KR101409761B1 - A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor - Google Patents

A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor Download PDF

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KR101409761B1
KR101409761B1 KR1020120066147A KR20120066147A KR101409761B1 KR 101409761 B1 KR101409761 B1 KR 101409761B1 KR 1020120066147 A KR1020120066147 A KR 1020120066147A KR 20120066147 A KR20120066147 A KR 20120066147A KR 101409761 B1 KR101409761 B1 KR 101409761B1
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red ginseng
fermented red
enzyme
concentrate
fermented
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KR20130142707A (en
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최일동
라제현
윤병훈
박현준
허철성
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주식회사한국야쿠르트
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Priority to JP2015518310A priority patent/JP5917772B2/en
Priority to PCT/KR2012/007053 priority patent/WO2014007427A1/en
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Abstract

본 발명은 홍삼을 효소전환과 유산균발효를 이용한 발효홍삼농축액의 제조방법에 관한 것으로서 4년 홍미삼 주정 추출물과 식용가능효소인 Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL 및 Rapidase C80Max 중 3종의 효소를 사용하며, 김치로부터 분리한 유산균 Lactobacillus sakei HY7802를 이용하여 발효하여 홍삼으로부터 소화율 및 흡수율이 증대된 화합물 K가 강화된 발효홍삼 농축액을 얻을 수 있다.The present invention relates to a method for preparing fermented red ginseng concentrate by converting red ginseng into an enzyme and fermenting lactic acid bacteria. The present invention relates to a method for producing fermented red ginseng concentrate using red ginseng extract, , And fermented red ginseng concentrate with fermentation of compound K enhanced fermentation efficiency and absorption rate from red ginseng can be obtained using lactic acid bacteria Lactobacillus sakei HY7802 isolated from kimchi.

Description

효소전환과 유산균 발효를 이용한 화합물 Ｋ의 함량이 강화된 발효홍삼 농축액의 제조방법 및 그 제조방법에 의해 제조된 발효홍삼 농축액을 유효성분으로 함유하는 제품{A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor}TECHNICAL FIELD The present invention relates to a method for preparing a fermented red ginseng concentrate having enhanced content of compound K using enzyme conversion and lactic acid fermentation, and a fermented red ginseng concentrate prepared by the method, enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng product as effective factor}

본 발명은 효소전환과 유산균 발효를 이용한 화합물 K의 함량이 강화된 발효홍삼 농축액의 제조방법 및 그 제조방법에 의해 제조된 발효홍삼 농축액을 유효성분으로 함유하는 제품에 관한 것으로서, 보다 상세하게는 홍미삼을 주정으로 추출하고 그 추출물을 자체 선정한 식용 가능한 복합효소로 전환한 후, 김치에서 분리한 유산균을 접종, 발효시킴으로서 다양한 기능의 생체유효성이 높은 화합물 K의 함량이 강화된 발효홍삼 농축액의 제조방법 및 그 제조방법에 의해 제조된 발효홍삼 농축액을 유효성분으로 함유하는 제품에 관한 것이다.
The present invention relates to a method for preparing fermented red ginseng concentrate having enhanced content of compound K using enzyme conversion and lactic acid fermentation and a product containing the fermented red ginseng concentrate prepared by the preparation method as an active ingredient, A method for preparing fermented red ginseng concentrate having enhanced contents of various physiologically effective compounds K by inoculating and fermenting lactic acid bacteria isolated from kimchi after converting the extract into a foodable complex enzyme, And a fermented red ginseng concentrate prepared by the production method as an active ingredient.

인삼 및 홍삼의 효능성분으로 알려진 것은 사포닌(saponin)이다. 인삼 및 홍삼의 사포닌은 배당체의 일종으로 다른 식물에서 발견되는 사포닌과는 다른 특이한 화학구조를 가지고 있으며, 그 효능도 큰 차이가 난다. 인삼 및 홍삼의 사포닌은 타 식물계 사포닌과 구별하기 위해서 인삼(ginseng) 배당체(glycoside)란 의미로 '진세노사이드(Ginsenoside)'라고 부른다. 최근 분석기술의 발달에 따라 지금까지 30여종의 진세노사이드 화학구조가 밝혀졌다. 이는 서양삼 14종, 중국의 전칠삼 15종에 비해 월등히 많은 종류가 들어있다. 진세노사이드는 dammarane 골격을 갖는 배당체에 일종의 화학구조상 R1, R2, R3자리에 어떤 종류의 당이 몇 개 붙느냐에 따라 디올계(PPD)와 트리올계(PPT)의 진세노사이드 등으로 분류된다. 가공과정 중 이러한 진세노사이드에 높은 압력을 가하거나 효소 첨가, 또는 가열을 하면 당이 일부 떨어져 나가기도 하고 새로운 이중결합이 생기면서 특이한 진세노사이드가 만들어진다. 따라서 인삼의 가공방법에 따라 여러 가지 특이 진세노사이드가 함유된 제품들이 출시되고 있다.
Saponin is known to be an effective ingredient of ginseng and red ginseng. Saponin of ginseng and red ginseng is a kind of glycosides, and has a unique chemical structure different from that of saponin found in other plants, and its effect is also greatly different. Saponin of ginseng and red ginseng is called 'ginsenoside' to distinguish it from ginseng saponin. With the recent development of analytical technology, over 30 types of ginsenoside chemical structures have been identified. There are far more species than 14 kinds of Western ginseng and 15 kinds of Chinese ginseng. Ginsenosides are divided into diols (PPD) and triosyl (PPT) ginsenosides, depending on the type of glycosides attached to the glycosides of the dammarane skeleton in the chemical structure of R1, R2 and R3. When high pressure is applied to these ginsenosides, added with enzymes, or heated during processing, the sugar is partially removed and new double bonds are formed, resulting in a unique ginsenoside. Therefore, products containing various specific ginsenosides have been released according to processing methods of ginseng.

인삼 사포닌이 체내에 흡수되기 위해서는 디올계 진세노사이드는 20(S)-프로토파낙사디올 20-O-β-D-글루코피라노사이드(20(S)-protopanaxadiol 20-O-β-D-glucopyranoside, FGM 1, 이하, '화합물 K'라 함)으로 그리고 트리올계 진세노사이드는 20(S)-프로토파낙사트리올 (20(S)-protopanaxatriol, FGM 4)로 분해된 후 체내에 흡수되면서 효능을 발휘하는데 열이나 산, 압력, 소화효소 등에 의해서는 소량만 전환, 흡수될 뿐이다. 최근 연구결과에 따르면 인삼 사포닌이 이러한 최종산물로 전환할 때 장내미생물이 필요하다고 밝혀졌다.
In order for ginsenoside to be absorbed into the body, the diol-based ginsenoside is 20-O-β-D-glucopyranoside 20 (O) -protopanaxadiol 20- glucopyranoside, FGM 1, hereinafter referred to as 'compound K') and the triol ginsenoside is decomposed into 20 (S) -protopanaxatriol (20 (S) -protopanaxatriol, FGM 4) , But only a small amount is converted and absorbed by heat, acid, pressure and digestive enzymes. Recent studies have shown that intestinal microbes are needed when ginseng saponin is converted into such a final product.

