KR101871169B1 - New kimchi lactic acid bacteria Weissella cibaria MFST and composition using concentrated extract of Ziziphus jujuba seed - Google Patents
New kimchi lactic acid bacteria Weissella cibaria MFST and composition using concentrated extract of Ziziphus jujuba seed Download PDFInfo
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- KR101871169B1 KR101871169B1 KR1020150190048A KR20150190048A KR101871169B1 KR 101871169 B1 KR101871169 B1 KR 101871169B1 KR 1020150190048 A KR1020150190048 A KR 1020150190048A KR 20150190048 A KR20150190048 A KR 20150190048A KR 101871169 B1 KR101871169 B1 KR 101871169B1
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Abstract
본 발명은 신규 김치유산균 Weissella cibaria MFST 균주 및 이를 이용한 대추씨 조성물에 관한 것으로, 상기 김치유산균 Weissella cibaria MFST 균주로 미강을 발효시킨 미강 발효 추출물은 높은 함량으로 GBAB를 생성하며, 우수한 항산화 활성을 갖는 바, 상기 미강 발효 추출물과 대추씨 추출 농축액을 포함한 대추씨 조성물은 기능성이 보다 향상된 대추씨 건강식품으로서 유용하게 사용될 수 있다. The present invention relates to a novel Kimchi lactic acid bacterium, Weissella cibaria MFST strain, and a dairy product composition using the same, wherein the fermented rice bran fermented with rice bran fermented with Weissella cibaria MFST strain produces high content of GBAB and exhibits excellent antioxidative activity , The rice husk seed composition including the fermented rice husk extract and the jujube extract concentrate may be useful as a jujube seed health food having improved functionality.
Description
본 발명은 신규 김치유산균 Weissella cibaria MFST 균주 및 이를 이용한 대추씨 조성물에 관한 것이다. The present invention relates to a novel Kimchi lactic acid bacterium, Weissella cibaria MFST strain, and a jujube seed composition using the same.
대추(Ziziphus jujuba)는 갈매나무과(Rhamnace)의 지지푸스(Zizyphus)속 낙엽, 활엽, 향목의 열매이다. 대추의 약용성분으로는 각종 스테롤, 알칼로이드, 사포닌, 비타민, 세로토닌(serotonin), 유기산, 지방산류, 폴리페놀, 히드록시메틸 푸프푸랄, 플라보노이드 및 아미노산류 등이 보고되어 있다.Jujubu (Ziziphus jujuba) is a fruit of leaves, leaves and aroma of Zizyphus in Rhamnace. Examples of medicinal components of jujube include various sterols, alkaloids, saponins, vitamins, serotonins, organic acids, fatty acids, polyphenols, hydroxymethylpuppurals, flavonoids and amino acids.
대추는 수천년 동안 한방에서 생약으로 소화, 완화제, 강장약으로 사용되어 왔고, 민간에선 잘 익은 대추를 말렸다가 달여 먹으면 열을 내리게 하고 변을 묽게 하여 변비를 없애며 기침도 멎게 하는 것으로 알려져 있으며, 생식하면 각성작용이있고 볶은 것은 최면작용이 있는 것으로 알려져 있다. 대추는 갖가지 독을 해독, 중화하고 몸의 염증을 없애 주며 허약한 체질을 튼튼하게 해 주는 효과가 높아 한방에서는 으뜸가는 보약의 하나로, 오늘날과 같이 공해 독이 범람하고 인스턴트식품이 넘쳐나는 시대에 건강을 지키고 우리 민족 고유의 입맛을 되찾는 데에 매우 적절한 식품이다. Jujube has been used as a digestive, emollient, and tonic medicine for thousands of years in herbal medicine. In the private sector, it is said that the jujube is dried, dried, and eaten for a long time to reduce fever, thinning the stomach to stop constipation and stop coughing. It has been known that hypnotic action is aroused and roasted. Jujube is effective in detoxifying and neutralizing all kinds of poisons, eliminating the inflammation of the body and strengthening the fragile constitution. It is one of the top preservatives in the oriental medicine. It is a very suitable food to protect and restore the taste of Korean people.
대추 열매는 식용이며, 대추의 다양한 부위가 다중의 약리적 특성, 예를 들어, 피임, 진통 및 항당뇨 특성을 지니고 있다. 일부 지역에서는 대추의 줄기 혼합물을 임신 방지에 사용한다. 한편, 대추씨의 효과에 대한 과학적 연구는 거의 없는 실정이며, 단지 전통 의약에서는 대추씨를 불면증 및 불안 해소에 이용하고 있다. Jujube fruit is edible and various parts of jujube have multiple pharmacological properties such as contraception, analgesic and antidiabetic properties. In some areas, jujube stem mixtures are used to prevent pregnancy. On the other hand, there is little scientific study on the effect of jujube seeds. In traditional medicine, jujube seeds are used to relieve insomnia and anxiety.
