KR101972838B1 - Fungicidal composition comprising the extract from Capsella bursa-pastoris and method for preparing the same - Google Patents
Fungicidal composition comprising the extract from Capsella bursa-pastoris and method for preparing the same Download PDFInfo
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- KR101972838B1 KR101972838B1 KR1020170093855A KR20170093855A KR101972838B1 KR 101972838 B1 KR101972838 B1 KR 101972838B1 KR 1020170093855 A KR1020170093855 A KR 1020170093855A KR 20170093855 A KR20170093855 A KR 20170093855A KR 101972838 B1 KR101972838 B1 KR 101972838B1
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- extract
- kctc
- lactic acid
- chitosan
- polylysine
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/10—Preserving against microbes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/50—Polysaccharides, gums
- A23V2250/51—Polysaccharide
- A23V2250/511—Chitin, chitosan
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
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- A23Y2220/00—
Abstract
본 발명은 냉이로부터 추출된 항진균 조성물 및 이를 포함하는 조성물의 제조방법에 관한 것으로, 보다 상세하게는 냉이추출물을 포함하는 항진균 조성물 및 그 제조방법에 관한 것이다. 본 발명의 조성물은 수분을 많이 함유하는 식품의 곰팡이를 예방 및 억제 효과를 나타낸다.The present invention relates to an antifungal composition extracted from cucumber and a method of preparing a composition containing the antifungal composition, and more particularly, to an antifungal composition comprising a cucumber extract and a preparation method thereof. The composition of the present invention exhibits an effect of preventing and inhibiting fungi in a food containing a large amount of water.
Description
본 발명은 냉이로부터 추출된 항진균 조성물 및 이를 포함하는 조성물의 제조방법에 관한 것으로, 보다 상세하게는 냉이추출물을 포함하는 항진균 조성물 및 그 제조방법에 관한 것이다.The present invention relates to an antifungal composition extracted from cucumber and a method of preparing a composition containing the antifungal composition, and more particularly, to an antifungal composition comprising a cucumber extract and a preparation method thereof.
보존료란 넓은 의미로 식품의 변질, 부패, 변색 및 화학변화를 방지하여 식품의 영양가와 신선도를 유지시키기 위해 사용되는 첨가물로서 살균제, 산화방지제가 포함되며, 좁은 의미로는 세균, 곰팡이, 효소 등 미생물의 증식을 억제하여 식품을 보존하는 첨가물을 말한다. 보존료는 합성보존료와 천연보존료로 구분이 되는데, 합성보존료는 식품의 미생물에 의한 부패를 방지할 목적으로 사용되는 식품첨가물 중 화학적 합성품을 말하며 안전성의 문제가 제기되고 있다. 많이 이용되는 합성보존료로 데히드로초산, 소르빈산, 안식향산 등이 있다. 천연보존료는 천연에서 유래한 항균제로 생리활성을 가지고 있어 항균효과와 생리활성 효과를 동시에 얻을 수 있다. 많이 이용되는 천연보존료로는 자몽종자추출물, 허브추출물 등이 있다.Preservatives are broadly used additives that are used to maintain the nutritional value and freshness of foods by preventing deterioration, decay, discoloration, and chemical change of foods. They include microbicides and antioxidants. In a narrow sense, microorganisms such as bacteria, fungi, And the food is preserved by suppressing the proliferation of the food. Preservatives are classified into synthetic preservatives and natural preservatives. Synthetic preservatives are chemical compounds among food additives that are used for the purpose of preventing microbial spoilage of food, and safety problems are raised. Synthetic preservatives commonly used include dehydroacetic acid, sorbic acid, and benzoic acid. Natural preservative is an antimicrobial agent derived from natural and has a physiological activity, so that it can obtain antimicrobial effect and physiological activity effect at the same time. Most commonly used natural preservatives include grapefruit seed extract and herbal extracts.
진균(곰팡이)은 균류 중 진균류에 속하는 미생물로, 실처럼 길고 가는 모양의 균사로 되어 있는 사상균을 말한다. 주로 온난다습한 환경에서 잘 자라며, 최적 온도는 30℃이다. 곰팡이는 일상생활에서 쉽게 접할 수 있으며, 떡이나 빵 등의 식품, 구두 등의 가죽제품, 주택의 벽면, 사람의 피부 등 다양한 곳에 존재한다. 식품에서는 건조, 염장, 냉장, 냉동 등의 방법으로 곰팡이 발생을 막고 있다.Fungus (fungus) is a microorganism belonging to fungi among fungi, and is a filamentous fungus which has long and thin hyphae like yarn. It grows well in warm and humid environments, and the optimum temperature is 30 ℃. Mold is easily accessible in everyday life, and it exists in various places such as food such as bread and bread, leather goods such as shoes, wall of house, skin of human being. In foods, molds are prevented by drying, salting, chilling, and freezing.
자몽종자추출물(Grapefruit seed extract, GSE)은 운향과 자몽의 종자를 물, 에탄올 또는 글리세린으로 추출하여 얻어지는 것으로 주 성분은 지방산과 플라보노이드이다. (식품첨가물공전, 2016) Grapefruit seed extract (GSE) is obtained by extracting mungbean and grape seeds with water, ethanol or glycerin. The main components are fatty acids and flavonoids. (Food Additives Revision, 2016)
자몽종자추출물에 함유되어 있는 성분 중 하나인 나린진(naringin)등의 플라보노이드 성분이 미생물의 세포벽 및 세포막의 기능을 약화시키고 효소 활성을 저해하여 DNA/RNA에서 비롯되는 세포증식기작을 억제한다. 또한 대장균, 포도상구균, 콜레라균 등 식중독 균에 모두 항균스펙트럼을 가지고 있어 널리 이용되고 있는 보존료이다. 균주에 따라 저해량이 다른데, 일반적으로 세균은 100~500ppm, 곰팡이는 1000ppm이다. 자몽종자추출물은 점성이 있고, 약간 특이한 냄새와 쓴맛을 가지고 있어 식품에 적용 시 제한적이다. A flavonoid component such as naringin, one of the components contained in the grapefruit seed extract, weakens the cell wall and cell membrane function of microorganisms and inhibits enzyme activity, thereby inhibiting the cell proliferation mechanism caused by DNA / RNA. It is also widely used as a preservative because it has an antimicrobial spectrum in food poisoning bacteria such as E. coli, Staphylococcus aureus, and cholera. The amount of inhibition varies depending on the strain. Generally, 100 to 500 ppm of bacteria and 1000 ppm of mold are present. Grapefruit seed extracts are viscous, have a somewhat unusual odor and bitter taste, and are limited in food applications.
