KR20050024872A - Fermented pollen extracts with antimicrobial activity - Google Patents
Fermented pollen extracts with antimicrobial activity Download PDFInfo
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- KR20050024872A KR20050024872A KR1020030062098A KR20030062098A KR20050024872A KR 20050024872 A KR20050024872 A KR 20050024872A KR 1020030062098 A KR1020030062098 A KR 1020030062098A KR 20030062098 A KR20030062098 A KR 20030062098A KR 20050024872 A KR20050024872 A KR 20050024872A
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- pollen
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- antimicrobial activity
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
- A23L21/20—Products from apiculture, e.g. royal jelly or pollen; Substitutes therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/16—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating loose unpacked materials
-
- 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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- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
유산균은 자연계에 널리 존재하며 탄수화물을 혐기적으로 이용하여 유산을 생산하는 미생물로서 직·간접적으로 식품에 첨가되어 이의 대사산물인 젖산에 의하여 식품의 저장성을 향상시키며, 식품의 향미와 조직을 개선한다. 발효식품을 통하여 섭취된 유산균은 장내로 유입된 후 장내 상피세포에 착생하게 되어 병원성 미생물의 저해 및 길항작용, 면역활성의 증진, 암 발생률의 감소, 그리고 발암원인성 효소의 감소 등 숙주 동물에 많은 도움을 준다고 알려져 있다. Lactobacillus is a microorganism that exists widely in nature and uses carbohydrates anaerobicly to produce lactic acid. It is added directly or indirectly to food to improve the shelf life of food by its metabolite, lactic acid, and improve the flavor and texture of food. . Lactobacillus ingested through fermented foods enters the intestinal epithelial cells and enters the intestinal epithelial cells, which are often used in host animals such as inhibiting and antagonizing pathogenic microorganisms, enhancing immune activity, reducing cancer incidence, and reducing carcinogenic enzymes. It is known to help.
식품을 자연에 방치하게 되면 곧바로 향미, 색조 등의 외관이 변화됨과 동시에 성분도 분해되어 영양가의 감소와 함께 식용으로서의 가치를 상실하게 되는데 여기에는 식품 속의 효소와 미생물의 작용에 의한다고 할 수 있다. 이 중에서 미생물의 증식에 의한 식품의 변질이 가장 많이 일어난다고 할 수 있다.When food is left in nature, the appearance of flavor, color, etc. changes immediately, and the ingredients are decomposed to reduce the nutritional value and lose the value as food. This is due to the action of enzymes and microorganisms in food. Among these, the deterioration of foods due to the growth of microorganisms can be said to occur the most.
유해 미생물의 생육을 억제하고, 식품의 부패를 방지하여 식품의 저장기간을 연장하기 위한 기존의 처리방안으로는 크게 2가지로 나누어 볼 수 있다. 즉, 고온, 고압 하에서의 살균, 자외선 및 방사선 조사 등의 방법으로서, 이는 식품에 함유되어 있는 위해한 세균, 곰팡이의 살균뿐만 아니라 각종 영양성분들이 파괴를 수반하는 문제점이 있다. 또 한가지는 상기의 효과를 위하여 식품에 첨가하는 약제인 식품 첨가제와 식품 보존제의 사용으로 종래의 식품보존제들은 안식향산, 솔빈산, 디하이드로 초산, 프로피온산 등의 화학합성 제제들로서 사용목적에 따라 최소량이 권장·사용되고 있으나 화학합성물로서의 보건위생상 안정성에 심각한 문제로 대두되고 있다.Existing treatment methods for inhibiting the growth of harmful microorganisms, preventing the corruption of food and extending the storage period of food can be divided into two types. That is, as a method of sterilization under high temperature, high pressure, ultraviolet rays and irradiation, there is a problem that not only harmful germs and molds contained in food, but also various nutrients are accompanied by destruction. Another is the use of food additives and food preservatives, which are agents added to foods for the above effects. Conventional food preservatives are chemical synthesis agents such as benzoic acid, sorbic acid, dihydroacetic acid and propionic acid. Although used, it is a serious problem in the health and hygiene stability as a chemical compound.
상기와 같은 인체에 유해한 독성과 자극성을 지닌 화학합성 제제의 사용을 줄이고 인체에는 무해하나 식품의 영양성분 파괴 없이 보존성 유지와 품질 유지가 가능한 무독성의 안전한 천연물질 개발의 필요성이 고조되고 있는 실정이다.There is a growing need for the development of non-toxic, safe natural materials that can reduce the use of chemical synthesis agents that are toxic and irritating to the human body and are harmless to the human body, but maintain preservation and quality without destroying nutrients in food.
화분은 꽃의 수술에서 나오는 일종의 생식세포로서 벌이 꽃에서 화밀을 수집하면서 수술의 꽃가루주머니에 있는 미세한 입자들을 꿀 및 타액과 함께 섞어 큰 입자로 만든 것으로, 벌의 생식 및 생육에 절대적인 단백질 공급원이 되고있을 뿐만 아니라 비타민, 미네랄, 지방산, 호르몬, 효소 및 조효소 등을 완벽하고 균형있게 함유하고 있어 완전식품으로 많이 이용되고 있다. 뿐만 아니라 세포의 기능을 원활하게 하여 산화된 세포에 활력을 주므로 만성 전립선염, 전립선 비대증, 당뇨, 변비, 비만, 위궤양, 갱년기 장애 등 질병의 예방 및 치료에 탁월한 효과를 나타낸다. Pollen is a type of germ cell from the stamen of flowers. The bee collects nectar from the flower and makes the fine particles in the stamen pollen bag mixed with honey and saliva into large particles. It is an absolute source of protein for bee reproduction and growth. In addition, it contains vitamins, minerals, fatty acids, hormones, enzymes and coenzymes, and is used as a perfect food. In addition, it functions to oxidize the cells by smoothing the function of the cells, showing excellent effects in the prevention and treatment of diseases such as chronic prostatitis, prostatic hyperplasia, diabetes, constipation, obesity, gastric ulcer, menopausal disorders.
