipid constituting sebum and changes it into free fatty acid. Acne bacterium stimulates the cells around the hair follicle walls or pores of the acne causing inflammation, and as a result, it is overshot, and at the moment clogs the initial cotton is produced. Free fatty acids produce enzymes that can amplify the inflammation, which causes the white blood cells to break down and the contents of the ruptured follicles enter the dermis, causing inflammation and causing acne.
아토피 피부염은 가려움증을 보이는 만성적이고, 재발성의 염증성 피부 상태를 말하며, 주로 유아와 소아에서 많이 발생하고, 유병기간은 1개월에서 44년으로, 평균 11.40±8.64년이며, 유병기간이 1개월~4년인 경우가 27%, 10년인 경우가 53.9%의 유병율을 가지고 있다고 보고되고 있다. 아토피 피부염의 임상적 특징들을 발육 시기에 따라 살펴보면 유아기에는 아토피 피부염의 첫 징후는 뺨에 나타나는 습진과 구진성 수포와 반점성 병소로 나타나고, 아동기에 습진성 병변(eczematous lesions), 보통 굽어지는 부분(antecubital fossae, 목, 팔목, 발꿈치)과 목의 뒷부분, 발등 그리고 손에 나타난다. 청소년과 성인에는 습진 병변이 아동기에서 청소년기, 성인기까지 계속되거나 그 질병이 성인기에 새로이 시작되면, 머리와 목뿐만 아니라 굽어지는 부분은 lichenified plaques와 관련되고, 아토피 피부염의 주요한 특징 중 하나는 표피에 의해 수분 상실의 증가에 의해 보이는 병변과 비병변 표피지역에 영향을 주는 건조증이며, 이러한 건조증은 아토피 피부염에서 피부기능 저하에 대한 여러 가지 기전이 제시되어 왔으며, 건조한 피부는 계속 문제가 되며, 겨울에 더욱 그러하다. Atopic dermatitis refers to a chronic, recurrent, inflammatory skin condition that is pruritic and usually occurs in infants and children. The duration of the disease is 1 month to 44 years, with an average of 11.40 ± 8.64 years. The prevalence rate is 27% for the year and 53.9% for the 10 years. Clinical features of atopic dermatitis are classified according to the developmental stage. In early childhood, the first signs of atopic dermatitis are eczema, acute blisters and spotty lesions on the cheeks, eczematous lesions in the childhood, antecubital fossae, neck, cuff, heel) and back of neck, foot and hands. When adolescent and adult eczema lesions continue from childhood to adolescence and adulthood, or when the disease is newly started in adulthood, the head and neck as well as the bending part are associated with lichenified plaques and one of the main features of atopic dermatitis is epidermal This dryness has been suggested as a mechanism of skin hypofunction in atopic dermatitis, dry skin is a problem, and in winter it is more problematic. It is true.
한편, 유산균은 포도당 등 당류를 에너지원으로 이용해 유산을 생성하는 균의 총칭으로 유해균 증식억제, 정장작용, 면역증강, 설사 및 변비 개선 작용을 가지고 있으며, 스트렙토코쿠스속 유산균, 페디오코크스속 유산균, 류코노스톡속 유산균, 락토바실러스속 유산균, 비피도박테리움속 유산균이여, 식품가공에 유산균을 이용하는 목적은 식품의 보존성 향상, 식품의 풍미향상 및 기능성 향상을 위한 것이다.On the other hand, lactic acid bacterium is a generic term for lactic acid producing bacteria using saccharides such as glucose as an energy source, and has a function of inhibiting the growth of harmful bacteria, improving the function of lactic acid bacteria, improving immunity, improving diarrhea and constipation, , Lactobacillus lactic acid bacteria, Lactobacillus sp. Lactic acid bacteria, and Bifidobacterium sp. Lactic acid bacteria. The purpose of using lactic acid bacteria in food processing is to improve the preservation of food, improve flavor of food, and improve functionality.
현재, 완전곡물을 기피하는 생활습관으로 인하여 현대인들에게 당뇨병, 비만 및 고혈압등 성인병이 크게 증가하고 있고, 이러한 성인병으로 고생하는 사람들은 잡곡을 섭취하려고 노력을 하지만, 소화흡수율이 낮아 그 효과가 크지 않다. 따라서, 잡곡류를 섭취하여 성인병을 예방하고 소화흡수율을 높이는 것이 필요하다. 급변하는 현대시대에 식생활의 간편성을 추구하는바 곡물류의 선식에 대한 관심이 나날이 높아져 가고 있다. 사람의 중요한 에너지원인 곡류를 이용하여 혼합곡류에서 잘 생장하는 유산균을 이용하여 곡물발효물을 제공함으로써 곡물의 소비를 촉진하여 농가의 경제적 문제해결에 도움이 될 뿐만 아니라 기능성이 우수한 곡물발효물을 제공하고자 한다.Currently, adult lifestyles such as diabetes, obesity and hypertension are increasing in modern people due to the lifestyle of avoiding whole grains. People suffering from these adult diseases try to eat grains, but the digestion rate is low and the effect is great not. Therefore, it is necessary to eat gravy to prevent adult disease and increase digestion rate. Interest in cereals is rising day by day in pursuit of simplicity of eating habits in the rapidly changing modern age. Using lactic acid bacteria that grow well in mixed cereals by using human cereal, which is an important energy source, it promotes the consumption of cereals by promoting the consumption of cereals, which not only helps to solve the economic problems of the farmers but also provides excellent fermented cereals I want to.
한국공개특허 제2017-0045050호에서는 '곡물 유산균 발효물 및 이의 제조방법'이 개시되어 있고, 한국공개특허 제2017-0048229호에서는 '효소 생산능이 우수한 전통 발효식품 유래 신규 균주 및 이를 이용하여 곡물 발효 효소식품을 제조하는 방법'이 개시되어 있으나, 본 발명에서와 같이 '항균활성이 우수한 발효곡물을 제조하기 위한 스타터용 페디오코커스 애시디락티시 SRCM 102024 균주 및 이의 용도'에 대해서는 밝혀진 바가 전혀 없다.Korean Patent Laid-Open Publication No. 2017-0045050 discloses a fermented product of lactic acid bacteria and a method for producing the fermented product. In Korean Patent Laid-Open Publication No. 2017-0048229, a novel strain derived from a traditional fermented food having excellent enzyme production ability, A method for producing enzyme food 'has been disclosed. However, as in the present invention, there has never been known a strain of Pediococcus acephaliticus SRCM 102024 for starter for producing a fermented grain having excellent antibacterial activity and its use .
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명에서는 과일 발효액으로부터 유산균을 분리하였는데, 상기 유산균은 피부염 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylcoccus epidermidis)에 대해 항균활성을 가지는 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주로 동정되었다.The present invention has been made in view of the above-mentioned needs. In the present invention, lactic acid bacteria are isolated from a fruit fermentation broth. The lactic acid bacteria are selected from the group consisting of Propionibacterium acnes, acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis) Phedi O Lactococcus her CD rakti when (Pediococcus with antibacterial activity against acidilactici ) SRCM 102024.
또한, 상기 균주를 곡물 발효를 위한 스타터 균주로 이용하여 발효곡물을 제조한 결과, 피부염 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylcoccus epidermidis)에 대해 항균활성을 가지며, 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 처리하지 않은 대조구에 비해 아미노태질소, 유기산, 유기당 및 유리아미노산 함량이 증가하고, α-글루코시다제 저해활성 및 프로테아제 활성이 증가되며, 유산균 총수가 증가된 기능성 발효곡물의 제조가 가능함을 확인함으로써, 본 발명을 완성하였다.In addition, as a result of using the strain as a starter strain for fermentation to prepare a fermented cereal grains, dermatitis causative agent of propionic sludge tumefaciens arc Ness (Propionibacterium acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis ), and the Pediococcus ( Pediococcus) acidic lactic acid, increased the content of amino acid nitrogen, organic acid, organic sugar and free amino acid, increased the inhibitory activity of alpha -glucosidase and protease activity and increased the total number of lactic acid bacteria, compared with the control without the acidilactici SRCM 102024 strain The present invention has been completed.
상기 과제를 해결하기 위해, 본 발명은 과일 발효액으로부터 분리한 피부염 원인균에 대한 항균활성을 가지며, 기탁번호가 KCCM12019P인 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 제공한다.In order to solve the above-mentioned problems, the present invention relates to a method for producing an antimicrobial agent having antimicrobial activity against a causative agent of dermatitis isolated from a fruit fermentation broth and having a deposit number of KCCM12019P ( Pediococcus acidilactici ) SRCM 102024 strain.
또한, 본 발명은 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 또는 이의 배양액을 유효성분으로 함유하는 발효곡물 제조용 스타터(starter) 조성물을 제공한다.In addition, the present invention provides the O Phedi Lactococcus her CD rakti when (Pediococcus acidilactici) SRCM 102024 fermentation for producing grain starter (starter) composition containing the strain or culture thereof as an active ingredient.
또한, 본 발명은 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 또는 이의 배양액을 스타터(starter) 균주로 이용하여 곡물을 발효하여 제조된 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 제공한다.The present invention also provides a fermented grain having antimicrobial activity against a causative organism of dermatitis produced by fermenting cereals using Pediococcus acidilactici strain SRCM 102024 or a culture thereof as a starter strain do.