유산균은 자연계에 널리 존재하며 탄수화물을 혐기적으로 이용하여 유산을 생산한다. 유산균이 발견되는 자연환경은 다양한데, 사람이나 동물의 장내에 존재할 뿐 아니라, 다양한 채소와 과일에서도 발견되며 요구르트, 우리나라의 김치 등 발효식품에서 중요한 역할을 담당한다. 특히 발효가 되면서 생산되는 박테리오신에 의한 항균작용이 알려져 있다. 유산균 종류 중 락토바실러스(Lactobacillus)속은 배양시간이 지날수록 점점 pH가 낮아진다. 페디코커스(Pediococcus)속의 경우 배양됨에 따라 pH 3.9~4.0인 반면 락토바실러스(Lactobacillus)속은 pH 3.7~3.8로 낮은 것을 실험을 통해 확인되었다. (김치의 저장성 향상을 위한 항균활성 우수 유산균 선발, 2014). 또한 비피도박테리움(Bifidobacterium)속과 항균활성을 비교하였을 때 락토바실러스 에시도필러스(L.acidophilus)가 가장 억제환이 컸으며, 락토바실러스 카제이(L.casei), 비피도박테리움 롱검(B.longum)순이었다. 실험을 통해 락토바실러스(Lactobacillus)속이 항균활성이 높다고 보고되었다. (유산균에 의한 에셰키리아 콜리(Escherichia coli ) 와 살모넬라 티피무리움(Salmonella typhimurium)의 생육억제에 관한 연구, 2002). Lactic acid bacteria are widely found in nature and produce carbohydrates by anaerobic use of carbohydrates. The natural environment in which lactic acid bacteria are found is not only present in the intestines of humans and animals, but also in various vegetables and fruits, and plays an important role in fermented foods such as yogurt and kimchi in Korea. Especially, antimicrobial action by bacteriocin produced by fermentation is known. Among Lactobacillus species, Lactobacillus species gradually become lower in pH over time. In the case of Pediococcus , the pH of the genus Lactobacillus was lowered to 3.7 ~ 3.8, while that of the genus Lactobacillus was 3.9 ~ 4.0 as cultured. (Selection of lactic acid bacteria with excellent antimicrobial activity for improving the storage stability of kimchi, 2014). In addition, L. acidophilus showed the highest inhibitory activity when compared with Bifidobacterium genus. Lactobacillus casei ( L. casei), Bifidobacterium longum ( B. longum ). The experiment Lactobacillus bacteria (Lactobacillus) reported trick high antimicrobial activity. ( Escherichia by lactic acid bacteria coli) and Salmonella typhimurium (A Study on the Growth Inhibition of Salmonella typhimurium), 2002).
기존에 알려진 천연보존료(키토산, 폴리리신 등)를 본 연구에서 발굴한 천연소재와 병합처리한 기술개발의 신규성을 제시하였다.We proposed the novelty of the development of the technology combining the natural preservatives (chitosan, polylysine, etc.) with the natural materials discovered in this study.
키토산은 천연(게, 새우, 갑각류)에 존재하는 키틴을 탈아세틸화시킨 물질로서 글루코사민이 β-1,4로 결합된 천연고분자 다당류이다. 키토산은 천연제로 키토산 자체가 항균력을 지니고 있는 것으로 널리 알려져 있으나 항균력이 일정하지 않고 약한 산성용액에 녹여야만 이용이 가능하다는 단점을 가지고 있다.Chitosan is a deacetylated chitin present in natural (crab, shrimp, crustaceans) and is a natural high-molecular polysaccharide in which glucosamine is bonded with β-1,4. Chitosan is widely known to have the antimicrobial activity of natural chitosan itself, but it has disadvantages that it can be used only if it is dissolved in a weak acidic solution.
폴리리신은 방선균(Streptomyces albulus)이 생성하는 강염기성 천연 물질로서 아미노산(L-Lysine)이 25~35개로 결합되어 있고, 양(+)이온의 전하를 띠고 있다. 폴리리신이 음(-)이온의 전하를 띠고 있는 미생물의 세포막과 이온 결합하여 미생물의 생육을 저해함으로서 항균력 기작을 가지고 있다.Polylysine is a strain of Streptomyces albulus) the amino acid as a strongly basic natural material that generates (L-Lysine) and is a combination of 25 to 35 pieces, are tinged with charges of positive ions. Polylysine has an antimicrobial mechanism by inhibiting the growth of microorganisms by ionic bonding with the cell membrane of a microorganism bearing a negative (-) ion charge.
냉이(Capsella bursa- pastoris)란 겨자과에 속하는 월년초로 들이나 밭에서 자라며, 온대지방에 분포한다. 성분은 수분 81.5%, 단백질 7.3%, 탄수화물 5.6%, 섬유 2.0%, 무기질 2.7%이며, 칼슘과 인은 각각 116mg, 104mg이 함유되어 있다. 생약명으로 제채, 향선채라고도 하며 뿌리를 포함한 전초를 약재로 사용하고 있다. (냉이의 식용부위별 휘발성 향기성분, 1996). 「증보신림경제」에서 냉이는 오장을 이롭게 하고 죽으로 먹으면 간에 이롭고 눈을 밝게 한다고 등재되어 있으며, 중국에서도 홍역의 예방을 위해 예부터 사용되어 왔다. (냉이 물 추출물의 항산화 활성과 Tyrosinase 및 Elastase 저해 효과, 2012). 냉이는 특유의 향이 있어 식욕을 돋구어 주는데, 냉이의 향기성분으로는 부위별로 상이한 것을 연구결과를 통해 밝혀졌다. 뿌리 부위의 경우 ethyl benzene, hexanal, cyclobexanone, ethyl acetate 등 함유되어 있고, 잎 부위는 α-pinene, pentadecane, trans-2-tridecenal 등이 함유되어 있다. 주로 냉이에서 나는 향은 잎에 함유되어 있는 향기 성분인 myrcene, terpinolene 등이다. Capsella bursa- pastoris grows in fields and fields in early May of the month belonging to the mustard family, and is distributed in temperate regions. Ingredients are water 81.5%, protein 7.3%, carbohydrate 5.6%, fiber 2.0%, inorganic 2.7%, calcium and phosphorus are 116mg and 104mg respectively. It is also known as a medicinal herb, a scent, and is used as a medicinal herb that contains roots. (Volatile flavor components of edible parts of cucumber, 1996). In the "Shinseng Shinryeong Economy", the mulberry is listed as beneficial to the people and beneficial to the eyes when it is eaten by the dead, and it has been used in China for preventing measles. (Antioxidant Activity and Tyrosinase and Elastase Inhibitory Effects of Cold Water Extracts, 2012). It has been found through research that the flavor components of cucumber are different depending on the site. Ethyl benzene, hexanal, cyclobexanone, and ethyl acetate are contained in the root part, and the leaf part contains α-pinene, pentadecane, and trans-2-tridecenal. Myrcene, terpinolene, which is the fragrance ingredient contained in the leaves, is the incense which mainly comes out from the cold.
본 발명은 수분을 많이 함유하는 식품의 곰팡이를 예방 및 억제 하는 항진균 조성물을 제공하고자 한다. 또한, 본 발명은 상기 조성물의 효과적인 제조방법을 제공하고자 한다.The present invention provides an antifungal composition for preventing and inhibiting fungi in food containing a large amount of water. The present invention also provides a method for effectively producing the composition.