그러나 화분은 세포 구조상 외피와 내피의 2중층으로 되어 있으며, 외피는 100℃ 이상의 고온에서, 단백질 분해효소에 의해서, 아세톤과 같은 유기용매 및 황산, 질산과 같은 강한 산에 의해서도 파괴되지 않아서 현재로서는 단지 기계적인 충격을 가해서 파쇄하는 방법만이 알려져 있으며 그 효과는 매우 미약하여 화분의 외피를 파쇄하지 않고 섭취할 경우 화분에 존재하는 우수한 영양성분의 이용률은 매우 낮다. 뿐만 아니라, 화분의 외피는 인체의 알레르기 발생원으로 작용하므로 외피가 파쇄되지 않은 화분을 섭취할 때는 알레르기가 유발될 수 있어서 화분을 이용한 식품제조 및 가공에 많은 제한점이 되고 있다. However, pollen is composed of two layers of skin and endothelium due to its cell structure, and the skin is not destroyed by proteolytic enzymes at high temperature of 100 ℃ or above, even by organic solvent such as acetone and strong acid such as sulfuric acid and nitric acid. Only the method of crushing by applying mechanical shock is known and its effect is so weak that when consumed without crushing the shell of pollen, the utilization rate of the excellent nutrients present in the pollen is very low. In addition, since the outer shell of the pollen acts as an allergen source of the human body, allergens may be induced when the pollen is ingested without the outer shell being broken, and thus, there are many limitations in manufacturing and processing food using the pollen.
그래서 화분의 유효성분을 효과적으로 이용하기 위한 화분의 처리와 내용물의 추출방안으로는 화분에 기계적인 충격을 가해서 화분입자를 파괴시켜 추출하는 방안 (대한민국 특허 공개번호 2001-0035445, 등록번호 1994-0010742, 등록번호 10-0309699, 등록번호 1991-0001407), 화분을 물에 분산시켜 고온에서 가열·추출하거나, 계면활성제 혹은 액체질소를 사용하는 방안 (등록번호 1987-0001617, 등록번호 1989-0003995), 자체내의 미생물에 의한 단순 발효, 숙성을 거쳐 초음파 처리에 의한 내용물의 추출법 (공개번호 1984-0000240), 락토바실러스속 미생물의 접종, 발효시켜 저분자의 화분 엑기스를 만든 후 맥아덱스트린 첨가하고 진공 건조시킨 저분자의 화분제조법 ( 공개번호 1991-0015248) 등이 고안되어 있다.Therefore, as a method of processing and extracting the contents of the pollen to effectively use the active ingredient of the pollen, a method of extracting by destroying pollen particles by applying a mechanical impact to the pot (Korean Patent Publication No. 2001-0035445, Registration No. 1994-0010742, Registration No. 10-0309699, Registration No. 1991-0001407), Dispersing pots in water to heat and extract at high temperatures, or use surfactants or liquid nitrogen (Registration No. 1987-0001617, Registration No. 1989-0003995), self Extraction of contents by sonication through simple fermentation and maturation by microorganisms (Publication No. 1984-0000240), inoculation and fermentation of Lactobacillus microorganisms to make pollen extracts of low molecular weight, followed by adding maladextrin and vacuum drying Pollen manufacturing methods (published number 1991-0015248) have been devised.
그러나 상기의 고안들은 복잡한 공정으로 비경제적이며, 제조과정 중 화분의 살균으로 유효성분들의 파괴와 함께 화분 추출물의 변색을 초래할 뿐만 아니라, 계면활성제 및 유기용매의 사용으로 인한 식품원료로의 사용이 불가능하게 한다. 또한 이와같은 고안들은 화분의 추출물을 화장품 원료, 의료용 치료제 및 건강보조식품의 원료로 사용하기 위해 고안된 기술로서 화분 발효물의 항균력을 이용한 식품의 원료 및 첨가물로 사용한 고안은 없다. However, the above designs are uneconomical in complex processes, and the disinfection of pollen extracts with the destruction of active ingredients by disinfection of pollen during the manufacturing process, as well as the use of food ingredients due to the use of surfactants and organic solvents is impossible. Let's do it. In addition, these designs are a technique designed to use the extract of pollen as a raw material of cosmetic raw materials, medical treatments and health supplements, there is no design used as a raw material and additives of food using the antibacterial activity of pollen fermentation.
현재 식품 보존에 이용 가능한 천연항균제로서는 자몽종자 추출물 (GFSE), 프로폴리스 (Propolis), 키토산 (Chitosan), 용균효소 (Lysozyme) 등을 들 수 있지만, 가격이 비싸고 항균효과를 기대하려면 상당히 많은 양을 첨가하여야만 하므로서 식품의 풍미나 물성에 심한 악영향을 초래하는 단점을 갖고 있다.Natural antimicrobials currently available for food preservation include grapefruit seed extract (GFSE), propolis, chitosan and lysozyme, but they are expensive and require significant amounts of antimicrobial effects. Since it must be added, it has the disadvantage of causing severe adverse effects on the flavor and physical properties of the food.
