KR101961903B1 - Beverage composition for anti-oxidation comprising chrysanthemum flower extract short-term fermented by lactic acid bacteria and the method of preparation thereof - Google Patents
Beverage composition for anti-oxidation comprising chrysanthemum flower extract short-term fermented by lactic acid bacteria and the method of preparation thereofInfo
- Publication number
- KR101961903B1 KR101961903B1 KR1020160159152A KR20160159152A KR101961903B1 KR 101961903 B1 KR101961903 B1 KR 101961903B1 KR 1020160159152 A KR1020160159152 A KR 1020160159152A KR 20160159152 A KR20160159152 A KR 20160159152A KR 101961903 B1 KR101961903 B1 KR 101961903B1
- Authority
- KR
- South Korea
- Prior art keywords
- lactic acid
- chrysanthemum
- acid bacteria
- chrysanthemum flower
- extract
- Prior art date
Links
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 104
- 235000007516 Chrysanthemum Nutrition 0.000 title claims abstract description 56
- 239000004310 lactic acid Substances 0.000 title claims abstract description 52
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 52
- 239000000284 extract Substances 0.000 title claims abstract description 43
- 241000894006 Bacteria Species 0.000 title claims abstract description 36
- 239000000203 mixture Substances 0.000 title claims abstract description 13
- 238000000034 method Methods 0.000 title abstract description 7
- 244000189548 Chrysanthemum x morifolium Species 0.000 title description 51
- 235000013361 beverage Nutrition 0.000 title description 3
- 238000002360 preparation method Methods 0.000 title description 3
- 230000003064 anti-oxidating effect Effects 0.000 title 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 241000723353 Chrysanthemum Species 0.000 claims abstract description 13
- 235000015097 nutrients Nutrition 0.000 claims abstract description 7
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- 239000012138 yeast extract Substances 0.000 claims description 6
- 241000194031 Enterococcus faecium Species 0.000 claims description 5
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Images
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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
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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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
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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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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Botany (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Non-Alcoholic Beverages (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 국화꽃의 물추출물에 대한 유산균 발효에 의해 항산화 활성이 증진된 천연 발효 조성물의 제조방법에 관한 것으로 국화꽃 물추출물에 영양원을 첨가하고 고온 단기 발효 특성이 있는 Enterococcus속 유산균주를 접종하여 항산화 활성이 증대된 국화꽃 물추출물의 유산균 발효액 및 이를 이용한 항산화 기능성 음료를 제공하는 뛰어난 효과가 있다. The present invention relates to a method for producing a natural fermentation composition in which the antioxidative activity is enhanced by fermentation of a water extract of Chrysanthemum flower by a fermentation with lactic acid bacteria. The fermentation method comprises adding a nutrient source to a chrysanthemum flower water extract and inoculating a Enterococcus sp. There is an excellent effect of providing a lactic acid fermentation broth of an extract of Chrysanthemum chrysanthemum which has increased antioxidant activity and an antioxidant functional beverage using the same.
Description
본 발명은 식용 가능한 국화꽃 발효 추출액을 함유하는 항산화 기능성 음료 조성물에 관한 것으로서, 더욱 상세하게는 단일 원료 국화꽃에 증류수를 첨가하여 그 추출물을 수득하고, 상기 국화꽃 물추출물에 고온 단시간 발효 특성을 갖는 유산균을 접종, 발효하여 얻은 유산균 발효액을 유효성분으로 함유하는 인체 항산화 활성 증진용 식품 조성물 및 그 제조방법에 관한 것이다.The present invention relates to an antioxidant functional beverage composition containing an extractable chrysanthemum flower fermented extract. More particularly, the present invention relates to an antioxidant functional beverage composition containing distilled water to a single raw chrysanthemum flower to obtain an extract thereof, Which is obtained by inoculating and fermenting a lactic acid bacterium having an antioxidant activity as an active ingredient, and a method for producing the same.