또한, 본 발명은 상기 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 유효성분으로 함유하는 여드름 또는 아토피 피부염의 예방 또는 개선용 건강기능식품 조성물을 제공한다.Also, the present invention provides a health functional food composition for preventing or ameliorating acne or atopic dermatitis containing an effective ingredient of a fermented grain having an antibacterial activity against the causative agent of dermatitis.
또한, 본 발명은 상기 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 유효성분으로 함유하는 여드름 또는 아토피 피부염의 개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for improving acne or atopic dermatitis containing, as an active ingredient, a fermented grain having an antibacterial activity against the causative agent of dermatitis.
본 발명에서 분리한 유산균인 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주는 피부염 원인균인 프로피오니박테리움 아크네스 및 스타필로코커스 에피더미디스에 대해 항균활성을 가지는 기능성 발효곡물의 제조가 가능함을 확인하였다.The strain Pediococcus acidilactici SRCM 102024, which is a lactic acid bacterium isolated in the present invention, can be used for the production of functional fermented grains having antimicrobial activity against the dermatitis causing bacteria such as Propionibacterium acnes and Staphylococcus epidermidis .
따라서, 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액은 여드름 및 아토피 피부염 치료제로서 이용가능하므로, 기능성 식품 및 화장료 조성물로 관련 산업에 유용하게 사용될 수 있다.Thus, the Pediococcus acidilactici ) SRCM 102024 can be used as a therapeutic agent for acne and atopic dermatitis, and thus can be usefully used in related industries as a functional food and cosmetic composition.
도 1은 본 발명에서 사용된 균주 분리 및 배양 방법을 나타낸다.
도 2는 본 발명에서 분리한 균주를 이용한 발효곡물의 제조 과정을 나타낸다.
도 3은 본 발명에서 분리한 균주를 이용한 곡물발효액으로부터 미생물을 분리하는 방법을 나타낸다.
도 4는 본 발명에서 분리한 균주의 16S rRNA 시퀀싱을 위해 사용된 프라이머 정보를 나타낸다.
도 5는 본 발명에서 분리한 유산균의 16S rRNA 염기서열을 기반으로 분석한 계통수를 나타낸다. Neighbor joining method를 이용하여 구축되었으며, Blast 결과와 가장 가까운 균주 10개와 비교하여 작성되었다.
도 6은 본 발명의 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액의 항균활성 결과이다. 이는 분리 균주들을 처리한 것으로, 세균류에서는 클리어 존이 형성되지 않았으며, 클리어 존이 형성된 왼쪽 플레이트 배지의 아래 두개의 홀(hole)은 유산균인 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 처리 결과인 것을 확인하였다.
도 7은 본 발명의 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액의 유기산 분석 결과이다. 1: 블랭크(Blank), 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
도 8은 본 발명의 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액의 유리당 분석 결과이다. 1: 블랭크(Blank), 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
도 9는 본 발명의 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액의 총 유리아미노산 분석 결과이다. 1: 블랭크(Blank), 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
도 10은 본 발명의 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 이용한 곡물발효액의 항산화 활성 분석 결과이다. 1: 블랭크(Blank), 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 1016771 shows a method for isolating and culturing a strain used in the present invention.
FIG. 2 shows a process for producing fermented grains using the strains isolated in the present invention.
FIG. 3 shows a method for separating microorganisms from a cereal fermentation broth using a strain isolated in the present invention.
Figure 4 shows the primer information used for 16S rRNA sequencing of strains isolated in the present invention.
FIG. 5 shows phylogenetic analysis based on the 16S rRNA nucleotide sequence of the lactic acid bacteria isolated in the present invention. Neighbor joining method was used and compared with 10 strains nearest to the blast result.
6 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 strain. It was treated with isolated strains. No clear zone was formed in the fungi. Two holes under the left plate culture medium with clear zone were formed by Pediococcus (Lactobacillus acidophilus) acidilactici ) SRCM 102024 strain.
7 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 was used for the analysis of organic acids in cereal fermentation broth. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stock Methenseeroides SRCM 101677, 9: Pediococcus acidiflactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
8 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 strain. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
9 is trying Phedi O Lactococcus according to the present invention when CD rakti (Pediococcus acidilactici ) SRCM 102024 strain of the present invention. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
10 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 was used for the analysis of antioxidant activity of cereal fermentation broth. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
본 발명의 목적을 달성하기 위하여, 본 발명은 과일 발효액으로부터 분리한 피부염 원인균에 대한 항균활성을 가지며, 기탁번호가 KCCM12019P인 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 제공한다.In order to accomplish the object of the present invention, the present invention provides a method for producing an antimicrobial agent having antimicrobial activity against a causative agent of dermatitis isolated from a fruit fermentation broth and having a deposit number of KCCM12019P ( Pediococcus acidilactici ) SRCM 102024 strain.
본 발명에서는 과일 발효액으로부터 유산균을 분리하였는데, 상기 유산균은 피부염 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylcoccus epidermidis)에 대해 항균활성을 가지는 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주로 동정되었으며, 상기 균주를 한국미생물보존센터(KCCM)에 2017년 4월 20일에 기탁하였다(기탁번호: KCCM12019P).In the present invention, lactic acid bacteria were isolated from a fruit fermentation broth. The lactic acid bacteria were selected from Propionibacterium acnes, acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis) Phedi O Lactococcus her CD rakti when (Pediococcus with antibacterial activity against acidilactici ) SRCM 102024, and the strain was deposited at the Korean Microorganism Conservation Center (KCCM) on April 20, 2017 (Accession No .: KCCM12019P).
"항균 (antimicrobial)"의 의미는 어떤 농도에서 미생물의 성장 또는 생존을 감소, 방지, 억제, 또는 제거하는 능력을 의미한다.By "antimicrobial" is meant the ability to reduce, prevent, inhibit, or eliminate microbial growth or survival at any concentration.
본 발명의 일 구현 예에 따른 균주에서, 상기 피부염 원인균은 여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 또는 아토피 원인균인 스타필로코커스 에피더미디스(Staphylcoccus epidermidis)일 수 있으나, 이에 제한되지 않는다.In the strain according to one embodiment of the present invention, the causative agent of dermatitis is Propionibacterium acnes causing acne It is acnes) or atopic pathogen is Staphylococcus epidermidis (Staphylcoccus epidermidis), but is not limited thereto.
또한, 본 발명은 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 또는 이의 배양액을 유효성분으로 함유하는 발효곡물 제조용 스타터(starter) 조성물을 제공한다.In addition, the present invention relates to the aforementioned Pediococcus acid starch composition for the production of fermented grains containing an acidilactici SRCM 102024 strain or a culture thereof as an active ingredient.
본 발명에 있어서, 발효곡물 제조용 스타터(starter)란 발효곡물 제조를 위해 발효에 관여하는 미생물을 포함하는 제제 또는 조성물을 의미한다. 발효곡물 제조 시에 첨가함으로써 발효된 곡물에서 생장할 수 있는 미생물 또는 우점종으로 생장할 수 있는 미생물을 제공하기 위하여 사용된다. 상기 곡물 발효용 스타터를 사용하여 발효곡물을 제조하는 경우, 상기 곡물 발효용 스타터에 포함된 미생물에 의하여, 발효곡물의 품질을 일정하게 조절하거나, 특정한 목적, 일 예로 발효곡물에서 이취를 발생시키지 않거나, 감소시키는 목적 또는 발효곡물에서 구수한 맛이나 단맛을 강화시키기 위한 목적을 달성할 수 있다. 본 발명에서는 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 또는 이의 배양액을 스타터 균주로 이용함으로써, 피부염 원인균에 대한 항균 활성이 있으며, 더욱이 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 처리하지 않은 대조구에 비해 아미노태질소, 유기산, 유기당 및 유리아미노산 함량이 증가하고, α-글루코시다제 저해활성 및 프로테아제 활성이 증가되며, 유산균 총수가 증가된 발효곡물을 제조할 수 있다.In the present invention, a starter for manufacturing fermented grain means a preparation or a composition containing microorganisms involved in fermentation for producing fermented grain. It is used to provide a microorganism capable of growing in fermented grains or a microorganism capable of growing into a dominant species by addition at the time of fermented grain production. When the fermented grain is produced using the starter for cereal fermentation, the quality of the fermented grain is controlled by the microorganisms contained in the starter for cereal fermentation, , The purpose of reducing, or the purpose of enhancing the flavor or sweetness imparted by the fermented grain. In the present invention, Pediococcus acidilactici SRCM 102024 or a culture thereof as a starter strain has antimicrobial activity against a causative agent of dermatitis and furthermore has an antibacterial activity against Pediococcus acidic , lactic acid, lactic acid bacteria, acidic lactic acid , SRCM 102024 strain, amino acid nitrogen, organic acid, organic sugar and free amino acid content, α-glucosidase inhibitory activity and protease activity were increased, Can be manufactured.
본 발명의 균주를 배양하는 방법은 당업계에 통상적으로 이용되는 방법에 따라 배양할 수 있으며, 특별한 방법에 한정되는 것은 아니다.The method for culturing the strain of the present invention can be carried out according to a method commonly used in the art, and is not limited to a specific method.
