KR20220059611A - Wild ginseng root by co-cultivating with ectomycorrhizal fungi and wood decay fungi and production method of the same - Google Patents
Wild ginseng root by co-cultivating with ectomycorrhizal fungi and wood decay fungi and production method of the same Download PDFInfo
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- mycelium
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Abstract
Description
본 발명은 유용한 버섯의 균사체가 감염된 산삼배양근 및 그 제조방법에 관한 것으로서, 더욱 상세하게는 산삼으로부터 산삼배양근을 유도하고 산삼배양근의 배양 중 식용 또는 약용버섯의 균사체를 배양중인 산삼배양근에 감염시켜 배양함으로써 종래의 산삼배양근의 배양만을 특징으로 하는 산삼 배양근 및 그 제조방법에서 한걸음 더 나아가 송이버섯을 비롯한 유용한 버섯의 균사체를 산삼배양근에 감염시켜 배양한 신규 산삼 배양근 및 그 배양방법에 관한 것으로 송이버섯을 비롯한 외생균근성 식용버섯 또는 목재부후성 약용 버섯 균사체와 산삼배양근의 공동배양에 의한 유용 버섯의 균사체를 산삼배양근에 감염시킨 후 공동 배양한 산삼배양근 및 그 제조방법에 관한 것이다.The present invention relates to a wild ginseng cultured root infected with a useful mushroom mycelium and a method for producing the same, and more particularly, a wild ginseng cultured root is induced from wild ginseng, and the mycelium of an edible or medicinal mushroom is infected with a wild ginseng cultured root cultured during culturing of wild ginseng cultured root. By doing so, it goes one step further from the conventional cultured wild ginseng root and its manufacturing method, which is characterized only by culturing wild ginseng root, and relates to a novel wild ginseng cultured root and a method for culturing the same, wherein the mycelium of useful mushrooms including matsutake mushroom is infected with the wild ginseng cultured root and cultured. It relates to a wild ginseng cultured root and a method for producing the same after infecting the mycelium of a useful mushroom by co-cultivation of an exogenous mycorrhizal edible mushroom or wood-rotting medicinal mushroom mycelium and wild ginseng cultured root.
산삼은 인삼과 같은 Panax과의 다년생 초본으로 인삼의 기원식물로써, 일반적으로 천종, 지종, 장뇌의 3종류로 분류되며, 천종과 지종은 국내 산양에서 자생하는 산삼으로 조류나 동물이 천종과 지종의 산삼종자를 식이 한 후 산양에 배설한 뒤 발아되어 생장한 것을 말하며, 장뇌산삼은 사람이 산삼의 종자를 채취하여 산야에 파종 후 야생의 조건으로 자라게 한 것을 말한다. 산삼은 오래 된 것 일수록 희소가치가 매우 크며 그 약성 또한 비례하여 높아진다고 알려져 있으나, 아직 명백한 자료는 존재하지 않는다.Wild ginseng is a perennial herb in the Panax family, such as ginseng, and as the origin of ginseng, it is generally classified into three types: Cheon, Ji, and Camphor. It refers to the germination and growth after excreting the seeds to the goats after feeding them. It is known that wild ginseng has a very high scarcity value as it gets older and its weakness also increases proportionally, but there is no clear data yet.
산삼의 약효성분과 약리적 효능을 탐구하기 위하여 광범위한 연구가 계속되고 있으며, 지금까지 과학적으로 밝혀진 대표적 효능으로는 신체 조절기능의 항상성 유지 작용이라 할 수 있으며, 이러한 작용에 근거하여 피로 및 스트레스에 효과적으로 작용하며, 항 당뇨작용, 혈압조절, 항암작용, 동맥경화 및 고혈압의 예방, 두되 기능의 강화, 위장기능 강화, 면역기능 강화 , 항 바이러스작용 등이 보고되고 있다. 또한 체내의 기능이 지나치게 항진된 경우에는 이를 저하시켜주고, 과도하게 저하된 경우에는 오히려 항진시켜줌으로써 생체를 정상으로 유지시켜주는 작용 즉, 적응소의 역할을 할 뿐 아니라 큰 부작용도 없는 것으로 알려져 있다.Extensive research is being conducted to explore the medicinal properties and pharmacological effects of wild ginseng, and the representative efficacy scientifically discovered so far is the maintenance of homeostasis of body control functions. Anti-diabetic action, blood pressure control, anticancer action, prevention of arteriosclerosis and high blood pressure, enhancement of the function of the stomach, strengthening of gastrointestinal function, enhancement of immune function, antiviral action, etc. have been reported. In addition, it is known that when the function of the body is excessively enhanced, it is lowered, and when it is excessively decreased, it is rather promoted, thereby maintaining the body in a normal state, that is, it serves as an adaptation site and has no major side effects.