경구 복용한 사포닌이 흡수되려면 장내 미생물에 의해 분해되어야 하는데 Privotella oris균이 진세노사이드를 사포닌을 대사하는 대표적 균이며 이외에 유산균 등의 여러 가지 유익균이 사포닌 대사에 관여한다. 일반인들 중 인분을 조사해 추정한 연구에서 정상인 중 30%는 사포닌을 대사할 수 있는 균이 전혀 없고, 비록 균이 있는 나머지 70% 중에도 대사 능력에 차이가 있어 디올계와 트리올계 사포닌을 모두 정상적으로 대사할 수 있는 사람은 소수에 지나지 않는다. 특히 지속적인 항암치료나 약제를 복용하는 경우에는 장내균총이 안정적이지 않으므로 사포닌 대사율이 더 떨어짐이 보고되었다. 이런 면에서 이미 외부에서 장내 미생물로 발효하여 개인의 신체 상황에 관계없이 인삼의 효능성분을 흡수할 수 있는 형태로 전환하여 누구나 효과를 볼 수 있도록 발효 홍삼을 개발하는 것은 매우 바람직하다고 볼 수 있다.
In order for saponin taken orally to be absorbed to be absorbed, it must be degraded by intestinal microorganisms. Privotella oris is a representative bacterium that metabolizes ginsenoside, and in addition, lactic acid bacteria and other beneficial bacteria participate in saponin metabolism. In a study that estimated the percentage of normal people, 30% of normal people had no bacteria capable of metabolizing saponin, and even in the remaining 70% of bacteria, there was a difference in metabolic ability, so that both diol and triol saponins were metabolized normally Only a handful of people can do it. Especially, when continuous chemotherapy or medication is used, the intestinal flora is not stable and the saponin metabolic rate is further decreased. In this regard, it is highly desirable to develop fermented red ginseng so that anyone can take advantage of it by transforming into a form capable of absorbing the effective ingredients of ginseng, regardless of the physical condition of the individual, by fermentation from the outside to intestinal microorganism.

지금까지 언급한 바와 같이 모든 인삼사포닌은 장내 미생물의 분해를 통해 최종 대사물로 대사되어야 흡수된다. 그러나 사람의 장내 미생물 균총은 균일하지도 않고 시시각각 변하는 것이어서 개인에 따라 인삼 사포닌 흡수율이 다름이 밝혀졌다. 따라서 이것을 극복할 수 있는 방법이 바로 인삼 및 홍삼을 장내 미생물로 발효시켜 직접 체내에 흡수시키는 방안이 있다. 발효 홍삼은 이러한 장내미생물의 차이로 인한 흡수와 효능의 차이를 줄이기 위해 홍삼을 미리 특유 미생물로 발효시킴으로써 흡수가 용이한 유용 사포닌으로 전환하여 개인차 민족차를 극복하여 인삼 효능의 표준화를 이룰 수 있다는 장점이 있다. 이러한 이유 때문에 국내에서 최근 발효 홍삼에 대한 제품이 출시되고 있으며, 이에 대한 효능이 연구되고 있다.
As mentioned above, all ginseng saponins are metabolized into end metabolites through degradation of intestinal microorganisms. However, the intestinal microflora of a human is not uniform and varies from time to time, so that the absorption rate of ginseng saponin differs depending on the individual. Therefore, there is a way to overcome this by directly fermenting ginseng and red ginseng as intestinal microorganisms and absorbing them directly into the body. Fermented red ginseng is fermented as a specific microorganism in advance to reduce the difference in absorption and efficacy due to differences in intestinal microorganisms, so that the fermented red ginseng can be converted into a useful saponin easily absorbed, thereby overcoming individual differences and achieving standardization of ginseng efficacy . For this reason, products for the recently fermented red ginseng have been launched in Korea, and the efficacy thereof has been studied.

발효 홍삼은 사포닌의 균형적인 대사에도 도움을 준다. 디올계 사포닌은 신체를 안정화 시키고 면역력을 높이며 지나친 것은 낮추어 정상화시키는 작용이 있다. 트리올계 사포닌의 경우 혈액순환을 좋게 하고 원기를 높이며 활력을 주는 서로 보완하며 상반되는 작용을 가지고 있다. 만약 장내 균총이 불안정하여 디올계와 트리올계 사포닌을 한 쪽만 대사시키거나 둘 다 대사시키지 못하면 사포닌 흡수에 불균형이 오고, 따라서 인삼을 복용하여도 효과가 떨어질 수 있으며 일부 사람에서는 열이 나거나 답답함 등의 불편한 경험을 하게 된다. 그러나 외부에서 디올계와 트리올계 사포닌을 균형적으로 대사시켜 신체에 흡수시켰을 경우에는 인삼 및 홍삼이 가지고 있는 본연의 효능을 모두 볼 수 있으며 그 외 부작용 등이 현저히 줄어든다는 연구결과들이 보고되고 있다.
Fermented red ginseng also helps to balance saponin metabolism. Diol saponin stabilizes the body and improves immunity. The triol saponins complement each other to enhance blood circulation, enhance the vitality and vitality, and have opposite functions. If the intestinal flora is unstable and the metabolism of both the diol and triol saponins is not metered or both are not metabolized, the saponin absorption will be unbalanced. Therefore, the effect may be reduced even by taking ginseng, and in some people, fever You will experience an uncomfortable experience. However, researchers have reported that when the diol system and the triol saponin are balancedly metabolized from the outside and absorbed into the body, all the effects of ginseng and red ginseng can be seen, and other side effects are remarkably reduced.

대한민국 공개번호 제10-2011-0052218호(발명의 명칭: 펙티넥스를 이용하여 화합물 K의 함량이 증가된 인삼 및 홍삼 농축액의 제조방법, 2011.05.18. 공개)Korean Patent Publication No. 10-2011-0052218 (the name of the invention: a method for producing ginseng and red ginseng concentrate whose content of compound K is increased using pectinex, published on May 18, 2011) 대한민국 공개번호 제10-2010-0069194호(발명의 명칭: 효소 및 초고압을 이용한 홍삼제조방법, 2010.06.24. 공개)Korean Patent Publication No. 10-2010-0069194 (Title of the invention: Method for producing red ginseng using enzymes and ultra-high pressure, published on June 24, 2010) 대한민국 공개번호 제10-2006-0001834호(발명의 명칭: 김치 유산균을 이용한 발효인삼 또는 발효홍삼의 제조 방법, 2006.01.06. 공개)Korean Patent Publication No. 10-2006-0001834 (the name of the invention: a method for producing fermented ginseng or fermented red ginseng using lactic acid bacteria of Kimchi, published on Jan. 1, 2006) 대한민국 공개번호 제10-2010-0089701호(발명의 명칭: 2단계 발효 방식에 의한 발효 홍삼 및 그 제조 방법, 2010.08.12. 공개)Korean Patent Publication No. 10-2010-0089701 (the name of the invention: fermented red ginseng by two-step fermentation method and its production method, disclosed on Aug. 12, 2010)

Biotransformation of Panax notoginseng saponins into ginsenoside compound K production by Paecilomyces bainier sp. 229.Biotransformation of Panax notoginseng saponins into ginsenoside compound K production by Paecilomyces bainier sp. 229. J Appl Microbiol. 2008 Mar;104(3):699-706.J Appl Microbiol. 2008 Mar; 104 (3): 699-706.

본 발명은 홍미삼으로부터 효소전환과 김치유래 유산균을 이용한 발효를 통하여 화합물 K의 함량이 강화된 발효홍삼 농축액을 제조하는 방법 및 그 제조방법에 의해 제조된 발효홍삼 농축액을 유효성분으로 함유하는 제품을 제공하는 것을 목적으로 한다.
The present invention provides a method for preparing a fermented red ginseng concentrate having enhanced compound K content through fermentation using red blood cells derived from red ginseng and fermentation using lactic acid bacteria derived from kimchi, and a product containing fermented red ginseng concentrate prepared by the preparation method as an active ingredient .