국화는 국화과(Compositae)에 속하는 다년생초본으로 꽃의 크기에 따라 대국, 중국, 소국으로 나누어지며 꽃이 피는 시기에 따라 계절별 국화로 구분된다. 국화는 식약처에서 지정한 식용 가능한 꽃으로 국화주, 국화전, 국화차 등 식용으로 이용가능하다. Chrysanthemum is a perennial plant belonging to Compositae. It is divided into large country, small country, small country according to the size of flower and it is classified into seasonal chrysanthemum according to the time of flowering. Chrysanthemum is an edible flower designated by the Ministry of Forestry, and can be used for food such as chrysanthemum, chrysanthemum, and chrysanthemum tea.
국화꽃의 주요성분으로는 플라보노이드 화합물인 luteolin, apigenin, apignin 7-O-β-D-glucoside 및 luteolin 7-O-β-D-glucoside 등이, sequiterpene lactone 화합물인 cu-mambrin A, cumambrin B, arteglasin A 및 angeloyljadin 등이 알려져 있다. 국화는 전통식품의 기능성 소재로서 위장, 감기, 두통, 현기증, 고혈압 등에 유효하며, 바이러스 억제효과, 항균활성 및 항암활성 등에 효과가 있는 것으로 보고되어 있다. Luteolin, apigenin, apignin 7-O-β-D-glucoside and luteolin 7-O-β-D-glucoside are the main components of chrysanthemum flowers. The major components of chrysanthemum flowers are catecholamines such as cu-mambrin A, cumambrin B, arteglasin A and angeloyljadin are known. Chrysanthemum is a functional material of traditional foods and is effective for gastrointestinal, cold, headache, dizziness, hypertension, etc. It has been reported to be effective for virus inhibitory effect, antimicrobial activity and anticancer activity.
최근에는 국화 지상부에서 항 곰팡이 활성, 항궤양 활성 및 면역증진활성 등이 보고되어 국화 꽃 및 잎은 건강기능식품의 원료로 각광받기 시작했다. In recent years, antifungal activity, anti-ulcer activity, and immunity-promoting activity have been reported in the chrysanthemum, and chrysanthemum flowers and leaves have become popular as health food ingredients.
천연물은 비타민, 무기물, 색소 및 각종 특성 성분 등의 중요한 영양소를 풍부하게 함유하고 있어 건강의 유지, 개선, 질병 예방에 크게 기여하는 것으로 알려져 있다. 그러나 저장성이 취약하기 때문에 저장성과 기능성 향상을 위한 식품 가공이 필요하다. Natural products are rich in important nutrients such as vitamins, minerals, pigments and various characteristic components, and are known to contribute to health maintenance, improvement, and disease prevention. However, because of poor storage stability, food processing is needed to improve storage and functionality.
식품을 발효하면 고분자 물질이 저분자 물질로 분해되어 소화 흡수율을 높이고, 새로운 풍미 생성으로 기호성이 증대되며, 생리활성을 갖는 성분이 생성되어 최근 발효식품에 대한 연구가 활발하게 진행되고 있다. When fermented food, the polymer substance is decomposed into a low molecular substance to increase the digestion and absorption rate, to increase the palatability due to the generation of new flavor, and to produce a component having physiological activity, and recent studies on fermented foods are actively proceeding.
유산균은 당류를 발효하여 에너지를 획득하고 다량의 유산(lactic acid)를 생성하며, 소비되는 당의 50% 이상을 대사 생산물로 생성하지 않고 부패를 방지하는 유익균이다. 유산균은 그람양성균으로 통성혐기성 또는 혐기성 미생물이며 운동성은 없고 대부분이 카탈라아제 음성이며 생육에는 각종 비타민, 아미노산, 펩티드 등을 요구한다. 일반적으로 유산균의 섭취는 장내 미생물들이 음식물의 소화, 흡수, 분해와 배설 등을 돕는 유익균의 증가를 유도하여 소화작용을 도와 소화기계 질환에 개선 효과를 나타낸다(Rolfe, 2000, J Nutr, 130:396-402). 또한, 유산균 발효를 통한 유용 물질의 합성에 의한 영양 및 건강 증진 효과가 있으며, 정장 작용, 면역증강 작용, 간경화 개선, 항암 작용, 피부미용 효과 등 영양학적인 효과 등 다양한 효능이 보고되어 있다.
