KR101822752B1 - Manufacturing method for enzymatic extracts of deer antlerdeer antler containing highly physiologically active ingredients of deer antler and enzymatic extracts of deer antlerdeer antler using the same - Google Patents
Manufacturing method for enzymatic extracts of deer antlerdeer antler containing highly physiologically active ingredients of deer antler and enzymatic extracts of deer antlerdeer antler using the same Download PDFInfo
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- KR101822752B1 KR101822752B1 KR1020160098701A KR20160098701A KR101822752B1 KR 101822752 B1 KR101822752 B1 KR 101822752B1 KR 1020160098701 A KR1020160098701 A KR 1020160098701A KR 20160098701 A KR20160098701 A KR 20160098701A KR 101822752 B1 KR101822752 B1 KR 101822752B1
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- antler
- deer
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- extract
- weight
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Abstract
Description
본 발명은 녹용의 생리활성 성분을 극대화시킨 단백질 및 탄수화물 가수분해 효소 추출물의 제조방법에 관한 것으로, 녹용 성분 함유 식품제조 시, 우론산, 시알산 및 콜라겐 등의 유효 생리활성 성분의 농도를 극대화하고, 항산화 성능이 우수한 녹용추출물 및 제조 방법에 관한 것이다.
The present invention relates to a method for producing a protein and a carbohydrate hydrolase extract which maximizes physiologically active ingredients of deer antler and maximizes the concentration of effective physiologically active ingredients such as uronic acid, sialic acid and collagen And antioxidant properties, and a process for producing the same.
녹용이란 숫사슴의 뿔이 딱딱하게 각질화되기 전 잘라서 약으로 사용하는 것을 말한다. 예로부터 녹용은 성장이 늦은 아이, 허약한 체질의 아이들의 발육 촉진, 골격을 튼튼하게 해주는 효과, 지능 발달 및 소화흡수를 높여주고, 면역력을 길러주는 효과가 있는 것으로 알려져 왔다. 본초강목에 따르면 녹용은 지속적으로 젊음의 생기를 부여하고, 기운을 돋으며 피부보양 효과가 있다고 하여 노화방지에 특효가 있는 것으로 알려졌다.It is said that the antlers are cut and used as a medicine before the horns of the stag are hardened. It has been known from the past that antler has the effect of promoting growth of children with late growth, children with fragile constitution, strengthening the skeleton, improving intelligence development and digestion and absorbing immunity. According to the herbal gangmok, it is known that antler antioxidant has a special effect for anti-aging because it constantly gives vitality to youthful, energizing and skin-protecting effect.
또한 녹용은 몸 속 장기들의 활동을 도와주어 피로회복, 숙면, 식욕을 돋구고, 강력한 항염증 작용으로 몸속에 있는 독소를 제거해주는 효과와 감기 등의 잔병치레를 막아주며 적혈구의 생성을 촉진시키는 효과가 있어 빈혈환자, 출산전후, 큰 수술을 한 뒤에 복용하면 도움이 된다고 한다. 뿐만 아니라 녹용은 근골을 강하게 하는 효과가 있어 허리와 무릎, 다리에 힘이 없을 때, 골다공증 등의 증상이 있을 때 도움이 되며 성호르몬의 분비를 촉진시켜주는 효과가 있어 정력이 안 좋을 때 발기부전일 때 그리고 여성의 배란작용을 활발하게 해 주는 효과가 있다.In addition, antler antioxidants help the organs of the body to recover from fatigue, improve sleep and appetite, and have a strong anti-inflammatory action that removes toxins from the body and prevents the warts from getting cold and promotes the production of red blood cells. Anemia patients, before and after childbirth, after a big surgery, it is helpful to take. In addition, antler has a strong effect on muscles, and when there is no strength in the back, knees, and legs, it helps when there are symptoms such as osteoporosis, and it stimulates the secretion of sex hormones. And the effect of stimulating the ovulation of women.
사슴의 뿔은 사슴의 종류와 년생에 따라 자르는 시기가 조금씩 차이는 있으나, 평균 5~8월 사이에 자르는데, 이것을 절각이라고 한다. 뿔을 절각한 후 머리에 남는 부위는 딱딱하게 굳어지게 되고 이듬해 2~3월 경에 저절로 떨어지게 되며 이것을 낙각이라고 한다. 낙각된 후에 그 자리에 새로운 뿔이 돋아나기 시작하며 낙각한 시기를 기준으로 절각을 하기도 하는데, 꽃사슴의 경우 낙각 후 65일, 엘크의 경우는 80일, 레드디어의 경우는 70일 후에 절각하여 녹용으로 사용한다.Deer horns vary slightly depending on the type and age of the deer, but they are cut from May to August on average. After turning the horns, the remaining part of the head is hardened and then falls off in the second or third month of the following year. The new horn starts to grow on the spot after it has been withdrawn, and it is folded on the basis of the declining time. In the case of the scarlet, 65 days after declination, 80 days for elk, 70 days for reddish, .
우리나라에서는 녹용 생산을 위하여 많은 사슴이 사육되고 있는데, 꽃사슴이 대다수를 이루고, 적록(赤鹿)에 속하는 뉴질랜드산과 대록(大鹿)에 속하는 아메리카 엘크 등이 사육되고 있으며 일부는 약용으로 유통되고 있다. 엘크사슴은 대형종의 붉은 사슴속의 사슴으로 현재 우리나라에 도입되어 사육되고 있는 사슴 중 가증 큰 품종으로, 캐나다, 미국북부, 러시아 등지에서 수입되어 사육되고 있다.In Korea, many deer are being raised for the production of antler, and the majority of the deer are formed, and the New Zealand mountains belonging to the red deer and the American elk belonging to the big deer are raised, and some are distributed as medicinal products. Elk deer is a large deer deer in red deer. It is introduced in Korea and is a large number of deer among breeders. It is imported from Canada, northern USA, and Russia.
엘크사슴은 녹용생산성과 자록 생산성이 뛰어나기 때문에 점차 사육이 늘어나고 있는 추세이다. 엘크사슴은 2~3월 사이에 묵은 뿔이 떨어지고 새 뿔이 돋아나기 시작하여 약 80~90일 사이에 녹용을 수확하여 이용하는데, 가지수가 많고 상대부분이 매우 발달되어 있기 때문에 좋은 품질의 녹용을 수확할 수 있다.Elk deer is increasingly breeding because of its excellent antler production and self-productivity. The elk deer has antlers that fall from February to March and the new horns begin to sprout. After about 80 to 90 days, the antlers are harvested and harvested. It can be harvested.
그러나 최근 금융 위기 등 복잡한 경제 정세로 인한 소비 위축의 영향으로 한약의 소비와 더불어 녹용 소비 역시 감소하였고, 이는 사슴 사육 두수의 감소로까지 이어져 축산 농가의 생계까지 위협하고 있다. 또한 녹용이 건강증진에 도움이 되는 많은 생리적 활성물질과 영양소를 함유하고 있음에도 열수추출 형태인 '보약'으로만 음용되는 실정이고, 녹용 특유의 비린 맛 때문에 높은 영양가치에도 불구하고 녹용함유 식품들이 활발하게 개발되고 있지 못한 실정이다.However, the consumption of herbal medicine as well as the consumption of antler have also decreased due to the consumption contraction caused by the complicated economic situation such as the recent financial crisis, which leads to a decrease in the number of deer hatching, which threatens the livelihood of the livestock farmers. Despite the fact that antler contains many physiologically active substances and nutrients that contribute to health promotion, it is only consumed with 'hot water extract', 'tablets.' Despite its high nutritional value due to the unique flavor of antler, It is not developed.