따라서 본 발명은 다양하고 많은 영양성분을 함유하고 있는 화분을 유산균 배양 배지로 하여 발효시켜서 인체에 유용한 생리활성물질의 이용성을 최대화시킬 뿐만 아니라 강력한 항균력으로 식품의 원료 및 첨가물로 사용할 수 있게 하였다.Therefore, the present invention is not only to maximize the usability of the bioactive material useful for the human body by fermenting the potted plant containing a variety of nutrients as a lactic acid bacteria culture medium, it can be used as a raw material and additives of food with strong antibacterial activity.
따라서 본 발명은 상기와 같은 문제점을 해결하기 위하여 구성된 것으로, 화분을 락토바실러스 람노수스로 발효시키거나, 이 발효물을 사카로마이세스 세르비시아로 재 발효시켜 얻은 화분 발효추출물을 식품보존제의 원료로 사용함으로서, 파쇄하지 않은 화분섭취에 의한 알레르기 요인을 미리 제거하고, 화분이 갖고있는 다양한 영양성분과 함께 항균활성으로 인하여 식품 고유의 맛과 색상 등에 영향을 미치지 않고 식품의 보존과 품질유지가 가능한 화분 발효추출물을 함유하는 식품의 원료 및 첨가물을 제공하는 것을 목적으로 한다.Therefore, the present invention is configured to solve the above problems, fermented pollen with Lactobacillus rhamnosus, or fermented extract obtained by re-fermenting the fermented product with Saccharomyces servicia as raw material of food preservative By using it, it is possible to eliminate allergic factors due to the ingestion of pollen that is not crushed in advance, and to preserve food and maintain the quality without affecting the food's unique taste and color due to the antibacterial activity with various nutrients that pollen has An object of the present invention is to provide raw materials and additives for foods containing pollen fermentation extracts.
상기와 같은 목적을 달성하기 위하여 본 발명은, 유아의 분변, 원유 및 김치류에서 얻은 48주의 미생물로부터 부패성 미생물에 대하여 항균활성이 가장 우수한 미생물을 선발하여 이의 미생물학적 성상을 분석한 결과, 락토바실러스 (Lactobacillus) 속에 속하는 람노수스 (rhamnosus) 임을 확인하고 이를 락토바실러스 람노수스 에스엔티피01 (Lactobacillus rhamnosus SNTP01)로 명명하고 2003년 7월 21일자로 한국농용미생물보존센타(KACC)에 기탁하여 KACC 91057로 수탁받았다.In order to achieve the above object, the present invention, 48 microorganisms obtained from feces, crude oil and kimchi of infants were selected from the microorganisms having the best antimicrobial activity against the decaying microorganisms and analyzed the microbiological characteristics thereof, Lactobacillus ( Lactobacillus rhamnosus belongs to the genus Lactobacillus (Lactobacillus rhamnosus SNTP01) and named it as July 21, 2003 deposited in the Korea Agricultural Microbial Conservation Center (KACC) and deposited as KACC 91057 received.
[제조예 1][Production Example 1]
건조된 화분을 20mesh 이하로 미세하게 분쇄하여 80℃에서 15분간 가열 살균한 후, 화분의 농도가 5%되게 멸균수에 용해시켜 pH를 6.8로 조정하고 미리 준비한 락토바실러스 람노수스 에스엔티피01을 0.5%되도록 접종하여 37℃에서 120시간 동안 발효시켰다. 이 발효물을 4℃, 8,000xg에서 20분간 원심분리하여 비교적 큰 고형물을 제거한 상층액을 다시 0.45㎛의 막여과기로 여과하여 화분 발효추출물을 제조하였다.The dried pollen is finely pulverized to 20 mesh or less and heated and sterilized at 80 ° C. for 15 minutes, and then dissolved in sterile water so that the concentration of the pot is 5%, the pH is adjusted to 6.8, and the previously prepared Lactobacillus rhamnosus SENTIP01 is 0.5. It was inoculated to the% fermentation for 120 hours at 37 ℃. The fermented product was centrifuged at 4 ° C. and 8,000 × g for 20 minutes, and the supernatant from which a relatively large solid was removed was filtered again with a membrane filter of 0.45 μm to prepare a pollen fermentation extract.
[제조예 2][Production Example 2]
제조예 1에서 화분의 농도를 15% 사용한 것 외에는 제조예 1과 동일하게 제조하였다. The preparation was carried out in the same manner as in Preparation Example 1, except that 15% of the concentration of the pollen was used in Preparation Example 1.
[제조예 3][Manufacture example 3]
제조예 1에서 화분의 농도를 30% 사용한 것 외에는 제조예 1과 동일하게 제조하였다. Production Example 1 was prepared in the same manner as in Production Example 1 except that the concentration of the pollen was used 30%.
[제조예 4][Production Example 4]
제조예 1에서 화분의 농도를 40% 사용한 것 외에는 제조예 1과 동일하게 제조하였다. The preparation was carried out in the same manner as in Preparation Example 1, except that 40% of the concentration of the pollen was used in Preparation Example 1.