이에, 대추씨를 이용한 제품군을 개발하기 위하여, 대추씨의 효과에 기능성을 더 부여할 수 있도록 하는 가공방법의 개발이 필요한 실정이다.Therefore, it is necessary to develop a processing method that can impart more functionality to the effect of jujube seeds in order to develop a product group using the jujube seeds.
따라서 본 발명은 대추씨의 기능성을 더욱 향상시킬 수 있는 신규 김치유산균을 스크리닝하며, 이를 이용하여 기능성이 향상된 대추씨 조성물을 제공하는 데에 있다. Accordingly, the present invention is to provide a jujube seed composition having improved functionality by screening novel Kimchi lactic acid bacteria that can further improve the functionality of jujube seeds.
상기 과제의 해결을 위하여, 본 발명은 숙성된 김치에서 분리되며, 감마-아미노부티르산(Gamma-amino butyric acid, GABA)을 생산하는 김치유산균 Weissella cibaria MFST 균주를 제공한다. In order to solve the above problems, the present invention provides a kimchi lactic acid bacteria strain, Weissella cibaria MFST, which is isolated from aged kimchi and produces gamma-amino butyric acid (GABA).
또한 본 발명은 상기 김치유산균 Weissella cibaria MFST 균주로 발효시킨 미강 발효 추출물을 제공한다. Also, the present invention provides a fermented rice bran extract fermented with the Kimchi lactic acid bacterium Weissella cibaria MFST strain.
또한 본 발명은 김치유산균 Weissella cibaria MFST 균주로 발효시킨 미강 발효 추출물 1 내지 10 중량%; 및 대추씨 추출 농축액 90 내지 99 중량%를 포함하는 대추씨 조성물을 제공한다. The present invention also relates to a fermented rice bran fermented by fermentation with a fermentation broth of Weissella cibaria MFST, 1 to 10% by weight; And 90 to 99% by weight of a jujube extract concentrate.
본 발명에서 새로 스크리닝 한 김치유산균 Weissella cibaria MFST 균주로 미강을 발효시킨 미강 발효 추출물은 높은 함량으로 GBAB를 생성하며, 우수한 항산화 활성을 갖는 바, 상기 미강 발효 추출물과 대추씨 추출 농축액을 포함한 대추씨 조성물은 기능성이 보다 향상된 대추씨 건강식품으로서 유용하게 사용될 수 있다. In the present invention, the fermented rice bran fermented with fermented rice bran as the Weissella cibaria MFST strain, which is newly screened in the present invention, produces GBAB at a high content and has an excellent antioxidative activity. As a result, the fermented extract of rice bran and the jujube seed extract Can be usefully used as a jujube seed health food having improved functionality.
도 1은 본 발명에 따른 다양한 미강 발효 추출물의 총 페놀성 화합물 함량을 분석한 것이고,
도 2는 본 발명에 따른 다양한 미강 발효 추출물의 DPPH radical 소거능을 측정한 것이고,
도 3은 본 발명에 따른 최종제품의 총 페놀성 화합물 함량을 분석한 것이고[추출A는 발효미강 효소(protease+viscozyme) 처리추출액의 72 시간 처리군, 추출B는 발효미강 효소(protease+viscozyme) 처리추출액의 48 시간 처리군, 추출C는 발효미강 효소(viscozyme) 처리추출액의 24 시간 처리군],
도 4는 본 발명에 따른 최종제품의 DPPH radical 소거능을 측정한 것이다[추출A는 발효미강 효소(protease+viscozyme) 처리추출액의 72 시간 처리군, 추출B는 발효미강 효소(protease+viscozyme) 처리추출액의 48 시간 처리군, 추출C는 발효미강 효소(viscozyme) 처리추출액의 24 시간 처리군].FIG. 1 is a graph showing the total phenolic compound content of various rice gherkin fermented extracts according to the present invention,
FIG. 2 is a graph showing DPPH radical scavenging activity of various rice fermented extracts according to the present invention,
FIG. 3 is an analysis of the total phenolic compound content of the final product according to the present invention, wherein [Extract A] is a 72 hour treatment group of a protease + viscozyme-treated extract, and B is a protease + viscozyme, Treated extract for 48 hours, extract C for 24 hours treated with fermented rice bran enzyme (viscozyme) treated extract]
FIG. 4 is a graph showing DPPH radical scavenging activity of the final product according to the present invention. [Extraction A is a 72 hour treatment group of protease + viscozyme-treated extract and extraction B is a treatment group of fermented rice bran enzyme (protease + viscozyme) , And extract C was treated with fermented rice bran enzyme (viscozyme) for 24 hours].