세포배양기법의 발달로 산삼세포를 배양하여 캘러스로 만들거나 캘러스에서 유래된 단세포를 배양하여 산삼의 기관을 분화 시킬 수 있게 되어 대중적으로 산삼을 연구하거나 이용하는 것이 가능하게 되었다. 이에 관련한 선행기술로는 산삼배양근의 제조방법에 관한 기존특허로는 무균적으로 처리된 산삼시료를 2,4-D를 함유한 WPM배지에서 배양하여 캘러스를 유도하고, 유도된 캘러스를 2∼5ppm의 IBA가 첨가된 WPM배지에서 배양하여 산삼 부정근을 유도하는 방법(대한민국 특허 10-031890)과 무균적으로 처리된 산삼시료를 2,4-D, Pichloram 및 NAA를 각각 함유한 MS배지에서 배양하여 캘러스를 유도하고, 유도된 캘러스를 2,4-D가 첨가된 MS배지에서 증식시킨 다음, 생장조절제로서 BSAA(benzo selenienyl acetic acid)가 함유된 배지에서 배양하는 방법(대한민국 특허10-0353636)등이 있다.With the development of cell culture techniques, wild ginseng cells can be cultured to make callus, or single cells derived from callus can be cultured to differentiate wild ginseng organs, making it possible to study or use wild ginseng publicly. As a prior art related to this, in the existing patent on a method for producing cultured wild ginseng root, callus is induced by culturing aseptically treated wild ginseng samples in WPM medium containing 2,4-D, and 2-5 ppm of the induced callus is used. A method of inducing wild ginseng root induction by culturing in WPM medium supplemented with IBA (Korean Patent 10-031890) and aseptically treated wild ginseng samples were cultured in MS medium containing 2,4-D, Pichloram and NAA, respectively. A method of inducing callus, proliferating the induced callus in MS medium to which 2,4-D is added, and then culturing in a medium containing BSAA (benzo selenienyl acetic acid) as a growth regulator (Korean Patent 10-0353636), etc. There is this.
버섯균사체 효능을 기술한 선행 연구자료는 상황버섯 균사체 추출물이 백혈구에서 분비하는 세포 사이를 중개하는 생리활성물질 Cytokine 분비생성능 효과가 확인되어 면역증진 효능(송호연, 2012)에 관한 연구결과와 팽이버섯, 큰느타리버섯, 동충하초 버섯 균사체로 발효시킨 복령과 후박의 항산화 및 항암효과등 상승효과를 기술한 연구자료(손미예,2007)가 있으며, 버섯 균사체로부터 분리된 생질활성물질은(Moradali,2007) 다당체로써 글리포란(면역조절), 렌티난(항종양), 시조필란(항바이러스)과 다당체-펩타이드 중합체중 PSP(항종양), PSK(항바이러스,항생)등이 있으며, 단백질로써 Fips(면역조절), Ganoderic acid·Ganoderiol(항-HIV, 항종양, 감염억제, 항히스타민), Lucidumols(항고혈압)등에 관해 기술하였다.Previous research data describing the efficacy of mushroom mycelium include the results of studies on the immune-enhancing efficacy (Song Ho-yeon, 2012) and enoki mushroom, There are research data (Miye Son, 2007) that describe synergistic effects such as antioxidant and anticancer effects of bokryeong and green peppermint fermented with mycelium of Cordyceps oyster mushroom and Cordyceps mushroom mycelium. As such, there are glyphoran (immune regulation), lentinan (antitumor), schizophyllan (antiviral), and PSP (antitumor) and PSK (antiviral, antibiotic) among polysaccharide-peptide polymers, and Fips (immunity) as a protein control), Ganoderic acid Ganoderiol (anti-HIV, anti-tumor, anti-infection, antihistamine), and Lucidumols (anti-hypertension) were described.