[제조예 5]Production Example 5
건조된 화분을 20mesh 이하로 미세하게 분쇄하여 80℃에서 15분간 가열 살균한 후, 화분의 농도가 5%되게 멸균수에 용해시켜 pH를 6.8로 조정하고 미리 준비한 락토바실러스 람노수스 에스엔티피01을 1.5%되도록 접종하여 37℃에서 120시간 동안 발효시켰다. 이 발효물들에는 화분이 갖고있는 당분을 상당히 함유하고 있어서 발효추출물의 산화적 갈변의 원인이 되므로 이를 방지하고, 점도 저하 및 색상을 옅게하기 위하여 각각의 발효물에 다시 사카로마이세스 세르비시아 (Saccharomyces cerevicia) KACC 30008 을 1%되도록 접종하여 30℃에서 72시간 동안 배양시킨 다음, 4℃, 8,000xg에서 20분간 원심분리하여 비교적 큰 고형물을 제거한 상층액을 다시 0.45㎛의 막여과기로 여과하여 화분 발효추출물을 제조하였다.The dried pollen is finely pulverized to 20 mesh or less and heated and sterilized at 80 ° C. for 15 minutes, and then dissolved in sterile water so that the concentration of the pot is 5%, the pH is adjusted to 6.8, and the previously prepared Lactobacillus rhamnosus SENTIP01 01 It was inoculated to the% fermentation for 120 hours at 37 ℃. These fermented products contain a significant amount of sugar in pollen, which causes oxidative browning of fermented extracts. To prevent this, the saccharomyces cerevisiae ( Saccharomyces cerevicia) KACC 30008 was inoculated to 1% and incubated for 72 hours at 30 ° C, centrifuged at 4 ° C and 8,000xg for 20 minutes, and the supernatant from which relatively large solids were removed was filtered again with a membrane filter of 0.45㎛. Fermented extract was prepared.
[제조예 6][Manufacture example 6]
제조예 5에서 화분의 농도를 15% 사용한 것 외에는 제조예 5와 동일하게 제조하였다. The preparation was carried out in the same manner as in Preparation Example 5, except that 15% of the concentration of the pollen was used in Preparation Example 5.
[제조예 7][Manufacture example 7]
제조예 5에서 화분의 농도를 30% 사용한 것 외에는 제조예 5와 동일하게 제조하였다. The preparation was carried out in the same manner as in Preparation Example 5, except that 30% of the concentration of the pollen was used in Preparation Example 5.
[제조예 8][Manufacture example 8]
제조예 1에서 화분의 농도를 40% 사용한 것 외에는 제조예 5와 동일하게 제조하였다. The preparation was carried out in the same manner as in Preparation Example 5, except that 40% of the concentration of the pollen was used in Preparation Example 1.
[실시예1] 화분 발효물의 물리적 성질Example 1 Physical Properties of Fermented Potted Plant
상기 제조예에서 얻어지는 화분 발효추출물의 물리적 성질을 다음과 같이 비교·조사하였다.The physical properties of the pollen fermentation extract obtained in the preparation example were compared and investigated as follows.
색상은 색차계 (Digital color measuring/difference calculating meter, Nippon Denshoku Kogyo Co. Ltd., ND-1001 DP, Japan)를 사용하여 Hunters L(Lightness). a(red/green). b(yellow/blue) 값을 측정하였으며 (표 1), 점도는 점도계 (RVF, Viscometer, Brook field Engineering Lab. Inc., USA)로 20±1℃에서, 당도는 당도계 (Hand Refractometer, Atago,Japan)로, 맛의 관능검사는 18명의 페널요원에 의해 평점법(7점)으로 조사되었다 (표 2).The color is Hunters L (Lightness) using a digital color measuring / difference calculating meter, Nippon Denshoku Kogyo Co. Ltd., ND-1001 DP, Japan. a (red / green). b (yellow / blue) values were measured (Table 1), and the viscosity was measured at 20 ± 1 ° C. using a viscometer (RVF, Viscometer, Brook field Engineering Lab. Inc., USA), and the sugar content was measured by Hand Refractometer, Atago, Japan. ), Taste sensory test was performed by 18 panelists by grading method (7 points) (Table 2).
[표 1] 화분 발효물의 색상[Table 1] Color of pollen fermented product
[표 2] 화분 발효물의 물리적 특성[Table 2] Physical properties of pollen fermentation
표 1과 2에서와 같이 화분 발효추출물의 물리적 성질에는 화분 발효물 조제시 사용된 화분의 량에는 크게 영향을 미치지 않고 거의 비슷하였으며, 화분의 사용량이 증가함에 따라 색상이 노랑색에서 붉은색으로 짙어지며, 당도와 점도가 증가하였다. 그러나 락토바실러스 람노수스 에스엔티피01를 접종하여 5일간 발효한 다음, 다시 사카로마이세스 세르비시아를 접종하여 3일간 발효하면 사카로마이세스 세르비시아를 접종하지 않은 발효 추출물에 비해 점도 및 당도가 상당히 떨어졌으며 또한 색상도 L값이 증가하여 연하고 밝은 노랑색을 띄었다.As shown in Table 1 and 2, the physical properties of pollen fermented extracts were almost the same without significantly affecting the amount of pollen used to prepare the fermented fermented products, and the color increased from yellow to red as the use of pollen increased. , Sugar and viscosity increased. However, when inoculated with Lactobacillus rhamnosus SENTIP01 and fermented for 5 days, and then inoculated with Saccharomyces servicia for 3 days, the viscosity and sugar content were higher than those of fermented extracts not inoculated with Saccharomyces servicia. Was significantly decreased, and the color was also lighter, with a higher L value.
[실시예 2] 화분 발효물의 색상변화Example 2 Color Change of Fermented Potted Plant
상기 제조예 2와 6에서 얻은 15%의 화분 발효추출물을 45일간 하기의 각 조건에서 보관한 다음, 각 추출물들의 색상을 실시예 1과 같이 측정하여 색상의 변화를 조사한 결과를 하기 표 3에 나타내었다. 즉, 화분 발효추출물의 보관 조건은 암소에서 밀봉하여 냉장(4℃)보관 (암소/밀봉/냉장), 암소에서 밀봉하여 상온(27℃)보관 (암소/밀봉/상온), 암소에서 개봉하여 냉장보관 (암소/개봉/냉장), 암소에서 개봉하여 상온보관 (암소/개봉/상온), 명소에서 밀봉하여 상온보관 (명소/밀봉/상온), 명소에서 개봉하여 상온보관 (명소/개봉/상온)이었다.The 15% pollen fermentation extracts obtained in Preparation Examples 2 and 6 were stored for 45 days at the following conditions, and then the color of each extract was measured as in Example 1, and the results of the color change are shown in Table 3 below. It was. In other words, the storage conditions of pollen fermented extracts are sealed in a cow (4 ℃) for storage (cow / sealing / refrigeration), sealed in a cow for storage at room temperature (27 ℃) (cow / sealing / room temperature), and opened in cow and refrigerated. Storage (cow / open / refrigerated), open at cow and store at room temperature (cow / open / room temperature), seal at spot and store at room temperature (spot / sealed / room temperature), open at sight (room / open / room temperature) It was.