Lactic acid bacteria are beneficial to ferment saccharides to obtain energy, to produce a large amount of lactic acid, and to prevent spoilage without producing more than 50% of the consumed sugar as a metabolic product. Lactic acid bacteria are Gram-positive bacteria and they are anaerobic or anaerobic microorganisms with no motility. Most of them are catalase-negative and require various vitamins, amino acids and peptides for growth. In general, the intake of lactic acid bacteria induces an increase in beneficial bacteria that help digestion, absorption, decomposition and excretion of food, thereby improving digestion and improving digestive system diseases (Rolfe, 2000, J Nutr, 130: 396 -402). In addition, there is a nutritional and health promoting effect by synthesis of a useful substance through fermentation of lactic acid bacteria, and various effects such as a dressing action, an immunity enhancing action, an improvement of liver cirrhosis, an anticancer action, a skin cosmetic effect and the like are reported.
따라서, 최근 천연 식품 소재를 발효시킴으로써 기능성 강화 특성을 향상시킬 뿐 아니라 생체이용률이 높은 유용성분으로 전환하는 기술이 지속적으로 개발되고 있다. Therefore, recently, technologies for converting functional ingredients into bioactive ingredients with high bioavailability by fermenting natural food materials have been continuously developed.
본 발명과 관련된 선행기술로서 대한민국 등록특허 제10-1005960호 이는 생국화꽃에 설탕을 1:1로 효모에 의해 자연발효 숙성시킨 후 추출에 관한 면역 증진용 국화차 음료에 관한 것이 그리고 10-2011-0117876에는 금은화+국화꽃+진달래꽃을 1:1:1중량비로 혼합하여 YM배지나 MRS배지에서 효모를 접종 발효시켜 되는 항염증 화장료 조성물이 개시되어 있다. 또, 대한민국 특허출원 10-2010-33573호에는 국화잎에 에탄올 등 유기용매 추출한 추출물의 아토피 개선 기능성 화장료가 그리고 특허출원 10-2016-75056호에는 금물초 꽃 + 원추리 꽃 + 능소화 꽃 + 해당화 꽃을 1:1:1:1 중량비로 혼합하여 수추출물을 얻고 유산균을 접종하여 8℃에서 7일간 빌효한 추출물의 피부 주름 개선 기능이 공지되어 있다.As a prior art related to the present invention, Korean Patent No. 10-1005960 discloses a chrysanthemum tea beverage for enhancing immunity related to extraction after natural fermentation and maturation of yeast with 1: -0117876 discloses an anti-inflammatory cosmetic composition which is prepared by fermenting yeast in a YM medium or MRS medium by mixing gold silver, chrysanthemum, and rhododendron flowers at a weight ratio of 1: 1: 1. Korean Patent Application No. 10-2010-33573 discloses a cosmetic composition for treating atopy of an extract of an extract of an organic solvent such as ethanol on a chrysanthemum leaf and a patent application No. 10-2016-75056 for a cosmetic composition having an antimicrobial flower, a primrose flower, a Jacob flower, 1: 1: 1: 1 weight ratio to obtain a water extract, and inoculating the lactic acid bacteria, the function of improving the skin wrinkles of the extract of Billy extract at 8 ° C for 7 days is known.
그러나 현재까지 국화꽃 단일 원료의 추출물에 단기 고온발효 특성을 갖는 유산균을 접종 발효시켜 항산화 활성을 증진시킨 기술에 관하여는 공지된 바 없다.However, there is no known technology for fermenting lactic acid bacteria having short-term high-temperature fermentation characteristics to an extract of Chrysanthemum flower monocotyledonous material to enhance antioxidant activity.
따라서 본 발명의 목적은 단일 원료 국화꽃의 물추출물에 고온 단기간 발효 특성이 있는 유산균을 접종하여 항산화 활성을 현저히 증진된 국화꽃 물추출물의 유산균 발효액을 제공하는 데 있다. Accordingly, it is an object of the present invention to provide a lactic acid fermentation broth of a chrysanthemum flower water extract which has been remarkably enhanced in antioxidative activity by inoculating a water extract of a single raw chrysanthemum flower with lactic acid bacteria having high-temperature short-term fermentation characteristics.