잣버섯 액체배양 다당체의 면역계 사이토카인 생산, 저령 균사체로부터 에르곤(Ergone)증진 및 에르곤의 암세포의 생장억제에 효과등과 담자균류등의 균류에서만 생합성되는 항산화물질인 에르로티오네인(Ergothionine)의 액체배양을 통한 함량증대에 대한 것은 버섯균사체의 액체배양 및 유용물질생산 보고서(이위영, 2008)에 공표된 바 있다. 그리고 도라지를 버섯균사체로 고체발효시켜 총페놀성화합물과 베타클루칸의 함량을 증가시키는 발효방법에 관한 특허(대한민국 특허 10-18842980000)에서는 표고균사체 및 차가버섯균사체로 도라지를 발효하는 방법을 제사한 바 있으나 이는 총페놀성 화합물이 비발효된 대조군보다 각각 1.7배 내지 2.5배 증가, 베타글루칸은 2.5배 내지 32배로 증가되고, 도라지의 사포닌중에서 platycodine D의 함량이 증가되는 기술이 확인된 바 있으며, 또, 홍국, 동충하초 및 꽃송이버섯의 균사체를 배지에 접종하여 배양한 버섯균사체 배양액을 함유하는 화장료조성물에 관한 특허로서 제조되는 버섯 균사체 배양액을 함유하는 화장료조성물은, 콜레스테롤 생합성 촉진 및 히알루로산 생성효소의 발현 촉진을 통하여 우수한 주름개선, 보습 및 피부재생 효능을 기술한 버섯균사체 배양액을 유효성분으로 함유하는 화장료 조성물(대한민국 특허 10-12148720000)이 선행기술로 확인된다.Ergothionine, an antioxidant that is biosynthesized only in fungi such as basidiomycetes, and has effects on the production of immune system cytokines from liquid cultured polysaccharides from pine mushroom, promotion of ergone from the old mycelium, and inhibition of the growth of cancer cells of ergon. The content increase through liquid culture of Mushroom Mycelium has been published in the report on liquid culture of mushroom mycelium and production of useful substances (Lee Wi-young, 2008). And in the patent on a fermentation method to increase the content of total phenolic compounds and beta-glucan by solid fermenting bellflower with mushroom mycelium (Korean Patent 10-18842980000), a method of fermenting bellflower with shiitake mycelium and chaga mycelium was offered. However, the total phenolic compounds increased by 1.7 to 2.5 times, respectively, compared to the non-fermented control group, beta-glucan increased by 2.5 to 32 times, and the technology of increasing the content of platycodine D in the saponins of bellflower was confirmed, In addition, a cosmetic composition containing a mushroom mycelium culture solution prepared as a patent for a cosmetic composition containing a mushroom mycelium culture solution cultured by inoculating a medium with mycelium of red yeast rice, cordyceps cordyceps and cypress mushroom, promotes cholesterol biosynthesis and hyaluronic acid synthase A cosmetic composition (Korean Patent 10-12148720000) containing a mushroom mycelium culture as an active ingredient, which describes excellent wrinkle improvement, moisturizing and skin regeneration effects through promoting the expression of
끝으로 동충하초 추출물을 이용하여 사포닌이 강화된 산삼배양근을 제조하는 방법으로서,밀리타리스동충하초(Codyceps militaris) 또는 눈꽃동충하초(Paecilom yces japonica)의 추출물을 산삼배양근 배지에 첨가하여 각각 배양하는 단계로 코디세핀 성분이 함유된 사포닌 강화 산삼배양근의 제조방법에 관한 특허가 출원되었으나 거절된 사례가 있다(대한민국 특허 출원 10-2015-0190999).Finally, as a method for producing a saponin-enhanced wild ginseng root by using a cordyceps cordyceps extract, it is a step of culturing each by adding an extract of Cordyceps militaris or Paecilom yces japonica to a cultured wild ginseng root medium. A patent has been applied for for a method for preparing saponin-enhanced wild ginseng root containing sepin component, but has been rejected (Korean Patent Application 10-2015-0190999).