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명자들은 보다 기능성이 향상된 대추씨를 이용한 식품을 개발하기 위하여 연구노력 한 결과, 다양한 균주 중에서도 특히 새로 스크리닝 한 김치유산균 Weissella cibaria MFST 균주로 미강을 발효시킨 미강 발효 추출물에서 매우 높은 함량으로 GBAB를 생성하며, 우수한 항산화 활성을 나타내는 것을 확인하였는 바, 상기 미강 발효 추출물과 대추씨 추출 농축액을 포함한 대추씨 조성물을 개발함으로써 본 발명을 완성하게 되었다.As a result of efforts to develop a food using the jujube seeds having improved functionality, the present inventors have found that GBAB is produced in a very high content in a fermented rice bran extract fermented with rice bran fermented with a newly screened Kimchi lactic acid bacterium Weissella cibaria MFST strain . As a result, the present inventors have completed the present invention by developing a jujube seed composition containing the fermented rice bran extract and the jujube extract concentrate.
본 발명은 숙성된 김치에서 분리되며, 감마-아미노부티르산(Gamma-amino butyric acid, GABA)을 생산하는 김치유산균 Weissella cibaria MFST 균주를 제공한다. The present invention provides a strain of Weissella cibaria MFST isolated from aged kimchi and producing gamma-aminobutyric acid (GABA).
상기 균주는 KCCM11760P로 수탁된 것이 바람직하다.The strain is preferably deposited with KCCM 11760P.
또한, 본 발명은 상기 김치유산균 Weissella cibaria MFST 균주로 발효시킨 미강 발효 추출물을 제공한다. 이때, 상기 미강 발효 추출물은 김치유산균 Weissella cibaria MFST 균주를 발효물에 대하여 1~2 중량% 접종하며 35℃에서 배양한 것일 수 있다.In addition, the present invention provides a fermented rice bran extract fermented with the Kimchi lactic acid bacterium Weissella cibaria MFST strain. At this time, the rice germinated fermented extract may be obtained by inoculating 1 to 2% by weight of Kimchi lactic acid bacterium Weissella cibaria MFST against the fermented product and culturing at 35 ° C.
또한, 본 발명은 상기 김치유산균 Weissella cibaria MFST 균주로 발효시킨 미강 발효 추출물 1 내지 10 중량%; 및 대추씨 추출 농축액 90 내지 99 중량%를 포함하는 대추씨 조성물을 제공한다.The present invention also relates to a fermented rice bran fermented by fermenting with the fermentation broth of Weissella cibaria MFST, 1 to 10% by weight; And 90 to 99% by weight of a jujube extract concentrate.
상기 대추씨 추출 농축액은 대추씨 총 100 중량부에 대하여 200 중량부의 물을 가하여 열수추출하고, 마이크로필터 여과 후 진공농축하여 50 내지 70 Brix로 농축한 농축액을 살균하여 제조할 수 있다.The juice extract concentrate can be prepared by adding 200 parts by weight of water to 100 parts by weight of jujube seeds, subjecting to hot extraction, filtering the microfilter, concentrating in vacuum, and concentrating the concentrate at 50 to 70 Brix.
일 실시예로서, 상기 대추씨 추출 농축액은 대추씨 총 100 중량부에 대하여 200 중량부의 물을 가하여 95℃에서 8시간 3회 열수추출하고, 25 마이크로필터 여과 후 진공농축하여 60 Brix로 농축한 농축액을 95℃에서 20분 동안 살균하여 제조할 수 있다.In one embodiment, 200 parts by weight of water is added to 100 parts by weight of the jujube seed extract, the mixture is subjected to hot water extraction at 95 ° C for 3 hours, filtered through 25 microfilters, concentrated in vacuo and concentrated to a concentration of 60 Brix Lt; RTI ID = 0.0 > 95 C < / RTI > for 20 minutes.
상기 조성물은 유자즙 또는 꿀에서 선택된 하나 이상을 더 포함할 수 있다.The composition may further comprise at least one selected from citron juice or honey.
보다 바람직하게는, 상기 조성물은 김치유산균 Weissella cibaria MFST 균주로 발효시킨 미강 발효 추출물 1 내지 5 중량%; 대추씨 추출 농축액 94 내지 97 중량%; 유자즙 0.05 내지 0.2 중량%; 및 꿀 0.5 내지 3 중량%를 포함할 수 있다. More preferably, the composition comprises 1 to 5% by weight of rice bran fermented extract fermented with a strain of Weissella cibaria MFST; 94 to 97% by weight of a jujube extract concentrate; 0.05 to 0.2% by weight of citron juice; And 0.5 to 3% by weight of honey.