본 발명의 다른 목적으로는 상기 국화꽃의 물추출물의 단기간 고온발효 특성의 유산균주 발효액을 유효성분으로 함유하는 건강 기능성 식품 조성물을 제공하는 데 있다.Another object of the present invention is to provide a health functional food composition containing, as an active ingredient, a lactic acid bacterium fermentation broth having high-temperature fermentation characteristics of the water extract of chrysanthemum flower for a short period.
본 발명의 상기 목적은 국화꽃을 그 중량의 15배에 해당하는 증류수에 침지시키는 제 1단계와; 항산화 활성이 높은 추출 조건 30~90℃에서 2시간 동안 수추출하는 제 2단계와; 상기 국화꽃 물추출물에 고온 단기간 발효 특성이 있는 GRAS(Generally Recognized As Safe)급 유산균 Enterococcs faecium KCTC 13225를 접종하고, 영양원으로 glucose 0~5%, yeast extract 0~5%를 첨가하는 제 3단계를 통하여 발효하여 단일 원료로 구성된 국화꽃 단일원료 추출물의 유산균 발효액을 제공하고 그 항산화 효과를 평가함으로써 달성하였다.The above object of the present invention is achieved by a method for producing a chrysanthemum flower comprising: a first step of immersing a chrysanthemum flower in distilled water corresponding to 15 times its weight; A second step of extracting water at a temperature of 30 to 90 ° C for 2 hours with high antioxidant activity; The third step is to inoculate GRC (Generally Recognized As Safe) lactic acid bacteria Enterococcus faecium KCTC 13225 with high temperature and short term fermentation characteristics and to add 0 ~ 5% of glucose and 0 ~ 5% of yeast extract as a nutrient source. The fermentation broth of fermented Lactobacillus japonica, which is fermented through a single raw material, was obtained and its antioxidative effect was evaluated.
본 발명의 국화꽃 물추출물의 유산균 발효액은 항산화 효과가 현저히 증진되어 있으며, 천연 유해성분만을 사용하기 때문에 인체에 부작용이 없어 건강 기능성 식품의 소재로 사용가능한 효과가 있을 뿐만 아니라 고온 단기간 발효에 의해 유기산이 포함되지 않은 발효액을 제공하는 뛰어난 효과가 있다. The lactic acid fermentation broth of the chrysanthemum flower water extract of the present invention has remarkably enhanced antioxidative effect and has no adverse effect on the human body due to the use of natural harmful nutrition, so that it can be used as a material for health functional food, There is an excellent effect of providing a fermentation liquid which does not contain the fermentation broth.
도 1은 본 발명에 따른 국화꽃 물추출물의 유산균 발효액의 제조방법의 개략적인 공정을 나타낸 다이어그램이다.
도 2는 본 발명에 따른 국화꽃의 추출 조건에 따른 각 DPPH radical 소거활성 측정결과를 나타낸 그래프이다.
도 3은 본 발명에 따른 국화꽃의 추출 조건에 따른 각 총 폴리페놀 함량에 대한 측정결과를 나타낸 그래프이다.
도 4는 본 발명에 따라 제조된 국화꽃 추출물의 유산균 발효액의 DPPH radical 소거능에 의한 항산화 활성의 결과를 나타낸 그래프이다.
도 5는 본 발명에 따라 제조된 국화꽃 추출물의 유산균 발효액의 항산화 활성을 총 폴리페놀 함량의 측정 결과를 나타낸 그래프이다.1 is a diagram showing a schematic process of a process for producing a lactic acid fermentation broth of Chrysanthemum morifolium extract according to the present invention.
2 is a graph showing the measurement results of DPPH radical scavenging activity according to the extraction conditions of the chrysanthemum flower according to the present invention.
FIG. 3 is a graph showing the results of measurement of total polyphenol contents according to the extraction conditions of chrysanthemum flowers according to the present invention. FIG.
4 is a graph showing the results of antioxidant activity of DPPH radical scavenging activity of lactic acid fermentation broth of Chrysanthemum morifolium extract prepared according to the present invention.