또한 종래의 녹용은 한약재로 주로 물에 다른 약재와 함께 넣고 오랫동안 끓인 열수추출방식으로 음용하여 왔고, 녹용을 포함하는 다양한 식품들이 개발되지 않아 그 이용방법이 협소한 문제가 있었다. 특히 고가의 녹용을 열수추출이라는 천편일률적인 방법으로 유효성분을 추출하는 방법을 사용함으로써 뛰어난 기능적인 활성성분을 다량 포함하고 있음에도 불구하고 충분하게 이용하지 못하는 문제가 있었다.In addition, conventional antler antler has been mainly used as a herbal medicine, and has been put into water with other medicines and consumed for a long time by boiling hot water extraction method, and various foods including antler have not been developed. Especially, the method of extracting an effective ingredient by using a high-priced antler extract called hot water extraction has been used, but it has not been used sufficiently although it contains a large amount of excellent functional active ingredients.
최근 과학의 발달로 천연물로부터 생리적 유효성분을 추출하는 방법이 다양화되면서, 추출방법에 따라 생리적 활성정도가 크게 달라지는 것으로 알려졌는데, 실제로 복분자의 경우 초음파 병행추출로 항암활성 및 면역활성 증진효과가 증가하는 것이 확인되었으며, 초임계 추출에 의하여 항산화 활성이 증가하는 것이 보고되었다. 따라서 고가인 녹용의 영양소 추출방법을 다양화하고, 이를 보다 용이하게 섭취할 수 있는 방법에 대한 연구가 필요하다.
In recent years, it has been known that the physiological activity level varies greatly depending on the extraction method, as the method of extracting the physiologically active ingredient from natural products has been diversified due to the development of science. In fact, in the case of bokbunja, the effect of enhancing the anticancer activity and immune activity , And it has been reported that the antioxidant activity is increased by supercritical extraction. Therefore, it is necessary to study the dietary methods of dietary antler extract and to make it easier to ingest.
본 발명은 종래의 열수추출방법으로 이용하던 녹용을 다양한 추출방법을 이용하여 고생리활성물질을 함유하도록 추출하는 방법과 이를 이용한 녹용의 고생리활성추출물을 제공하고, 녹용 추출물의 생리활성을 극대화시키고 섭취를 용이하게 할 수 있는 부재료의 배합비율을 제공하는 데에 목적이 있다.
The present invention provides a method for extracting antler extract used as a conventional hot water extraction method so as to contain a physiologically active substance using various extraction methods and a hyperglycemic extract extract of the antler extract using the method and maximizing the physiological activity of the antler extract It is an object of the present invention to provide a compounding ratio of a sub-ingredient which can facilitate ingestion.
본 발명은 생녹용과 물을 1 : 4의 무게비로 혼합한 후, 95 ℃에서 7시간 끓여 감압 농축하여 60~70 ㅀBrix 이 되도록 녹용농축액을 제조하는 단계 (1), 상기 (1) 단계의 녹용농축액에 가수분해효소를 질량비 100:1로 혼합하고, pH, 온도를 맞춘 후 24시간 반응시키는 단계(2), 상기 (2) 단계의 가수분해물을 여과하고, 효소활성을 제거한 후, pH를 7로 맞추어, 동결건조시키는 단계(3);로 이루어진 것을 특징으로 하는 녹용의 고생리활성성분 함유 가수분해 효소추출물의 제조방법을 제공한다.
The present invention relates to a process for producing a deer-friendly concentrate (1) comprising mixing a raw green bean and water at a weight ratio of 1: 4, boiling at 95 ° C for 7 hours and concentrating under reduced pressure to obtain a deer concentration of 60 to 70 ㅀ Brix, (2) mixing the hydrolyzed enzyme with the hydrolysis enzyme at a mass ratio of 100: 1, adjusting the pH and temperature, and allowing to react for 24 hours, filtering the hydrolyzate of step (2) And (3) lyophilization of the dextrin-active component of the deer antler.
본 발명에 따른 녹용의 고생리활성성분 함유 가수분해 효소추출물의 제조방법을 통하여 녹용 추출물을 제조함으로써, 고가의 녹용에서 생리활성 성분의 추출을 배가시키고 이를 식품제조에 이용함으로써 현대인의 건강증진과 식품산업에 기여할 수 있다.
The present invention relates to a method for manufacturing a hydrolytic enzyme-containing extract of deer antler according to the present invention, which enables to extract a physiologically active ingredient from an expensive deer antler, It can contribute to industry.
도 1은 녹용 부위별로 제조한 녹용 농축액의 HPLC chromatogram을 나타낸 것이다. (A : 상대, B: 중대, C: 하대)
도 2는 가수분해효소 처리과정의 개략도를 나타낸 것이다.
도 3은 녹용의 상대, 중대, 하대의 가수분해 효소 추출과정을 보여주는 사진이다.
도 4는 녹용의 각 가수분해효소 산물내의 Uronic acid의 함량을 나타낸 그래프이다.
도 5은 녹용의 각 가수분해효소 산물내의 Sialic acid의 함량을 나타낸 그래프이다.
도 6은 녹용의 각 가수분해효소 산물내의 글루코스아미노글리칸의 함량을 나타낸 그래프이다.
도 7은 녹용의 각 가수분해효소 산물내의 총 탄수화물의 함량을 나타낸 그래프이다.
도 8은 녹용의 각 가수분해효소 산물내의 콜라겐의 함량을 나타낸 그래프이다.
도 9는 녹용의 각 가수분해효소 산물내의 DPPH 라디칼 제거활성을 나타낸 것이다. (A: DPPH 라디칼 제거활성도 그래프. B: 60 μM DPPH의 메타놀 용액에서 각 농도에 따른 녹용 상대에서의 flavourzyme 처리 추출물의 ESR spectra 결과. C: ESR spectra)
도 10은 녹용의 각 가수분해효소 산물내의 하이드록실 라디칼 소거활성을 나타낸 것이다. (A: 하이드록실 라디칼 제거활성도 그래프. B: Fenton reaction system에서 각 농도에 따른 녹용에서의 flavourzyme 처리 추출물의 ESR spectra 결과)
도 11은 녹용의 각 가수분해효소 산물내의 알킬 라디칼 소거활성을 나타낸 것이다. (A: 알킬 라디칼 제거활성도 그래프. B: 4-POBN 처리된 AAPH 인큐베이션 중 관찰되는 녹용 상대의 acalase 처리 추출물에서 각 농도에 따른 ESR spectra 결과)
도 12는 식물추출물별 DPPH 라디칼 소거활성을 나타낸 그래프이다.FIG. 1 is an HPLC chromatogram of a deer antler prepared by the antler part. (A: Relative, B: Company, C: Inferior)
Figure 2 shows a schematic diagram of a hydrolytic enzyme treatment process.