상기의 목적을 달성하기 위하여 본 발명은 냉이를 에탄올로 추출한 추출물을 포함하는 항진균 조성물을 제공한다. 이때, 상기 에탄올은 농도가 70%인 것이 바람직하다. 또한, 상기 항진균 조성물에 유산균, 키토산 및 폴리리신을 더 포함하는 것이 바람직하다 아울러, 상기 유산균은 락토바실러스 파라카제이(Lactobacillus paracasei)(KCTC 333071)인 것이 바람직하다.In order to achieve the above object, the present invention provides an antifungal composition comprising an extract obtained by extracting cold water with ethanol. At this time, the concentration of the ethanol is preferably 70%. In addition, the antifungal composition preferably further comprises lactic acid bacteria, chitosan and polylysine. In addition, the lactic acid bacterium is preferably Lactobacillus paracasei (KCTC 333071).
본 발명은 냉이를 에탄올로 추출한 추출물을 포함하는 항진균 조성물의 제조방법을 제공한다. 이때, 상기 에탄올은 농도가 70%인 것을 특징으로 한다. 상기 제조방법은 냉이추출물에 유산균, 키토산 및 폴리리신을 더 포함하는 것이 바람직하다. 아울러, 상기 유산균은 락토바실러스 파라카제이(Lactobacillus paracasei)(KCTC 333071)인 것이 바람직하다.The present invention provides a method for producing an antifungal composition comprising an extract obtained by extracting cold water with ethanol. At this time, the ethanol has a concentration of 70%. Preferably, the method further comprises adding lactic acid bacteria, chitosan and polylysine to the cold extract. In addition, the lactic acid bacterium is preferably Lactobacillus paracasei (KCTC 333071).
본 발명에 따른 조성물은 냉이 에탄올 추출물을 포함함으로써, 곰팡이의 증식을 억제하는 항진균 조성물을 제공한다. 본 발명에 따른 상기 조성물 제조방법은 냉이 에탄올 추출물, 유산균, 키토신 및 폴리리신을 포함하는 조성물을 한번에 보다 효율적으로 제조할 수 있다.The composition according to the present invention contains an extract of cold ethanol to provide an antifungal composition that inhibits the growth of fungi. The composition according to the present invention can more efficiently prepare a composition containing cold ethanol extract, lactic acid bacterium, chitosan and polylysine at once.
도 1은 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 농도에 따른 각 균주별 최소억제농도(MIC) 측정결과를 나타낸 것이다.
도 2는 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 Rhizopus oryzae KCTC 6940 균주에 대한 최소억제농도(MIC)의 측정결과를 pH별로 나타낸 것이다.
도 3은 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 Aspergillus niger KCTC 6971 균주에 대한 최소억제농도(MIC)의 측정결과를 pH별로 나타낸 것이다.
도 4는 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 Penicillium citrinum KCTC 6990 균주에 대한 최소억제농도(MIC)의 측정결과를 pH별로 나타낸 것이다.
도 5는 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 Debaryomyces hansenii KCTC 27008 균주에 대한 최소억제농도(MIC)의 측정결과를 pH별로 나타낸 것이다.
도 6은 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물의 Pichia menbranitaciens KCTC 27403 균주에 대한 최소억제농도(MIC)의 측정결과를 pH별로 나타낸 것이다.
도 7은 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물과 냉이 에탄올 추출물의 농도에 따른 식빵의 곰팡이 발생일수를 관찰한 결과를 나타낸 것이다.
도 8은 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물과 냉이 에탄올 추출물의 농도에 따른 떡의 곰팡이 발생일수를 관찰한 결과를 나타낸 것이다.
도 9는 냉이 에탄올 추출물, 유산균발효액, 키토산, 폴리리신을 포함하는 복합물과 냉이 에탄올 추출물의 농도에 따른 시럽의 곰팡이 발생일수를 관찰한 결과를 나타낸 것이다.FIG. 1 shows the results of measurement of the minimum inhibitory concentration (MIC) of each strain according to the concentration of the complex containing ethanol extract, lactic acid fermentation broth, chitosan and polylysine.
FIG. 2 is a graph showing the results of measurement of the minimum inhibitory concentration (MIC) of Rhizopus oryzae KCTC 6940, a combination of cold ethanol extract, lactic acid fermentation broth, chitosan and polylysine, by pH.
FIG. 3 is a graph showing the results of measurement of the minimum inhibitory concentration (MIC) of Aspergillus niger KCTC 6971, a combination of cold ethanol extract, lactic acid fermentation broth, chitosan and polylysine.
FIG. 4 is a graph showing the results of measurement of the minimum inhibitory concentration (MIC) of
FIG. 5 shows the results of measurement of the minimum inhibitory concentration (MIC) of the Debaryomyces hansenii
FIG. 6 shows the results of measurement of the minimum inhibitory concentration (MIC) of Pichia menbranitaciens
FIG. 7 shows the results of observing the number of days of fungal development in the bread according to the concentration of cold ethanol extract, lactic acid fermentation broth, chitosan, polylysine-containing complex and cold ethanol extract.
FIG. 8 shows the results of observation of the number of days of mold development of rice cakes according to the concentrations of the ethanol extract, the fermentation broth of lactic acid bacteria, the chitosan, and polylysine, and the ethanol extract.
FIG. 9 shows the result of observing the number of days of fungal development in the syrup depending on the concentrations of cold ethanol extract, lactic acid fermentation broth, chitosan, polylysine, and cold ethanol extract.
본 발명은 다양한 변환을 가할 수 있고 여러가지 실시예를 가질 수 있는 바, 특정 실시 예들을 도면에 예시하고 상세한 설명에서 상세하게 설명하고자 한다. 그러나, 이는 본 발명을 특정한 실시 형태에 대해 한정하려는 것이 아니며, 본 발명의 사상 및 기술 범위에 포함되는 모든 변환, 균등물 내지 대체물을 포함하는 것으로 이해 되어야 한다. 본 발명을 설명함에 있어서 관련된 공지 기술에 대한 구체적인 설명이 본 발명의 요지를 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다.BRIEF DESCRIPTION OF THE DRAWINGS The present invention is capable of various modifications and various embodiments, and specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.
본 출원에서 사용한 용어는 단진 특정한 실시예를 설명하기 위해 사용된 것으로, 본 발명을 한정하려는 의도가 아니다. 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 본출원에서, "포함하다" 또는 "가지다" 등의 용어는 명세서상에 기재된 특징, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것이지, 하나 또는 그 이상의 다른 특징들이나 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해 되어야 한다. The terminology used in this application is used to describe a particular embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprises" or "having" and the like are used to specify that there is a feature, a number, a step, an operation, an element, a component or a combination thereof described in the specification, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.
본 발명의 일측면은, 냉이 에탄올 추출물을 이용해 고수분식품에서의 항진균 조성물을 제공한다. 냉이 에탄올 추출물과 유산균을 함께 포함하는 조성물은 냉이에탄올 추출물을 단독으로 포함한 조성물에 비해 항진균 활성이 뛰어나므로 곰팡이를 효과적으로 억제할 수 있다.One aspect of the present invention provides an antifungal composition in a highly water-soluble food using a cold ethanol extract. The composition comprising both the cold ethanol extract and the lactic acid bacteria is more effective than the composition containing the cold ethanol extract alone so that the mold can be effectively inhibited.