표 3TABLE 3
상기 표 3에서 보듯이, 본 발명의 화분 발효추출물은 보관기간에 따라 색상이 노랑색에서 갈색으로 변하였다. 화분 발효추출물의 색상은 햇빛이나 온도에는 큰 영향을 받지 않았으나, 공기와의 접촉에 의해 색상의 옅은 노랑색에서 짙은 갈색으로 큰 변화를 관찰하였다. 그러나 사카로마이세스 세르비시아로 다시 발효한 추출물은 보관조건에 큰 영향을 받지 않고 제조직후의 초기색상에 큰 변화없이 매우 안정했다. 따라서 화분 발효추출물의 색상 안정화에는 사카로마이세스 세르비시아의 접종, 발효과정이 매우 유효하다고 판단된다.As shown in Table 3, the pollen fermentation extract of the present invention was changed from yellow to brown color depending on the storage period. The color of pollen fermented extract was not significantly affected by sunlight or temperature, but a big change was observed from light yellow to dark brown by contact with air. However, the extract fermented again with Saccharomyces servicia was not affected by storage conditions and was very stable without significant change in the initial color immediately after manufacture. Therefore, the inoculation and fermentation process of Saccharomyces servicia is very effective for stabilizing the color of fermented fermentation extracts.
[실시예 3]Example 3
상기 제조예에서 얻은 화분 발효추출물의 부패성 미생물에 대한 항균활성을 다음과 같이 비교·조사하였으며, 사용된 공시균주, 배양 및 배지조건은 표 4와 같다.The antibacterial activity of the pollen microorganisms of the pollen fermentation extract obtained in the preparation example was compared and investigated as follows, and the used strains, culture and culture conditions are shown in Table 4.
전배양은 상기의 균주를 적정의 배양조건에서 24시간 배양하여 각각을 104 cell/ml되게 미생물의 수를 조정하였으며, 항균력의 측정은 미국의 NCCLS법 (National Committee for Clinical Laboratory Standards. 1990)에 따라 액체배지 희석법 (broth dilution method)로 측정하였다. 즉, 전배양한 균액를 공시배지에서 일정농도의 화분 발효추출물과 함께 공시온도에서 24시간 동안 배양하고 이를 600nm에서 흡광도의 측정으로 조제한 화분발효 추출물의 항균력을 조사하였으며, 공시균주의 생육이 90% 저해되는 농도인 최소생육저해농도-90 (Minimum inhibitory concentration, MIC90) 및 공시균주의 생육이 50% 저해되는 농도인 최소생육저해농도-50 (Minimum inhibitory concentration, MIC50)으로 각각 표기하였다. The preculture was incubated for 24 hours under the appropriate culture conditions, and the number of microorganisms was adjusted to 104 cells / ml, and the antimicrobial activity was measured according to the National Committee for Clinical Laboratory Standards (1990). It was measured by the broth dilution method. In other words, the cultured fungi were incubated for 24 hours at the indicated temperature with a certain concentration of pollen fermentation extracts in a test medium, and the antimicrobial activity of the fermented extract prepared by measuring the absorbance at 600 nm was investigated, and the growth of the test strain was inhibited by 90%. The minimum growth inhibitory concentration (MIC90) and the minimum inhibitory concentration (MIC50), which are 50% inhibition of the growth of test strains, are indicated.
[실시예 4] 5% 화분 발효추출물의 항균력Example 4 Antibacterial Activity of 5% Pollen Fermented Extract
상기 제조예 1에서 제조한 5% 화분 발효추출물의 항균력을 측정한 결과를 하기 표 5에 나타내었다.The results of measuring the antimicrobial activity of the 5% pollen fermentation extract prepared in Preparation Example 1 are shown in Table 5 below.
표 4의 균주의 배양 및 배지조건에서 5% 화분 발효추출물을 각각 무첨가, 4%, 3.8%, 3.6%, 3.4%, 3.2% 및 3.0%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 5% 화분 발효추출물의 항균력은 9종의 시험균에 대하여 MIC90은 3.8%, MIC50은 3.4∼3.6% 정도였다.Under the culture and culture conditions of the strains of Table 4, 5% pollen fermented extracts were added to the medium so that no addition, 4%, 3.8%, 3.6%, 3.4%, 3.2% and 3.0% were incubated for 24 hours, and then at 600 nm. The antimicrobial activity was investigated by measuring absorbance. The antimicrobial activity of 5% pollen fermented extracts was about 3.8% for MIC90 and 3.4 ~ 3.6% for MIC50 against 9 species.
표 5Table 5
[실시예 5] 15% 화분 발효추출물의 항균력Example 5 Antibacterial Activity of 15% Pollen Fermented Extract
상기 제조예 2에서 제조한 15% 화분 발효추출물의 항균력을 측정한 결과를 하기 표 6에 나타내었다.The antimicrobial activity of the 15% pollen fermented extract prepared in Preparation Example 2 is shown in Table 6 below.