FIG. 3 is a photograph showing the hydrolytic enzyme extraction process of the counterpart, middle, and bottom of antler.
4 is a graph showing the content of Uronic acid in each hydrolytic enzyme product of antler.
FIG. 5 is a graph showing the content of sialic acid in each hydrolytic enzyme product of antler.
FIG. 6 is a graph showing the content of glucose aminoglycan in each hydrolytic enzyme product of antler.
7 is a graph showing the content of total carbohydrates in each hydrolytic enzyme product of antler.
FIG. 8 is a graph showing the content of collagen in each hydrolytic enzyme product of antler.
Fig. 9 shows DPPH radical scavenging activity in each antagonist product of antler. (A: graph of DPPH radical scavenging activity B: ESR spectra of flavorzyme treated extracts in antifoaming agent according to each concentration in 60 μM DPPH methanol solution C: ESR spectra)
Fig. 10 shows the hydroxyl radical scavenging activity in each antagonist product of deer antler. (A: Graph of hydroxyl radical scavenging activity, B: ESR spectra of flavorzyme extracts in antler of each concentration in Fenton reaction system)
Fig. 11 shows the alkyl radical scavenging activity in each hydrolytic enzyme product of antler. (A: graph of alkyl radical scavenging activity B: ESR spectra results of each concentration in acerase-treated extracts of antler antagonist observed in 4-POBN-treated AAPH incubation)
12 is a graph showing DPPH radical scavenging activity of each plant extract.
본 발명은 녹용의 생리활성 성분을 극대화시킨 단백질 및 탄수화물 가수분해 효소 추출물의 제조방법 및 이를 이용한 녹용 가수분해 효소추출물에 관한 것이다. 이하 본 발명을 구체적인 실시예를 들어 자세히 설명한다.
The present invention relates to a process for producing protein and carbohydrate hydrolase extracts maximizing physiologically active ingredients of deer antler, and to an antler dehydrogenase extract using the same. Hereinafter, the present invention will be described in detail with reference to specific examples.
I. 발명에 사용한 재료 및 방법I. Materials and Methods Used in the Invention
1. 녹용을 이용한 식품소재 (녹용 분말 및 녹용 농축액)의 제조1. Manufacture of food ingredients (deer antler and deer antler) using antler
(1) 녹용 분말: 녹용 분말은 생녹용을 분쇄하여 동결건조하였다.(1) Deer antler powder: The deer antler powder was pulverized and lyophilized.
(2) 녹용 농축액: 가전 약탕기(JH-D40, 휴먼플러스)를 이용하여 생녹용 1 Kg에 물 4L를 넣은 후 95 ℃에서 7시간 추출 후 감압 농축하여 60~70 ㅀBrix 이상이 되도록 제조하였다.
(2) Deer antler: 4 Kg of water was added to 1 Kg of raw green tea using a household electric kettle (JH-D40, Human Plus), and then extracted at 95 ° C for 7 hours and concentrated under reduced pressure to have a Brix of 60 to 70. Brix.
2. 녹용 가공 소재별 (녹용 분말 및 녹용 농축액) 일반 성분 및 유효성분 분석2. Analysis of general ingredients and effective ingredients of antler processing materials (antler grains and deer antler)
(1) 녹용 가공 소재별 일반성분 분석 : 일반성분은 AOAC법 (2005)에 따라 수분함량은 105℃ 상압건조법, 조단백질 함량은 Kjeldahl법, 조지방 함량은 Soxhlet법으로 분석하였다.(1) Analysis of general components according to the processed materials: The moisture content was 105 ℃, the crude protein content was measured by Kjeldahl method and the crude fat content by Soxhlet method according to AOAC method (2005).
(2) 녹용 가공 소재별 유효성분 분석(2) Analysis of effective ingredients of each antler
가) 우론산 (Uronic acid) 분석: 녹용탕 시료를 4℃에서 0.5M EDTA 2Na(pH 7.4)로 탈칼슘화하였다. Crude Papein을 5mM EDTA와 5mM Cystein HCL을 포함하는 0.1M Phosphate buffer (pH 6.5)와 혼합하여 65℃에서 30분간 Papein을 활성화시켰다. 그 후 활성화된 Papein으로 65℃에서 16시간 동안 단백질을 분해시켜서 Carbazole 반응에 의해 530㎚에서 측정하였다(Scott, 1960 ; Kosakai & Yoshizawa, 1979). 이때 함량은 녹용탕 시료에 대한 중량%로 표기하였다.A) Uronic acid analysis: A sample of the antler was decalcified at 4 ° C with 0.5M EDTA 2Na (pH 7.4). Crude Papein was mixed with 0.1 M phosphate buffer (pH 6.5) containing 5 mM EDTA and 5 mM Cystein HCl, and Papein was activated for 30 minutes at 65 ° C. Protein was then digested with activated Papein at 65 ° C for 16 hours and then measured at 530 nm by Carbazole reaction (Scott, 1960; Kosakai & Yoshizawa, 1979). At this time, the content was expressed as% by weight with respect to the sample of the antler.
나) 시알산 (Sialic acid) 분석: 녹용탕 시료를 80℃에서 1시간 동안 0.1N Sulfuric acid (H2SO4)로 가수 분해한 후 Warren방법 (Warren, 1959)에 의해서 549㎚에서 측정한다. 이때 함량은 녹용탕 시료에 대한 중량%로 표기하였다.Sialic acid analysis: Nerotonic acid samples are hydrolyzed with 0.1 N sulfuric acid (H2SO4) at 80 ° C for 1 hour and measured at 549 nm by the Warren method (Warren, 1959). At this time, the content was expressed as% by weight with respect to the sample of the antler.
다) 글리코사미노글리칸 (Glycosaminnoglycan)류의 분석: 우론산 (Uronic acid)에서 papein으로 분해시킨 것을 Dimethylmethylene blue dye binding 방법에 의해 540㎚에서 측정함. 이때 함량은 녹용탕 시료에 대한 %로 표기하였다.C) Analysis of Glycosaminoglycan: The degradation of papein in uronic acid was measured at 540 nm by the method of dimethymeethylene blue dye binding. At this time, the content was expressed as% with respect to the antler sample.
라) 총 당량 분석: 샘플 A, B 그리고 C용액을 200배와 400배로 희석시켰다. glucose standard 100㎍/㎖와 Fructose standard 100㎍/㎖를 0, 10, 20, 30, 40, 50의 농도로 준비하고 5% 황산과 95.5%의 H2SO4를 준비하였다. standard 용액을 테스트 튜브에 2㎖ 넣고 페놀용액을 1㎖ 첨가한 후 서둘러 황산을 5㎖ 넣고 튜브를 voltexing 하여 10분간 방치하고 물속에 15분간 넣어두고, 준비된 standard용액을 microplate에 200uL씩 나눠 담은 후, 산소와 반응하기 전에 absorbance를 측정하였다. 이 때 standard의 absorbance결과는 0.97이상이 되어야 하며, 0.97이상이 되면 같은 방법으로 샘플 용액을 실험하였다.Total equivalence analysis: Samples A, B and C were diluted 200 and 400 times. glucose standard 100 μg / ml and fructose standard 100 μg / ml were prepared at concentrations of 0, 10, 20, 30, 40 and 50, and 5% sulfuric acid and 95.5% of H 2 SO 4 were prepared. standard solution is added to 2 ml of test tube, 1 ml of phenol solution is added, then 5 ml of sulfuric acid is added in a hurry, the tube is vortexed and left for 10 minutes, placed in water for 15 minutes, Absorbance was measured before reacting with oxygen. At this time, the absorbance of the standard should be more than 0.97, and when it is more than 0.97, the sample solution was tested in the same way.