본 발명의 다른 일측면은 냉이 에탄올 추출물과 유산균, 키토산, 폴리리신을 모두 포함하는 조성물을 제조하는 제조방법을 제공한다. 본 발명의 다른 일측면에서 상기 방법은 냉이를 70%농도에 에탄올에 추출하는 단계를 포함할 수 있다. 냉이 에탄올 추출물을 단독으로 준비하는 방법에 비해 본 발명에 따른 방법은 보다 효율적으로 본 발명의 일측면에 따른 조성물을 제조할 수 있다.Another aspect of the present invention provides a method for producing a composition comprising both a cold ethanol extract and a lactic acid bacterium, chitosan, and polylysine. In another aspect of the present invention, the method may comprise extracting cucumber into ethanol at a concentration of 70%. The method according to the present invention can more efficiently prepare the composition according to one aspect of the present invention in comparison with the method in which the cold ethanol extract is prepared singly.
본 명세서에서 "냉이 추출물(Capsella bursa- pastoris extract)"은 넓게는 냉이에서 비롯된 추출물을 모두 포함하는 개념으로, 구체적으로는 냉이를 70%농도의 에탄올으로 추출한 액을 포함한다. In the present specification, " Capsella bursa- pastoris extract " The concept includes all extracts derived from cucumber, and specifically includes cucumber extracts with 70% ethanol concentration.
본 명세서에서 "유산균"은 당류를 발효하여 에너지를 획득하고 다량의 락트산(lactic acid)을 생성하는 세균을 총칭한다. 일반적으로 유산균은 락토바실러스(Lactobacillus), 락토코커스(Lactococcus), 류코노스톡(Leuconostoc), 페디오코커스(Pediococcus), 및 비피도박테리움(Bifidobacterium)속 균을 포함한다.In the present specification, " lactic acid bacteria " is generically referred to as bacteria that ferment saccharides to obtain energy and produce a large amount of lactic acid. Generally, the lactic acid bacteria include Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, and Bifidobacterium.
본 명세서에서 키토산은 갑각류에 존재하는 키틴을 탈아세틸화 시킨 물질을 총칭한다. 본 발명에서는 항균력이 있는 유화제 역할을 기대할 수 있고, 약산성에 녹는 성질 때문에 유산균 상등액과 함께 사용할 경우 용해도가 높아지는 효과가 있다.In the present specification, chitosan is collectively referred to as a substance in which chitin present in crustacea is deacetylated. In the present invention, an emulsifier having antimicrobial activity can be expected, and its solubility increases when it is used together with a lactic acid bacteria supernatant because of its melting property in a weak acidity.
본 명세서에서 폴리리신은 방선균(Streptomyces albulus)이 생성하는 강염기성 천연 물질로서 아미노산(L-Lysine)이 25-35개로 결합되어 있고, 양(+)이온 전하를 띠고 있는것을 포함한다. In the present specification, polylysine is a strong basic natural substance produced by Streptomyces albulus, which contains 25-35 amino acids (L-Lysine) and has positive (+) ion charge.
본 발명의 일측면에서 유산균은 락토바실러스 플란타리움(Lactobacillus plantarum) (KCCM11322),락토바실러스 사케이(Lactobacillus sakei) (KCCM40264), 락토바실러스 살리바리어스 서브스페이스. 살리시니어스(Lactobacillus salivarius subsp. salicinius) (KCCM40998), 락토바실러스 플란타리움 SJC 001(Lactobacillus plantarum SJC 001) (KCCM40962), 락토바실러스 컬바투스 C3(Lactobacillus curvatus C3) (KCCM43009), 락토바실러스 사케이 WIKIM50(Lactobacillus sakei WIKIM50) (KCCM43213), 락토바실러스 파라카제이(Lactobacillus paracasei) (KCTC33071), 보유특허균 (KACC91459P)중에서 선택된 하나 이상을 포함한다.In one aspect of the present invention, the lactic acid bacterium is selected from the group consisting of Lactobacillus plantarum (KCCM 11322), Lactobacillus sakei (KCCM 40264), Lactobacillus salivarius subsp. Lactobacillus salvarius subsp. Salicinius (KCCM 40998), Lactobacillus plantarum SJC 001 (KCCM 40962), Lactobacillus curvatus C3 (KCCM 43009), Lactobacillus saccharum WIKIM50 Lactobacillus sakei WIKIM50) (KCCM43213), Lactobacillus paracasei (KCTC33071), and the patented bacteria (KACC91459P).
본 발명의 일측면은, 고수분 식품에 자주 발생하는 곰팡이를 조사하여 제어대상으로 리조푸스 오리제(Rhizopus oryzae) (KCTC 6940), 아스퍼질러스 나이거(Aspergillus niger) (KCTC 6971), 페니실리움 시트리눔(Penicillium citrinum) (KCTC 6990), 디바리오마이세스 한세니(Debaryomyces hansenii) (KCTC 27008), 피치아 멘브레니타시엔(Pichia menbranitaciens) (KCTC 27403) 다음과 같은 5종의 곰팡이를 선정하고, 이들 5종의 균주에 대해 곰팡이 저해실험을 진행해 대조군과 개발된 시료를 사용하여 곰팡이가 생기는 시간, 개수, 클린존의 크기 등을 비교하여 효과를 검증했다. 최소저해농도(MIC:Minimum Inhibitory Concentration)을 측정해 그 결과를 확인했다. 또한 활성 안정성 평가를 위해서 추출물과 상기 5종의 균주에서의 pH안정성 평가를 진행 하였다.One aspect of the present invention relates to a method for controlling fungi that frequently occur in highly watered foods, which comprises controlling Rhizopus oryzae (KCTC 6940), Aspergillus niger (KCTC 6971), penny Such as Penicillium citrinum ( KC TC 6990), Debaryomyces hansenii (KCTC 27008), Pichia menierebrenitacien menbranitaciens) (K CTC 27403) following the selection of the mold 5 species such, these proceeded to mold inhibition experiments for the strains of 5 kinds of time caused a mold by using the sample D and the control group, the number, a clean zone size, etc. And the effect was verified. The minimum inhibitory concentration (MIC) was measured and the results were confirmed. In order to evaluate the stability of the enzyme, the pH stability of the extracts and the 5 strains were evaluated.
이하, 실시예를 들어 본 발명의 구성 및 효과를 보다 구체적으로 설명한다. 그러나 이들 실시예는 본 발명의 이해를 돕기 위해 예시의 목적으로만 제공된 것일 뿐 본 발명의 범주가 그에 의해 제한되는 것은 아니다.Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to examples. However, these embodiments are provided for illustrative purposes only to facilitate understanding of the present invention, and the scope of the present invention is not limited thereto.
실시예1Example 1 : 천연 추출물 제조 : Production of natural extract
소재를 물로 깨끗이 세척하고 24시간동안 40℃에서 저온건조 하였다. 추출물 제조를 위해 냉이와 용매(DW, 70% 에탄올)를 1:5의 비율로 24 시간 동안 저온추출하고 저온추출한 추출물에 다시 동일량의 용매를 가하여 60℃에서 4시간 진탕 추출하였다. 제조된 추출물은 감압여과하고 실린지 필터(syringe filter)로 제균하였다.The material was cleaned with water and dried at 40 ° C for 24 hours. For the preparation of extracts, low temperature extraction of cold water and solvent (DW, 70% ethanol) at a ratio of 1: 5 for 24 hours, low temperature extraction, and the same amount of solvent were added again at 60 ℃ for 4 hours. The extract was filtered under reduced pressure and sterilized with a syringe filter.