상기와 같은 목적을 달성하기 위하여, 본 발명은 a)4년근 홍미삼에 50% 주정(酒精)을 첨가하여 홍미삼 주정추출물을 제조하는 단계; b)상기 a)단계의 홍미삼 주정추출물을 냉각한 후 여과하는 단계; c)상기 b)단계의 여과액을 25~30브릭스(Brix)까지 농축하여 주정(酒精)을 제거하는 단계; d)상기 c)단계의 농축액을 희석하여 Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL 및 Rapidase C80Max로 이루어진 군으로부터 선택된 3종의 효소의 조합농도 1~3중량%의 범위로 50℃에서 48~72시간 동안 처리하여 사포닌을 전환시키는 단계; e)상기 d)단계의 효소 반응액에 락토바실러스 사케이(Lactobacillus sakei) HY7802 균주를 접종하여 발효시키는 단계; f)상기 e)단계의 발효액을 가열하여 d)단계의 효소를 불활성화 시키는 단계; g)상기 f)단계의 효소가 불활성화 된 발효액을 70~80브릭스(Brix)까지 감압 농축하는 단계; 및 h)상기 g)단계의 농축액을 살균하는 단계를 포함하는 화합물 K가 강화된 발효홍삼 농축액의 제조방법을 제공하는 것을 특징으로 한다.
In order to accomplish the above object, the present invention provides a method for producing a Korean red ginseng extract comprising: a) preparing a red ginseng extract by adding 50% ginseng to 4 year old red ginseng; b) cooling the red ginseng extract of step (a) and filtering; c) concentrating the filtrate from step b) to 25 to 30 Brix to remove alcohol; (d) diluting the concentrate of step (c) to a concentration of 1 to 3% by weight of three enzymes selected from the group consisting of Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL and Rapidase C80Max, ≪ / RTI > for 72 hours to convert saponin; e) inoculating Lactobacillus sakei strain HY7802 into the enzyme reaction solution of step d) and fermenting the same; f) heating the fermentation broth of step e) to deactivate the enzyme of step d); g) Concentrating the fermentation broth in which the enzyme of step f) is inactivated under reduced pressure to 70 to 80 Brix; And h) sterilizing the concentrate of step g). The present invention also provides a method for producing a fermented red ginseng concentrate.

상기 a)단계의 홍미삼 주정추출물의 제조는 홍미삼 무게의 20배의 50% 주정을 이용하여 80℃에서 6시간 동안 3회 추출하는 것이 바람직하다. 일반적으로 홍삼 추출시 10~20배의 주정을 사용한다고 알려져 있어 수율을 최대로 하기 위해서 홍삼 무게의 20배로 사용하여 3회 추출한다.
The preparation of the red ginseng extract of step (a) is preferably carried out three times at 80 ° C for 6 hours using a 20% 50% solution of red ginseng. Generally, it is known to use 10 ~ 20 times of alcohol when extracting red ginseng. It is extracted three times by using 20 times the weight of red ginseng in order to maximize the yield.

상기 b)단계의 홍미삼 주정추출물의 냉각은 40~50℃까지 하며, 여과는 퍼라이트(perlite)를 이용하는 것이 바람직하다. 퍼라이트 여과는 일반적으로 미세한 현탁물이나 콜로이달 입자를 제거하기 위함으로 진세노사이드 성분은 여과에 의해서 함량의 변화가 없기 때문이다. 40~50℃로 냉각하는 이유는 추출온도인 80℃에서는 추출물의 용해도 등의 원인으로 여과 효과가 반감되기 때문이다.The cooling of the red ginseng extract of step (b) is preferably performed at 40 to 50 ° C, and perlite is preferably used for filtration. The purite filtration is generally used to remove fine suspension or colloidal particles, and the ginsenoside component has no change in content by filtration. The reason for cooling to 40 to 50 ° C is that the filtration effect is reduced by half due to the solubility of the extract at 80 ° C as the extraction temperature.

또한 상기 b)단계의 여과액을 25~30브릭스(Brix)까지 농축하여 주정(酒精)을 제거하는 것이 바람직하다. 최소한 25브릭스(Brix)까지 농축을 하지 않으면 남아 있는 주정이 효소반응을 저해할 수 있다.
It is also preferable that the filtrate of step b) is concentrated to 25 to 30 Brix to remove the alcohol. If not concentrated to at least 25 Brix, the remaining alcohol may inhibit the enzyme reaction.

또한 상기 d)단계의 3종의 효소의 조합은 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max의 효소 조합인 것이 바람직하다.
Also, it is preferable that the combination of the three enzymes of step d) is an enzyme combination of Cytolase PCL5, Sumizyme AC and Rapidase C80Max.

상기 f)단계의 효소의 불활성화는 90℃에서 10분 동안 실시하는 것이 바람직하다. 효소를 불활성화 해서 다음 단계인 유산균 발효 시 유산균의 생육저해의 가능성을 없애기 위함이다.
The inactivation of the enzyme in step f) is preferably carried out at 90 DEG C for 10 minutes. And inactivating the enzyme to eliminate the possibility of inhibiting the growth of the lactic acid bacteria in the next step, lactic acid fermentation.

상기 h)단계의 살균은 90℃에서 2시간 동안 실시하는 것이 바람직하다.
The sterilization in step h) is preferably carried out at 90 ° C for 2 hours.

한편, 본 발명의 화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 함유하는 기능성 음료는 본 발명의 화합물 K가 강화된 발효홍삼 농축액과 혼합과즙시럽 및 물을 일정한 비율로 조합하여 150bar에서 균질한 후 10℃ 이하로 냉각한 후 유리병, 패트병 등 소포장 용기에 포장하여 제조한다.
Meanwhile, the functional beverage containing the compound K-enriched fermented red ginseng concentrate of the present invention as an active ingredient is prepared by combining the concentrated fermented red ginseng concentrate with the compounded fruit juice syrup and water of the present invention at a constant ratio and homogenizing at 150 bar After cooling to below 10 ° C, it is packaged in small containers such as glass bottles and plastic bottles.

또한, 본 발명의 화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 함유하는 건강기능식품은 화합물 K가 강화된 발효홍삼 농축액을 포함하는 것 이외에 영양보조성분으로서 비타민 B1, B2, B5, B6, E 및 초산에스테르, 니코틴산 아미드, 올리고당 등이 첨가될 수 있으며 여타의 식품 첨가물이 첨가되어도 무방하다.
In addition, the health functional food containing the compound K-enriched fermented red ginseng concentrate of the present invention as an active ingredient contains not only the fermented red ginseng concentrate with compound K but also vitamin B 1 , B 2 , B 5 , B 6 , E and acetic acid esters, nicotinic acid amides, oligosaccharides and the like may be added, and other food additives may be added.

삭제delete

본 발명의 화합물 K가 강화된 발효홍삼 농축액은 효소전환과 유산균 발효과정을 통하여 진세노사이드를 변환시킴으로써 인체 내에서 소화 흡수율이 증대된 화합물 K를 다량 함유하고 있는 건강기능식품 또는 식품소재로서 활용할 수 있다.
The compound K-enriched fermented red ginseng concentrate of the present invention can be used as a health functional food or a food material containing a large amount of a compound K having increased digestion and absorption rate in the human body by converting the ginsenoside through enzyme conversion and lactic acid fermentation have.

도 1은 본 발명에서 화합물 K의 전환 활성을 가지는 효소를 선발하기 위해서 반응물을 TLC 전개한 사진이다.
도 2는 본 발명에서 베타-글루코시다아제(beta-glucosidase) 활성을 보이는 균주들 중 진세노사이드 전환능을 보이는 균주를 선발하기 위해서 반응물을 TLC 전개한 사진이다.
도 3은 본 발명의 효소전환 및 유산균 발효에 의한 발효홍삼 농축액 제조방법의 바람직한 실시예를 보인 공정도이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph of a reaction product developed by TLC in order to select an enzyme having the activity of converting compound K in the present invention. FIG.
FIG. 2 is a photograph showing a TLC development of a reaction product in order to select a strain exhibiting a ginsenoside-converting ability among strains showing beta-glucosidase activity in the present invention.
FIG. 3 is a process diagram showing a preferred embodiment of a method for producing fermented red ginseng concentrate by enzyme conversion and lactic acid fermentation according to the present invention.