마) 콜라겐 (Collagen) 분석: Bergman and Loxley (1963)를 사용하여 Hydroxyproline량으로 정량하였다. 시료 0.1g을 screw cap tube에 넣고 6N염산 10㎖을 첨가하여 110℃에서 24시간 이상 가수분해 한 후, 가수분해 된 용액을 감압 농축하여 염산을 완전히 제거하고 구연산/초산완충용액 100㎖을 넣어 syringe filter로 filtering을 한 후 구연산/초산완충용액 900μg에 시료 100㎕를 희석하여 Hydroxyprolin량 측정시료로 사용하였다. 측정용 시료 300μl와 isopropanol 600μl을 혼합한 후 산화용액 300μl 첨가하여 vortexing 후 4분간 정지시켜 Ehrlich's시약 4㎖을 첨가하여 60℃에서 25분간 가열 처리한 후 흐르는 물에 2-3분간 냉각함. 냉각시킨 후 4시간 이내에 558nm에서 비색 정량한 후, 정량된 Hydroxyproline 함량에서 collagen계수 7.52를 곱하여 collagen량으로 환산하였다.Collagen analysis: Bergman and Loxley (1963) were used to quantify the amount of hydroxyproline. Add 0.1 g of the sample to a screw cap tube, add 10 ml of 6N hydrochloric acid, and hydrolyze at 110 ° C for more than 24 hours. The hydrolyzed solution is concentrated under reduced pressure to remove hydrochloric acid completely and 100 ml of citric acid / After filtering, 100 μl of the sample was diluted in 900 μg of citric acid / acetic acid buffer solution and used as a sample for measuring the amount of hydroxyproline. 300 μl of the sample for measurement and 300 μl of isopropanol are added, and 300 μl of the oxidizing solution is added. After vortexing, 4 ml of Ehrlich's reagent is added, and the sample is heated at 60 ° C. for 25 minutes and then cooled in flowing water for 2-3 minutes. After cooling, colorimetric determination was made at 558 nm within 4 hours, and the amount of collagen was calculated by multiplying collagen coefficient 7.52 by quantified hydroxyproline content.
바) 아미노산 (Amino acid) 분석: 시료는 산안정 전처리를 실시하였다. 즉, 약 0.08g을 칭량한 후, 6N HCl 10ml을 넣고 혼합하여 질소로 치환한 후, 밀봉하여 110℃에서 24시간 동안 가수분해시켰다. 이 가수분해물을 여과 (0.45 nm syringe fiter, whatman)한 후, 55~60℃에서 감압 여과한 다음 0.2N sodium citrate buffer (pH 2.2)로 10ml(Y)가 되게 정용한다. 기기분석은 아미노산 자동분석기 (L-8500A, Hitachi) 이용하여 캡슐에 20㎍(Z)를 주입하여 시료자동주입기에 주입하여 Ninhydrin법으로 440nm와 570nm로 분석하며 계산식은 다음과 같다.F) Amino acid analysis: The samples were subjected to acid stabilization pretreatment. That is, about 0.08 g was weighed, and 10 ml of 6N HCl was added thereto, followed by mixing and replacing with nitrogen, followed by sealing and hydrolysis at 110 ° C for 24 hours. The hydrolyzate is filtered (0.45 nm syringe fiter, whatman), filtered under reduced pressure at 55 to 60 ° C, and adjusted to 10 ml (Y) with 0.2 N sodium citrate buffer (pH 2.2). Instrumental analysis was performed using an automatic amino acid analyzer (L-8500A, Hitachi), injected into a capsule with 20 μg (Z), injected into an automatic injector and analyzed by Ninhydrin method at 440 nm and 570 nm.
Amino acid content(%)=M*B/1,000,000*100Amino acid content (%) = M * B / 1,000,000 * 100
M(ng)=A 10(cystine 5)*MW(Mol.Wt)M (ng) = A10 (cystine 5) * MW (Mol.Wt)
A(area)=sample area/standard areaA (area) = sample area / standard area
B(희석배수)=(1,000/X)*(Y/Z)B (dilution factor) = (1,000 / X) * (Y / Z)
Z(ml)=(주입량㎕(Z)/희석배수(B))/1,000
Z (ml) = (injection amount? (Z) / dilution multiple (B)) / 1,000
3. 녹용 가공 소재별 (녹용 분말 및 녹용 농축액) 항산화능 분석: DPPH, hydroxyl, alkyl 및 superoxide radical 소거활성3. Analysis of antioxidant activity of antler seeds (antler grains and antler extract concentrates): DPPH, hydroxyl, alkyl and superoxide radical scavenging activity
가) DPPH 라디칼 소거활성분석: DPPH 라디칼 소거활성은 Nanjo 등 (1996)에 의한 방법으로 측정하였다. 각각의 녹용탕 추출물에 60ul의 에탄올 용액, 60ul의 DPPH 용액 (0.06 mM)을 혼합한 후 그 중 50ul를 100ul의 Teflon 모세관에 옮겨 담아 ESR spectrophotometer 분석에 사용하였다. (측정조건 : central field, 3475g ; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 5mW; gain, 6.3×105 ; temperature, 298 K)A) Analysis of DPPH radical scavenging activity: DPPH radical scavenging activity was measured by Nanjo et al. (1996). 60ul of ethanol solution and 60ul of DPPH solution (0.06mM) were mixed with each antler extract and 50ul was transferred into 100ul Teflon capillary tube and used for ESR spectrophotometer analysis. (Measurement condition: central field, 3475 g; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 5 mW; gain, 6.3 × 10 5 ;
나) Hydroxyl 라디칼 소거활성분석: Hydroxyl 라디칼 소거활성은 Haber-Weiss의 촉매 이온 방법을 사용하였다. 5,5-Dimethyl-1-pyrroline-N-oxide(DMPO) 스핀 트랩을 이용하여 hydroxyl 라디칼을 발생시키고, 전자스핀공명 기기(electron spin resonance)를 사용하여 분석하였다. 각 효소에 의한 가수분해물 (0.2ml)을 0.2ml의 DMPO(0.3M), 0.2ml의 FeSO4 (10mM) 그리고 0.2ml의 H2O2(10mM)를 phosphate 완충용액(pH 7.2)과 혼합하여 Teflon 모세관에 100ul를 옮긴 다음 2분 30초 후에 전자스핀공명 기기를 이용하여 측정하였다. (측정조건: central field, 3475 g; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 1mW; gain 6.3×105 및 temperature, 298 K)B) Hydroxyl radical scavenging activity analysis: Hydroxyl radical scavenging activity was measured by Haber-Weiss's catalytic ion method. Hydroxyl radicals were generated using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) spin trap and analyzed using electron spin resonance. The hydrolyzate (0.2 ml) of each enzyme was mixed with 0.2 ml of DMPO (0.3 M), 0.2 ml of FeSO 4 (10 mM) and 0.2 ml of H 2 O 2 (10 mM) in