또한, 본 발명은 상기 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주 또는 이의 배양액을 스타터(starter) 균주로 이용하여 곡물을 발효하여 제조된 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 제공한다.The present invention also provides a fermented grain having antimicrobial activity against a causative organism of dermatitis produced by fermenting cereals using Pediococcus acidilactici strain SRCM 102024 or a culture thereof as a starter strain do.
본 발명의 일 구현 예에 따른 발효곡물에서, 상기 피부염 원인균은 여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 또는 아토피 원인균인 스타필로코커스 에피더미디스(Staphylcoccus epidermidis)일 수 있으나, 이에 제한되지 않는다.In the fermented grain according to one embodiment of the present invention, the dermatitis causing bacteria are selected from the group consisting of Propionibacterium acnes causing acne acnes) or atopic pathogen is Staphylococcus epidermidis (Staphylcoccus epidermidis ). < / RTI >
본 발명의 일 구현 예에 따른 발효곡물에서, 상기 발효곡물은 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024 균주를 처리하지 않은 대조구에 비해 아미노태질소, 유기산, 유기당 및 유리아미노산 함량이 증가하고, α-글루코시다제 저해활성 및 프로테아제 활성이 증가되며, 유산균 총수가 증가된 것일 수 있으나, 이에 제한되지 않는다.In a fermented grain according to an embodiment of the present invention, the fermented grain is selected from the group consisting of Pediococcus acidilactici) SRCM increases the amino nitrogen, organic acids, organic sugars and free amino acid content than the control not treated 102 024 the strain and, α- glucosidase, and the increase in the inhibitory activity and protease activity, may be a lactic acid bacteria is increased, but the total number , But is not limited thereto.
또한, 본 발명은 상기 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 유효성분으로 함유하는 여드름 또는 아토피 피부염의 예방 또는 개선용 건강기능식품 조성물을 제공한다.Also, the present invention provides a health functional food composition for preventing or ameliorating acne or atopic dermatitis containing an effective ingredient of a fermented grain having an antibacterial activity against the causative agent of dermatitis.
또한, 본 발명의 상기 건강기능식품은 여드름 또는 아토피 피부염을 예방하거나 개선하기 위해 섭취할 수 있는 것이면 특별히 제한되지 않는다.The health functional food of the present invention is not particularly limited as long as it can be ingested to prevent or ameliorate acne or atopic dermatitis.
본 발명의 상기 발효곡물을 식품첨가물로 사용하는 경우, 상기 발효곡물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 발효곡물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 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.When the fermented grain of the present invention is used as a food additive, the fermented grain can be directly added or 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 its intended use (prevention, health or therapeutic treatment). Generally, the fermented grains of the present invention are added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, in the production of food or beverage. 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 range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.
본 발명의 건강 음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.
상기 외에 본 발명의 추출물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the extract of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
또한, 본 발명은 상기 피부염 원인균에 대한 항균활성을 가지는 발효곡물을 유효성분으로 함유하는 여드름 또는 아토피 피부염의 개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for improving acne or atopic dermatitis containing, as an active ingredient, a fermented grain having an antibacterial activity against the causative agent of dermatitis.
본 발명의 여드름 또는 아토피 피부염 개선용 화장료 조성물에 있어서, 상기 화장료 조성물은 피부외용연고, 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 아이크림, 모이스처 크림, 핸드 크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어지는 군으로부터 선택된 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형의 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다.In the cosmetic composition for improving acne or atopic dermatitis according to the present invention, the cosmetic composition may be at least one selected from the group consisting of an ointment for external use, a cream, a softening agent, a nutrient lotion, a pack, an essence, a hair tonic, a shampoo, a rinse, a hair conditioner, Skin lotion, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, But are not limited to, formulations selected from the group consisting of cleansing lotions, cleansing creams, body lotions and body cleansers. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산아연 등이 이용될 수 있다. When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation 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. In the case of a spray, in particular, / Propane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 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 present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.
When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.
재료 및 방법Materials and methods
1. 영농법인 가인 과일 발효액으로부터 균주 분리 및 동정1. Isolation and Identification of Strain from Fruit Fermentation Liquor
(1) 미생물 분리(1) Microbial separation
실험재료는 전라북도 순창군에 위치한 영농법인 가인에서 여러 가지 과일을 이용하여 제조한 과일발효액을 사용하였고, 발효액은 5년 발효한 것으로부터 미생물을 분리하였다. 미생물 분리 방법은 각 시료 10g을 멸균생리식염수 90 ml에 희석하여 균질화시킨 후 순차적으로 희석하고, 희석액을 NA(nutrient agar, Difco), MRS 배지에 도말하여 37℃에서 24시간, 30℃, 48~72시간 배양하였다. 이때 형성된 집락 중 세균, 유산균으로 추정되는 특정한 콜로니를 선발한 후 순수분리 하였다. 균주 분리 및 배양 방법은 도 1의 모식도와 같이 수행하였다. 이 균주들은 동정의뢰 분석하였으며, 10% 글리세롤 스탁하여 시료로 사용하였다.
Experimental materials were fruit fermented liquid prepared by various fruits at Cain farming in Sunchang - gun, Jeollabuk - do. The fermentation broth was fermented for 5 years and isolated microorganisms. For microbial separation, 10 g of each sample was diluted in 90 ml of sterilized physiological saline, homogenized, and sequentially diluted. The diluted solution was applied to NA (nutrient agar, Difco) and MRS medium and incubated at 37 ° C. for 24 hours, And cultured for 72 hours. At this time, specific colonies which are presumed to be bacteria and lactic acid bacteria were selected and purified. The method for isolating and culturing the strain was performed as shown in the schematic diagram of FIG. These isolates were identified and analyzed using 10% glycerol stock.
(2) 미생물 분리 균주 동정(2) Identification of microorganism-isolated strains
분리된 균주는 마크로젠에 의뢰하여 16S rRNA 시퀀싱 및 BLAST 테스트를 실시하여 동정결과를 도출하였다.
The isolated strains were subjected to 16S rRNA sequencing and BLAST test by Macrogen, and the identification results were obtained.
(3) 분리균주 항균활성(3) Antibacterial activity of isolate strain
Agar well diffusion assay를 이용하여 위해 미생물 3종(표 1)에 대한 항균활성을 확인하였다. NA 액체배지 5 mL에 순수 분리균을 배양하여 상등액을 13,000 rpm, 4℃ 30분간 원심분리하여, 0.85% 소프트 아가를 만든 후 각 웰에 상등액을 90 ㎕씩 분주하고, 무균상태에서 자연건조를 거친 후 37℃에서 24시간 배양하였다. 배양 후, 웰 주위에 형성된 저해 환의 지름(Cm)을 측정하여 항균활성의 유무를 확인하였다.Agar well diffusion assay was used to confirm antimicrobial activity against the three microorganisms (Table 1). NA liquid medium, and the supernatant was centrifuged at 13,000 rpm for 30 minutes at 4 ° C to prepare 0.85% soft agar. 90 μl of the supernatant was dispensed into each well, and the supernatant was naturally dried in an aseptic state And then cultured at 37 ° C for 24 hours. After culturing, the diameter (Cm) of the inhibitory circle formed around the well was measured to confirm the presence of antibacterial activity.
2. 발효액 일반성분 분석2. Analysis of general components of fermentation broth
(1) pH (1) pH
시료 5g, 증류수 45g을 교반하여 총 50g을 상온에서 자동 적정기(Excellence Titrator T50M, Switzerland)를 이용하여 3회 반복 측정한 뒤 평균값을 구하였다.
5 g of the sample and 45 g of distilled water were stirred and a total of 50 g was measured at room temperature three times using an automatic titrator (Excellence Titrator T50M, Switzerland), and the average value was determined.
(2) 산도(2) pH
시료 5g, 증류수 45g을 교반하여 총 50g을 상온에서 자동 적정기(Excellence Titrator T50M, Switzerland)를 이용하여 3회 반복 측정한 뒤 평균값을 구하였다.
5 g of the sample and 45 g of distilled water were stirred and a total of 50 g was measured at room temperature three times using an automatic titrator (Excellence Titrator T50M, Switzerland), and the average value was determined.
(3) 당도(3) sugar content
시료 1g을 10배 희석하여 당도계(Master-2M, 1M. ATAGO Co., LTD. Japan)를 사용하여 측정하였다.
1 g of the sample was diluted 10-fold and measured using a sugar meter (Master-2M, 1M ATAGO Co., LTD.
3. 곡물발효 실험 방법3. Grain fermentation experiment method
1) 실험재료1) Experimental material
곡물 발효에 사용하는 곡물로는 발아보리, 발아현미, 보리, 현미 등을 사용하였고, 발아보리와 현미, 발아현미와 보리, 발아보리와 발아현미를 각각 제조하여 원료를 다르게 하여 균주를 접종하였다. 균주는 영농법인 가인 과일발효액에서 분리된 균주 페디오코커스 애시디락티시(Pediococcus acidilactici) 1종과 발효미생물산업진흥원에 보유하고 있는 유산균 균주 류코노스톡 메센테로이데스(Leuconostoc mesenteroides) 2종, 락토바실러스 플란타룸(Lactobacillus plantarum) 2종을 분양받아 실험을 진행하였다.
Germination barley, germinated brown rice, barley and brown rice were used for grain fermentation, and germinated barley, brown rice, germinated rice, barley, germinated barley and germinated brown rice were prepared. The strain was isolated from a fermentation broth of a farming cain, Pediococcus acidilactici ) and lactobacillus strains belonging to the fermentative microorganism industry promotion institute ( Leuconostoc mesenteroides ) 2 species, Lactobacillus plantarum ( Lactobacillus plantarum ) was distributed and tested.