표 4의 균주의 배양 및 배지조건에서 15% 화분 발효추출물을 각각 무첨가, 3.4%, 3.2%, 3.0%, 2.8%, 2.6% 및 2.4%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 15% 화분 발효추출물의 항균력은 9종의 시험균에 대하여 MIC90은 2.8∼3.0%, MIC50은 2.6%로 5% 화분 발효추출물의 항균력보다는 훨씬 더 강하였다..Under the culture and culture conditions of the strains of Table 4, 15% pollen fermented extracts were added to the medium so that no addition, 3.4%, 3.2%, 3.0%, 2.8%, 2.6% and 2.4% were incubated for 24 hours, and then at 600 nm. The antimicrobial activity was investigated by measuring absorbance. The antimicrobial activity of 15% pollen fermented extract was 2.8-3.0% for MIC90 and 2.6% for MIC50, which was much stronger than that of 5% pollen fermented extract.
표 6Table 6
[실시예 6] 30% 화분 발효추출물의 항균력Example 6 Antibacterial Activity of 30% Pollen Fermented Extract
상기 제조예 3에서 제조한 30% 화분 발효추출물의 항균력을 측정한 결과를 하기 표 7에 나타내었다.The antimicrobial activity of the 30% pollen fermented extract prepared in Preparation Example 3 is shown in Table 7 below.
표 4의 균주의 배양 및 배지조건에서 30% 화분 발효추출물을 각각 무첨가, 3.0%, 2.8%, 2.6%, 2.4%, 2.2% 및 2.0%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 30% 화분 발효추출물의 항균력은 9종의 시험균에 대하여 MIC90은 2.6∼2.8%, MIC50은 2.2%였다.Under the culture and culture conditions of the strains of Table 4, 30% pollen fermented extracts were added to the medium so that no addition, 3.0%, 2.8%, 2.6%, 2.4%, 2.2% and 2.0% were incubated for 24 hours, and then at 600 nm. The antimicrobial activity was investigated by measuring absorbance. The antimicrobial activity of the 30% pollen fermented extracts was 2.6-2.8% for MIC90 and 2.2% for MIC50 against 9 species.
표 7TABLE 7
[실시예 7] 40% 화분 발효추출물의 항균력Example 7 Antibacterial Activity of 40% Pollen Fermented Extract
상기 제조예 4에서 제조한 40% 화분 발효추출물의 항균력을 측정한 결과를 하기 표 8에 나타내었다.The antimicrobial activity of the 40% pollen fermented extract prepared in Preparation Example 4 is shown in Table 8 below.
표 4의 균주의 배양 및 배지조건에서 40% 화분 발효추출물을 각각 무첨가, 3.0%, 2.8%, 2.6%, 2.4%, 2.2%, 2.0% 및 1.8%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 40% 화분 발효추출물의 항균력은 9종의 시험균에 대하여 MIC90은 2.4∼2.8%, MIC50은 2.0%로 30% 화분 발효추출물의 항균력과 비슷하였다.Under the culture and culture conditions of the strains of Table 4, 40% pollen fermented extracts were added to the medium so that no addition, 3.0%, 2.8%, 2.6%, 2.4%, 2.2%, 2.0% and 1.8%, respectively, was incubated for 24 hours. , The antimicrobial activity was examined by measuring the absorbance at 600nm. The antimicrobial activity of 40% pollen fermented extracts was 2.4-2.8% for MIC90 and 2.0% for MIC50 for 9 species, similar to that of 30% pollen fermented extracts.
표 8Table 8
[실시예 8] 5% 화분발효물에 사카로마이세스 세르비시아의 접종 효과Example 8 Inoculation Effect of Saccharomyces servicia on 5% Pollen Fermentation
상기 제조예 5에서 제조한 5% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 항균력을 측정한 결과는 하기 표 9에 나타내었다.The antimicrobial activity of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia to the 5% pollen fermented product prepared in Preparation Example 5 is shown in Table 9 below.
표 4의 균주의 배양 및 배지조건에서 무첨가, 4.0%, 3.8%, 3.6%, 3.4%, 3.2% 및 3.0%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사한 바, 9종의 시험균에 대하여 MIC90이 모두 3.8%, MIC50은 3.4∼3.6%로 사카로마이세스 세르비시아의 처리는 항균력에는 영향을 주지 않았다.In the culture and culture conditions of the strains of Table 4, added to the medium to be added, 4.0%, 3.8%, 3.6%, 3.4%, 3.2% and 3.0% incubated for 24 hours, and then examined the antimicrobial activity by measuring the absorbance at 600nm The MIC90 was 3.8% and the MIC50 was 3.4-3.6% for the nine test bacteria. Saccharomyces servicia treatment did not affect the antimicrobial activity.
표 9Table 9
[실시예 9] 15% 화분발효물에 사카로마이세스 세르비시아의 접종 효과Example 9 Inoculation Effect of Saccharomyces servicia on 15% Pollen Fermentation
상기 제조예 5에서 제조한 15% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 항균력을 측정한 결과는 하기 표 10에 나타내었다.The antimicrobial activity of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia on the 15% pollen fermented product prepared in Preparation Example 5 is shown in Table 10 below.