다) Alkyl 라디칼 소거활성분석: Alkyl 라디칼은 AAPH 처리에 의해 발생시켰다. 10mmol/L의 AAPH 10mmol/L의 4-POBN 그리고 적정농도의 시료를 phosphate- bufferedsaline (PBS, pH 7.4)와 혼합하여 항온수조에서 37℃로 30분 동안 반응시킨 후, Telflon capillary tube 에 100ul를 옮겨 담아 ESR spectrophotometer 분석에 사용하였다. (측정조건 : central field, 3475 g; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 10mW; gain, 6.3×105; temperature, 298 K)C) Analysis of Alkyl radical scavenging activity: Alkyl radicals were generated by AAPH treatment. After 10 mmol /
라) Superoxide 라디칼 소거활성분석: Superoxide 라디칼은 자외선 조사된 리보플라빈/EDTA 계에 의해 측정하였다. 365nm 의 자외선에서 1분 동안 조사된 각기 다른 농도의 샘플과 0.3mM의 리보플라빈, 5.0 mM의 EDTA, 0.1M의 DMPO를 혼합하여 반응시켰다. 반응물을 Telflon capillary tube에 100ul 를 옮겨 담아 ESR spectrophotometer 분석에 사용하였다. (측정조건 ; central field, 3475 g; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 4mW; gain, 6.3×105; temperature, 298 K)
D) Superoxide radical scavenging activity: Superoxide radicals were measured by ultraviolet irradiated riboflavin / EDTA system. The samples were incubated for 1 min at 365 nm in ultraviolet light and 0.3 mM riboflavin, 5.0 mM EDTA, and 0.1M DMPO were mixed and reacted. The reactants were transferred to a Telflon capillary tube and used for ESR spectrophotometer analysis. (Measurement condition: central field, 3475 g; modulation frequency, 100 kHz; modulation amplitude, 2 g; microwave power, 4 mW; gain, 6.3 x 10 5 ;
4. 녹용의 유효성분 손실을 최소화시킬 수 있는 최적의 녹용 소재제형 및 기능성 향상을 위한 부원료 배합비율 확립하였다.
4. Formulation of optimal antler material to minimize the loss of effective ingredient of antler and the ratio of sub ingredient to improve functionality were established.
II. 결과II. result
1. 녹용을 이용한 식품소재 (녹용 분말 및 녹용 농축액)의 제조1. Manufacture of food ingredients (deer antler and deer antler) using antler
(1) 엘크 녹용의 부위별 일반 성분(1) general components of elk antler
엘크 녹용의 각 부위별 (상대, 중대, 하대) 영양성분을 분석한 결과는 표 1과 같다. 각 부위의 건물 함량은 27~38%의 범위를 보였다. 조단백, 조지방 및 총 당 함량의 경우 상대, 중대, 하대의 순으로 낮아지는 결과를 보였다. 반면 회분의 경우 하대가 37.6%로 가장 높은 값을 보였다. 또한 회분의 증가와 동일하게 콜라겐, 칼슘과 인의 값도 하대로 갈수록 증가하는 결과를 보였다. 이와는 반대로 녹용의 생리활성물질로 알려져 있는 Glycosaminoglycan, uronc acid 및 sialic acid의 함량은 상대로 갈수록 뚜렷하게 증가하는 결과를 보였다.Table 1 shows the results of nutrient analysis for each part of elk antler (relative, major, and lower). The building content of each site was in the range of 27 ~ 38%. Crude protein, crude fat and total sugar content were lower in order of relative, major, and inferior. On the other hand, in the case of ash, the lowest value was 37.6%. In addition, the values of collagen, calcium and phosphorus increased as the ash increased. On the other hand, the contents of Glycosaminoglycan, uronc acid and sialic acid, which are known as physiologically active substances of antler, show a marked increase.
acid (mg/mL)Uronic
acid (mg / mL)
acid (mg/mL)Sialic
acid (mg / mL)
(g/mL)Ca
(g / mL)
(g/mL)P
(g / mL)
(2) 엘크 녹용의 부위별 아미노산 조성(2) Amino acid composition of each part of elk antler
녹용의 부위를 달리하여 제조한 녹용 농축액의 아미노산 조성 분석 결과는 표 2와 도 1에 나타내었다. 녹용 농축액 내 유리 아미노산들 가운데 특히 proline과 glycine의 경우는 녹용 중대나 하대를 첨가한 경우 다소 증가하는 경향을 보였다. 일반적으로 녹용 내의 collagen은 Glycine-Proline-Hydroxyproline 구조로 되어 있으며 상대나 중대에 비해 하대에 collagen 함량이 높았던 결과와 일맥상통함을 확인할 수 있었다.Table 2 and FIG. 1 show the analysis results of the amino acid composition of the antler range concentrate prepared by varying the antler part. Especially proline and glycine among the free amino acids in the antler juice showed a tendency to increase with the addition of the antler of the antler. Generally, the collagen in the antler is composed of Glycine-Proline-Hydroxyproline structure, which is consistent with the result of the collagen content in the lower part of the collagen compared to the other collagen.
(3) 엘크 녹용의 단백질 및 탄수화물 가수분해 효소 추출물의 제조공정(3) Production process of protein and carbohydrate hydrolase of elk antler
본 발명에서는 단백질 및 탄수화물 가수분해 효소를 이용하여 엘크 녹용의 각 부위별 효소적 가수분해 추출물을 제조하였으며, 본 발명에 사용한 효소 및 가수분해 공정은 표 3 및 도 2와 같다.In the present invention, an enzymatic hydrolysis extract of each part of elk antler was prepared using protein and carbohydrate hydrolase, and the enzymes and the hydrolysis process used in the present invention are shown in Table 3 and FIG.
xylanase, pentosanase, arabanase-glucanase, cellulase
xylanase, pentosanase, arabinase
hemicellulase, zylanase, arabanase-glucanase, cellulase
hemicellulase, zylanase, arabinase
(4) 엘크 녹용의 단백질 및 탄수화물 가수분해 효소 추출물의 수율(4) Yield of protein and carbohydrate hydrolase extracts of elk antler
본 발명에서는 단백질 및 탄수화물 가수분해 효소를 이용하여 엘크 녹용의 각 부위별 효소적 가수분해 추출물을 제조하였으며, 각 부위별 추출물의 수율은 표 4에 나타내었다.In the present invention, an enzymatic hydrolysis extract of each part of elk antler was prepared using protein and carbohydrate hydrolase, and the yield of each extract was shown in Table 4.