이때, 상기 미강 발효 추출물은 주로 미강 유래 생리활성물질인 GABA 및 페놀성 화합물을 제공하는 역할을 수행하며, 상기 대추씨 추출 농축액은 제품 물성의 안정성을 부여하는 역할을 수행하며, 상기 유자즙은 산미를 부여하는 역할을 수행하며, 상기 꿀은 물성을 개선하는 역할을 수행하며, 상기 원료들이 상기 범위를 벗어나 배합되면 물성안정 및 맛의 최적조건 유지에 문제가 야기될 수 있다. The fermented extract of rice bran further serves as a physiologically active substance of Ganoderma lucidum, GABA and a phenolic compound. The concentrate of jujube extract concentrates the stability of the physical properties of the product, The honey has a role of improving the physical properties. If the ingredients are out of the above range, stability of the physical properties and maintenance of the optimum condition of the taste may be caused.
본 발명의 조성물은 식품 조성물로 활용될 수 있으며, 특히 GABA 함량이 증가되어 뇌기능 개선용 건강기능식품으로 사용될 수 있다. "건강기능식품"이라 함은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The composition of the present invention can be utilized as a food composition, and in particular, it can be used as a health functional food for improving brain function by increasing GABA content. "Health functional food" refers to foods prepared and processed using raw materials and ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods. "Functional" It is intended to take nutrients for the purpose of controlling nutrients and obtaining effects that are useful for health use such as physiological action.
본 발명의 식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, "식품 첨가물"로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안정청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다. The food composition of the present invention may contain conventional food additives and, unless otherwise specified, the suitability of the food additives as "food additives" is to be determined by the Food and Drug Administration in accordance with General Regulations and General Test Methods approved by the Food and Drug Administration. Shall be determined according to the relevant standards and standards.
본 발명의 식품 조성물은 뇌기능 개선용 식품 및 음료 등에 다양하게 이용될 수 있으며, 예컨대, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강기능성 보조 식품, 식품 첨가제 등에 사용될 수 있고, 분말, 과립, 정제, 캡슐 또는 음료인 형태로 사용할 수 있다. 상기 기재한 것 외에도 모든 식품 형태일 수 있다.The food composition of the present invention can be used for various foods and beverages for improving brain function and can be used for various foods, beverages, gums, tea, vitamin complex, health functional supplement, food additive, , Tablets, capsules or beverages. May be in any food form other than those described above.
본 발명의 식품 조성물은 보조성분으로 비타민 B, C, E나 베타카로틴, Ca, Mg, Zn 등의 미네랄 함유 화합물이나 레시틴 등의 인지질 또는 말톨, 올리고당, 아미노산 등의 성분 등을 추가로 포함할 수 있으며, 특히, 비타민 C, E 나 베타카로틴, 말톨 등의 성분 2-3개를 혼합하여 사용하면 생체 활성효과를 보강할 수 있어 바람직하다. The food composition of the present invention may further contain minerals containing compounds such as vitamin B, C, E, beta-carotene, Ca, Mg and Zn, phospholipids such as lecithin or components such as maltooligosaccharides and amino acids as auxiliary components In particular, when two or three components such as vitamin C, E, beta-carotene and maltol are mixed and used, the bioactive effect can be reinforced.
또한, 상기 성분에 추가로 미각을 향상시키기 위하여, 공지의 첨가제로써, 매실, 레몬향, 파인애플향 또는 허브향과 같은 천연향료나 천연과즙, 플로로필린, 플로보노이드 등의 천연색소 및 감미성분인 과당, 당알코올, 설탕 등, 구연산, 구연산나트륨 같은 산미제를 첨가할 수 있다. In addition to the above ingredients, in order to further improve the taste, known additives include natural flavors such as plum, lemon flavor, pineapple flavor or herb flavor, natural flavors such as natural fruit juice, florophylline, Fructose, sugar alcohol, sugar, citric acid, and sodium citrate may be added.
또한, 본 발명의 식품 조성물이 음료형태인 경우, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토오스, 수크로오스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.When the food composition of the present invention is in the form of a drink, it may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, 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.
또한, 본 발명의 식품 조성물은 뇌기능 개선을 위한 목적으로, 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다.The food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of improving brain function.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.
<실시예 1> 미강 발효 우수 균주 분리 및 활성 검토<Example 1> Isolation and activity of rice bran fermentation isolate
1. 미강 발효 우수 균주 스크리닝 및 분리1. Screening and separation of rice bran fermentation
숙성된 김치를 시료로 사용하여 0.02% 소듐 아자이드(sodium azide)를 함유한 MRS Agar(Difco, Detroit, USA)에 도말하고, 30℃에서 24시간 배양 후 콜로니 주위에 노란색 환을 생성하는 균주들을 선정하였고, 이후 상기 배지에서 3번 반복 도말함으로서 순수 분리하였다. 선별된 콜로니는 배양방법을 달리하여 균의 스크리닝 실험을 실시하였다. The fermented kimchi was applied to MRS Agar (Difco, Detroit, USA) containing 0.02% sodium azide and cultured at 30 ° C for 24 hours to produce yellow circles around the colonies And then purely separated by repeating 3 times in the medium. The selected colonies were screened for bacteria by different culture methods.