표 4의 균주의 배양 및 배지조건에서 무첨가, 3.4%, 3.2%, 3.0%, 2.8%, 2.6% 및 2.4%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 15% 화분 발효추출의 항균력은 9종의 시험균에 대하여 MIC90은 2.8%, MIC50은 2.6%로 항균력은 사카로마이세스 세르비시아에 의해 약간 강화되었다.In the culture and culture conditions of the strains of Table 4, added to the medium to no addition, 3.4%, 3.2%, 3.0%, 2.8%, 2.6% and 2.4% incubated for 24 hours, and then investigated the antibacterial activity by measuring the absorbance at 600nm did. The antimicrobial activity of 15% pollen fermentation extract was 2.8% for MIC90 and 2.6% for MIC50 against 9 test bacteria, and the antimicrobial activity was slightly enhanced by Saccharomyces serbia.
표 10Table 10
[실시예 10] 30% 화분발효물에 사카로마이세스 세르비시아의 접종 효과Example 10 Inoculation Effect of Saccharomyces servicia on 30% Pollen Fermentation
상기 제조예 5에서 제조한 30% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 항균력을 측정한 결과는 하기 표 11에 나타내었다.The antimicrobial activity of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia on the 30% pollen fermented product prepared in Preparation Example 5 is shown in Table 11 below.
표 4의 균주의 배양 및 배지조건에서 무첨가, 3.0%, 2.8%, 2.6%, 2.4%, 2.2% 및 2.0%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 30% 화분 발효추출의 항균력은 9종의 시험균에 대하여 MIC90은 2.6%, MIC50은 2.2%로 항균력은 사카로마이세스 세르비시아의 처리로 약간 향상되었다.In the culture and culture conditions of the strains of Table 4, added to the medium at no addition, 3.0%, 2.8%, 2.6%, 2.4%, 2.2% and 2.0% incubated for 24 hours, and then investigated the antibacterial activity by measuring the absorbance at 600nm did. The antibacterial activity of the 30% pollen fermentation extract was 2.6% for MIC90 and 2.2% for MIC50 against 9 test bacteria, and the antimicrobial activity was slightly improved by treatment with Saccharomyces servisia.
표 11Table 11
[실시예 11] 40% 화분발효물에 사카로마이세스 세르비시아의 접종 효과Example 11 Inoculation Effect of Saccharomyces servicia on 40% Pollen Fermentation
상기 제조예 5에서 제조한 40% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 항균력을 측정한 결과는 하기 표 12에 나타내었다.The results of measuring the antimicrobial activity of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia to the 40% pollen fermented product prepared in Preparation Example 5 are shown in Table 12 below.
표 4의 균주의 배양 및 배지조건에서 무첨가, 2.8%, 2.6%, 2.4%, 2.2%, 2.0% 및 1.8%되게 배지에 첨가하여 24시간동안 배양한 다음, 600nm에서 흡광도의 측정으로 항균력을 조사했다. 40% 화분 발효추출의 항균력은 9종의 시험균에 대하여 MIC90은 2.4∼2.6%, MIC50은 2.0%로 사카로마이세스 세르비시아의 처리로 항균력이 약간 향상하였다.In the culture and culture conditions of the strains of Table 4, added to the medium at no addition, 2.8%, 2.6%, 2.4%, 2.2%, 2.0% and 1.8%, and incubated for 24 hours, and then investigated the antimicrobial activity by measuring the absorbance at 600nm did. The antimicrobial activity of the 40% pollen fermentation extract was 2.4-2.6% for MIC90 and 2.0% for MIC50 against 9 test bacteria, and the antimicrobial activity was slightly improved by treatment with Saccharomyces servicia.
표 12Table 12
이상의 결과를 종합하면, 화분 발효추출물의 항균력은 에스케르시아 콜라이를 비롯한 9종의 공시균주에 대하여 모두 항균효과를 나타내 보였으며, MIC50이 2.0%∼3.6% 정도의 항균력을 나타내어 식품보존제로의 사용가능성을 확인할 수 있었다.Taken together, the antibacterial activity of the fermented extract of pollen showed antibacterial effect against 9 species including Escherichia coli, and MIC50 showed 2.0 ~ 3.6% of antimicrobial activity. The possibility was confirmed.
[실시예 12] 항균력의 지효성Example 12 Sustainability of Antimicrobial Activity
상기 제조예 5에서 얻은 화분 발효추출물의 부패성 미생물에 대한 항균력의 지효성을 다음과 같이 조사하였으며, 사용된 공시균주는 표 4과 같다.The effectiveness of the antimicrobial activity against the decaying microorganisms of the pollen fermentation extract obtained in Preparation Example 5 was investigated as follows.
항균력의 지효성은 우선 121 에서 15분간 멸균시키고 50 로 맞춘 뉴트리언트 한천배지에 에스케르시아 콜라이 (Escherichia coli) KCTC 2441의 전배양액 0.2ml를 넣고 잘 혼합하여 직경 8.5cm의 페트리 디쉬에 부어서 고형화 시킨 다음, 그 위에 제조예 5의 화분 발효추출물을 50㎍을 직경 1.0cm의 종이 디스크(Whatman No.2) 위에 주입하고 37℃에서 3일간 배양한 다음 15일간 보관하면서 생장저해환(inhibition zone)의 크기 및 변화를 관찰하여 화분 발효추출물의 항균력의 지효성을 측정하였으며, 그 결과를 하기 그림 1에 나타내었다.Sustainability of antimicrobial activity is first sterilized at 121 to 15 minutes, 0.2 ml of the preculture of Escherichia coli KCTC 2441 in a nutritic agar medium adjusted to 50, mixed well, poured into a petri dish of 8.5 cm in diameter, and solidified. , 50 μl of the pollen fermentation extract of Preparation Example 5 was injected onto a 1.0 cm diameter paper disk (Whatman No. 2), incubated at 37 ° C. for 3 days, and then stored for 15 days to suppress the growth zone (inhibition zone) size. And the change was observed to determine the sustainability of the antibacterial activity of fermented extract of pollen, the results are shown in Figure 1 below.