모든 가수분해 추출물에 있어서 전반적으로 상대의 추출 수율이 가장 높게 나타났으며, 이후 중대, 하대의 순으로 나타났다. 효소가수분해 추출물과 열수 추출 및 에탄올 추출물의 수율은 큰 차이를 보이지 않았으며 효소가수분해 추출물의 경우, 상대는 11.27~26.67%, 중대의 경우 6.49~17.38%, 하대의 경우 4.52~11.28%의 수율 범위를 보였다. 녹용의 분획별 가수분해도의 차이를 나타내는 것은 녹용의 조단백질의 함량이 상대가 높고 하대가 낮은 경향이 있기 때문인 것으로 판단된다. 또한, 효소는 기질과 결합하여 기질을 생성물로 전환시키는 활성부위(active site)를 가지고 있는데, 보통 활성부위는 효소분자의 외부에 갈라진 틈을 형성하고 있으므로 기질의 구조와 상보적인 형태로 되어 있다. 효소별 가수분해도의 차이는 이와 같은 효소의 단백질에 대한 선택적 기질특이성에 따른 것으로 판단된다. The extraction yields of all hydrolyzed extracts were the highest in all, followed by the order of major and minor. The yields of hydrolyzed and hydrolyzed hydrolyzed extracts were not significantly different from those of hydrolyzed hydrolyzed hydrolysates. The hydrolyzed hydrolyzate yields were 11.27 ~ 26.67%, 6.49 ~ 17.38%, and 4.52 ~ 11.28% Range. The difference in the degree of hydrolysis of the fractions of antler is believed to be due to the high relative content of crude protein and the low tendency of antler. In addition, the enzyme has an active site that binds to the substrate to convert the substrate to the product. Usually, the active site forms a gap in the outer part of the enzyme molecule, and thus has a complementary form to the structure of the substrate. The difference in the degree of hydrolysis by enzymes is considered to be due to the selective substrate specificity of these enzymes.
(5) 엘크 녹용의 단백질 및 탄수화물 가수분해 효소추출물의 생리활성성분(5) Physiologically active components of protein and carbohydrate hydrolase of elk antler
엘크 녹용의 단백질 및 탄수화물 가수분해 효소 추출물의 생리활성 성분 결과는 표 5~9 및 도 4~8에 나타내었다. 엘크 녹용의 다양한 효소가수 분해물 중 uronic acid 함량의 경우 전반적으로 하대에 비해 상대와 중대에서 높은 함량을 보였으며 특히 녹용 중대의 Termamyl과 Protamex 가수분해물에서 가장 높은 값을 보였다 (표 5, 도. 4). 녹용 효소 가수분해 추출물에서 Sialic acid 함량의 경우 대부분 상대에서 가장 높은 값을 보였으며, 특히 단백질 가수분해 효소인 Flavourzyme 및 탄수화물 가수분해 효소인 Ultroflo 추출물에서 가장 높은 값을 보였다 (표 6, 도. 5).The results of physiologically active components of protein and carbohydrate hydrolase extracts of elk antler are shown in Tables 5 to 9 and FIGS. 4 to 8. The uronic acid content of the various enzymatic hydrolysates of elk antler was higher in the opponent and medium-sized group than in the lower group, and the highest value was observed in the termamyl and protamex hydrolysates of the antler group (Table 5, Fig. 4) . Sialic acid content was the highest in the relative antioxidant extract, especially in the Flavourzyme protein hydrolytic enzyme and Ultroflo carbohydrate hydrolyzate (Table 6, Fig. 5) .
표 7과 도. 6에서 보는 바와 같이 Glycosaminoglycan 함량의 경우도 상대 및 중대 추출물이 하대 추출물에 비해 높은 함량을 보였으며, 특히 열수 추출물과 Viscozyme 추출물을 비롯하여 대부분의 단백질 가수분해 효소 추출물 (Alcalase, Neutrase, Flavourzyme, Protamex)에서 높은 함량을 나타내었다. 반면 총 당 함량은 전반적으로 단백질 가수 분해 추출물에 비해 탄수화물 가수분해 추출물 (AMG, Termamyl, Ultroflo)에서 높은 값을 보였다 (표 8, 도. 7). 한편 단백질 가수분해효소인 Flavourzyme과 pepsin 가수분해물의 경우 중대 및 하대에서 높은 함량을 보였다. Collagen 함량의 경우는 전반적으로 상대에서 하대로 갈수록 증가하였으며, 특히 탄수화물 가수분해 효소인 Cellucalst, AMG 추출물과 단백질 가수분해 효소인 Flavourzyme, Alcalase 및 Neutrase 추출물의 중대와 하대 추출물에서 높은 함량을 보였다(표 9, 도. 8).Table 7 also. As shown in Fig. 6, the content of Glycosaminoglycan was higher than that of the other extracts. Especially, most of the protein hydrolytic enzyme extracts (Alcalase, Neutrase, Flavourzyme, Protamex) including hot water extract and Viscozyme extract Respectively. On the other hand, total sugar content was higher in carbohydrate hydrolyzed extract (AMG, Termamyl, Ultroflo) than protein hydrolyzed extract (Table 8, Fig. On the other hand, the content of hydrolysates of Flavourzyme and pepsin, which are protein hydrolytic enzymes, was high in medium and low. The content of collagen was increased from the other to the lower part. Especially, the contents of Cellucalst, AMG extract and protein hydrolytic enzymes Flavourzyme, Alcalase and Neutrase extracts were high in carbohydrate hydrolysis enzymes (Table 9 , Fig. 8).
(6) 엘크 녹용의 단백질 및 탄수화물 가수분해 효소 추출물의 라디칼 소거활성(6) Radical scavenging activity of protein and carbohydrate hydrolase of elk antler
녹용 효소 가수분해 추출물의 항산화 활성은 ESR spectrophotometer를 사용하여 분석하였으며, 그 결과는 표 10 및 도 9~11에 나타내었다. 녹용의 효소 가수분해물별 항산화 활성은 DPPH, hydroxyl 및 alkyl radical scavenging activity를 분석하여 각 radical에 대해 50%의 radical 억제하는 농도인 IC50 (mg/mL)로 나타내었다.The antioxidative activity of hydrolyzate of antler starch was analyzed by ESR spectrophotometer. The results are shown in Table 10 and FIGS. 9 to 11. Antioxidative activities of antler starch hydrolysates were analyzed by DPPH, hydroxyl and alkyl radical scavenging activity and expressed as IC 50 (mg / mL), which is a radical inhibitory concentration of 50% for each radical.
모든 radical에 대한 항산화효능은 상대가 가장 높았으며 중대, 및 하대로 갈수록 낮아지는 결과를 보였다. 또한 모든 radical scavenging activity는 농도에 비례하여 증가하였다. 특히 DPPH radical scavenging activity는 녹용 상대에서 protamex (0.96 mg/mL), Flavourzyme (1.28 mg/mL), alcalase (1.71 mg/mL) 및 termamyl (1.77 mg/mL) 추출물의 항산화 효능이 우수한 것으로 나타났다. The antioxidative effect of all radicals was the highest in the relative group, and the antioxidative effect was lower in the major and the lower group. In addition, all radical scavenging activity increased in proportion to the concentration. Especially, DPPH radical scavenging activity showed antioxidant activity of protamex (0.96 mg / mL), Flavourzyme (1.28 mg / mL), alcalase (1.71 mg / mL) and termamyl (1.77 mg / mL)
DPPH는 짙은 자색을 띄는 비교적 안정한 프리 라디칼로서 다양한 천연소재로부터 항산화 물질을 검색하는데 많이 사용되고 있다. 자유 라디칼은 인체 내에서 지질 또는 단백질 등과 결합하여 노화를 일으키기 쉬운데 페놀성 화합물의 경우 프리 라디칼을 환원시키거나 상쇄시키는 능력이 강해 인체 내에서 프리 라디칼에 의한 노화를 억제하는 척도로 이용할 수 있다.DPPH is a relatively stable free radical with a deep purple color and is widely used to search for antioxidants from various natural materials. Free radicals are likely to cause aging by binding to lipids or proteins in the body. In the case of phenolic compounds, they have a strong ability to reduce or offset free radicals, which can be used as a measure to inhibit free radical aging in the body.