상기와 동일한 방법으로 선별된 균주 6종(이하, K-1 내지 K-6; 이 중 K-1이 Weissella cibaria MFST), 미강에서 30℃에서 24시간 동안 배양된 균주 2종(이하, G-1 내지 G-2), MRS broth 9ml에 멸균 미강 1 g을 첨가하여 혼합 분리한 균주 4종(이하, MG-1 내지 MG-4)을 포함한 총 12개의 균을 채집하여 실험에 사용하였으며, 대조군은 일반 미강을 배양하여 동일한 방법으로 실험하였다. Six strains (hereinafter, K-1 to K-6; K-1 of Weissella cibaria MFST) selected in the same manner as above were cultured in rice bran at 30 ° C for 24 hours (hereinafter referred to as G- (MG-1 to MG-4) obtained by mixing and separating 1 g of sterilized rice bran into 9 ml of MRS broth were collected and used in the experiment, Were cultured in the same manner.
선별된 균주들을 배양 시 최적 조건인 35℃에서 9시간 배양한 후. 3000rpm에서 10분간 원심분리하여 상등액을 버리고 인산완충액(pH 7.0)으로 세척한 후 사용하였다. 미강과 미강 중량의 200%(v/w)의 증류수, 1%(v/w) 선별 균주들을 각각 첨가한 다음 37℃에서 24시간 발효하였다. 이후 각각의 발효 미강에 5배의 증류수를 첨가하여 65℃의 water bath에서 12시간 동안 열수추출하였고, 추출물은 여과 후 감압하여 농축 후 동결 건조하여 시료로 사용하였다.The selected strains were incubated at 35 ℃ for 9 hours. After centrifugation at 3000 rpm for 10 minutes, the supernatant was discarded and washed with phosphate buffer (pH 7.0). 200% (v / w) distilled water and 1% (v / w) selection strains of rice bran and rice bran were added respectively and fermented at 37 ° C for 24 hours. After 5 times of distilled water was added to each fermented rice bran, the extracts were subjected to hot water extraction for 12 hours in a water bath at 65 ° C. The extract was filtered, concentrated under reduced pressure, and lyophilized to use as a sample.
선별 균주들의 항산화 작용에 기여하는 총 폴리페놀의 함량을 종래 알려진 방법에 따라 측정한 결과, 1 mg/mL의 농도에서 Weissella cibaria MFST 군(K-1, 547.439 μg/mL), G-1군(467.674 μg/mL), MG-4군 (519.731 μg/mL), MG-1군(514.494 μg/mL), MG-3군 (508.396 μg/mL), 대조군(452.141 μg/mL) 순으로 나타났다. W. cibaria MFST 군(K-1)의 경우 페놀 화합물 함량이 가장 낮은 K-5(289.252 μg/mL)보다 2배 높은 활성을 나타냈다. The contents of total polyphenols, which contribute to the antioxidant activity of the selective strains, were measured according to a conventional method. As a result, the Weissella cibaria MFST group (K-1, 547.439 μg / (508.396 μg / mL) and control (452.141 μg / mL), respectively, in the order of MG-1 and MG-4. The activity of W. cibaria MFST group (K-1) was two times higher than that of K-5 (289.252 μg / mL), which has the lowest phenolic compound content.
그리고, 선별균주들의 DPPH radical 소거능 결과, 1 mg/mL 농도에서 W.cibaria MFST 군(84.0715%)이 가장 높은 활성을 나타냈으며, 각 실험군의 DPPH radical 소거능은 70.164% - 84.071%로 높은 활성을 나타내었다. 이를 토대로 W.cibaria MFST을 최종 선별 분리균주로 선택하였다.As a result, DPPH radical scavenging activity of the selective strains showed the highest activity of W. cibaria MFST group (84.0715%) at 1 mg / mL concentration. DPPH radical scavenging activity of each experimental group was as high as 70.164% - 84.071% . Based on this, W. cibaria MFST was selected as the final screening isolate.