FIG. 5 is a graph showing the results of measurement of total polyphenol content of antioxidant activity of lactic acid bacteria fermentation broth of Chrysanthemum morifolium extract prepared according to the present invention. FIG.
본 발명은 국화꽃으로부터 물추출한 국화꽃 추출액에 유산균을 접종한 후 발효시켜 제조된 국화꽃 물추출물의 유산균 발효액과 이를 유효성분으로 함유하는 항산화 활성 증진용 기능성 식품 조성물을 제공한다. The present invention provides a lactic acid fermentation broth of chrysanthemum flower water extract prepared by inoculating a chrysanthemum flower extract extracted from a chrysanthemum flower with water and fermenting the chrysanthemum flower extract, and a functional food composition for enhancing antioxidative activity containing the extract as an active ingredient.
본 발명에 따르면 사용할 수 있는 바람직한 유산균은 Enterococcus속 유산균주이나 가장 바람직하게는 Enterococcus faecium KCTC 13225이다.Preferred lactic acid bacteria that can be used according to the present invention are those of the genus Enterococcus or most preferably those of Enterococcus faecium KCTC 13225.
발효적성이 우수한 Lactobacullus속의 균주는 저온 내지 중온에서 발효기간이 길고 유기산 냄새로 바람직하지 않다.The strain Lactobacullus having excellent fermentation suitability has a long fermentation period at a low temperature to a middle temperature and is not preferable as an organic acid smell.
또한, 본 발명에 따른 국화꽃 추출물의 유산균 발효 조성물의 제조방법은 (a) 발효원료인 국화꽃의 경우 생화 또는 건조된 국화꽃을 파쇄하여 사용될 수 있다. (b) 국화꽃 물추출물은 국화꽃에 증류수를 15배 첨가하여 30~90℃에서 1~3시간, 가장 바람직하게는 60℃에서 2시간 추출하는 단계가 포함된다. (c) 상기 단계에서 얻은 국화꽃 추출액에 본 발명에 따른 상기 유산균을 1%(w/w) 접종한 후 영양원으로 glucose 0~5%, yeast extract 0~5%를 첨가하는 단계를 포함한다. (d) 상기 a) 및 b) 단계 후에 배지조성물에 Enterococcus faecium(KCTC 13225) 균주를 접종하여 35~40℃에서 24~72시간, 가장 바람직하게는 37℃에서 48시간 동안 발효하는 단계를 포함한다. Also, the method for producing the fermented lactic acid bacteria composition of the present invention can be used by (a) chrysanthemum as a raw material for fermentation by crushing flowers or dried chrysanthemum flowers. (b) Chrysanthemum flower water extract comprises extracting distilled water 15 times at 30 to 90 ° C for 1 to 3 hours, most preferably at 60 ° C for 2 hours by adding distilled water to the chrysanthemum flower. (c) inoculating 1% (w / w) of the lactic acid bacterium according to the present invention to the chrysanthemum flower extract obtained in the above step, adding 0 to 5% of glucose and 0 to 5% of yeast extract as a nutrient source. (d) adding the Enterococcus strain to the medium composition after steps a) and b) faecium (KCTC 13225) to 24 ~ 72 hours to inoculated strains at 35 ~ 40 ℃, most preferably comprising the step of fermenting at 37 ℃ for 48 hours.
본 발명은 또한, 국화꽃의 유산균 발효액의 DPPH radical 소거활성, 총 페놀 함량 측정을 통해 항산화 효과를 평가하는 단계가 포함된다.
The present invention also includes a step of evaluating the antioxidative effect by measuring DPPH radical scavenging activity and total phenol content of lactic acid fermentation broth of Chrysanthemum flowers.
또한, 본 발명은 국화꽃을 유산균으로 발효한 발효액이 항산화 효과가 있는 발효물에 함유되어 있는 유산균수 측정 결과를 제공한다.
The present invention also provides a result of measuring the number of lactic acid bacteria contained in a fermented product having an antioxidative effect by fermentation broth fermented with lactic acid bacteria.
이하, 본 발명의 구체적인 내용을 실시예를 들어 상세하게 설명한다. 하기 실시예는 본 발명을 예시하기 위한 것으로 본 발명의 권리범위가 이에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail with reference to examples. The following examples are intended to illustrate the invention and are not to be construed as limiting the scope of the invention.