2) 곡물 발효조건2) Grain Fermentation Condition
(1) 유산균 균주 배양 및 곡물발효 조건 (1) Culture of lactic acid bacteria and fermentation conditions of cereals
본 실험에서 사용한 균주는 발명자의 분리 균주와 발효미생물산업진흥원에서 분양받은 균을 포함하여 5 종류의 유산균 균주를 사용하여 실험을 진행하였다. 분양균주는 락토바실러스 MRS 아가 (Difco) 배지에 도말하여 35℃, 48시간 1차 배양한 후 락토바실러스 MRS 배지 35℃, 24시간 2차 배양하여 사용하였다.The strains used in this experiment were five strains of lactic acid bacteria including the strains isolated from the inventors' isolates and the fermented microorganism Industry Promotion Agency. The pre-cultured strain was streaked on Lactobacillus MRS agar (Difco) medium, and then primary cultured at 35 ° C for 48 hours, followed by secondary culturing for 24 hours at 35 ° C in Lactobacillus MRS medium.
(2) 곡물발효 원료선정(2) Selection of raw materials for cereal fermentation
곡물 원료는 보리, 발아보리, 현미, 발아현미를 선정하고 각각을 조합하여 R(발아보리+현미), G(발아보리+발아현미), Y(발아현미+보리)로 하고 분양받은 균주를 각각 또는 병용 접종하여 발효를 진행하였다. 전 과정을 도식화하여 도 2에 나타내었다. The raw materials of cereals are selected from barley, germinated barley, brown rice and germinated brown rice, and the strains which have been sold as R (germinated barley + brown rice), G (germinated barley + germinated brown rice) and Y Or in combination with each other. The whole process is schematically shown in Fig.
(3) 곡물발효 수분 첨가량 설정(3) Setting of grain fermentation water addition amount
발아보리와 현미, 발아보리와 발아현미, 발아현미와 보리를 각각 1:1로 무게를 측정하였다. 측량한 곡물에 수분을 20%, 30%, 40%, 50%, 60% (W/W)로 제조하여 증자한 후 분양받은 균주를 발효 조건별로 107 CFU/㎖ 접종하여 발효를 진행하였다.Germinated barley and brown rice, germinated barley and germinated brown rice, germinated rice and barley were weighed 1: 1. Moisture contents of 20%, 30%, 40%, 50% and 60% (W / W) were added to the cultured cereal grains and the fermented broth was inoculated with 10 7 CFU / ㎖ of fermentation conditions.
3) 3) 곡물발효물의Grain fermented 특성 평가 실험방법 Characteristic evaluation experiment method
(1) 수분(1) Moisture
시료 5g을 적외선 수분기(Kette,720)를 이용하여 측정하였다.
5 g of the sample was measured using an infrared water flow meter (Kette, 720).
(2) pH (2) pH
시료 5g, 증류수 45g을 교반하여 총 50g을 상온에서 자동 적정기(Excellence Titrator T50M, Switzerland)를 이용하여 3회 반복 측정한 뒤 평균 값을 구하였다.
5 g of sample, and 45 g of distilled water were stirred, and a total of 50 g was repeatedly measured three times using an automatic titrator (Excellence Titrator T50M, Switzerland) at room temperature, and the average value was determined.
(3) 산도(3) pH
시료 5g, 증류수 45g을 교반하여 총 50g을 상온에서 자동 적정기(Excellence Titrator T50M, Switzerland)를 이용하여 3회 반복 측정한 뒤 평균 값을 구하였다.
5 g of sample, and 45 g of distilled water were stirred, and a total of 50 g was repeatedly measured three times using an automatic titrator (Excellence Titrator T50M, Switzerland) at room temperature, and the average value was determined.
(4) 아미노태질소(4) Amino nitrogen
아미노태질소 함량은 자동 적정기(Excellence Titrator T50M, Switzerland)를 이용하여 시료 1g을 취하여 증류수 50 mL를 가하고 1시간 동안 교반한 후 0.1 N NaOH 용액으로 pH 8.4까지 적정하였다. 여기에 중성 포르말린 용액 20 mL를 가하고 다시 0.1 N NaOH 용액으로 pH 8.4가 되도록 적정하였다.
Amino nitrogen content was measured by using an automatic titrator (Excellence Titrator T50M, Switzerland), taking 1 g of sample, adding 50 mL of distilled water, stirring for 1 hour, and titrating to pH 8.4 with 0.1 N NaOH solution. To this was added 20 mL of neutral formalin solution and titrated to pH 8.4 with 0.1 N NaOH solution.
(5) 유기산 및 유리당 (5) Organic acids and free sugars
유기산과 유리당의 분석은 시료 5g에 증류수 45 mL를 가하여 1시간 동안 균질화시킨 후 0.45㎛ 멤브레인 필터를 통과시킨 후 표 5의 분석조건으로 HPLC로 동시에 분석하였다. To analyze the organic acids and free sugars, 45 mL of distilled water was added to 5 g of the sample, homogenized for 1 hour, passed through a 0.45 μm membrane filter, and simultaneously analyzed by HPLC under the analysis conditions shown in Table 5.
(6) 유리아미노산(6) free amino acids
유리아미노산은 시료 2g을 취하여 3차 증류수 30 mL 넣고 교반한 후 50 mL로 정용한수 초음파를 이용하여 20분간 추출한 후 원심분리(3,000 rpm, 10분)한 다음 상등액 2mL에 5% TCA 2mL를 넣은 후 원심분리(10,000 rpm, 10분)한 후 상등액을 취하여 0.02 N HCL로 희석한 후 0.2㎛ 시린지 필터에 통과시킨 후 표 6의 분석조건으로 아미노산 분석기로 분석하였다For free amino acids, take 2 g of sample, add 30 mL of third-order distilled water, stir with 50 mL of distilled water for 20 minutes, centrifuge at 3,000 rpm for 10 minutes, add 2 mL of 5% TCA to 2 mL of supernatant After centrifugation (10,000 rpm, 10 min), the supernatant was diluted with 0.02 N HCl, passed through a 0.2 μm syringe filter, and analyzed with an amino acid analyzer under the analysis conditions of Table 6
Hitachi 4.6×40mm (Ammonia filtering)Hitachi 4.6 x 60 mm (speration)
Hitachi 4.6 × 40 mm (Ammonia filtering)
(7) DPPH 라디칼 소거능 측정(7) Measurement of DPPH radical scavenging ability
DPPH(1,1-diphenyl-2-picrylhydrazyl) 라디칼 소거능은 전자공여작용 (electron donationg abilities, EDA)에 대한 효과로 각 시료의 환원력을 측정하였다. 즉, 시료 150 ㎕에 0.2 mM DPPH 용액 (99% MeOH에 용해) 150 ㎕를 가한 후 30분간 상온에서 방치한 후 분광광도계를 사용하여 흡광도 517 nm에서 흡광도의 변화를 측정하였다. 각 시료의 라디칼 소거능은 아래의 식에 의해 시료 첨가구 및 무 첨가구간의 흡광도 차이를 백분율로 나타내었다.DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured by the effect of electron donation abilities (EDA). That is, 150 μl of a 0.2 mM DPPH solution (dissolved in 99% MeOH) was added to 150 μl of a sample, and the mixture was allowed to stand at room temperature for 30 minutes. Then, the absorbance at 517 nm was measured using a spectrophotometer. The radical scavenging activity of each sample was expressed as a percentage of the absorbance difference between the sample addition and the no addition region by the following equation.
전자공여능(%)=(1-시료 첨가구의 흡광도/무처리 구의 흡광도) × 100
Electron donating ability (%) = (1-absorbance of sample added / absorbance of non-treated) x 100
(8) α-글루코시다제 (α-glucosidase) 저해 활성(8)? -Glucosidase inhibitory activity
AGI (α-glucosidase inhibitory) 저해활성은 다음과 같은 방법으로 측정하였다. 즉, 시료 50 ㎕에 0.3 U/mL α-글루코시다제 효소액 50 ㎕(in 0.1 M PBS, pH 6.8)을 혼합하여 37℃에서 10분 동안 전배양하였다. 3 mM ρ-NPG (ρ-nitro-phenyl-α-glucopyranoside, in 0.1 M PBS, pH 6.8) 100 ㎕을 가한 후 37℃에서 10분간 반응시킨 후 0.1 M Na2CO3 100㎕를 가하여 반응을 정지시켰다. 이때 생성된 ρ-니트로페놀의 양을 분광 광도계(ELISA reader, Infinite 200 TECAN)를 사용하여 흡광도 405 nm에서 측정하였다. 이때 대조구는 배양액 대신 0.1 M PBS (pH 6.8)을 사용하였으며, blank로는 균주 배양액 및 효소액 대신 0.1 M PBS (pH 6.8)을 사용하였다.AGI (α-glucosidase inhibitory) inhibitory activity was measured by the following method. That is, 50 μl of 0.3 U / ml α-glucosidase enzyme solution (50 μl in 0.1 M PBS, pH 6.8) was mixed with 50 μl of the sample and pre-cultured at 37 ° C for 10 minutes. 3 mM ρ-NPG (ρ- nitro-phenyl-α-glucopyranoside, in 0.1 M PBS, pH 6.8) was added to 100 ㎕ After 10 min at 37 ℃ 0.1 M Na 2 CO 3 Was added to stop the reaction. The amount of ρ-nitrophenol produced at this time was measured at an absorbance of 405 nm using a spectrophotometer (ELISA reader, Infinite 200 TECAN). As a control, 0.1 M PBS (pH 6.8) was used instead of the culture, and 0.1 M PBS (pH 6.8) was used instead of the culture medium and enzyme solution as the blank.