이하, 실시예를 통하여 본 발명을 보다 상세하게 설명한다. 그러나 다음의 실시예는 본 발명의 범위를 한정하는 것은 아니며, 본 발명의 기술적 사상의 범위 내에서 당업자에 의한 통상적인 변화가 가능하다.
Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following embodiments are not intended to limit the scope of the present invention, and ordinary variations by those skilled in the art within the scope of the technical idea of the present invention are possible.

<실시예 1>&Lt; Example 1 >

홍삼 원료의 선정Selection of raw materials for red ginseng

풍기인삼농협으로부터 구입한 6년근 본삼, 6년근 미삼, 4년근 본삼, 4년근 미삼을 구입하여 사용하였다.6 - year old ginseng, 6 - year old ginseng, 4 - year old ginseng and 4 - year old ginseng purchased from Punggi Ginseng Nonghyup were purchased and used.

6년근 홍삼의 본삼 100g, 6년근 홍삼의 미삼 100g, 4년근 홍삼의 본삼 100g, 4년근 홍삼의 미삼 100g 각각에 70% 주정 2ℓ를 넣고 80℃에서 환류추출하여 감압농축 후 동결건조하였다.2 ℓ of 70% alcohol was added to 100 g of 6 years old red ginseng root, 100 g of 6 years old red ginseng root, 100 g of 4 year old red ginseng root, and 100 g of 4 years old red ginseng root respectively and the mixture was refluxed at 80 ° C. and concentrated under reduced pressure.

상기 방법에 의해 얻어진 주정추출물에서 진세노사이드(ginsenoside) Rb1 및 Rg1의 함량을 측정하여 결과를 하기 표 1에 나타내었다.
The content of ginsenosides Rb1 and Rg1 in the alcohol extract obtained by the above method was measured and the results are shown in Table 1 below.

Figure 112012049075563-pat00001
Figure 112012049075563-pat00001

상기 표 1에서 확인할 수 있는 바와 같이, 본삼에 비해서 미삼에 화합물 K의 주요 기질이 되는 Rb1의 함량이 많았으며, 6년근 보다 4년근에서 함량이 더 많았다. 따라서, 화합물 K의 함량이 강화된 발효홍삼 농축액의 원료로써 4년근 홍미삼을 선택하였다.
As shown in Table 1, the content of Rb1, which is a major substrate of compound K, was higher in the ginseng than in the ginseng, and the content was higher in the 4-year-old muscle than in the 6-year-old muscle. Therefore, 4 years old red ginseng was selected as a raw material of the fermented red ginseng concentrate having enhanced compound K content.

<실시예 2>&Lt; Example 2 >

사용 효소의 선정Selection of Enzymes

화합물 K의 주요 기질이 되는 진세노사이드 Rb1을 사용한 효소반응을 통하여 식용 가능한 효소 13종(Sumizym AC, TG-B, Lumicell YX1, Cellulase KN, Rapidase TF, Pectinase UltraSP, Cytolase PCL5, Filtrase Premium, Multifect Pectinase FE, Crystalzyme APXL, Rapidase C80MAx, GC220, Multifect CXXL)으로부터 화합물 K를 생성하는 효소 7종을 선별하였다.
Bacterial enzymes (Sumizym AC, TG-B, Lumicell YX1, Cellulase KN, Rapidase TF, Pectinase UltraSP, Cytolase PCL5, Filtrase Premium, Multifect Pectinase FE, Crystalzyme APXL, Rapidase C80MAx, GC220, Multifect CXXL).

상기의 반응은 1%(w/v)의 진세노사이드 Rbl 수용액 1㎖에 13종의 효소 각각을 총 반응 부피 1㎖의 5%인 0.05㎖를 첨가하여 50℃에서 200rpm으로 24시간 반응하여 총 반응 부피 1㎖의 동량의 수포화 부탄올(butanol)로 진세노사이드를 추출한 후 TLC(thin layer chromatography)에 점적 및 전개하여 화합물 K의 생성여부를 확인하였다.In the above reaction, 1 ml of 1% (w / v) ginsenoside Rb1 aqueous solution was added with 13 kinds of enzymes, 0.05 ml of 5% of the total reaction volume of 1 ml, and reacted at 50 ° C and 200 rpm for 24 hours, The ginsenosides were extracted with an equal volume of water saturated butanol in a reaction volume of 1 ml and then dotted and developed on TLC (thin layer chromatography) to confirm the formation of compound K. [

그 결과를 도 1에 나타내었다.The results are shown in Fig.

세로축의 "C-K"는 "화합물 K"를 나타낸다. "AC"는 "Sumizym AC"를 나타낸다.
"CK" in the vertical axis indicates "compound K &quot;."AC" represents "Sumizym AC".

도 1에서 확인할 수 있는 바와 같이, Cytolase PCL5(제조사: DSM, 효소 종류: pectinase, pectin lyase, polygalacturonase and endoarabinase, 효소 생산 미생물: Aspergillus niger), Multifect Pectinase FE(제조사: Genencor, 효소종류: pectinase, cellulase, henmicellulase and beta-glucosidase, 효소 생산 미생물: Aspergillus niger), Sumyzyme AC(제조사: ShinNippon, 효소 종류: cellulase, beta-glucosidase and hemicellulase, 효소 생산 미생물: Aspergillus niger), Cellulase KN(제조사: ShinNippon, 효소 종류: cellulase, hemicellulase, pretease and naringinase, 효소 생산 미생물: Aspergillus niger), Rapidase TF(제조사: DSM, 효소 종류: pectinase and hemicellulase, 효소 생산 미생물: Aspergillus niger), Crystalzyme APXL(제조사: DSM, 효소 종류: pectin esterase, pectin depolymerase, cellulase and hemicellulase, 효소 생산 미생물: Aspergillus niger) 및 Rapidase C80Max(제조사: YC International, 효소 종류: pectinase, 효소 생산 미생물: Aspergillus niger)의 총 7종의 화합물 K로의 전환 효소를 선별하였다.
As can be seen from FIG. 1, Cytolase PCL5 (manufacturer: DSM, enzyme type: pectinase, pectin lyase, polygalacturonase and endoarabinase, enzyme producing microorganism: Aspergillus niger), Multifect Pectinase FE (manufacturer: Genencor, enzyme type: pectinase, Aspergillus niger), Sumyzyme AC (manufacturer: ShinNippon, enzyme type: cellulase, beta-glucosidase and hemicellulase, enzyme producing microorganism: Aspergillus niger), Cellulase KN (manufacturer: ShinNippon, enzyme type Aspergillus niger), Rapidase TF (manufacturer: DSM, enzyme type: pectinase and hemicellulase, enzyme producing microorganism: Aspergillus niger), Crystalzyme APXL (manufacturer: DSM, enzyme type: pectin esterase, pectin depolymerase, cellulase and hemicellulase, enzyme producing microorganism: Aspergillus niger) and Rapidase C80Max (manufactured by YC International, enzyme type: pectinas e, enzyme producing microorganism: Aspergillus niger).