실시예2Example 2 : 추출물의 곰팡이 생장 최소 억제 농도 평가( : Evaluation of Minimum Inhibitory Concentration of Fungal Growth of Extracts MICMIC ))
곰팡이 포자 현탁액을 제조하기 위하여 감자즙 배지(potato dextrose agar, PDA)에서 배양해 놓은 곰팡이를 백금이로 긁어 멸균생리식염수에 현탁하여 포자 현탁액을 제조하였다. 제조한 현탁액과 염색약을 1:1로 염색하고 헤마사이토메타(haemacytometer)에 주입하여 현미경으로 포자수를 계수하여 포자수를 (1~9)X105로 맞추었다.In order to prepare mold spore suspension, molds cultured in potato dextrose agar (PDA) were scraped with platinum and suspended in sterile physiological saline to prepare spore suspension. The resulting suspension and dye were stained with a 1: 1 dye, injected into a haemacytometer, and the number of spores was counted with a microscope to adjust the number of spores to (1 to 9) × 10 5 .
각 곰팡이에 대한 최소억제농도(MIC:minimum inhibitory concentration)는 96 well plate 내에서 시료 추출물, 곰팡이 포자 현탁액, 감자 추출 배지(potato dextrose broth, PDB)를 96 well plate 1 : 1 : 1로 주입한 후 25℃에서 배양하면서 마이크로플레이트 리더(microplate reader)로 12시간마다 420 nm에서 흡광도를 측정하여 곰팡이의 성장이 나타나지 않는 시료의 가장 낮은 농도로 결정하였다.The minimum inhibitory concentration (MIC) for each fungus was determined by injecting a 1: 1: 1 96-well plate with a sample extract, a fungal spore suspension, and a potato dextrose broth (PDB) Absorbance was measured at 420 nm every 12 hours with a microplate reader while incubating at 25 ° C to determine the lowest concentration of the sample that did not exhibit mold growth.
Rhizopus
oryzae Rhizopus oryzae
Rhizopus
oryzae
Aspergillus
nigerAspergillus or Lee
Aspergillus
nige r
Penicillium citrinum Penicillium citrinum
Penicillium citrinum
Debaryomyces
hansenii Diva Mayes Hanseyini
Debaryomyces
hansenii
Pichia
menbranitaciens Pichia Menbrenitaxi
Pichia
menbranitaciens
DWBlueberries
DW
EtOHBlueberries
EtOH
상기 [표 1]에서 본 발명의 항진균 효능은 냉이 70% ethanol 추출물에서 넓은 균주에 대하여 상당한 항진균력이 있는 것으로 보고 냉이 추출물을 선정 하였다.In Table 1, the antifungal activity of the present invention was determined to have a considerable antifungal activity against a broad strain in a 70% ethanol extract of cyanobacterium.
실시예3Example 3 : 유산균 선정 실험 : Lactic acid bacteria selection experiment
앞서 선정한 냉이 추출물과 병합처리에 의한 항진균 활성의 상승 효과를 위해 항균 물질인 박테리오신을 생성하는 유산균을 선별하였다.Lactic acid bacteria producing bacteriocin, an antimicrobial substance, were selected for the synergistic effect of antifungal activity by the combination of the previously selected cold extract and the combined treatment.
실험에 사용한 유산균 배양액 상등액은 액체배양된 유산균을 원심분리기에 5000rpm에서 10분간 돌려 상등액을 분리하여 유산균체는 버리고 발효산물이 남은 상등액만 사용하였다.The supernatant of the lactic acid bacteria culture liquid used in the experiment was obtained by centrifuging the liquid cultured lactic acid bacteria in a centrifuge at 5,000 rpm for 10 minutes to separate the supernatant and discard the lactic acid bacteria and use only the supernatant remaining the fermented product.
각 곰팡이에 대한 최소억제농도(MIC:minimum inhibitory concentration)는 96 well plate 내에서 냉이 추출물, 곰팡이 포자 현탁액, 감자 추출 배지(potato dextrose broth, PDB)를 96 well plate에 1 : 1 : 1로 주입한 후 25℃에서 배양하면서 마이크로플레이트 리더(microplate reader)로 12시간마다 420 nm에서 흡광도를 측정하여 곰팡이의 성장이 나타나지 않는 시료의 가장 낮은 농도로 결정하였다.The minimum inhibitory concentration (MIC) for each fungus was determined by 1: 1: 1 injection of cold extract, fungal spore suspension, and potato dextrose broth (PDB) in 96 well plates After incubation at 25 ° C, the absorbance at 420 nm was measured with a microplate reader every 12 hours to determine the lowest concentration of the sample that did not show mold growth.
( Rhizopus
oryzae ) Rhizopus oryzae
( Rhizopus
oryzae )
( Aspergillus
niger)Aspergillus or Lee
( Aspergillus
nige r)
시트리눔
( Penicillium citrinum) Penicillium
Citrinum
( Penicillium citrinum)
( Debaryomyces
hansenii ) Diva Mayes Hanseyini
( Debaryomyces
hansenii )
멘브레니타시엔
( Pichia
menbranitaciens ) Pichia
Memblenitasien
( Pichia
menbranitaciens )
(Lactobacillus plantarum )
(KCCM11322)Lactobacillus plantarum
(Lactobacillus plantarum )
(KCCM11322)
(Lactobacillus sakei )
(KCCM40264)Lactobacillus SA
(Lactobacillus sakei )
(KCCM40264)
서브스페이스. 살리시니어스
(Lactobacillus salivarius
subsp . salicinius )
(KCCM40998)Lactobacillus salivarius
Subspace. Salicornese
(Lactobacillus salivarius
subsp . salicinius )
(KCCM40998)
(Lactobacillus plantarum SJC 001)
(KCCM40962)Lactobacillus plantarum SJC 001
(Lactobacillus plantarum SJC 001)
(KCCM40962)
(Lactobacillus curvatus C3)
(KCCM43009)Lactobacillus Culbatatus C3
(Lactobacillus curvatus C3)
(KCCM43009)
(Lactobacillus sakei WIKIM50)
(KCCM43213)Lactobacillus casei WIKIM50
(Lactobacillus sakei WIKIM50)
(KCCM43213)
(Lactobacillus paracasei )
(KCTC33071)Lactobacillus paracasei
(Lactobacillus paracasei )
(KCTC33071)
(KACC91459P)Patented microorganism
(KACC91459P)
상기 [표 2]에서 본 발명의 항진균 효능은 락토바실러스 파라카제이(Lactobacillus paracasei) (KCTC 33071)가 넓은 균주에 대하여 상당한 항진균력이 있는 것으로 보고 락토바실러스 파라카제이(Lactobacillus paracasei) (KCTC 33071)를 선정 하였다.In Table 2, the antifungal effect of the present invention was confirmed by comparing Lactobacillus paracasei (KCTC 33071) with Lactobacillus paracasei (KCTC 33071) Respectively.