효소저해능(%) = (1-반응구의 ρ-니트로페놀 생성량/무처리구의 ρ-니트로페놀 생성량) × 100
Enzyme inhibitory activity (%) = (1-p-nitrophenol formation amount in reaction section / p-nitrophenol production amount in non-processing section) x 100
(9) 단백질 및 전분 분해 효소활성 분석(9) Analysis of protein and starchase activity
선별 균주의 세포 외 효소들 중 단백질, 섬유소, 전분 분해효소의 활성을 측정하기 위하여 각 효소와 특이적으로 반응할 기질 성분을 포함한 평판배지를 제조하였다. 단백질 분해효소(protease)의 활성은 2% 스킴밀크 (DifcoTM, USA)에 1.5% 아가(Junsei Chemical Co. Ltd., Japan)를 첨가하여 스킴밀크 아가를 제조하였으며, 효소활성을 측정하기 위하여 각각의 배지에 5 mm의 구멍을 뚫었다. 선별된 유산균은 MRS 액체배지에서 37℃, 24시간 진탕배양하였고, 선별균주 배양액은 15,000xg로 10분간 원심분리하여 상등액 취하고 0.45㎛ 멤브레인 필터 (Sartorius, Germany)로 제균한 뒤 100㎕씩 각각의 세포 외 효소 활성 측정 배지의 구멍에 분주하여 37℃에서 24시간 반응시킨 후 웰 주위로 생기는 환의 크기를 측정하였다.
In order to measure the activity of protein, fiber, and starch hydrolytic enzymes in the extracellular enzymes of the selective strains, a plate medium containing substrate components that specifically react with each enzyme was prepared. The activity of the protease was determined by adding 1.5% agar (Junsei Chemical Co. Ltd., Japan) to 2% skim milk (Difco ™, USA) A 5 mm hole was drilled in the medium. Was selected lactobacillus culture MRS broth ℃ 37, 24 hours with shaking at, screening strain culture solution is taken back by each supernatant was centrifuged for 10 minutes to 15,000x g in a filtering 0.45㎛ membrane filter (Sartorius, Germany) 100㎕ After extracellular enzymatic activity was measured, the cells were reacted at 37 DEG C for 24 hours, and the size of the circle around the wells was measured.
(10) 곡물발효 추출물의 항균 활성 (10) Antimicrobial activity of grain fermented extract
식품의 부패를 막기 위해 수분활성도, 염장, 당장 방법을 이용하고 있지만, 적용범위가 한정되어 있기 때문에 인공합성보존료를 사용하여 저장성을 높이고 있다. 아토피에 대한 소비자의 건강지향적 욕구가 증가하여 아토피를 발생하는 위해미생물을 선정하여, 한국미생물보존센터(KCCM)에서 3종을 분양받아 실험에 사용하였고, 이에 대한 항균력을 검증하고자 하였다.Water activity, salting, and direct method are used to prevent food corruption. However, because of limited application range, artificial synthetic preservatives are used to improve storage stability. In order to increase the consumer 's health - oriented desire for atopy, we selected the microorganisms which cause atopy, and the three kinds of microorganisms were distributed in KCCM to test the antibacterial activity against them.
(11) 곡물발효 후 유산균 총 균수 측정 (11) Total bacteria count of lactic acid bacteria after cereal fermentation
시료 25g을 무균적으로 취하여 멸균수 225 mL을 넣어 Bag mixer로 균질화하여, 각 시료를 10 희석법으로 희석한 후 희석액 100 ㎕를 락토바실러스 MRS 아가 (Difo) 배지에 도말하여 35℃, 48시간 배양하여 콜로니 수를 계수(CFU/mL)하였다.
25 g of the sample was aseptically taken, 225 mL of sterilized water was added, and homogenized with a bag mixer. Each sample was diluted by 10 dilution, 100 μL of the diluted solution was plated on a Lactobacillus MRS agar (Difo) medium and cultured at 35 ° C. for 48 hours The number of colonies was counted (CFU / mL).
실시예Example 1. 발효액 이화학적 분석 1. Physicochemical analysis of fermentation broth
1) pH, 산도, 당도 측정1) pH, acidity and sugar content
영동법인 가인의 과일발효액인 원재료의 일반성분 분석결과는 표 8과 같다. 일반성분 분석 결과에서는 pH는 3.57±0.02, 산도는 7.44±1.34, 당도는 42.97±0.05의 결과를 보였다. Table 8 shows the results of the analysis of the general components of raw materials that are fruit juices of Cain in Yeongdong corporation. As a result of the general composition analysis, the pH was 3.57 ± 0.02, the acidity was 7.44 ± 1.34, and the sugar content was 42.97 ± 0.05.
발효액 원재료
Raw material of fermentation liquid
3.57±0.02
3.57 + 0.02
7.44±1.34
7.44 ± 1.34
42.97±0.05
42.97 ± 0.05
2) 발효액 미생물 분석2) Analysis of microorganisms in fermentation broth
발효액의 총 균수 측정 및 대장균군을 분석하여 결과를 표 9에 정리하였다. 미생물 분석결과는 총균수, 대장균군, B.C에서 음성으로 나타났다. The total number of bacteria in the fermentation broth was measured and the coliform group was analyzed. The results are summarized in Table 9. Microbial analysis results were negative for total bacteria, E. coli and B.C.
3) 발효액으로부터 미생물 분리 및 동정 3) Isolation and identification of microorganisms from fermentation broth
균주 분리는 도 1과 같이 실시하였으며 분리된 균주는 마크로젠에 의뢰하여 16S rRNA 시퀀싱 및 BLAST 테스트를 실시하여 동정결과를 도출하였다. 분석시 사용한 프라이머는 도 4와 같다. 상기 균주의 동정 과정에서 얻어진 염기서열(서열번호 1)을 NCBI에 대입한 결과 NR_042057.1로 염기서열의 96%가 리딩되었고 99%가 일치하였으며 페디오코커스 애시디락티시(Pediococcus acidilactici) 균주로 동정되었다. 신뢰성을 나타내는 E-Value는 0.0을 나타내 신뢰성이 높은 결과임이 확인되었다. 신규 분리 동정 균의 계통수 분석을 실시한 결과는 도 5에 나타내었다.The isolation of the strain was carried out as shown in Fig. 1, and the isolated strains were subjected to 16S rRNA sequencing and BLAST test by Macrogen, and the results were identified. The primers used in the analysis are shown in Fig. As a result of substituting the nucleotide sequence (SEQ ID NO: 1) obtained in the step of identification of the above strain into NCBI, 96% of the nucleotide sequences were read and 99% were identical to NR_042057.1 and Pediococcus acidilactici . The E-value indicating reliability was 0.0, confirming that the reliability was high. The results of the phylogenetic analysis of the newly isolated isolates were shown in Fig.
이 균의 특성으로는 페디오코커스는 락토바실라세아(Lactobacillaceae) 과(科)에 속하는 그람 양성 유산균의 한 속(屬)이며 이들은 보통 쌍 혹은 두 쌍으로 발생하며 대칭성을 유지하며 두 개로 분리된다. 이 균은 류코노스톡(Leuconostoc) 그리고 락토바실러스(Lactobacillus)와 같은 다른 유산균처럼 양배추를 사우어크라우트(Sauerkraut)로 바꾸는 발효에 대해 그 원인이 되는 균으로 알려져 있는 균이다.
Pediococcus is a genus of gram-positive lactic acid bacteria belonging to Lactobacillaceae and family, which usually occur in pairs or pairs and maintain symmetry and are separated into two . These bacteria are bacteria that flow Stock Kono (Leuconostoc) and other lactic acid bacteria such as Lactobacillus for the cabbage such as (Lactobacillus) in fermented sauerkraut to change (Sauerkraut) bacteria that are known as the most likely cause.
4) 발효액의 항균 활성4) Antimicrobial activity of fermentation broth
발효액을 대상으로 실험한 항균활성 분석결과는 표 10과 도 6과 같다. 항균활성에서는 분리 균주 중에는 세균류에서 항균활성을 나타내지 않았으며, 유산균인 스타필로코커스 에피더미디스 KCCM35494에서 높은 항균활성을 나타내었다.The results of the antimicrobial activity analysis of the fermentation broth are shown in Table 10 and FIG. In the antimicrobial activity, the isolates did not exhibit antimicrobial activity in the fungi, and lactic acid bacteria Staphylococcus epidermidis KCCM35494 showed high antimicrobial activity.