<실시예 3>&Lt; Example 3 >

선별 효소의 최적 조합 선정Selection of optimal combination of selective enzymes

선별된 상기 실시예 2의 7종의 효소 중 Multifect Pectinase FE는 Genencor사의 단종으로 인해 제품 생산에 적용이 불가능하여 제외하였으며, Rapidase TF와 Rapidase C80Max 중 Rapidase C80Max를 선정함에 따라 총 5종의 효소 Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL 및 Rapidase C80Max를 가지고 최적 조합을 찾는 실험을 진행하였다.
Of the seven enzymes selected in Example 2, Multifect Pectinase FE was excluded due to the discontinuation of Genencor Inc., and Rapidase TF and Rapidase C80Max were selected as Rapidase C80Max. Thus, 5 kinds of enzyme Cytolase PCL5 , Sumizyme AC, Cellulase KN, Crystalzyme APXL and Rapidase C80Max.

상기 실시예 1의 10%(w/v)의 4년근 홍미삼 주정추출물 10㎖에 상기 5종류의 효소 중 3종류의 효소의 조합을 각각 1%씩 4년근 홍미삼 주정추출물 총 부피 10㎖의 3%인 0.3㎖ 첨가하여 50℃에서 200rpm으로 24시간 반응하여 반응물의 화합물 K를 HPLC를 통하여 정량하였다.The 10% (w / v) 4-year old red ginseng extract of Example 1 was mixed with 10% of the 4-year-old red ginseng extract in a total volume of 10 ml of 3% And the reaction product was reacted at 50 DEG C and 200 rpm for 24 hours to quantify the compound K of the reaction product through HPLC.

그 결과를 하기 표 2에 나타내었다.
The results are shown in Table 2 below.

효소의 조합Enzyme combination 화합물 K의 함량
(mg/g)The content of compound K
(mg / g) Crystalzyme APXLCrystalzyme APXL Cytolase PCL5Cytolase PCL5 Sumizym ACSumizym AC 1.321.32 Crystalzyme APXLCrystalzyme APXL Cytolase PCL5Cytolase PCL5 Cellulase KNCellulase KN 1.171.17 Crystalzyme APXLCrystalzyme APXL Cytolase PCL5Cytolase PCL5 Rapidase C80MaxRapidase C80Max 1.191.19 Crystalzyme APXLCrystalzyme APXL Sumizym ACSumizym AC Cellulase KNCellulase KN 1.321.32 Crystalzyme APXLCrystalzyme APXL Sumizym ACSumizym AC Rapidase C80MaxRapidase C80Max 1.401.40 Crystalzyme APXLCrystalzyme APXL Cellulase KNCellulase KN Rapidase C80MaxRapidase C80Max 0.680.68 Cytolase PCL5Cytolase PCL5 Sumizym ACSumizym AC Cellulase KNCellulase KN 1.521.52 Cytolase PCL5Cytolase PCL5 Sumizym ACSumizym AC Rapidase C80MaxRapidase C80Max 1.861.86 Cytolase PCL5Cytolase PCL5 Cellulase KNCellulase KN Rapidase C80MaxRapidase C80Max 0.860.86 Sumizym ACSumizym AC Cellulase KNCellulase KN Rapidase C80MaxRapidase C80Max 1.001.00

상기 표 2에서 확인할 수 있는 바와같이, 화합물 K의 생성능이 가장 좋은 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max의 효소조합을 선정하였다.
As shown in Table 2, enzyme combinations of Cytolase PCL5, Sumizyme AC, and Rapidase C80Max, which have the best ability to produce Compound K, were selected.

<실시예 4><Example 4>

선별 효소 조합의 최적 조건 결정Determination of Optimum Condition of Screening Enzyme Combination

상기 실시예 3에서 선별된 3종 효소조합, Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max의 최적 전환조건을 설정하기 위하여 기질인 상기 실시예 1의 4년근 홍미삼 주정추출물의 농도를 5%, 10% 및 15%, 상기 3종 효소 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max의 조합 농도를 1%(Cytolase PCL5 0.33%, Sumizyme AC 0.33% 및 Rapidase C80Max 0.34%), 2%(Cytolase PCL5 0.67%, Sumizyme AC 0.66% 및 Rapidase C80Max 0.67%) 및 3%(Cytolase PCL5 1%, Sumizyme AC 1% 및 Rapidase C80Max 1%), 반응시간을 24시간, 48시간 및 72시간 중 선택된 조건으로 50℃에서 200rpm으로 진행하였으며 반응표면분석법(response surface methology)을 이용하여 하기 표 3과 같이 최적전환 조건을 결정하였다.
In order to set optimal conversion conditions of the three enzyme combinations, Cytolase PCL5, Sumizyme AC, and Rapidase C80Max selected in Example 3, the concentration of the 4-year-old Chinese red ginseng extract of Example 1 was 5%, 10%, and 15% (Cytolase PCL5 0.33%, Sumizyme AC 0.33% and Rapidase C80Max 0.34%), 2% (Cytolase PCL5 0.67%, Sumizyme AC 0.66%, and the combination of the three enzymes Cytolase PCL5, Sumizyme AC and Rapidase C80Max) Rapidase C80Max 0.67%) and 3% (Cytolase PCL5 1%, Sumizyme AC 1% and Rapidase C80Max 1%) and the reaction time was selected at 24 hours, 48 hours and 72 hours at 50 ° C and 200 rpm, the optimal conversion conditions were determined using the response surface methodology as shown in Table 3 below.

Sample Sample 반응조건
(4년근 홍미삼 주정추출물/
반응시간/
3종 효소조합 농도)Reaction conditions
(4 years old red ginseng extract /
Reaction time /
3 enzyme combination concentration) 화합물 K 생성량
(mg/g)Compound K yield
(mg / g) 1One 10%/3d/3%10% / 3d / 3% 2.592.59 22 5%/2d/3%5% / 2d / 3% 7.217.21 33 15%/1d/2%15% / 1d / 2% 0.30.3 44 10%/1d/3%10% / 1d / 3% 0.90.9 55 15%/2d/3%15% / 2d / 3% 1.71.7 66 5%/2d/1%5% / 2d / 1% 2.632.63 77 10%/1d/1%10% / 1d / 1% 0.360.36 88 10%/2d/2%10% / 2d / 2% 0.430.43 99 10%/2d/2%10% / 2d / 2% 0.660.66 1010 10%/3d/1%10% / 3d / 1% 0.560.56 1111 5%/3d/2%5% / 3d / 2% 5.195.19 1212 15%/2d/1%15% / 2d / 1% 0.330.33 1313 15%/3d/2%15% / 3d / 2% 1.431.43 1414 10%/2d/2%10% / 2d / 2% 1.071.07 1515 5%/1d/2%5% / 1d / 2% 2.762.76

상기의 표 3에서 확인할 수 있는 바와 같이, 4년근 홍미삼 주정추출물 5%, 효소조합농도 3중량% 및 반응시간 48시간(샘플 2)의 경우에 최대의 화합물 K의 생성량을 보였다.As can be seen from the above Table 3, the maximum amount of compound K was shown in the case of 5% of 4-year-old ginseng extract, 3% by weight of enzyme combination concentration and 48 hours of reaction time (sample 2).

이후 72시간 동안 반응하여 화합물 K의 생성량을 측정한 결과 48시간 반응과 비교했을 때 그 생성량이 7.43 mg/g으로 다소 증가하는 결과를 나타내었다.After 72 hours, the amount of Compound K was measured. As a result, the amount of Compound K was increased to 7.43 mg / g compared with that of 48 hours reaction.

따라서 발효홍삼농축액 제조시에 반응 조건을 4년근 홍미삼 주정추출물 5%, 효소조합농도 3중량% 및 반응시간 72시간으로 결정하였다.
Therefore, the reaction conditions for the fermented red ginseng concentrate were determined as 5% of 4 - year - old ginseng extract, 3% by weight of enzyme combination concentration and 72 hours of reaction time.