실시예4Example 4 : 유산균 복합물과 천연보존료의 병합으로 인한 항진균 증진 효과 : Enhancement of Antifungal Activity by Combination of Lactic Acid Bacteria Complex and Natural Preservative
냉이추출물과 락토바실러스 파라카제이(Lactobacillus paracasei) (KCTC 33071)의 발효 상등액, 천연 첨가물로 인정되어 있는 폴리리신 그리고 천연 항균제인 키토산을 병합처리할 경우 항진균력이 강화되는지 여부를 검토해 보았다.We investigated whether the antifungal activity of Lactobacillus paracasei (KCTC 33071) fermentation supernatant, polylysine, which is a natural additive, and chitosan, a natural antimicrobial agent, were enhanced by the addition of cold extracts and Lactobacillus paracasei (KCTC 33071).
복합물(이하, 조성물과 균등한 개념으로 사용함) 소재Composite (hereinafter, used as a concept equivalent to the composition) Material
가. 항진균 소재 추출물 : 냉이 70% Ethanol 추출물 end. Antifungal material extract: 70% Ethanol extract
나. 사용 유산균 균주 I. Use lactic acid bacteria strain
유산균 스크리닝 중 가장 높은 항진균력을 가지는 락토바실러스 파라카제이(Lactobacillus paracasei) (KCTC 33071)를 사용하여 실험하였다. Lactobacillus paracasei (KCTC 33071), which has the highest antifungal activity among lactic acid bacteria screening, was used.
다. 키토산 All. Chitosan
키토산은 천연(게, 새우, 갑각류)에 존재하는 키틴을 탈아세틸화시킨 물질로서 글루코사민이 β-1,4로 결합된 천연고분자 다당류이다. 키토산은 천연제로 키토산 자체가 항균력을 지니고 있는 것으로 널리 알려져 있으나 항균력이 일정하지 않고 약한 산성용액에 녹여야만 이용이 가능하다는 단점을 가지고 있다. 본 실험에서는 처방에 사용할 경우 항균력이 있는 유화제 역할을 기대할 수 있고 약산성에 녹는 성질 때문에 유산균 상등액과 함께 사용할 경우 용해도가 높아지는 장점이 있다. ㈜키토라이프가 생산하는 키토산분말(KL-245)을 사용하였다.Chitosan is a deacetylated chitin present in natural (crab, shrimp, crustaceans) and is a natural high-molecular polysaccharide in which glucosamine is bonded with β-1,4. Chitosan is widely known to have the antimicrobial activity of natural chitosan itself, but it has disadvantages that it can be used only if it is dissolved in a weak acidic solution. In this experiment, antimicrobial activity of antimicrobial agent can be expected to be expected when used in prescription. Due to its melting property in weak acidity, it has an advantage of increasing solubility when used in combination with lactic acid bacteria supernatant. Chitosan powder (KL-245) produced by KITO LIFE Co., Ltd. was used.
라. 폴리리신 la. Polylysine
폴리리신은 방선균(Streptomyces albulus)이 생성하는 강염기성 천연 물질로서 아미노산(L-Lysine)이 25~35개로 결합되어 있고, 양(+)이온의 전하를 띠고 있다. 폴리리신이 음(-)이온의 전하를 띠고 있는 미생물의 세포막과 이온 결합하여 미생물의 생육을 저해함으로서 항균력 기작을 가지고 있다. 본 발명에서는 ㈜신승하이켐이 생산하는 폴리리신(50P)을 사용하였다.Polylysine is a strain of Streptomyces albulus) the amino acid as a strongly basic natural material that generates (L-Lysine) and is a combination of 25 to 35 pieces, are tinged with charges of positive ions. Polylysine has an antimicrobial mechanism by inhibiting the growth of microorganisms by ionic bonding with the cell membrane of a microorganism bearing a negative (-) ion charge. In the present invention, polylysine (50P) produced by Shin Seung Hee Chem Co., Ltd. was used.
A : 냉이추출물(고형분 20brix)A: Cold extract (solid content 20brix)
B : 유산균 발효액(107 pH 3.6)B: Lactic acid fermentation broth (10 7, pH 3.6)
C : 키토산(1%)C: chitosan (1%)
D : 폴리리신(2%)D: polylysine (2%)
상기 [표 3]의 배합비에 따라 최소억제농도(MIC)를 측정 하였다.The minimum inhibitory concentration (MIC) was measured according to the compounding ratio shown in [Table 3].
( Rhizopus oryzae ) Rhizopus oryzae
( Rhizopus oryzae )
( Aspergillus niger)Aspergillus or Lee
( Aspergillus niger r)
( Penicillium citrinum ) Penicillium citrinum
( Penicillium citrinum )
( Debaryomyces hansenii ) Diva Mayes Hanseyini
( Debaryomyces hansenii )
( Pichia menbranitaciens ) Pichia Menbrenitaxi
( Pichia menbranitaciens )
상기 [표 4]에서 곰팡이 리조푸스 오리제(Rhizopus oryzae) (KCTC 6940), 아스퍼질러스 나이거(Aspergillus niger) (KCTC 6971), 페니실리움 시트리눔(Penicillium citrinum) (KCTC 6990), 디바리오마이세스 한세니(Debaryomyces hansenii) (KCTC 27008) 및 피치아 멘브레니타시엔(Pichia menbranitaciens) (KCTC 27403) 5종에 대하여 항진균력이 모두 나타나지만 그 중에서도 A, B, C, D를 모두 넣은 4번의 배합비의 항진균력이 뛰어났다.In Table 4, fungi Rhizopus oryzae oryzae (KCTC 6940), Aspergillus niger (KCTC 6971), Penicillium citrinum ( KC TC 6990), Debaryomyces The antifungal activity of 5 hansenii (KCTC 27008) and Pichia menbranitaciens ( K CTC 27403) was observed, but the antifungal activity of 4 combinations of A, B, C and D It was excellent.
본 발명의 항진균 소재는 냉이 추출물, 유산균 상등액, 키토산, 폴리리신의 복합물(25:25:25:25)이 총 5종의 곰팡이에 대한 광범위한 항진균력을 갖는 것으로 판단된다. The antifungal material of the present invention is believed to have a broad spectrum of antifungal activity against all five fungi (25: 25: 25: 25) in combination with cold extract, lactic acid supernatant, chitosan and polylysine.
이 복합물을 SLCP 복합물이라 칭하고 이후의 실험에 사용하였다.This composite was called an SLCP composite and was used in subsequent experiments.
실시예5Example 5 : 선정된 복합물( : Selected Complexes ( SLCPSLCP ) ) MICMIC 측정 Measure
상기에서 제조한 복합물의 농도에 따른 항진균력 여부를 검토해 보았다.Antifungal activity was evaluated according to the concentration of the composite prepared above.