(할로크기 (cm), 평균±표준편차)Antimicrobial activity
(Halo size (cm), mean ± standard deviation)
KCCM40416
S. epidermidis
KCCM40416
KCCM40416
pH7.0 S. epidermidis
KCCM40416
pH 7.0
S. epidermidis KCCM35494
pH7.0 S. epidermidis KCCM40416
pH 7.0
5) 유산균 분리균주 항균활성5) Antibacterial activity of isolate of lactic acid bacteria
유산균 분리균주의 항균활성 분석결과는 표 11과 같다. 영농조합법인 가인에서 제공한 발효액에서 분리동정한 페디오코커스 애시디락티시 SRCM 102024 균주는 유산균인 스타필로코커스 에피더미디스 KCCM35494에 대해 할로크기가 3.8cm로 나타나 다른 균주들에 비해 높은 항균활성을 나타내었다. Table 11 shows the results of the antimicrobial activity analysis of the isolates of the lactic acid bacteria. The strain Pediococcus aceyltactii SRCM 102024 isolated and isolated from the fermentation broth provided by Cain, the Agricultural Cooperatives Act, is a strain of Staphylococcus epidermidis The halo size of KCCM35494 was 3.8cm, which showed higher antimicrobial activity than other strains.
(할로크기 (cm), 평균±표준편차)Antimicrobial activity
(Halo size (cm), mean ± standard deviation)
S. epidermidis KCCM40416
S. epidermidis KCCM35494
P. acnes KCCM41747
애시디락티시Pedio Caucus
Ashidi Laktishi
플란타룸Lactobacillus
Flora Room
플란타룸Lactobacillus
Flora Room
메센테로이데스Ryukono Stock
Messeneroides
메센테로이데스Ryukono Stock
Messeneroides
실시예Example 2. 곡물 발효 조건 2. Grain fermentation conditions
1) 유산균 선정1) Selection of lactic acid bacteria
본 실험에서는 발효미생물산업진흥원에 보유하고 있는 류코노스톡 메센테로이데스(Leuconostoc mesenteroides) 2종과 락토바실러스 플란타룸(Lactobacillus plantarum) 2종을 포함하여 4종의 유산균 균주를 분양받아 실험을 진행하였고, 영농법인 가인 과일발효액에서 분리된 본 발명의 균주 페디오코커스 애시디락티시(Pediococcus acidilactici) 1종을 더하여 모두 5종을 배양하여 실험을 진행하였다. In this experiment, the fermented microorganism Industry Promotion Agency has leuconostoc mesenteroides ( Leuconostoc mesenteroides ) Two species Lactobacillus ( Lactobacillus) plantarum ), which was isolated from a fermentation broth of a farming corporation, and the strain Pediococcus acidilactici ) were added to each of the five cultivars.
2) 곡물발효 원료선정2) Selection of raw materials for cereal fermentation
발아보리와 현미, 발아보리와 발아현미, 발아현미와 보리를 각각 제조하여 곡물의 원료를 다르게 한 후 115℃, 20분 증자 과정을 하였다. 분양받은 균주별로 접종하여 발효를 진행하였으며, 유산균 균주를 (107 CFU/㎖) 접종하여, 37℃에서 발효를 진행하였다. 그 결과 발아보리와 현미가 우수한 결과를 나타내었다.
Germinated barley and brown rice, germinated barley and germinated brown rice, germinated rice and barley were prepared, respectively. Fermentation was carried out at 37 ℃ by inoculation with lactic acid bacteria strain (10 7 CFU / ㎖). As a result, germinated barley and brown rice showed excellent results.
실시예Example 3. 곡물 발효 특성 평가 결과 3. Evaluation results of grain fermentation characteristics
(1) 수분(1) Moisture
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 수분 분석 결과는 표 13과 같다. 본 연구에서는 곡물발효하지 않은 대조구가 0일차에서는 42.55%값을 보였으며, 7일차에서는 39.10%로 감소하는 경향을 보였다. 단일균주 접종 발효가 진행된 곡물도 0일차에서는 42.56%에서 발효가 진행될수록 38.65%, 39.07%, 39.51%, 41.25%, 40.48%로 감소하는 경향을 보였고, 복합균주로 진행된 곡물도 0일차에서는 42.56%에서 발효가 진행될수록 40.21%, 37.97%, 39.92%로 감소하는 경향을 보였다. 이처럼 유의적인 차이를 보이지 않았다.Table 13 shows the results of moisture analysis of the fermented cereal fermented with different strains. In this study, the untreated control was 42.55% at day 0 and decreased to 39.10% at
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(2) pH (2) pH
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 pH 분석은 표 14와 같다. 본 연구에서는 곡물발효하지 않은 대조구가 0일차에서는 6.42 값을 보였으며, 7일차에서는 4.12로 감소하는 경향을 보였다. 단일균주 접종 발효가 진행된 곡물도 0일차에서는 6.45에서 발효가 진행될수록 5.85, 4.81, 4.05, 4.71, 4.60%로 감소하는 경향을 보였고, 복합균주로 진행된 곡물도 0일차에서는 6.45에서 발효가 진행될수록 4.77, 4.33, 4.95로 감소하는 경향을 보였다. 대조구를 포함한 9 균주 중 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024가 pH가 가장 낮은 값을 보였다. 이는 발효되는 과정에서 생성되는 유산균과 미생물이 유기산을 생성하여 pH는 낮아지는 것으로 보여진다. Table 14 shows the pH analysis of the fermented cereal fermented with a single strain and a complex strain. In this study, the control value of cereal fermentation showed a value of 6.42 at the 0th day and decreased to 4.12 at the 7th day. As the fermentation progressed at 6.45, the fermentation progressed to 5.85, 4.81, 4.05, 4.71 and 4.60% at 0 day, and the fermentation progressed to 4.45 , 4.33, and 4.95, respectively. Among 9 strains including the control, Pediococcus acidilactic ) SRCM 102024 showed the lowest pH value. These results suggest that lactic acid bacteria and microorganisms formed during fermentation produce organic acids and pH is lowered.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(3) 산도(3) pH
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 적정산도는 표 15와 같다. 본 연구에서는 곡물발효하지 않은 대조구가 0일차에서는 0.28%값을 보였으며, 7일차에서는 0.30%로 증가하는 경향을 보였지만 유의적인 차이를 보이지는 않았다. 단일균주 접종 발효가 진행된 곡물도 0일차에서는 0.27%에서 발효가 진행될수록 1.59%, 1.16%, 2.55%, 1.20%, 1.40%로 증가하는 경향을 보였고, 복합균주로 진행된 곡물도 0일차에서는 0.27%에서 발효가 진행될수록 1.50%, 1.70%, 1.90% 증가하는 경향을 보였다. 9개 균주 중 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024가 적정산도가 가장 높은 값을 보였고, 이는 다른 균주들보다 유산균 발효가 잘된 것이라고 판단된다. 다른 균주들은 유의적인 차이를 보이지 않았다.Table 15 shows the titratable acidity of the fermented cereal fermented with different strains. In the present study, the control value of the unfermented grain was 0.28% at the 0th day and increased to 0.30% at the 7th day, but the difference was not significant. As the fermentation progressed from 0.27% on the 0th day, the number of grains increased to 1.59%, 1.16%, 2.55%, 1.20% and 1.40%, respectively. , Fermentation progressed by 1.50%, 1.70% and 1.90%, respectively. Of the nine strains, Pediococcus acidilactic ) SRCM 102024 showed the highest titratable acidity, indicating that lactic acid fermentation was better than other strains. No significant differences were found in other strains.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(4) 아미노태질소(4) Amino nitrogen
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 아미노태질소는 표 16과 같다. 본 연구에서는 곡물발효하지 않은 대조구가 0일차에서는 24.00mg%값을 보였으며, 7일차에서는 22.30mg%로 감소하는 경향을 보였지만 유의적인 차이 보이지는 않았다. 단일균주 접종 발효가 진행된 곡물도 0일차에서는 24.10mg%에서 발효가 진행될수록 35.30mg%, 32.10mg%, 77.00mg%, 50.40mg%, 58.50mg%로 증가하는 경향을 보였고, 복합균주로 진행된 곡물도 0일차에서는 24.10mg%에서 발효가 진행될수록 45.30mg%, 49.50mg%, 37.66mg% 증가하는 경향을 보였다. 9개 균주 중 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024가 아미노태질소가 가장 높은 값을 보였고, 이는 다른 균주들보다 발효가 가장 잘된 것이라고 판단된다.Amino nitrogen content of the fermented cereal fermented by different strains of the single strain and the complex strain is shown in Table 16. In the present study, 24.00 mg% of the control group without grain fermentation was observed at day 0, and decreased to 22.30 mg% at 7 days, but the difference was not significant. As the fermentation progressed from 24.10mg% on the 0th day, the grains inoculated with single strains tended to increase to 35.30mg%, 32.10mg%, 77.00mg%, 50.40mg% and 58.50mg%, respectively. In the day 0, the fermentation progressed from 24.10mg% to 45.30mg%, 49.50mg% and 37.66mg%, respectively. Of the nine strains, Pediococcus acidilactic ) SRCM 102024 showed the highest value of amino nitrogen, which is considered to be the best fermentation than other strains.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(5) 유기산 및 유리당(5) Organic acids and free sugars
시료는 실험방법에 따른 전처리 후 HPLC 분석은 옥살산(Oxalic acid), 시트르산(Citric acid), 말산(Malic acid), 숙신산(Succinic acid), 젖산(Lactic acid), 포름산(Formic acid), 아세트산(Acetic acid) 7종이 분석되었다. 이 중 시트르산(citric acid)은 측정되지 않았으며 젖산(lactic acid)이 가장 많이 검출되었고 아세트산, 숙신산, 말산, 포름산 및 옥살산 순으로 검출되었다. 도 7에는 총 유기산 함량을 나타내었으며 3종 균이 함께 발효된 5일 발효 조건의 복합균주 발효에서 유기산이 가장 높게 측정되었다. 유리당은 시료 전처리 후 제시된 HPLC 조건에서 프럭토스, 글루코스, 수크로스, 말토스가 분석되었으며 프럭토스, 글루코스보다 수크로스, 말토스가 검출되었으며 말토스가 50~90%를 차지하는 것으로 분석되었다. 도 8에는 시료별 총 유리당 함량을 나타내었으며 5일 발효 조건의 3종 균의 복합 발효 조건에서 가장 높게 측정되었다.