<실시예 5>&Lt; Example 5 >

5-1. 김치유산균으로부터 진세노사이드 전환 유산균의 분리5-1. Isolation of Lactic Acid Bacteria Converting Ginsenoside from Kimchi Lactic Acid Bacteria

여러 지역의 가정집 및 음식점으로부터 여러 종의 김치를 수집하여 각 김치의 김치액을 멸균수를 이용하여 희석한 후 MRS Agar 배지에 도말하여 30℃에서 배양한 후 균주모양, 색, 투명도, 크기, 외형구조 등을 육안으로 관찰하여 김치 유산균을 1차 선발하였다.
Several kinds of kimchi were collected from various local households and restaurants, and the kimchi liquid of each kimchi was diluted with sterilized water, and then spread on MRS agar medium and cultured at 30 ° C. The shape, color, transparency, size, The structure of Kimchi lactic acid bacteria was firstly selected by visual observation.

상기의 방법을 통하여 1차 선발된 108 종의 균주를 베타-글루코시다아제(β-glucosidase) 활성, 진세노사이드 전환능 및 홍삼 추출액에서의 성장능을 통하여 최고의 전환능 및 성장능을 보이는 1종의 유산균을 최종 선발하였다.
Through the above-mentioned method, 108 strains selected first were classified into one kind of β-glucosidase activity, ginsenoside conversion ability and growth ability in red ginseng extract, Of lactic acid bacteria were finally selected.

먼저, 베타-글루코시다아제 활성을 갖는 유산균을 선발하기 위하여 1차 선발된 유산균 배양물(균체포함) 0.2㎖ 각각에 최종 농도가 10mM이 되도록 p-Nitrophenyl β-D-glucoside 용액 0.2㎖를 첨가하여 37℃에서 1시간 동안 반응하였다. 1.6㎖의 0.5M 탄산나트륨(Na2CO3)을 첨가하여 반응을 종료하고 420nm에서 흡광도를 측정하여 OD420 값이 1.5 이상인 균주 8종을 선발하였다. First, 0.2 ml of p-Nitrophenyl β-D-glucoside solution was added to each 0.2 ml of the cultured lactic acid bacteria (including cells) selected first to select lactic acid bacteria having β-glucosidase activity so that the final concentration was 10 mM And reacted at 37 DEG C for 1 hour. 1.6 ml of 0.5 M sodium carbonate (Na 2 CO 3 ) was added to terminate the reaction and the absorbance at 420 nm was measured to select eight strains having an OD 420 value of 1.5 or more.

그 결과를 하기 표 4에 나타내었다.
The results are shown in Table 4 below.

Figure 112012049075563-pat00002
Figure 112012049075563-pat00002

상기 표 4의 균주 8종을 대상으로 진세노사이드 Rb1의 전환능을 측정하기 위하여 1%(w/v)의 진세노사이드 Rbl 수용액 1㎖에 1㎖의 상기 8종의 유산균 배양물 각각을 첨가하여 37℃에서 200rpm으로 24시간 반응하여 TLC에 점적 및 전개한 후 진세노사이드 전환능을 보인 4종의 균주를 선발하였다. To determine the ability to convert ginsenoside Rb1 into 8 strains of Table 4, 1 ml of each of the above 8 kinds of lactic acid bacteria cultures was added to 1 ml of 1% (w / v) ginsenoside Rb1 aqueous solution And then reacted at 37 ° C for 24 hours at 200 rpm. The strain was then spotted and expanded on TLC, and four strains showing ginsenoside conversion ability were selected.

그 결과를 도 2에 나타내었다.
The results are shown in Fig.

도 2에서 확인할 수 있는 바와 같이, 진세노사이드 전환능을 보인 4종의 균주는 #25, #31, #35, #66임을 알 수 있었다.
As can be seen from Fig. 2, it was found that the four strains showing the ginsenoside conversion ability were # 25, # 31, # 35, and # 66.

상기 선발된 4종의 균주를 10% 홍미삼 추출물에 접종한 후 37℃에서 48시간 배양 후 성장능을 관찰하였다. The selected four strains were inoculated into 10% red ginseng extract and cultured at 37 ° C for 48 hours.

그 결과를 하기의 표 5에 나타내었다.
The results are shown in Table 5 below.

Figure 112012049075563-pat00003
Figure 112012049075563-pat00003

상기 표 5에서 확인할 수 있는 바와 같이, 최고의 성장능을 보인 균주 1종 #35을 최종 선발하였다.
As shown in Table 5, the strain # 1 having the best growth ability was finally selected.

5-2. 분리된 김치유산균의 동정5-2. Identification of Isolated Kimchi Lactic Acid Bacteria

상기 5-1의 선별된 균주 #35를 MRS 배지에서 배양한 후 게놈 DNA를 분리하였다. 16rDNA 부위는 시발체(primer)로 PCR(polymerase chain reaction)을 이용하여 증폭시켰다. 시발체 서열은 5`-AGA GTT TGA TCC TGG CTC AG-3`(순방향)과 5`-GGT TAC CTT GTT ACG ACT T-3`(역방향)을 사용하였다. PCR 반응을 위하여 추출한 게놈 DNA 50ng, 각 dNTPs 100μM, 시발체 각 0.2μM, 1× enzyme buffer, Taq polymerase 2unit를 넣고 증류수로 전체 50μl 부피가 되도록 추가하였다. 증폭반응은 초기 94℃에서 90초 동안 변성한 후, 28사이클로 94℃에서 30초, 42℃에서 60초, 72℃에서 60초 실시하였고, 추가로 72℃에서 5분 연장하였다. PCR 산물은 1.5% 아가로스 겔(agarose gel)에서 분리한 후, 용출하여 (주)코스모진텍에 염기 서열 분석을 의뢰하여 결과를 얻었다. 그 얻은 결과를 BLSAT Search를 통하여 분석할 결과 기존에 데이터베이스(Data Base)에 올려져 있는 락토바실러스 사케이(Lactobacillus sakei) 균주들과 99% 이상의 상동성을 나타내었다.
Genomic DNA was isolated after culturing 5-1 selected strains # 35 in MRS medium. The 16 rDNA region was amplified by PCR (polymerase chain reaction) as a primer. The primer sequences were 5'-AGA GTT TGA TCC TGG CTC AG-3` (forward) and 5'-GGT TAC CTT GTT ACG ACT T-3` (reverse direction). For the PCR reaction, 50 ng of extracted genomic DNA, 100 μM of each dNTPs, 0.2 μM of each primer, 1 × enzyme buffer and 2 units of Taq polymerase were added and added to the total volume of 50 μl with distilled water. The amplification reaction was carried out at 94 ° C for 30 seconds, at 42 ° C for 60 seconds, at 72 ° C for 60 seconds, and further extended at 72 ° C for 5 minutes. The PCR product was separated from the 1.5% agarose gel, eluted, and subjected to sequencing analysis by Cosmosintech Co., Ltd. to obtain the results. As a result of BLSAT search, the result was 99% or more homologous with the Lactobacillus sakei strain in database.

본 발명에 따른 유산균의 특성은 다음과 같다.The characteristics of the lactic acid bacteria according to the present invention are as follows.