실시예 1. 본 발명 국화꽃 추출물 제조Example 1. Production of chrysanthemum flower extract of the present invention
본 발명에 사용되는 국화꽃은 건조된 국화꽃을 분쇄하여 사용하였으나, 생화가 사용될 수 있다. 국화꽃 물추출물은 건조된 국화꽃 분말에 그 중량의 15배에 해당하는 증류수를 첨가하여 60℃에서 2시간 추출하였다. 60℃에서 2시간 동안 추출한 경우 항산화 활성 및 총 폴리페놀 함량이 가장 높았다(도2 및 도3). 추출 후 감압여과를 통해 회수된 여액으로 국화꽃 물추출물을 공시재료로 사용하였다.
The chrysanthemum flower used in the present invention is used by pulverizing dried chrysanthemum flower, but the flower can be used. The water extract of Chrysanthemum morifolium was extracted with dried distilled water at 60 ° C for 15 hours. The antioxidant activity and total polyphenol content were the highest when extracted at 60 ° C for 2 hours (FIGS. 2 and 3). After extraction, the water extract of Chrysanthemum chrysanthemum was used as a test material for the filtrate recovered through vacuum filtration.
실시예 2. 국화꽃 물추출물의 유산균 발효액 제조 Example 2. Preparation of lactic acid fermented broth of chrysanthemum flower water extract
상기 실시예 1에서 얻은 공시재료 국화꽃 물추출물에 이와 별도로 배양된 Enterococcus faecium KCTC 13225를 106 cfu/mL로 접종하고, 여기에 yeast extract 0~5%, glucose 0~5% 질소 및 탄소 영양원을 첨가하여 37℃에서 48시간 유산균의 고온, 40시간 발효하였다.
Enterococcus faecium KCTC 13225, separately cultivated, was added to the chrysanthemum flower water extract obtained in Example 1 at 10 6 cfu / mL, supplemented with 0 to 5% of yeast extract, 0 to 5% of nitrogen and carbon nutrient And fermented at 37 DEG C for 48 hours at a high temperature of lactic acid bacteria for 40 hours.
실험예 1. DPPH소거능에 의한 항산화 활성Experimental Example 1. Antioxidative activity by DPPH scavenging activity
DPPH(α,α-diphenyl-β-picrylhydrazyl)는 화합물 내 질소 중심의 안정화된 구조의 라디컬(radical)로 존재하는데 이 안정화된 radical의 소거능 정도에 따라 항산화 활성을 측정할 수 있다. 본 발명에서 DPPH 라디칼 소거능의 측정은 Biosis 등(1958)의 방법에 의해 측정하였다. 상기 실시예 1과 실시예 2에서 얻은 각 시료 0.5mL에 DPPH 5mL을 넣고 실온, 암소에서 30분 동안 반응시킨 후 517nm에서 흡광도를 측정하여 하기 수학식1에 의해 저해율을 계산하였다. 양성대조군으로는 vitamin C를 사용하였다.
DPPH (α, α-diphenyl-β-picrylhydrazyl) exists as a radical of the stabilized structure of nitrogen center in the compound. Antioxidant activity can be measured according to the degree of scavenging of the stabilized radical. In the present invention, the DPPH radical scavenging activity was measured by Biosis et al. (1958). 5 mL of DPPH was added to 0.5 mL of each of the samples obtained in Example 1 and Example 2, incubated at room temperature and dark for 30 minutes, and the absorbance at 517 nm was measured and the inhibition rate was calculated by the following equation (1). Vitamin C was used as a positive control.
실험 결과, 도4에서 알 수 있듯이 DPPH 소거능에 의한 항산화 활성은 실시예 1의 국화꽃 물추출물 시료에 비해 실시예2의 유산균 발효를 한 시료가 DPPH 소거능이 증가되어 항산화 활성이 증대되었다.As a result, as shown in FIG. 4, the antioxidative activity by the DPPH scavenging activity was enhanced by the DPPH scavenging activity of the lactic acid fermented sample of Example 2 as compared with the chrysanthemum flower water extract sample of Example 1.