After pretreatment according to the experimental method, samples were analyzed for oxalic acid, citric acid, malic acid, succinic acid, lactic acid, formic acid, acetic acid acid) were analyzed. Among them, citric acid was not detected, lactic acid was detected the most, and acetic acid, succinic acid, malic acid, formic acid and oxalic acid were detected in the order. FIG. 7 shows the total organic acid content and the highest concentration of organic acid was obtained in the fermentation of the combined strains of the 5-day fermentation conditions in which the three strains were fermented together. Glucose was analyzed by fructose, glucose, sucrose and maltose after HPLC pretreatment. Fructose, glucose and maltose were detected in maltose and maltose, respectively. FIG. 8 shows the total free sugar content of each sample and was highest in the combined fermentation conditions of three microorganisms under the 5-day fermentation condition.
(6) 유리아미노산(6) free amino acids
발효 일수 별 균주별 발효 후 HPLC에 의해 유리아미노산을 분석한 결과 5일 발효 조건에서 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 유리아미노산 총량이 가장 높게 검출되었다. 또한 아미노산 중 GABA가 5일 발효에서 상승하는 결과를 보였으며 이는 곡물발효의 특성을 잘 나타내는 바 제품의 특성으로 제시될 수 있을 것이다. 도 9에는 실험 조건별 총 유리아미노산의 결과 값을 나타내었다.
The free amino acids were analyzed by HPLC after fermentation by strain according to the number of days of fermentation. As a result, the fermentation conditions of Pediococcus acidilactic ) In SRCM 102024, the total amount of free amino acids was the highest. In addition, GABA in amino acid was increased during 5 days fermentation, which could be suggested as a characteristic of grain fermentation. FIG. 9 shows the results of the total free amino acids according to the experimental conditions.
(7) DPPH 라디칼 소거능 측정(7) Measurement of DPPH radical scavenging ability
항산화 활성으로는 DPPH 라디칼 소거 반응을 실험하였으며 그 결과는 도 10에 나타내었다. 이는 물 추출물에서는 항산화 효과가 미비하여 70% 에탄올 추출에 의한 추출물로 실험하였다. 발효 일수 3일/5일/7일에 따라 균주별 발효 조건에 따라 DPPH 라디칼 소거능의 변화가 미비하였으므로 발효에 따른 영향을 받지 않는 것으로 해석된다.The DPPH radical scavenging reaction was tested for antioxidative activity and the results are shown in FIG. The water extracts showed little antioxidative effect and were tested with 70% ethanol extract. The changes in DPPH radical scavenging ability were not affected by fermentation conditions according to the fermentation conditions according to
전자공여능(%)=(1-시료 첨가구의 흡광도/무처리 구의 흡광도) × 100
Electron donating ability (%) = (1-absorbance of sample added / absorbance of non-treated) x 100
(8) α-글루코시다제(α-glucosidase) 저해 활성(8)? -Glucosidase inhibitory activity
AGI (α-glucosidase inhibitory) 저해활성은 사용된 균주의 상등액과 곡물발효의 물 추출물을 시료로 하여 측정되었다. 반응 후 실험 결과는 다음과 같은 식에 대입하여 산출하였다.AGI (α-glucosidase inhibitory) inhibitory activity was measured by using the supernatant of the strain used and the water extract of the fermentation of grain. The experimental results after the reaction were calculated by substituting the following equation.
효소저해능(%) = (1-반응구의 ρ-nitrophenol 생성량/무처리 구의 ρ-nitrophenol 생성량) × 100
Enzyme inhibitory activity (%) = (1-p-nitrophenol production amount in the reaction vessel / p-nitrophenol production amount in the untreated sphere) × 100
선정된 단일 균주 5종 중 락토바실러스 플란타룸(Lactobacillus plantarum) SRCM 101502, 락토바실러스 플란타룸(Lactobacillus plantarum) SRCM 101503, 그리고 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 AGI 활성이 가장 높게 나타났으며 그 결과는 다음의 표 17과 같다. 락토바실러스 플란타룸(Lactobacillus plantanum) 2종과 페디오코커스 애시디락티시(Pediococcus acidilactici)에서 높은 AGI 활성 정도가 비슷하게 나타났으며 류코노스톡 메센테로이데스(Leuconostoc mesenteroides)에서는 거의 활성이 나타나지 않았다. Single strain of Lactobacillus selected five kinds Planta Room (Lactobacillus plantarum) SRCM 101502, Lactobacillus Planta Room (Lactobacillus plantarum ) SRCM 101503, and Pediococcus acidilactic ) The highest activity of AGI activity was observed in SRCM 102024 and the results are shown in Table 17 below. Two kinds of Lactobacillus plantanum and Pediococcus acidilactici showed a high degree of AGI activity and showed almost no activity in Leuconostoc mesenteroides .
2; 락토바실러스 플란타룸 SRCM 101502, 3; 락토바실러스 플란타룸 SRCM 101503, 4; 페디오코커스 애시디락티시 SRCM 102024, 5; 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677
2; Lactobacillus
(9) 단백질 분해효소 활성 분석(9) Proteolytic enzyme activity assay
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 단백질 분해효소 활성 결과는 표 18과 같다. 0일차인 대조구에서는 단백질 분해효소 활성 나타나지 않았으나, 균주와 발효기간별로 단백질 분해효소 활성 실험한 각기 다른 결과를 다르게 나타내었다. 그 중 단일균주로 발효한 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 2.0 cm로 가장 높은 단백질 분해효소 활성 나타내었다. 대조구에서는 나타나지 않았으며, 다른 균주들은 유의적인 차이를 보이지 않았다.The results of proteolytic enzyme activity of the fermented cereal fermented with different strains of the single strain and the complex strain are shown in Table 18. The protease activity was not detected in the control group of 0 day old, but the different results of the protease activity test by the strain and fermentation period were shown differently. Among them, Pediococcus acidilactici SRCM 102024 fermented with a single strain showed the highest protease activity at 2.0 cm. And the other strains did not show any significant difference.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(10) 아토피와 여드름 균에 대한 항균활성(10) Antimicrobial activity against atopy and acne
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 항균활성 결과는 표 19와 같다. 0일차인 대조구에서는 항균활성이 나타나지 않았으나, 균주별로 발효기간별로 항균활성결과를 다르게 나타내었다. 그 중 아토피 균주인 프로피오니박테리움 아크네스 KCCM 41747 균주에서 단일균주로 5일 발효한 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 1.7cm로 가장 높은 항균력을 나타내었다. Table 19 shows the results of the antimicrobial activity of the fermented cereal fermented with a single strain and a complex strain. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. Among them, Propionibacterium acnes Five the date of entry into force of a single strain KCCM 41747 strain Lactococcus Pedy five kids CD rakti City (Pediococcus acidilactic ) SRCM 102024 showed the highest antibacterial activity at 1.7 cm.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 항균활성 결과는 표 20과 같다. 0일차인 대조구에서는 항균활성이 나타나지 않았으나, 균주별로 발효기간별로 항균활성결과를 다르게 나타내었다. 그 중 여드름 균주인 스타필로코커스 에피더미디스(Staphylcoccus epidermidis) KCCM40416 균주에서 단일균주로 발효한 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 2.9cm로 가장 높은 항균력을 나타내었다. Table 20 shows the results of the antimicrobial activity of the fermented cereal fermented with a single strain and a complex strain. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. Among them, acne strain Staphylococcus Epidermidis epidermidis) was trying Pedy five caucuses in the fermentation of a single strain KCCM40416 strain Sidi rakti City (Pediococcus acidilactic ) SRCM 102024 showed the highest antimicrobial activity as 2.9 cm.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 항균활성 결과는 표 21과 같다. 0일차인 대조구에서는 항균활성이 나타나지 않았으나, 균주별로 발효기간별로 항균활성결과를 다르게 나타내었다. 그 중 여드름 균주인 스타필로코커스 에피더미디스(Staphylcoccus epidermidis) KCCM35494 균주에서 단일균주로 발효한 페디오코커스 애시디락티시(Pediococcus acidilactici) SRCM 102024에서 3.8cm로 가장 높은 항균력을 나타내었고, 발효 5일차에 가장 높은 항균력을 보였다. 이는 스타필로코커스 에피더미디스 KCCM40416 균주의 2.8cm보다 훨씬 높은 항균력을 나타내었다. Table 21 shows the results of the antimicrobial activity of the fermented cereal grains fermented with different strains. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. As the star of acne strains Philo Caucus epidermidis (Staphylcoccus epidermidis) was trying Pedy five caucuses in the fermentation of a single strain KCCM35494 strain Sidi rakti City (Pediococcus acidilactici ) SRCM 102024 showed the highest antimicrobial activity at 3.8cm and showed the highest antimicrobial activity at the 5th day of fermentation. This may be due to the fact that Starfilacococcus epidermidis The antimicrobial activity of KCCM40416 strain was much higher than 2.8cm.