Hydroxyl 라디칼은 DMPO 트랩을 이용하여 Fe2+/H2O2 계에서 발생되어졌다. DMPO-OH의 정형적인 1:2:2:1 ESR 시그널이 관찰되었다. Hydroxyl radical에 대한 항산화 효능도 DPPH radical scavenging activity과 유사한 결과를 보였는데, Flavourzyme (0.89 mg/mL), protamex (1.37 mg/mL), pepsin (1.63 mg/mL), celluclast (1.95 mg/mL) 및 viscozyme (1.95 mg/mL) 추출물 순으로 hydroxyl radical scavenging activity가 큰 것으로 나타났다.Hydroxyl radicals were generated in the Fe 2+ / H 2 O 2 system using DMPO trap. A typical 1: 2: 2: 1 ESR signal of DMPO-OH was observed. Antioxidant efficacy against hydroxyl radicals was similar to DPPH radical scavenging activity. Flavourzyme (0.89 mg / mL), protamex (1.37 mg / mL), pepsin (1.63 mg / mL), celluclast hydroxyl radical scavenging activity was higher in the order of viscozyme (1.95 mg / mL).
Alkyl 라디칼은 37℃에서 30분 동안 AAPH 처리된 4-POBN/자유기로부터 측정되었다. Alkyl radical 소거활성의 경우도 녹용 단백질 가수분해물 중 상대가 가장 활성이 크고 중대, 상대로 내려올수록 활성이 작아지는 결과를 보였다. 효소 추출물별 alkyl radical scavenging activity는 녹용 상대에서 protamex (0.37 mg/mL), pepsin (0.42 mg/mL), alcalase (0.46mg/mL), viscozyme (0.79 mg/mL), termamyl (1.34 mg/mL), Neutrase (1.55 mg/mL) 및 Flavourzyme (1.64 mg/mL) 추출물의 항산화 효능이 우수한 것으로 나타났다.The Alkyl radical was measured from the 4-POBN / freezer treated with AAPH for 30 min at 37 < 0 > C. In the case of Alkyl radical scavenging activity, the relative activity of the antler protein hydrolyzate was the most active and the activity was decreased as the activity decreased. The alkyl radical scavenging activities of enzyme extracts were similar to those of protamex (0.37 mg / mL), pepsin (0.42 mg / mL), alcalase (0.46 mg / mL), viscozyme (0.79 mg / mL) , Neutrase (1.55 mg / mL) and Flavourzyme (1.64 mg / mL) were superior in antioxidant activity.
이러한 결과들로 미루어 녹용 단백질 가수분해물의 항산화능은 효소의 단백질에 대한 선택적 절단특이성에 의해 생성된 펩타이드의 종류에 따라 다를 것으로 판단된다. 단백질 가수분해물의 항산화능은 구성 펩타이드의 성상에 따라 차이가 있으며, 이는 가수분해에 의하여 생성된 펩타이드의 말단 아미노산에 의한 펩타이드의 입체구조 변화 및 분자량과 관련이 있다고 사료된다.
These results suggest that the antioxidant capacity of the antler protein hydrolyzate may be different depending on the type of peptide produced by the selective cleavage of the enzyme to the protein. The antioxidant capacity of the protein hydrolyzate varies depending on the characteristics of the constituent peptide, which is considered to be related to the change in the stereostructure and the molecular weight of the peptide due to the terminal amino acid of the peptide produced by hydrolysis.
2. 녹용의 유효성분 손실을 최소화시킬 수 있는 최적의 녹용 소재제형 및 기능성 향상을 위한 부원료 배합비율 확립2. Formulation of the best antler material to minimize the loss of effective ingredient of antler and establishment of the ratio of additives to improve functionality
본 발명에서 부원료로 활용하기 위한 식물 추출물의 DPPH radical 소거활성을 분석한 결과는 도 12와 같다. 즉 본 발명에서는 녹용을 이용한 다양한 가공제품 개발에 활용하고자 각 종 문헌을 참고로 하여 항산화성이 큰 것으로 알려져 있는 10종의 한약재를 선택한 후 각 식물 추출물의 항산화 활성을 측정하기 위해 electron spin resonance (ESR) spectrometer를 이용하여 DPPH radical 소거활성을 측정하였다. DPPH radical 소거활성을 측정하여 각각의 IC50 (mg/ml) 값을 비교해 본 결과, 각 식물추출물의 DPPH radical에 대한 IC50 값은 0.12~3.87 mg/ml의 범위를 보였으며, 그 중 백작약의 IC50 값이 0.12 mg/ml로 가장 큰 항산화능을 보였으며, 천궁, 당귀, 감초, 숙지황의 순서로 항산화기능이 뛰어난 것으로 나타났다. 반면 황기의 IC50 값이 3.87 mg/ml로 항산화능이 가장 적은 결과를 보였으며, 대추의 경우에는 항산화능이 거의 없는 것으로 나타났다.The results of analyzing the DPPH radical scavenging activity of the plant extract for use as an additive in the present invention are shown in FIG. In other words, in order to utilize in the development of various processed products using antler, the present invention selects ten kinds of herbal medicines known to have high antioxidative properties with reference to each kind of literature, and then, in order to measure the antioxidative activity of each plant extract, ) spectrometer was used to measure DPPH radical scavenging activity. IC 50 values for the DPPH radical of the result, each plant extract, compare the respective IC 50 (mg / ml) value by measuring the DPPH radical scavenging activity showed a 0.12 ~ 3.87 mg / ml range, decursiva of the The highest antioxidant activity was obtained with IC 50 value of 0.12 mg / ml. On the other hand, the IC 50 value of Hwanggi was 3.87 mg / ml, which showed the lowest antioxidant ability. Jujube showed almost no antioxidant ability.
상기의 식물추출물의 항산화 활성 측정 결과를 바탕으로 녹용의 효소 가수분해 추출물을 이용한 기능성 제품 개발을 위한 부원료 배합비율 예를 표 11에 나타내었으며, 이를 토대로 식물혼합추출물을 제조하였다. 구체적인 실시예는 다음과 같다. Based on the results of the antioxidative activity measurement of the above plant extracts, examples of mixing ratios for developing a functional product using the enzymatic hydrolysis extract of Deer antler are shown in Table 11, and a plant mixed extract was prepared based on the results. Specific examples are as follows.
<실시예 1>≪ Example 1 >
1) 인삼 14.9 중량%, 백출 15.1 중량%, 백복령 15.2 중량%, 당귀 4.2 중량%, 천궁 2.5 중량%, 숙지황 7.8 중량%, 백작약 0.5 중량%, 황기 16.9 중량%, 육계 16.6 중량%, 감초 6.2 중량%이 되도록 식물을 혼합한 후, 식물혼합물과 동량의 물을 섞는다. 1) ginseng 14.9% by weight, white 15.1% by weight, white bell 15.2% by weight, Angelica gigantis 4.2% by weight, Angelica gigantis 2.5% by weight, %, Then mix the plant mixture with an equal volume of water.