2. 미강 발효 추출물의 아미노산 분석2. Amino acid analysis of fermented rice extract
선별된 W.cibaria MFST을 이용한 미강 발효 추출물의 GABA(γ-aminobutyric acid) 함량 측정을 위해 동결건조된 시료 0.15g을 100배 희석하여 15mL을 만든 후 20% TCA(trichloroacetic acid) 15mL을 가여 3000rpm에서 원심분리 후 침전된 단백질을 제거하고 나온 상등액에 30mL 에틸에테르를 혼합하여 TCA와 지용성 물질을 제거한 후 수분층을 감압 농축하였다. 농축액에 0.2M 구연산 완충액을 가하여 5 mL로 정용한 후 필터를 사용하여 여과하여 아미노산 자동분석기(L-8800,Hitachi Co.LTD, JAPAN)로 분석하였다. To determine the content of GABA (γ-aminobutyric acid) in fermented rice bran extracts using selected W. cibaria MFST, 0.15 g of a freeze-dried sample was diluted 100-fold to 15 mL, and 15 mL of 20% TCA (trichloroacetic acid) After centrifugation, the precipitated proteins were removed, and 30 mL of ethyl ether was added to the supernatant to remove TCA and lipid-soluble substances, followed by concentration of the aqueous layer under reduced pressure. The concentrate was diluted to 5 mL with 0.2 M citric acid buffer, filtered through a filter, and analyzed with an automatic amino acid analyzer (L-8800, Hitachi Co.LTD, JAPAN).
그 결과, 표 1과 같이 총 아미노산 함량은 W.cibaria MFST 군이 4565.57 mg%이며 대조군은 4267.24 mg% 나타났으며, 대조군에서 Aspartic acid, Glycine이 W.cibaria MFST군보다 높게 나타났다. W.cibaria MFST군은 GABA, Glutamic acid, Alanine이 주 아미노산으로 전체 아미노산의 42%를 차지하였으며, 그 중 GABA는 15.698 mg%로 현저히 높게 나타났다. 총 아미노산 중 GABA가 높게 나타난 것은 분리 젖산균이 미강 발효 시 GABA의 활성을 높이는 것으로 사료된다.As a result, total amino acid content of W. cibaria MFST group was 4565.57 mg% and that of control group was 4267.24 mg% as shown in Table 1. In the control group, Aspartic acid and Glycine were higher than those of W. cibaria MFST group. In W. cibaria MFST group, GABA, glutamic acid and alanine were the major amino acids and accounted for 42% of total amino acids, among which GABA was remarkably high as 15.698 mg%. The highest GABA among the total amino acids seems to increase the activity of GABA when fermenting rice bran.
3. 미강 발효 추출물의 총 페놀성 화합물 함량 분석3. Analysis of Total Phenolic Compounds in Fermented Rice Mushroom Extracts
선별된 W. cibaria MFST 군의 항산화작용에 기여하는 총 폴리페놀 함량을 확인하기 위해 대조군과 젖산균 발효군인 W. cibaria MFST을 이용한 미강 발효 추출물을 분석하였다. 이때, 총 폴리페놀 함량은 Folin-Dennis법을 이용하였으며, 갈산(Gallic acid)로 표준 검정곡선을 구하여 시료의 총 폴리페놀 함량을 계산하였고 3회 반복적 실험을 수행하였다. To determine the total polyphenol content of the selected W. cibaria MFST group, the fermented extract of rice bran fermented with W. cibaria MFST was analyzed. The total polyphenol content of the sample was calculated by Folin-Dennis method and the standard curve was calculated with Gallic acid. The total polyphenol content of the sample was calculated and repeated experiment was performed three times.
총 폴리페놀 함량을 측정한 결과, 도 1과 같이 W.cibaria MFST 군이 609.611%로 대조군 460.375%보다 약 1.5배 높은 총 폴리페놀 함량을 나타냈다. 이는 젖산균 발효 과정에서 생성된 발효산물에 의해 페놀성 화합물의 함량이 증가하여 항산화 활성에 영향을 미친 것으로 판단된다. The total polyphenol content of the W. cibaria MFST group was 609.611% as shown in FIG. 1, and the total polyphenol content was about 1.5 times higher than that of the control group 460.375%. It was concluded that the content of phenolic compounds was increased by the fermentation products produced during the fermentation process of lactic acid bacteria.
4. 미강 발효 추출물의 DPPH radical 소거능 측정4. Measurement of DPPH radical scavenging ability of rice bran fermented extract
선별된 W.cibaria MFST를 이용한 미강 발효 추출물의 DPPH radical 소거능을 Bios의 DPPH 분석을 이용하여 측정하였으며, 그 결과 도 2와 같이 W.cibaria MFST 군의 DPPH radical 소거능이 대조군에 비해 우수한 것으로 나타났다.The DPPH radical scavenging activity of the fermented rice bran extract using the selected W. cibaria MFST was measured using Bios DPPH analysis. As a result, DPPH radical scavenging activity of W. cibaria MFST group was superior to that of the control group as shown in FIG.