농도에 따라 곰팡이 포자 현탁액을 사용하여 감자추출배지(PDB)를 넣고 배양하면서 시간 경과에 따른 흡광도를 측정하여 곰팡이의 최소저해농도인 MIC를 측정한 결과를 도 1에 나타내었다. 도 1을 살펴보면, 리조푸스오리제(Rhizopus oryzae) (KCTC 6940)를 대상으로 한 복합물 MIC값은 1650 ppm으로 나타났다. 아스퍼질러스 나이거 (Aspergillus niger) (KCTC 6971)를 대상으로 한 복합물 MIC값은 800 ppm으로 나타났다. 페니실리움 시트리눔(Penicillium citrinum) (KCTC 6990)를 대상으로 한 복합물 MIC값은 800 ppm으로 나타났다. 디바리오마이세스 한세니(Debaryomyces hansenii) (KCTC 27008)를 대상으로 한 복합물 MIC값은 800ppm으로 나타났다. 피치아 멘브레니타시엔(Pichia menbranitaciens ) (KCTC 27403)를 대상으로 한 복합물 MIC값은 800ppm으로 나타났다.FIG. 1 shows the result of measuring the minimum inhibitory concentration (MIC) of the fungus by measuring the absorbance over time while inserting the potato extract medium (PDB) using a mold spore suspension according to the concentration. Referring to FIG. 1, the composite MIC value for Rhizopus oryzae (KCTC 6940) was 1650 ppm. The composite MIC value for Aspergillus niger (KCTC 6971) was 800 ppm. The composite MIC value for Penicillium citrinum (KCTC 6990) was 800 ppm. The composite MIC value for Debaryomyces hansenii (KCTC 27008) was 800 ppm. Pichia The composite MIC value for menbranitaciens ( KCTC 27403) was 800 ppm.
상기 상술한 바와 같이 SLCP 복합물의 리조푸스 오리제(Rhizopus oryzae) (KCTC 6940), 아스퍼질러스 나이거(Aspergillus niger) (KCTC 6971), 페니실리움 시트리눔(Penicillium citrinum) (KCTC 6990), 디바리오마이세스 한세니(Debaryomyces hansenii) (KCTC 27008) 및 피치아 멘브레니타시엔(Pichia menbranitaciens) (KCTC 27403)에 대한 최소저해농도인 MIC는 800~1650 ppm으로 나타났다.As described above, the SLCP complex Rhizopus oryzae ( Rhizopus oryzae (KCTC 6940), Aspergillus niger (KCTC 6971), Penicillium citrinum ( KC TC 6990), Debaryomyces The minimum inhibitory concentration (MIC) for hansenii (KCTC 27008) and Pichia menbranitaciens ( K CTC 27403) was 800 to 1650 ppm.
실시예6Example 6 : 선정된 복합물( : Selected Complexes ( SLCPSLCP ) ) MICMIC 측정 Measure
SLCP 복합물을 식품에 적용할 경우 식품의 pH 변화에 따른 안정성을 조사하였다.When the SLCP complex was applied to food, the stability of the food according to the pH change was investigated.
SLCP 복합물의 pH를 0.1N NaOH, 0.1N HCl을 이용하여 pH 2, 4, 6, 8, 10으로 조정하고 24시간 이후에 본래 pH로 맞춘 다음 항진균 실험을 진행하여 pH 변화에 따라 항진균력에 변화가 있는지 알아보았다.The pH of the SLCP composite was adjusted to
도 2는 리조푸스오리제 (Rhizopus oryzae ) (KCTC 6940)를 대상으로 한 pH 안정성 실험으로 산성에서 알칼리(pH 2, 4, 6, 10) 조건까지 전체적으로 안정한 경향을 나타냈다.Fig. 2 is a graph showing the results of a comparison between Rhizopus oryzae ( KCTC 6940), the overall stability was stable from acidic to alkaline (
도 3은 아스퍼질러스 나이거 (Aspergillus niger) (KCTC 6971)를 대상으로 한 pH 안정성 실험으로 중성에서 알칼리(pH 6, 8, 10) 조건까지 안정한 경향을 나타냈다.FIG. 3 shows a stable tendency from neutral to alkaline (
도 4는 페니실리움 시트리눔(Penicillium citrinum ) (KCTC 6990)를 대상으로 한 pH 안정성 실험으로 산성에서 알칼리(pH 2, 4, 6, 8, 10) 조건까지 안정한 경향을 나타냈다.Figure 4 is a graph showing the effect of Penicillium < RTI ID = 0.0 > citrinum ) ( KCTC 6990), it showed a stable trend from acid to alkaline (
도 5는 디바리오마이세스 한세니(Debaryomyces hansenii ) (KCTC 27008)를 대상으로 한 pH 안정성 실험으로 산성에서 알칼리(pH 2, 4, 6, 8, 10) 조건까지 안정한 경향을 나타냈다.FIG. 5 is a graphical representation of the < RTI ID = 0.0 > Debaryomyces & hansenii ) ( KCTC 27008) showed stable tendency from acid to alkaline (
도 6은 피치아 멘브레니타시엔(Pichia menbranitaciens ) (KCTC 27403)을 대상으로 한 pH 안정성 실험으로 산성에서 알칼리(pH 2, 4, 6, 8, 10) 조건까지 안정한 경향을 나타냈다.Figure 6 is a graphical representation of Pichia < RTI ID = 0.0 > menbranitaciens) at acidic pH in stability experiments with the target (KCTC 27403) to alkaline conditions (
상기 실험결과를 통해 SLCP 복합물은 리조푸스 오리제(Rhizopus oryzae) (KCTC 6940), 아스퍼질러스 나이거(Aspergillus niger) (KCTC 6971), 페니실리움 시트리눔(Penicillium citrinum) (KCTC 6990), 디바리오마이세스 한세니(Debaryomyces hansenii) (KCTC 27008) 및 피치아 멘브레니타시엔(Pichia menbranitaciens) (KCTC 27403) 다음과 같은 곰팡이 5종에 대하여 산성에서 알칼리 조건까지 광범위한 pH에 안정한 것으로 판단된다.As a result of the above experiment, the SLCP composite was found to have a Rhizopus oryzae (KCTC 6940), Aspergillus niger (KCTC 6971), Penicillium citrinum ( KC TC 6990), Debaryomyces hansenii (KCTC 27008) and Pichia menbranitaciens ( K CTC 27403) are believed to be stable over a wide range of pH from acidic to alkaline conditions for the following five fungi:
실시예7 : 최종 항진균 복합물 시제품 적용실험-식빵Example 7: Test application of a final antifungal compound composition sample -
대표적인 고수분 식품 중 하나인 식빵에 제조 시 SLCP 복합물을 첨가하여 식빵의 보존성 향상에 미치는 영향을 검토하였다. SLCP composites were added to bread, which is one of the typical high-water foods, and the effect on the improvement of preservability of bread was investigated.
식빵을 제조 시 SLCP 복합물을 원료 중량 대비 0~2% 되도록 첨가하고 다음과 같은 배합비로 제조하였다. 완성된 식빵은 폴리에틸렌(PE) 재질의 지퍼백에 담아 보관하여 25℃ 배양기에서 곰팡이 증식 정도를 육안으로 관찰하였다.In preparing the bread, the SLCP composite was added in a proportion of 0 to 2% based on the weight of the raw material, and the blend ratio was as follows. The finished bread was stored in a polyethylene (PE) zipper bag, and the degree of fungal growth was visually observed at 25 ° C in an incubator.