본 발명에 따라 제조된 국화꽃 물추출물의 유산균 발효액은 glucose 5%, yeast extract 5%가 첨가된 배지에서 발효되어 고온 단기 발효 특성을 갖는 유산균 발효에 의해 항산화 활성이 30% 이상 증대되었으나, 영양원의 첨가 범위가 상기 5%로 제한되는 것은 아니다.
The lactic acid fermentation broth of Chrysanthemum morifolium prepared according to the present invention was fermented in a medium supplemented with 5% glucose and 5% yeast extract, and the antioxidative activity was increased by more than 30% by lactic acid fermentation with high temperature and short term fermentation characteristics. The range of addition is not limited to the above 5%.
실험예 2. 총 폴리페놀 함량에 의한 항산화 활성Experimental Example 2: Antioxidant activity by total polyphenol content
총 폴리페놀 함량은 Folin-Denis 법으로 측정하였다(Zhou et al., 1980). 상기 실시예 1과 실시예 2의 각 시료 1mL에 1N Folin-Ciocalteu's reagent 1mL을 가하고 3분간 정치시킨 후 10% Na2CO₃용액 1mL을 가하여 혼합한 다음 실온에서 1시간 정치시킨 후 700nm에서 흡광도를 측정하여 총 폴리페놀 함량을 수치화하였다. Total polyphenol content was determined by the Folin-Denis method (Zhou et al., 1980). 1 mL of each Folin-Ciocalteu's reagent was added to 1 mL of each of the samples of Example 1 and Example 2, and the mixture was allowed to stand for 3 minutes. 1 mL of 10% Na 2 CO 3 solution was added to the mixture and allowed to stand at room temperature for 1 hour. The polyphenol content was quantified.
실험 결과, 도5에서 알 수 있듯이 총 폴리페놀 함량은 실시예 1의 국화꽃 물추출물 시료에 비해 실시예2에 따른 유산균 발효를 수행한 시료에서 증대되었다.As a result, as shown in FIG. 5, the total polyphenol content was increased in the samples of the lactic acid bacteria fermented according to Example 2, compared with the sample of the chrysanthemum flower water extract of Example 1.
본 발명에 따라 제조된 국화꽃 물추출물의 유산균 발효물은 glucose 5%, yeast extract 5%가 첨가된 배지에서 본 발명 고온 단기 발효 특성을 갖는 유산균 발효에 의해 항산화 활성이 65% 이상 증대되었으나, 영양원의 첨가 범위가 반드시 5%로 제한되는 것은 아니다.
The antioxidant activity of the lactic acid bacterium fermented by the lactic acid bacteria produced according to the present invention was increased by 65% in the medium supplemented with glucose 5% and yeast extract 5% Is not necessarily limited to 5%.
실험예 3. pH 및 유산균수 측정 Experimental Example 3. Measurement of pH and number of lactic acid bacteria
본 발명의 상기 실시예1에서 얻은 국화꽃 물추출물과 실시예2에서 얻은 국화꽃 물추출물의 유산균 발효 조성물의 pH를 분석하고 유산균수 측정을 위해 실시예 2의 시료를 0.85% 멸균 생리 식염수로 10배 연속 희석법으로 희석한 후 MRS agar 배지에 접종한 다음 집락수를 계측하는 방법으로 유산균수를 산출하였다. The pH of the chrysanthemum flower water extract obtained in Example 1 of the present invention and the chrysanthemum flower water extract obtained in Example 2 were analyzed. To measure the number of lactic acid bacteria, the sample of Example 2 was suspended in 0.85% sterile physiological saline solution The number of lactic acid bacteria was calculated by measuring the number of colonies after inoculation on the MRS agar medium after dilution by serial dilution method.
실험결과 하기 표1에서 알 수 있듯이 실시예 1 시료의 pH는 5.02이고, 실시예 2 시료의 pH는 4.21로 측정되었다. 이를 통해 실시예2에서 얻은 국화꽃 물추출물의 유산균 발효물의 경우 유산균 발효로 유기산이 생성되어 산도가 pH가 4.5 이하로 감소하는 것을 알 수 있었다. 또한, 실시예2의 유산균 발효를 수행한 시료의 유산균수 측정결과 1.4 x 109 cfu/mL로 유산균 발효에 의해 109 이상의 유산균수의 증가가 측정되었다.