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 101677
1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
(11) 곡물발효 후 유산균 총 수(11) Total number of lactic acid bacteria after grain fermentation
단일균주와 복합균주를 달리하여 발효한 곡물발효물의 유산균수는 표 22와 같다. 본 연구에서는 곡물발효하지 않은 대조구가 0일차에서는 ND값을 보였으며, 7일차에서는 ND로 보였다. 단일균주 접종 발효가 진행된 곡물도 0일차에서는 1.3×107 CFU/㎖에서 발효가 진행될수록 4.6×107 CFU/㎖, 2.1×107 CFU/㎖, 9.5×107 CFU/㎖, 3.3×107 CFU/㎖, 2.1×107 CFU/㎖로 증가하는 경향을 보였고, 복합균주로 진행된 곡물도 0일차에서는 1.3×107 CFU/㎖에서 발효가 진행될수록 2.5×107 CFU/㎖, 7.0×107 CFU/㎖, 3.8×107 CFU/㎖ 증가하는 경향을 보였다. 9개 균주 중 페디오코커스 애시디락티시 SRCM 102024가 유산균수가 가장 높은 값을 보였다. 곡물발효시 발효 3일차에서 유산균수가 1.4×108 CFU/㎖로 증가하다가 5일차에 4.6×108 CFU/㎖로 가장 많이 증가하다가 7일차에는 9.5×107 CFU/㎖로 감소하는 경향을 보였다.Table 22 shows the number of lactic acid bacteria in the fermented cereal fermented with a single strain and a complex strain. In this study, the control value of the untreated control was ND at day 0 and ND at
1: 블랭크(Blank), 2: 락토바실러스 플란타룸 SRCM 101502, 3: 락토바실러스 플란타룸 SRCM 101503, 4: 페디오코커스 애시디락티시 SRCM 102024, 5: 류코노스톡 메센테로이데스 SRCM 101665, 6: 류코노스톡 메센테로이데스 SRCM 101677, 7: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503, 8: 페디오코커스 애시디락티시 SRCM 102024 + 류코노스톡 메센테로이데스 SRCM 101677, 9: 페디오코커스 애시디락티시 SRCM 102024 + 락토바실러스 플란타룸 SRCM 101503 + 류코노스톡 메센테로이데스 SRCM 1016771: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stock Methenseeroides SRCM 101677, 9: Pediococcus acidiflactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
<110> Microbial Institute for Fermentation Industry SHIN, Yong Sung LEE, Im Sook <120> Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses thereof <130> PN17191 <160> 5 <170> KopatentIn 2.0 <210> 1 <211> 1552 <212> DNA <213> Pediococcus acidilactici <400> 1 ttttcggggt tcccctgtac gacttcaccc taatcatctg tcccacctta gacggctagc 60 tcctaaaagg ttaccccacc ggctttgggt gttacaaact ctcatggtgt gacgggcggt 120 gtgtacaagg cccgggaacg tattcaccgc ggcatgctga tccgcgatta ctagcgattc 180 cgacttcgtg taggcgagtt gcagcctaca gtccgaactg agaatggttt taagagatta 240 gctaaacctc gcggtttcgc aactcgttgt accatccatt gtagcacgtg tgtagcccag 300 gtcataaggg gcatgatgat ttgacgtcgt ccccaccttc ctccggtttg tcaccggcag 360 tctcactaga gtgcccaact gaatgctggc aactagtaat aagggttgcg ctcgttgcgg 420 gacttaaccc aacatctcac gacacgagct gacgacaacc atgcaccacc tgtcattctg 480 tccccgaagg gaacgcctaa tctcttaggt tggcagaaga tgtcaagacc tggtaaggtt 540 cttcgcgtag cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 600 ttttgagttt caaccttgcg gtcgtactcc ccaggcggat tacttaatgc gttagctgca 660 gcactgaagg gcggaaaccc tccaacactt agtaatcatc gtttacggca tggactacca 720 gggtatctaa tcctgttcgc tacccatgct ttcgagcctc agcgtcagtt acagaccaga 780 cagccgccct tcgccactgg tgttcttcca tatatctacg catttcaccg ctacacatgg 840 agttccactg tcctcttctg cactcaagtc tcccagtttc caatgcactt cttcggttga 900 gccgaaggct ttcacattag acttaaaaga ccgcctgcgc tcgctttacg cccaataaat 960 ccggataacg cttgccacct acgtattacc gcggctgctg gcacgtagtt agccgtggct 1020 ttctggttaa ataccgtcac tgggtgaaca gttactctca cccacgttct tctttaacaa 1080 cagagcttta cgagccgaaa cccttcttca ctcacgcggc gttgctccat cagacttgcg 1140 tccattgtgg aagattccct actgctgcct cccgtaggag tctgggccgt gtctcagtcc 1200 caatgtggcc gattaccctc tcaggtcggc tacgcatcat cgccttggtg agccgttacc 1260 tcaccaacta gctaatgcgc cgcgggtcca tccagaagtg atagcagagc catcttttaa 1320 aagaaaacca ggcggttttc tctgttatac ggtattagca tctgtttcca ggtgttatcc 1380 cctgcttctg ggcaggttac ccacgtgtta ctcacccgtc cgccactcac ttcgtgttaa 1440 aatctcattc agtgcaagca cgtcatgatc aattaacgga agttcgttcg acttgcatgt 1500 attaggcacg ccgccagcgt tcatcctgag cagagtttta aaatctataa gg 1552 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 ggattagata ccctggta 18 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 agagtttgat cmtggctcag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ccgtcaattc mtttragttt 20 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 tacggytacc ttgttacgac tt 22 <110> Microbial Institute for Fermentation Industry SHIN, Yong Sung LEE, Im Sook <120> Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses the <130> PN17191 <160> 5 <170> Kopatentin 2.0 <210> 1 <211> 1552 <212> DNA <213> Pediococcus acidilactici <400> 1 ttttcggggt tcccctgtac gacttcaccc taatcatctg tcccacctta gacggctagc 60 tcctaaaagg ttaccccacc ggctttgggt gttacaaact ctcatggtgt gacgggcggt 120 gtgtacaagg cccgggaacg tattcaccgc ggcatgctga tccgcgatta ctagcgattc 180 cgacttcgtg taggcgagtt gcagcctaca gtccgaactg agaatggttt taagagatta 240 gctaaacctc gcggtttcgc aactcgttgt accatccatt gtagcacgtg tgtagcccag 300 gtcataaggg gcatgatgat ttgacgtcgt ccccaccttc ctccggtttg tcaccggcag 360 tctcactaga gtgcccaact gaatgctggc aactagtaat aagggttgcg ctcgttgcgg 420 gacttaaccc aacatctcac gacacgagct gacgacaacc atgcaccacc tgtcattctg 480 tccccgaagg gaacgcctaa tctcttaggt tggcagaaga tgtcaagacc tggtaaggtt 540 cttcgcgtag cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 600 ttttgagttt caaccttgcg gtcgtactcc ccaggcggat tacttaatgc gttagctgca 660 gcactgaagg gcggaaaccc tccaacactt agtaatcatc gtttacggca tggactacca 720 gggtatctaa tcctgttcgc tacccatgct ttcgagcctc agcgtcagtt acagaccaga 780 cagccgccct tcgccactgg tgttcttcca tatatctacg catttcaccg ctacacatgg 840 agttccactg tcctcttctg cactcaagtc tcccagtttc caatgcactt cttcggttga 900 gccgaaggct ttcacattag acttaaaaga ccgcctgcgc tcgctttacg cccaataaat 960 ccggataacg cttgccacct acgtattacc gcggctgctg gcacgtagtt agccgtggct 1020 ttctggttaa ataccgtcac tgggtgaaca gttactctca cccacgttct tctttaacaa 1080 cagagcttta cgagccgaaa cccttcttca ctcacgcggc gttgctccat cagacttgcg 1140 tccattgtgg aagattccct actgctgcct cccgtaggag tctgggccgt gtctcagtcc 1200 caatgtggcc gattaccctc tcaggtcggc tacgcatcat cgccttggtg agccgttacc 1260 tcaccaacta gctaatgcgc cgcgggtcca tccagaagtg atagcagagc catcttttaa 1320 aagaaaacca ggcggttttc tctgttatac ggtattagca tctgtttcca ggtgttatcc 1380 cctgcttctg ggcaggttac ccacgtgtta ctcacccgtc cgccactcac ttcgtgttaa 1440 aatctcattc agtgcaagca cgtcatgatc aattaacgga agttcgttcg acttgcatgt 1500 attaggcacg ccgccagcgt tcatcctgag cagagtttta aaatctataa gg 1552 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 ggattagata ccctggta 18 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 agagtttgat cmtggctcag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ccgtcaattc mtttragttt 20 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 tacggytacc ttgttacgac tt 22
Claims (7)
Improvement of acne or atopic dermatitis comprising as an active ingredient fermented cereals comprising germinated barley and brown rice having antimicrobial activity against Propionibacterium acnes or Staphylococcus epidermidis of claim 4 / RTI >
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