상기 [표 5]의 배합비로 1 kg기준 배합으로 제조하여 계량하였고 원료 중량을 기준으로 배합비를 계산하여 물 대신 SLCP 복합물을 넣어 반죽하였다. 대조군으로 냉이 추출물을 각각 1% 및 2% 첨가한 식빵을 제조하였다.The mixture was weighed at a mixing ratio of 1 kg in the above Table 5 and weighed. The mixture ratio was calculated based on the weight of the raw material, and SLCP composite was added to knead the mixture. As a control group, breads containing 1% and 2% crude extracts were prepared.
도 7에 나타난 바와 같이, 무첨가 식빵의 경우 3일 후 곰팡이가 관찰되었고 냉이 추출물 1%는 4일, 냉이 추출물 2%와 SLCP 1%는 5일 이후 곰팡이가 발생하였으나 냉이 2%추출물의 경우 관능적으로 떨어지기 때문에 SLCP복합물이 시제품 적용에 용이하다. As shown in FIG. 7, the mold was observed 3 days after the addition of the unsweetened bread. The molds were observed after 4 days in 1% cold extract, 2% cold extract and 1% SLCP in 5 days, SLCP composites are easy to apply to prototypes because they fall.
실시예8Example 8 : 최종 항진균 복합물 시제품 적용실험-떡 : Application of final antifungal compound compound prototype - rice cake
대표적인 고수분 식품 중 하나인 떡볶이 떡에 첨가하여 떡볶이 떡의 보존성 향상에 미치는 영향을 검토하였다. The effect of rice cakes on the preservation of rice cakes was investigated.
제조 후 보존료 처리를 위하여 농도에 따른 SLCP 복합물 수용액에 30초간 침지한 후 겉면을 건조한 다음 폴리에틸렌(PE) 재질의 지퍼백에 담아 보관하며 25℃ 배양기에서 곰팡이 증식 정도를 육안으로 관찰하였다. 대조군으로는 떡에 보통 사용하는 주정 95%와 보존료의 대조군으로 자몽종자추출물을 사용하였다.After the preparation, it was immersed for 30 seconds in aqueous solution of SLCP according to the concentration, and then the surface was dried and stored in a polyethylene (PE) zipper bag, and the degree of fungal growth was visually observed at 25 ° C incubator. As a control group, grapefruit seed extract was used as a control group of 95% alcohol and preservative commonly used in rice cakes.
도 8에 나타난 바와 같이, 무첨가 떡볶이 떡과 자몽종자 추출물 첨가 떡볶이 떡의 경우 4일 후 곰팡이가 관찰되었고 각각 주정 95%, 냉이 추출물 1%, 0.1%, 0.01%, SLCP 1%, 0.1% 6일, SLCP 0.01% 5일 이후에 곰팡이가 발생하는 것으로 보아 유통기한이 연장되었다.As shown in FIG. 8, molds were observed after 4 days in the case of rice cakes with no added rice cakes and grapefruit seed extracts, and fungi were observed at 4 days after sowing with 1%, 0.1%, 0.01%,
실시예9Example 9 : 최종 항진균 복합물 시제품 적용실험- : Application of the final antifungal compound composition prototype - 초코시럽Chocolate Syrup
대표적인 고수분 식품 중 하나인 식빵에 제조 시 SLCP 복합물을 첨가하여 식빵의 보존성 향상에 미치는 영향을 검토하였다. SLCP composites were added to bread, which is one of the typical high-water foods, and the effect on the improvement of preservability of bread was investigated.
초코소스를 제조 시 SLCP 복합물을 원료 중량 대비 1% 되도록 첨가하고 다음과 같이 제조하였다. 완성된 초코소스는 실제 제품에 사용하고 있는 HDPE제질의 통에 담아 보관하여 25℃ 배양기에서 곰팡이 증식 정도를 관찰하였다.In the preparation of chocolate sauce, the SLCP composite was added in an amount of 1% based on the weight of the raw material and prepared as follows. The finished chocolate sauce was stored in a HDPE bag used for the actual product, and the degree of fungal growth was observed at 25 ° C in an incubator.
도 9에 나타난 바와 같이, 무첨가 초코소스의 경우 8일 후, 소르빈산칼륨 1%는 9일 후 곰팡이가 발생하였다. 자몽종자추출물 1%, 냉이 1%, SLCP 1% 첨가 초코소스는 10일 후 곰팡이가 발생하여 유통기한이 연장되었다.As shown in Fig. 9, after 8 days in the case of the no-added chocolate source, 1% of the potassium sorbate produced mold after 9 days. The shelf life of the chocolate sauce added 1% of grapefruit seed extract, 1% of cold syrup, and 1% of SLCP was prolonged after 10 days due to fungi.
Claims (8)
상기 항진균 조성물은 상기 냉이를 70% 농도의 에탄올로 추출한 냉이추출물, 락토바실러스 파라카제이(Lactobacillus paracasei)(KCTC 333071), 키토산 및 폴리리신을 중량비 기준 1:1:1:1의 비율로 혼합한 것을 특징으로 하는 항진균 조성물.The method according to claim 1,
The antifungal composition was prepared by mixing the cold extract, Lactobacillus paracasei (KCTC 333071), chitosan and polylysine extracted at a concentration of 70% ethanol in a weight ratio of 1: 1: 1: 1 ≪ / RTI >
상기 항진균 조성물은 리조푸스 속(Rhizopus sp.), 아스퍼질러스 속(Aspergillus niger sp.), 페니실리움 속(Penicillium citrinum), 디바리오마이세스 속(Debaryomyces sp.) 또는 피치아 속(Pichia sp.)진균에 대한 항진균 활성을 가지는 것을 특징으로 하는 항진균 조성물.The method according to claim 1,
The antifungal composition may be selected from the group consisting of Rhizopus sp. , Aspergillus niger sp ., Penicillium citrinum , Debaryomyces sp. Or Pichia sp. . ) An antifungal activity against fungi.
상기 리조푸스 속(Rhizopus sp.)진균은 리조푸스 오리제(Rhizopus oryzae)인 것을 특징으로 하는 항진균 조성물. The method of claim 3,
Wherein the Rhizopus sp. Fungus is Rhizopus oryzae .
상기 아스퍼질러스 속(Aspergillus niger sp.)진균은 아스퍼질러스나이거(Aspergillus niger)인 것을 특징으로 하는 항진균 조성물The method of claim 3,
The genus Aspergillus (Aspergillus niger sp.) Fungi antifungal composition, characterized in that Aspergillus seuna of this (Aspergillus niger)
상기 페니실리움 속(Penicillium citrinum)진균은 페니실리움 시트리눔(Penicillium citrinum)인 것을 특징으로 하는 항진균 조성물.The method of claim 3,
Wherein the fungus of the genus Penicillium citrinum is Penicillium citrinum .
상기 디바리오마이세스 속(Debaryomyces sp.)진균은 디바리오마이세스 한세니(Debaryomyces hansenii)인 것을 특징으로 하는 항진균 조성물. The method of claim 3,
Wherein the fungus belonging to the genus Divariomyces is Debaryomyces hansenii .
상기 피치아 속(Pichia sp.)진균은 피치아 멘브레니타시엔(Pichia menbranitaciens)인 것을 특징으로 하는 항진균 조성물.
The method of claim 3,
Wherein said Pichia sp. Fungus is Pichia menbranitaciens. ≪ RTI ID = 0.0 > 8. < / RTI >
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