2) 85에서 2~3시간 끓여 고형분 15 중량% 이상이 될 때까지 농축 추출하여 식물혼합추출물을 제조한다.2) boil for 2 to 3 hours at 85 to concentrate and extract until the solid content reaches 15% by weight or more to prepare plant mixed extract.
3) 상기 식물혼합추출물을 발하고자 하는 제품의 제형에 맞게 농축액 (액상) 또는 동결 건조하여 분말형태로 제조한다.
3) The plant mixture extract is prepared in powder form by concentrating (liquid) or freeze-drying according to the formulation of the product to be extracted.
<실시예 2>≪ Example 2 >
(액상 혹은 동결건조 시료)Hydrolyzed extract of antler
(Liquid or freeze-dried sample)
1) 식물혼합추출물 (인삼 14.9 중량%, 백출 15.1 중량%, 백복령 15.2 중량%, 당귀 4.2 중량%, 천궁 2.5 중량%, 숙지황 7.8 중량%, 백작약 0.5 중량%, 황기 16.9 중량%, 육계 16.6 중량%, 감초 6.2 중량%)1) A mixture of plant extracts (14.9% by weight of ginseng, 15.1% by weight of white ginseng, 15.2% by weight of white ginseng, 4.2% by weight of angelica ginseng, 2.5% by weight of ginseng root, 7.8% by weight of sorghum, 0.5% , 6.2% by weight licorice)
2) 각 원료는 식품공전 및 첨가물 공전상의 적합한 원료를 구입하여 사용하였다. 검수된 적합한 원료를 배합비율에 맞게 칭량하였다.2) Each raw material was purchased by using suitable raw materials for food revolution and additives. A suitable raw material that was inspected was weighed to the blend ratio.
3) 인삼 14.9 중량%, 백출 15.1 중량%, 백복령 15.2 중량%, 당귀 4.2 중량%, 천궁 2.5 중량%, 숙지황 7.8 중량%, 백작약 0.5 중량%, 황기 16.9 중량%, 육계 16.6 중량%, 감초 6.2 중량%을 배합비율에 맞게 혼합추출기에 물과 함께 넣은 후 고형분이 15중량% 이상이 될 때까지 농축, 추출하여 식물혼합추출물을 제조하였다.3) Ginseng 14.9% by weight, White 15.1% by weight, Baekbok 15% by weight, Angelica gangrene 4.2% by weight, Angelica giganteum 2.5% by weight, Sulfur sulfur 7.8% by weight, % Were mixed with water in a mixing extractor in accordance with the blending ratio, and concentrated and extracted until the solid content exceeded 15% by weight to prepare a plant mixed extract.
4) 식물혼합추출물과 녹용 효소가수분해 추출물 및 기타 버섯추출액을 배합비율에 맞게 혼합, 농축기에 넣고 85로 고형분이 10중량%가 될 때까지 농축하였다.4) Plant mixture extract, hydrolyzed hydrolyzate of antler, and other mushroom extract were mixed in a mixing ratio, and the mixture was put into a concentrator and concentrated to 85% by weight to obtain a solid content of 10% by weight.
5) 4)의 농축액에 자몽종자추출물을 넣고 85에서 고형분이 최종 8중량%가 될 때까지 농축, 여과한 후 90에서 1시간 동안 살균처리 하여 최종 제품개발을 위한 원료로 사용하였다.5) The grapefruit seed extract was added to the concentrate of 4), and the solid content was concentrated to 85% by weight, filtered and sterilized at 90 for 1 hour to be used as a raw material for final product development.
6) 최종 원료는 개발하고자 하는 제품의 제형에 맞게 농축액 (액상) 혹은 동결 건조하여 사용하였다.
6) The final raw material was used as a concentrate (liquid) or lyophilized according to the formulation of the product to be developed.
본 발명에 따른 녹용의 고생리활성성분 함유 가수분해 효소추출물의 제조방법을 통하여 녹용 추출물을 제조함으로써, 고가의 녹용에서 생리활성 성분의 추출을 배가시키고 이를 식품제조에 이용함으로써 현대인의 건강증진과 식품산업에 기여할 수 있으므로 산업상 이용가능성이 있다.The present invention relates to a method for preparing a hydrolytic enzyme-containing extract of deer antler according to the present invention, which enables to produce a deer antler extract, It can be used industrially because it can contribute to the industry.
Claims (9)
상기 (1)단계의 녹용농축액에 Celluclast, Termamyl, AMG, Ultraflo, Viscozyme으로 이루어진 군으로부터 선택되는 하나 이상의 가수분해효소를 질량비 100:1로 혼합하여, 24시간 반응시키는 단계(2);
상기 (2)단계의 가수분해물을 여과하고, 효소활성을 제거한 후, pH를 7로 맞추어, 동결건조시키는 단계(3)로 이루어진 것을 특징으로 하는 녹용의 고생리활성성분 함유 가수분해 효소추출물의 제조방법.
(1) mixing the mixture of green, middle, and lower green, or a mixture thereof with water at a weight ratio of 1: 4, boiling at 95 ° C for 7 hours, and concentrating under reduced pressure to obtain a deer concentration of 60 to 70 ° Brix;
(2) mixing at least one hydrolytic enzyme selected from the group consisting of Celluclast, Termamyl, AMG, Ultraflo and Viscozyme in a mass ratio of 100: 1 to the deer antler of step (1) for 24 hours;
And (3) filtering the hydrolyzate of step (2), removing the enzyme activity, and adjusting the pH to 7, followed by lyophilization, to prepare a hydrolytic enzyme-containing extract of deer antler containing active ingredient Way.
나) 인삼 14.9 중량%, 백출 15.1 중량%, 백복령 15.2 중량%, 당귀 4.2 중량%, 천궁 2.5 중량%, 숙지황 7.8 중량%, 백작약 0.5 중량%, 황기 16.9 중량%, 육계 16.6 중량%, 감초 6.2 중량%의 식물혼합물을 물과 동일 중량으로 혼합 후, 85℃에서 고형분이 15 중량%가 될 때까지 농축시킨 식물혼합추출물을 제조하는 단계;
다) 상기 가)단계의 가수분해 효소추출물과 상기 나)단계의 식물혼합추출물을 혼합하는 단계로 이루어지는 것을 특징으로 하는 녹용의 고생리활성 성분 함유 가수분해 효소추출물의 제조방법.
(A) preparing a hydrolytic enzyme-containing extract containing a hyperactive component of antler according to (1);
B) 16.9% by weight of white ginseng, 16.6% by weight of white ginseng, 6.2% by weight of licorice, 6.2% by weight of white ginseng, 15.1% by weight of white ginseng, 15.2% by weight of white ginseng, % Of the plant mixture is mixed with water at the same weight, and then concentrated at 85 캜 until the solid content reaches 15% by weight.
(C) mixing the hydrolytic enzyme extract of step (a) and the plant mixed extract of step (b) above to prepare a hydrolytic enzyme-containing extract of deer antler.
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