그림 1. 화분 발효추출물의 항균력의 지효성Figure 1.Long-effect of antibacterial activity of pollen fermented extracts
상기의 그림에서 보는바와 같이 화분 발효추출물은 15일동안 보관을 하여도 생장 저해환의 크기에 전혀 변화없이 처음의 항균력을 그대로 유지되고 있어서 매우 안정하고 효력의 지속성을 확인하였다.As shown in the above figure, the fermented extract of pollen was kept very stable and sustained its effect as it retained its first antimicrobial activity without any change in the size of growth inhibiting ring even if stored for 15 days.
[실시예 13] 열에 대한 항균력의 안정성Example 13 Stability of Antimicrobial Activity Against Heat
상기 제조예 5에서 제조한 15% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 열에 대한 항균력의 안정성은 80℃와 100℃에서 1시간, 121℃에서 20분동안 각각 열처리하고 이를 2.6% 및 3.0%되게 뉴트리언트 배지에 첨가하여 에스케르시아 콜라이 (Escherichia coli) KCTC 2441에 대한 액체배지 희석법으로 항균력을 측정하여 열처리하기 전의 항균력과 서로 비교하여 그 결과를 하기의 표 13에 나타내었다.The stability of the antimicrobial activity against the heat of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia to the 15% pollen fermented product prepared in Preparation Example 5 was for 1 hour at 80 ° C. and 100 ° C., and 20 minutes at 121 ° C. The antimicrobial activity was measured by the liquid medium dilution method for Escherichia coli KCTC 2441 by adding heat to the nutrient medium at 2.6% and 3.0%, respectively. Table 13 shows.
표 13Table 13
[실시예 14] pH에 대한 항균력의 안정성Example 14 Stability of Antimicrobial Activity against pH
상기 제조예 5에서 제조한 15% 화분 발효물에 다시 사카로마이세스 세르비시아를 접종·발효하여 얻은 발효추출물의 pH에 대한 항균력의 안정성은 발효추출물의 pH를 1N KOH로 미리 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 및 10.0으로 조정한 다음, 뉴트리언트 배지에 2.6%, 3.0%되게 첨가하여 에스케르시아 콜라이 (Escherichia coli) KCTC 2441의 생육도를 흡광도 값으로 표시하여 항균력을 측정하고, pH를 조정하기 전의 항균력과 서로 비교함으로써 조사하였다. 그 결과를 하기의 표 14에 나타내었다.The stability of the antimicrobial activity against the pH of the fermented extract obtained by inoculating and fermenting Saccharomyces servicia to the 15% pollen fermented product prepared in Preparation Example 5 was previously determined to be 4.0, 4.5, Growth of Escherichia coli KCTC 2441 by adjusting to 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 and 10.0, and then adding 2.6% and 3.0% to the nutrient medium. The degree of antimicrobial activity was measured by displaying absorbance values, and compared with the antimicrobial activity before adjusting pH. The results are shown in Table 14 below.
표 14Table 14
상기 표 13 및 14에서 보듯이, 본 발명의 화분 발효추출물의 항균 원인물질은 모든 온도에서와 모든 pH범위에서도 항균력의 소실이 거의 없이 안정하였다.As shown in Tables 13 and 14, the antibacterial agent of the pollen fermentation extract of the present invention was stable at almost all temperatures and all pH ranges with little loss of antibacterial activity.
본 발명의 항균활성을 갖는 화분 발효물은 화분을 파쇄하지 않고 화분을 저온에서 순간 살균하여 유산균의 접종과 배양에 의한 발효과정으로 제조되므로 화분의 다양하고도 많은 영양성분 뿐만 아니라 인체에 유용한 생리활성물질의 이용성을 최대화시킴과 동시에 유산균이 생산·분비하는 젖산 및 박테리오신으로 인한 강력한 항균력으로 식품의 원료 및 첨가물로 이용될 수 있다.Pollen fermentation product having an antimicrobial activity of the present invention is prepared by fermentation process by inoculation and culture of lactic acid bacteria by instant sterilization of pollen at low temperature without crushing the pollen, so that the physiological activity useful for human body as well as various nutrients of pollen It can be used as raw materials and additives in foods by maximizing the usability of the substance and at the same time the strong antibacterial activity of lactic acid and bacteriocin produced and secreted by lactic acid bacteria.
도 1은 본 발명의 화분 발효추출물의 에스케르시아 콜라이에 대한 항균력의 지속성을 비교한 사진이다.1 is a photograph comparing the persistence of the antimicrobial activity against Escherichia coli of the pollen fermentation extract of the present invention.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101926428A (en) * | 2010-08-02 | 2010-12-29 | 山东省农业科学院农产品研究所 | Probiotic gingko pollen freeze-dried powder and preparation method thereof |
| CN104958771A (en) * | 2015-07-01 | 2015-10-07 | 杨俊� | Large-scale produced fermented pollen sterilization method |
| KR102282350B1 (en) * | 2020-11-17 | 2021-07-27 | 주식회사삼조에스피피 | Vongole pasta sauce and manufacturing method for thereof containing spicy clam stock base |
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2003
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101926428A (en) * | 2010-08-02 | 2010-12-29 | 山东省农业科学院农产品研究所 | Probiotic gingko pollen freeze-dried powder and preparation method thereof |
| CN104958771A (en) * | 2015-07-01 | 2015-10-07 | 杨俊� | Large-scale produced fermented pollen sterilization method |
| KR102282350B1 (en) * | 2020-11-17 | 2021-07-27 | 주식회사삼조에스피피 | Vongole pasta sauce and manufacturing method for thereof containing spicy clam stock base |
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