본 발명은 상기한 실정을 고려하여 발명한 것으로서 그 목적은 산삼으로부터 산삼배양근을 유도하고 산삼배양근의 배양 중 식용 또는 약용버섯의 균사체를 배양중인 산삼배양근에 감염시켜 공동배양함으로써 기존의 산삼배양근의 배양을 특징으로 하는 제조방법에서 더 나아가 유용한 버섯의 균사체를 산삼배양근에 감염시켜 배양하는 방법을 특징으로 하는 송이버섯 균사체등 유용한 버섯의 균사체를 감염시켜 생산하는 버섯균사체가 감염된 산삼배양근을 제공하는 데 있다.The present invention was invented in consideration of the above circumstances, and its purpose is to induce cultured wild ginseng from wild ginseng and infect the mycelium of edible or medicinal mushrooms during the cultivation of cultured wild ginseng to the cultured wild ginseng cultured root, thereby culturing the existing cultured wild ginseng root. It is to provide a wild ginseng cultured root infected with a mushroom mycelium produced by infecting a useful mushroom mycelium, such as a matsutake mushroom mycelium, characterized by a method of culturing and further infecting a useful mushroom mycelium with a wild ginseng cultured root. .
본 발명의 다른 목적은 제조공정이 간단하여 제조에 소요되는 시간 및 비용의 단축으로 저렴한 가격으로 단순하게 제조함으로써 경제성이 탁월한 송이버섯을 비롯한 외생균근성 식용버섯 또는 목재부후성 약용 버섯 균사체와 산삼배양근의 공동배양에 의한 유용한 버섯균사체와 감염배양시킨 산삼배양근의 제조방법을 제공하는 데 있다.Another object of the present invention is that the manufacturing process is simple and the time and cost required for manufacturing are reduced, so that the economical efficiency is excellent by manufacturing simply at a low price, including matsutake mushrooms, exogenous mycorrhizal edible mushrooms or wood-rotting medicinal mushroom mycelium and wild ginseng cultured root To provide a method for producing a useful mushroom mycelium and cultured wild ginseng root by co-culture of
본 발명의 상기 목적은 송이버섯을 포함하는 비롯한 외생균근성 식용버섯이나 목재부후성 약용 버섯의 균사체와 산삼배양근의 공동배양에 의한 유용한 버섯의 균사체를 공동 감염시켜 공동 배양된 산삼배양근의 제조방법으로서 산삼주근의 무균화 및 호르몬이 첨가된 혼용배지에서 치상된 산삼주근 조직으로부터 산삼 캘러스 및 부정근을 유도하는 산삼 부정근 유도단계(S1단계)와; 상기에서 유도시킨 산삼부정근으로부터 공기부양 생물반응기에서 대량 배양하는 액체배양단계(S2단계)와; 상기 단계에서 생장이 완료된 산삼배양근 배양과 달리 산삼액체 배양시작시 또는 배양중 산삼배양근의 생물배양기 내로 균근성버섯 또는 부후성 버섯의 균사체 접종시 멸균된 배양근의 배양액으로 균질기(Homogenizer)를 이용하여 균사체를 분쇄한 후 감염원을 준비하는 접종원 준비단계(S3단계)와; 상기 단계에서 생물반응기를 이용하여 산삼배양근의 접종 및 외생균근성 버섯 균사체를 동시접종하거나 버섯 수확전 0 ∼ 20일전 접종구를 통해 접종 또는 목재 부후성버섯 균사체를 버섯 수확전 0 ∼ 15일전 접종구를 통해 버섯 균사체를 산삼 배양근에 접종하는 접종단계(S4단계)로 이루어지는 것을 특징으로 한다.The above object of the present invention is a method for producing a co-cultured wild ginseng root by co-infecting the mycelium of a useful mushroom by co-culturing wild ginseng cultured root with the mycelium of an exogenous mycorrhizal edible mushroom or wood-rotting medicinal mushroom including matsutake mushroom. Wild ginseng root induction step (step S1) of sterilization of wild ginseng main root and inducing wild ginseng callus and irregular root from dentate wild ginseng main root tissue in a mixed medium to which hormones are added; A liquid culture step (step S2) of mass culturing in an air-lift bioreactor from wild ginseng root induced above; Unlike cultured wild ginseng root that has been grown in the above step, at the start of culturing wild ginseng liquid or when inoculating the mycelium of mycorrhizal fungi or decaying mushroom into the biological incubator of cultured