<실시예 2> 미강 발효 추출물과 대추씨 추출 농축액을 블랜딩한 제품 개발≪ Example 2 > Development of a product blended with a fermented rice bran extract and a jujube extract concentrate
앞서 실시예 1에서 선별한 GABA 생성이 우수한 김치유래 젖산균 W.cibaria MFST을 이용한 미강 발효 추출물과, 대추씨 추출 농축액을 블랜딩하여 고부가가치 대추 엑기스 제품을 개발하였다. 대추씨 추출 농축액은 대추 부산물인 대추씨에 2배수의 물을 가하여 95℃에서 8시간 3회 열수추출 하고 25 마이크로필터 여과 후 진공농축하여 60 Brix 이상으로 감압농축한 대추농축액을 사용하였으며 고흥 천연 유자과즙(한성푸드 영농조합법인)을 첨가하여 향과 맛을 보완해 주었다. 유자즙 첨가비율은 최종 제품의 물성을 고려하여 첨가하며 다음 표 2와 같이 제조하였다.A high value-added jujube extract product was developed by blending the fermented rice bran extract and the jujube extract concentrate using the lactic acid bacteria W. cibaria MFST, which is superior in GABA production, The juice extract of the jujube was extracted from the jujube by two times of water at 95 ℃ for 8 hours, filtered through 25 microfilters, concentrated in vacuo and concentrated under reduced pressure to 60 Brix. The juice (Hansung Foods Farming Association) was added to complement the fragrance and flavor. The citrate juice addition ratio was added considering the physical properties of the final product and prepared as shown in Table 2 below.
추출물 (중량%)Rice bran fermentation
Extract (wt%)
추출농축액 (중량%)Daechu
Extract concentrate (wt%)
<실시예 3> 대추씨 추출 농축액 블랜딩 제품의 관능평가≪ Example 3 > Sensory Evaluation of Blended Products of Jujube Extract Concentrate
실시예 2에 따른 대추씨 추출 농축액 블랜딩 제품의 관능평가를 위하여, 성인 30명을 대상으로 향, 단맛, 신맛, 쓴맛, 질감 및 종합적 기호도에 대하여 7점 척도법으로 기호도를 평가하였다. 이때 관능평점은 1점(아주나쁨)에서 7점(아주 좋음)으로 평가하였으며, 평가항목에 대한 관능검사 결과를 평균하여 다음 표 3에 나타내었다.For the sensory evaluation of the jujube extract concentrate according to Example 2, the degree of preference was evaluated by the 7 point scale for flavor, sweetness, sour taste, bitter taste, texture and overall preference for 30 adults. The sensory scores were evaluated from 1 point (very bad) to 7 points (very good), and the sensory evaluation results for the evaluation items were averaged and shown in Table 3 below.
그 결과, C군이 향을 제외한 모든 항목에서 가장 높은 값을 받았으며, 향과 단맛이 과하지 않으며 신맛, 쓴맛이 자극적이지 않다는 평을 받았다. 전체적으로 제품에 대한 기호도가 긍정적이었으나, 관능평가 결과 선호도가 가장 우수한 C군을 최종제품으로 제조하였다. As a result, group C received the highest value in all items except fragrance, and it was evaluated that flavor and sweetness were not excessive, and sour taste and bitter taste were not stimulating. As a whole, the preference of the product was positive, but the best product of the sensory evaluation result was produced as the final product.
<실시예 3> 대추씨 추출 농축액 최종제품의 품질특성 분석<Example 3> Analysis of quality characteristics of the final product of jujube extract concentrate
1. 최종제품의 pH, 당도, 색도1. pH, sugar content, color
실시예 2에서 선정한 최종제품의 pH, 당도 및 색도를 측정하였다. pH와 당도는 시료를 10배 희석하여 각각 pH 미터기(METTER TOLEDO)와 당도계(refractometer HI 96801, HANNA instruments)로 측정하였다. 색도는 표준백판(L=98.06, a=0.11, b=0.94)으로 보정된 색차계(CR-300 Colorimate, MINOLTA, Japan)를 사용하여 Hunter color인 명도(lightness, L), 적색도(redness, a) 및 황색도(yellowness, b) 값을 측정하였고 그 결과는 다음 표 4와 같다. The pH, the sugar content and the chromaticity of the final product selected in Example 2 were measured. The pH and sugar content were measured with a pH meter (METTER TOLEDO) and a sugar meter (refractometer HI 96801, HANNA instruments), respectively. The chromaticity was determined by Hunter color lightness (L), redness (R), and redness (R) using a colorimeter (CR-300 Colorimate, MINOLTA, Japan) calibrated with a standard white plate (L = 98.06, a = 0.11, b = 0.94) a) and yellowness (b) were measured. The results are shown in Table 4 below.