1)균의 형태1) Types of bacteria

엠알에스(MRS) 한천평판배지에서 37℃, 2일간 배양했을 때 균의 특성The characteristics of the bacteria when cultured on an MRS agar plate medium at 37 ° C for 2 days

① 세포의 형태: 간균    ① Cell shape: Bacillus

② 운동성: 없음    ② Mobility: None

③ 포자형성능: 없음    ③ Spore forming ability: None

④ 그람(Gram) 염색: 양성    ④ Gram staining: positive

2)균락의 형태2) Morphology

엠알에스(MRS) 한천평판배지에서 37℃, 2일간 배양했을 때 균락의 형태When cultured on MRS agar plate medium at 37 ° C for 2 days,

① 형상: 원형    ① Shape: Circular

② 융기: 볼록    ② Bump: convex

③ 표면: 매끈(smooth)    ③ Surface: smooth

3)생리적 성질3) Physiological properties

① 최적 생육온도: 36~38℃     ① Optimal growth temperature: 36 ~ 38 ℃

② 최적 생육 pH: 5.0~7.0    ② Optimum growth pH: 5.0 ~ 7.0

③ 산소에 대한 영향: 통성혐기성    ③ Effect on oxygen: Tumor anaerobic

4)카탈라제: -4) Catalase: -

5)가스형성여부: -5) Whether gas is formed: -

6)15℃에서 생육: -6) Growth at 15 ℃: -

7)45℃에서 생육: +7) Growth at 45 ° C: +

8)인돌생산: -8) Indole production: -

9)젖산생산: +
9) Lactic acid production: +

이상에서와 같은 본 발명의 신균주의 형태학적 특성, 생리적 및 생장 특성과 16S rDNA 분석을 통한 분자유전학적인 방법에 근거하여 동정한 결과, 본 발명의 신균주는 락토바실러스 사케이(Lactobacillus sakei)에 속한다는 것을 알 수 있었다. 따라서 본 발명자들은 본 발명의 신균주를 락토바실러스 사케이(Lactobacillus sakei) HY7802로 명명하고, 한국생명공학연구원 생물자원센터에 2011년 12월 6일자로 기탁하였다(기탁번호:KCTC 12097BP).
Based on the morphological characteristics, physiological and growth characteristics of the novel strain of the present invention, and the molecular genetic analysis by 16S rDNA analysis, the new strain of the present invention was identified as Lactobacillus sakei I could see that it belonged. Therefore, the present inventors named the new strain of the present invention as Lactobacillus sakei HY7802 and deposited it on December 6, 2011 (Accession No .: KCTC 12097BP) to the BRC Center of the Korea Biotechnology Research Institute.

<실시예 6>&Lt; Example 6 >

발효홍삼 농축액의 제조Preparation of Fermented Red Ginseng Concentrate

도 3의 공정도에서와 같이 본 발명의 발효홍삼 농축액의 제조방법은 다음과 같다.As shown in the process chart of FIG. 3, the method for producing the fermented red ginseng concentrate of the present invention is as follows.

(1)원료삼 계량(1) Triple weighing of raw materials

풍기인삼농협에서 구입한 4년근 홍미삼 100kg을 사용하였다.100kg of 4 years old red ginseng purchased from Punggi Ginseng Nonghyup was used.

(2)추출(2) Extraction

상기 홍미삼에 식품원료로 사용할 수 있는 50% 주정 2000ℓ씩(주정 1000ℓ+정제수 1000ℓ) 첨가하여 80℃에서 6시간 동안 3회 추출하였다.2000 liters of 50% alcohol (1000 liters of alcohol and 1000 liters of purified water), which can be used as a food material, was added to the red ginseng and extracted three times at 80 ° C for 6 hours.

(3)여과(3) Filtration

상기 추출물을 50℃로 냉각한 후 퍼라이트(perlite)를 이용하여 여과하였다.The extract was cooled to 50 DEG C and filtered using perlite.

(4)농축(4) Concentration

상기 여과액을 30브릭스(Brix)로 감압농축하였다.The filtrate was concentrated under reduced pressure to 30 Brix.

(5)효소전환(5) Enzyme conversion

상기 감압농축액에 정제수를 첨가하여 5브릭스(Brix)가 되게 희석한 후 여기에 효소조합농도 3중량%의 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max 효소조합을 첨가하여 50℃에서 72시간 동안 반응하였다.Purified water was added to the vacuum concentrate to dilute to 5 Brix. Then, a combination of Cytolase PCL5, Sumizyme AC, and Rapidase C80Max was added to the mixture at a concentration of 3% by weight, and the mixture was reacted at 50 ° C for 72 hours.

(6)유산균발효(6) Fermentation of lactic acid bacteria

상기 효소 반응액에 김치유래 유산균 락토바실러스 사케이(Lactobacillus sakei) HY7802를 접종하여 37℃에서 발효하였다. Lactobacillus sakei derived from kimchi was added to the enzyme reaction solution, HY7802 was inoculated and fermented at 37 占 폚.

(7)효소실활(7) Enzymatic inactivation

상기 발효액을 90℃에서 10분 동안 가열하여 상기 효소 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max를 불활성화 하였다.The fermentation broth was heated at 90 DEG C for 10 minutes to inactivate the enzymes Cytolase PCL5, Sumizyme AC, and Rapidase C80Max.

(8)농축(8) Concentration

상기 효소가 불활성화된 발효액을 75브릭스(Brix)가 되게 감압농축하였다.The enzyme-inactivated fermentation broth was concentrated under reduced pressure to a Brix of 75.

(9)살균 및 포장(9) Sterilization and packaging

상기 농축액을 90℃에서 2시간 동안 살균한 후 포장하여 본 발명의 화합물 K가 강화된 발효홍삼 농축액을 제조하였다.
The concentrate was sterilized at 90 DEG C for 2 hours and then packed to prepare a fermented red ginseng concentrate enhanced with Compound K of the present invention.

<실시예 7>&Lt; Example 7 >

화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 함유하는 기능성 음료의 제조Preparation of functional beverage containing compound K-enriched fermented red ginseng concentrate as an active ingredient

본 발명의 화합물 K가 강화된 발효홍삼 농축액과 혼합과즙시럽으로 구성된 기능성 음료를 제조하는 방법은 다음과 같다.A method for preparing a functional beverage composed of the compound K-enriched fermented red ginseng concentrate and the mixed fruit juice syrup of the present invention is as follows.

먼저, 혼합과즙시럽은 액상과당 13중량%, 백설탕 2.5중량%, 갈색설탕 2.5중량%, 혼합과즙농축액 56Brixo 10.9중량%, 펙틴 1.0중량%, 후레쉬 후르츠 믹스 에센스 0.1중량% 및 정제수 70중량%를 30~35℃에서 교반하여 혼합한 후 UHT 열처리(135℃에서 2초간 살균)한 후 냉각하여 제조하였다.
First, a mixed juice syrup was 13% by weight of liquid fructose, white sugar, 2.5 wt%, brown sugar 2.5% by weight, mixed juice concentrate 56Brix o 10.9% by weight of pectin, 1.0% by weight, fresh 0.1% by weight fruit mix essence and purified water 70% The mixture was stirred at 30 to 35 ° C, mixed, and heat-treated by UHT (sterilized at 135 ° C for 2 seconds) and cooled.

그리고 상기의 방법으로 제조된 혼합과즙시럽 30.4중량%와 상기 실시예 6의 화합물 K가 강화된 발효홍삼 농축액 0.1중량% 및 나머지 정제수 69.5중량%를 조합하여 150bar에서 균질한 후 10℃ 이하로 냉각한 후 이를 유리병, 패트병 등 소포장 용기에 포장하여 본 발명의 화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 함유하는 기능성 음료를 제조하였다.
Then, 30.4% by weight of the mixed fruit juice syrup prepared by the above method, 0.1% by weight of the fermented red ginseng condensed with the compound K of Example 6 and 69.5% by weight of the remaining purified water were combined and homogenized at 150 bar, And then packed in small containers such as glass bottles and plastic bottles to produce a functional beverage containing the compound K-enriched fermented red ginseng concentrate of the present invention as an active ingredient.