Experimental Results As shown in Table 1, the pH of the sample of Example 1 was 5.02 and the pH of the sample of Example 2 was 4.21. As a result, lactic acid fermented by the lactic acid bacteria fermentation of the chrysanthemum flower water extract obtained from Example 2 produced organic acid, and the pH value was decreased to below 4.5. In addition, the result of measurement of the number of the lactic acid bacteria of the sample subjected to the lactic acid fermentation of Example 2 showed an increase in the number of lactic acid bacteria of 10 9 or more by fermentation with lactic acid bacteria at 1.4 x 10 9 cfu / mL.
KCTC 13225 Enterococcus faecium
KCTC 13225
제조예1. 국화꽃 유산균 발효 음료 제조Production Example 1 Chrysanthemum flower lactic acid fermented beverage manufacturing
본 발명 실시예2에 따라 수득한 국화꽃 물추출물의 유산균 발효물이 함유된 음료를 하기 표2와 같이 제조하였다.The beverage containing the lactic acid bacteria fermented product of the water extract of Chrysanthemum morifolium obtained according to Example 2 of the present invention was prepared as shown in Table 2 below.
상기 제조예에 따라 제조된 음료에 대하여 무작위 추출한 남녀 각 5명을 판넬로하여 관능검사를 실시하였다. 대조구는 국화꽃 물추출물로 하였다. 관능 검사는 7점 척도법을 이용하였고, 실험 결과는 하기 표3과 같다.The sensory evaluation was performed on each of 5 male and female randomly selected beverages according to the above Preparation Example using a panel. The control was chrysanthemum flower water extract. The sensory test was performed using a 7-point scaling method. The experimental results are shown in Table 3 below.
실험 결과 색, 맛, 종합기호도에 있어서 대조구에 비하여 Enterococcus속 균주에 의해 발효된 유산균 발효액이 가장 우수한 것으로 평가되었다. Experimental results showed that fermented Lactobacillus fermented by Enterococcus spp. Was the best in color, taste and overall acceptability.
이상 설명한 바와 같이 본 발명은 국화꽃으로부터 물추출물을 얻고 이에 영양원을 첨가한 후 고온 단기 발효 특성을 갖는 유산균을 접종시켜 발효하여 항산화 활성이 증진된 발효액과 이를 이용한 식품을 제공하는 뛰어난 효과가 있으므로 건강 기능성 식품 산업상 매우 유용한 발명인 것이다. As described above, the present invention provides a fermentation broth in which a water extract is obtained from a chrysanthemum flower, a nutrient source is added thereto, fermented by inoculating lactic acid bacteria having high-temperature short-term fermentation characteristics, It is a very useful invention in the functional food industry.
Claims (1)
상기 제1단계에서 얻은 국화꽃 물추출물에 Enterococcus faecium KCTC13225 유산균주를 접종하는 제2단계(S2)와;
상기 제2단계에서 유산균을 접종하여 얻은 배지 조성액에 영양원으로서 glucose와 yeast extract를 첨가 혼합하여 37℃에서 40시간 고온 단기 발효시켜 유산균수 1.4x 109 cfu/mL 이상이며 pH4.5 이하인 제3단계(S3)로 제조된 국화꽃물의 유산균 발효 추출액.
A first step (S1) of crushing the dried chrysanthemum flower and extracting it at 60 DEG C for 2 hours to obtain water extraction;
A second step (S2) of inoculating Enterococcus faecium KCTC13225 lactic acid bacteria into the chrysanthemum flower water extract obtained in the first step;
Glucose and yeast extract were added as a nutrient source to the culture medium obtained by inoculating the lactic acid bacterium in the second step, and the mixture was fermented at 37 ° C. for 40 hours at a high temperature for a short period of time to produce a lactic acid bacterium having a lactic acid number of 1.4 × 10 9 cfu / (S3) fermented extract of Lactobacillus plant.
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