2. 최종제품의 아미노산 분석2. Amino acid analysis of the final product
실시예 2에서 선정한 최종제품과 추출방법을 달리한 발효미강 추출물 그룹 추출A, 추출B 및 추출C의 아미노산 분석은 아미노산 자동분석기(L-8800, Hitachi Co.LTD, JAPAN)로 분석하였다. 추출 방법을 달리하여 제조한 미강발효 추출물와 대추씨 추출물을 블랜딩하여 만든 최종제품에도 상당량의 GABA가 포함되어 있음을 표 5를 통해 확인하였다. 이 때, 추출A는 본 실험실에서 제조한 발효미강을 72시간 효소(protease+viscozyme, novozymes) 처리한 추출액이고, 추출B는 발효미강을 48시간 효소(protease+viscozyme, novozymes) 처리한 추출액이며, 추출C는 발효미강을 24시간 효소(viscozyme, novozymes) 처리한 추출액이다. Amino acid analysis of Extract A, Extract B and Extract C of fermented rice bran extract group, which was different from the final product selected in Example 2, was analyzed with an amino acid automatic analyzer (L-8800, Hitachi Co.LTD, JAPAN). Table 5 shows that GABA was contained in the final product, which was prepared by blending the fermented rice bran extract and the jujube extract prepared by different extraction methods. Extraction A is an extract solution obtained by treating the fermented rice bran produced in this laboratory for 72 hours with an enzyme (protease + viscozyme, novozymes). Extraction B is an extract solution obtained by treating the fermented rice bran with enzyme (protease + viscozyme, novozymes) Extraction C is an extract solution obtained by treating fermented rice bran with an enzyme (viscozyme, novozymes) for 24 hours.
3. 최종제품의 총 폴리페놀 함량 분석3. Analysis of the total polyphenol content of the final product
실시예 2에서 선정한 최종제품과 추출방법을 달리한 발효미강 추출물 그룹 추출A, 추출B 및 추출C의 총 폴리페놀 함량 분석은 Folin-Dennis법을 이용하였으며, 각각의 동결 건조한 시료를 농도별로 증류수에 녹여 사용하였다. 시료 0.2 mL에 폴린-시오칼토 시약(Folin-Ciocalteu reagent, Sigma, USA) 1 mL을 넣은 후 실온에 3분간 반응시킨 후 그 반응액에 10% Na2CO3 0.8 mL을 넣은 뒤 실온에서 1시간 반응시킨 후 765nm에서 흡광도를 측정하였다. 갈산(Gallic acid)으로 표준 검정곡선을 구하여 시료의 총 폴리페놀 함량을 계산하고 3회 반복적 실험을 수행하였다. 그 결과는 도 3과 같이 실시예에서 제조한 최종제품의 총 폴리페놀 함량이 추출A, 추출B 및 추출C보다 탁월하게 많은 것을 확인하였다.The Folin-Dennis method was used to analyze the total polyphenol content of Extract A, Extract B and Extract C of fermented rice bran extract group, which was different from the final product selected in Example 2, and each freeze-dried sample was added to distilled water Respectively. After adding 1 mL of Folin-Ciocalteu reagent (Sigma, USA) to 0.2 mL of the sample, react at room temperature for 3 minutes, add 0.8 mL of 10% Na 2 CO 3 to the reaction solution, After the reaction, absorbance was measured at 765 nm. The total polyphenol content of the sample was calculated by taking a standard calibration curve with gallic acid, and the experiment was repeated three times. As a result, it was confirmed that the total polyphenol contents of the final products prepared in the examples as shown in Fig. 3 were much higher than those of Extract A, Extract B and Extract C.
4. 최종제품의 DPPH radical 소거능 측정4. Measurement of DPPH radical scavenging ability of final product
실시예 2에서 선정한 최종제품과 추출방법을 달리한 발효미강추출물 추출A, 추출B 및 추출C은 Bios의 방법에 따라 1,1-diphenyl-2-picrylhydrazyl(DPPH, Sigma) radical 소거능을 측정하였다. 각 농도의 시료 1mL에 0.15mM DPPH용액 1.5mL을 가하고 실온에서 30분간 반응 후 517nm에서 흡광도를 측정하여, 시료첨가구와 비첨가구의 차이를 백분율로 나타내었다. 그 결과는 도 4와 같이 실시예에서 제조한 최종제품(5 mg/ml)의 DPPH radical 소거능이 가장 우수한 것으로 확인되었다.The 1,1-diphenyl-2-picrylhydrazyl (DPPH, Sigma) radical scavenging activity was measured according to the Bios method for Extract A, Extract B and Extract C of fermented rice bran extract, which was different from the final product selected in Example 2. To 1 mL of each concentration, 1.5 mL of 0.15 mM DPPH solution was added. After 30 minutes of reaction at room temperature, the absorbance was measured at 517 nm, and the difference between the sample addition and non-addition was expressed as a percentage. As a result, it was confirmed that DPPH radical scavenging ability of the final product (5 mg / ml) prepared in the example was the best as shown in FIG.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시예일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다. While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
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