<실시예 8>&Lt; Example 8 >

화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 함유하는 건강기능식품의 제조Production of a health functional food containing the compound K-enriched fermented red ginseng concentrate as an active ingredient

상기 실시예 6의 화합물 K가 강화된 발효홍삼 농축액 0.1중량%에 영양보조성분(비타민 B1, B2, B5, B6, E 및 초산에스테르, 니코틴산 아미드) 및 올리고당을 상기 상기 실시예 6의 화합물 K가 강화된 발효홍삼 농축액 100중량부에 대하여 10중량부가 되도록 첨가하여 고속회전 혼합기에서 혼합하였다. 상기 혼합물 100중량부에 대하여 멸균 정제수 10중량부를 첨가, 혼합하고 직경 1~2mm의 과립상으로 성형하였다. 상기 성형된 과립은 40~50℃의 진공건조기에서 건조시킨 후 12~14 메쉬(mesh)를 통과시켜 균일하게 과립을 제조하였다. 상기와 같이 제조된 과립은 적당량씩 압출 성형되어 정제 또는 분말로 되거나 경질캡슐에 충전되어 경질캡슐제품으로 제조하였다.
(Vitamin B 1 , B 2 , B 5 , B 6 , E and acetic acid ester, nicotinic acid amide) and oligosaccharide were added to 0.1 wt% of the concentrated fermented red ginseng concentrate of Compound 6 of Example 6, Was added in an amount of 10 parts by weight based on 100 parts by weight of the enriched fermented red ginseng concentrate, followed by mixing in a high-speed rotary mixer. To 100 parts by weight of the mixture, 10 parts by weight of sterilized purified water was added and mixed to form granules having a diameter of 1 to 2 mm. The molded granules were dried in a vacuum drier at 40 to 50 DEG C and then passed through 12 to 14 mesh to prepare uniform granules. The granules thus prepared were extruded in suitable amounts to be purified or powdered or filled into hard capsules to prepare hard capsule products.

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한국생명공학연구원Korea Biotechnology Research Institute KCTC12097BPKCTC12097BP 2011120620111206

Claims (9)

a)4년근 홍미삼에 50% 주정(酒精)을 첨가하여 홍미삼 주정추출물을 제조하는 단계;
b)상기 a)단계의 홍미삼 주정추출물을 냉각한 후 여과하는 단계;
c)상기 b)단계의 여과액을 25~30브릭스(Brix)까지 농축하여 주정(酒精)을 제거하는 단계;
d)상기 c)단계의 농축액을 희석하여 Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL 및 Rapidase C80Max로 이루어진 군으로부터 선택된 3종의 효소의 조합 농도 1~3중량%의 범위로 50℃에서 48~72시간 동안 처리하여 사포닌을 전환시키는 단계;
e)상기 d)단계의 효소 반응액에 락토바실러스 사케이(Lactobacillus sakei) HY7802 균주(기탁번호: KCTC 12097BP)를 접종하여 발효시키는 단계;
f)상기 e)단계의 발효액을 가열하여 d)단계의 효소를 불활성화 시키는 단계;
g)상기 f)단계의 효소가 불활성화 된 발효액을 70~80브릭스(Brix)까지 감압 농축하는 단계; 및
h)상기 g)단계의 농축액을 살균하는 단계를 포함하는 것을 특징으로 하는 화합물 K가 강화된 발효홍삼 농축액의 제조방법.
a) preparing a red ginseng extract by adding 50% ginseng to 4 year old red ginseng;
b) cooling the red ginseng extract of step (a) and filtering;
c) concentrating the filtrate from step b) to 25 to 30 Brix to remove alcohol;
(d) diluting the concentrate of step (c) to a concentration of 1 to 3% by weight of three enzymes selected from the group consisting of Cytolase PCL5, Sumizyme AC, Cellulase KN, Crystalzyme APXL and Rapidase C80Max, &Lt; / RTI &gt; for 72 hours to convert saponin;
e) Inoculating Lactobacillus sakei strain HY7802 (accession number: KCTC 12097BP) into the enzyme reaction solution of step d) and fermenting;
f) heating the fermentation broth of step e) to deactivate the enzyme of step d);
g) Concentrating the fermentation broth in which the enzyme of step f) is inactivated under reduced pressure to 70 to 80 Brix; And
h) sterilizing the concentrate of step g). &lt; RTI ID = 0.0 &gt; 8. &lt; / RTI &gt;
제1항에 있어서,
상기 a)단계의 홍미삼 주정추출물의 제조는 홍미삼 무게의 20배의 50% 주정(酒精)을 이용하여 80℃에서 6시간 동안 3회 추출하는 것을 특징으로 하는 발효홍삼 농축액의 제조방법.
The method according to claim 1,
The method for preparing a fermented red ginseng concentrate according to the above a), wherein the fermented red ginseng extract is extracted three times at 80 ° C for 6 hours using 50% of fermented ginseng at 20 times the weight of the fermented red ginseng.
제1항에 있어서,
상기 b)단계의 홍미삼 주정추출물의 냉각은 40~50℃까지 하며, 여과는 퍼라이트(perlite)를 이용하는 것을 특징으로 하는 발효홍삼 농축액의 제조방법.
The method according to claim 1,
Wherein the cooling of the red ginseng extract of step (b) is carried out at 40 to 50 ° C, and the filtration is performed using perlite.
제1항에 있어서,
상기 d)단계의 3종의 효소의 조합은 Cytolase PCL5, Sumizyme AC 및 Rapidase C80Max의 효소 조합인 것을 특징으로 하는 발효홍삼 농축액의 제조방법.
The method according to claim 1,
Wherein the combination of the three enzymes of step d) is an enzyme combination of Cytolase PCL5, Sumizyme AC, and Rapidase C80Max.
제1항에 있어서,
상기 f)단계의 효소의 불활성화는 90℃에서 10분 동안 실시하는 것을 특징으로 하는 발효홍삼 농축액의 제조방법.
The method according to claim 1,
Wherein the inactivation of the enzyme in step f) is carried out at 90 DEG C for 10 minutes.
제1항에 있어서,
상기 h)단계의 살균은 90℃에서 2시간 동안 실시하는 것을 특징으로 하는 발효홍삼 농축액의 제조방법.
The method according to claim 1,
Wherein the step (h) is sterilized at 90 DEG C for 2 hours.
제1항의 방법으로 제조된 화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 포함하는 기능성 음료.
A functional beverage comprising an active fermented red ginseng concentrate prepared by the method of claim 1 as an active ingredient.
제1항의 방법으로 제조된 화합물 K가 강화된 발효홍삼 농축액을 유효성분으로 포함하는 건강기능식품.
A health functional food containing an active fermented red ginseng concentrate prepared by the method of claim 1 as an active ingredient.
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KR1020120066147A 2012-06-20 2012-06-20 A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor KR101409761B1 (en)

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KR1020120066147A KR101409761B1 (en) 2012-06-20 2012-06-20 A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor
JP2015518310A JP5917772B2 (en) 2012-06-20 2012-09-03 A method for producing fermented red ginseng concentrate with enhanced content of compound K using enzyme conversion and lactic acid bacteria fermentation, and a product containing fermented red ginseng concentrate produced by the production method as an active ingredient.
PCT/KR2012/007053 WO2014007427A1 (en) 2012-06-20 2012-09-03 Method using enzyme conversion and lactobacillus fermentation for preparing concentrated fermented red ginseng liquid having enhanced compound k content, and product containing concentrated fermented red ginseng liquid prepared by said preparation method as active ingredient

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