wild ginseng during culturing, using a homogenizer as a culture solution of sterilized cultured roots. An inoculum preparation step of preparing an infectious agent after crushing the mycelium (step S3); In the above step, using a bioreactor, inoculation of wild ginseng cultured root and exogenous mycorrhizal mycelium simultaneously, or inoculation through an inoculation port 0 to 20 days before mushroom harvest, or inoculation of wood decay mushroom mycelium 0 to 15 days before mushroom harvest It is characterized in that it consists of an inoculation step (step S4) of inoculating mushroom mycelium into wild ginseng cultured roots through
본 발명은 산삼으로부터 산삼배양근을 유도하고 산삼배양근의 배양 중 식용 또는 약용버섯의 균사체를 배양중인 산삼배양근에 감염배양함으로써 기존의 산삼배양근의 배양을 특징으로 하는 제조방법에서 더 나아가 유용버섯 균사체를 산삼배양근에 감염 배양방법을 특징으로 하는 송이버섯 균사체를 포함하는 유용한 버섯 균사체를 감염시켜 제조되는 산삼배양근의 제조방법을 제공하는 효과가 있으며, 이를 식품재료로 하는 일반식품, 화장품 조성물, 기능성식품, 건강 기능성식품등에 포함하여 새로운 기능성 소재를 제공하는 효과가 있을 뿐만 아니라, 제조에 소요되는 시간 및 비용을 획기적으로 단축하여 저렴한 가격으로 대량 제조공급함으로써 경제성이 탁월한 각별한 효과가 있다.The present invention is a method of inducing wild ginseng cultured roots from wild ginseng and culturing the mycelium of edible or medicinal mushrooms during the cultivation of wild ginseng cultured roots to infect and culturing wild ginseng cultured roots. It has an effect of providing a method for producing wild ginseng cultured root prepared by infecting a useful mushroom mycelium including a matsutake mushroom mycelium, which is characterized by an infection culture method in cultured root, and general food, cosmetic composition, functional food, health using this as a food material Not only is it effective in providing new functional materials including in functional foods, but it also has a special effect with excellent economic feasibility by remarkably shortening the time and cost required for manufacturing and mass manufacturing and supplying at a low price.
도 1은 본 발명 버섯균이 감염된 산삼배양근의 대량 제조방법을 실행하는 순서도,
도 2는 산삼뿌리주근조직으로 부터 유도된 부정근을 보여주는 사진,
도 3은 생물반응기를 이용한 산삼배양근의 액체 배양 결과를 나타낸 사진,
도 4는 송이버섯 균사체를 감염시켜 배양된 산삼배양근 사진,
도 5는 송이버섯 균사체로 감염시킨 후 공동 배양된 산삼배양근의 확대사진이다.1 is a flowchart of executing the method for mass production of wild ginseng cultured root infected with mushroom bacteria of the present invention;
2 is a photograph showing an irregular root derived from wild ginseng root main muscle tissue;
3 is a photograph showing the liquid culture result of wild ginseng cultured root using a bioreactor;
Figure 4 is a photograph of wild ginseng cultured by infecting the mycelium of matsutake mushroom;
5 is an enlarged photograph of co-cultured wild ginseng cultured roots after infection with matsutake mushroom mycelium.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명하다할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.
도 1은 본 발명 버섯균이 감염된 산삼배양근의 대량 제조방법을 실행하는 순서도, 도 2는 산삼뿌리주근조직으로 부터 유도한 부정근을 보여주는 사진, 도 3은 생물반응기를 이용한 산삼배양근의 액체 배양 사진, 도 4는 송이버섯 균사체로 감염시켜 공동 배양된 산삼배양근 사진, 도 5는 송이버섯 균사체로 감염시킨 후 공동 배양된 산삼배양근의 확대사진으로써, 본 발명 송이버섯을 포함하는 외생균근성 버섯 또는 목재부후성 약용 버섯 균사체와 산삼배양근의 공동배양에 의한 유용 버섯균사체로 공동 감염시켜 공동 배양된 산삼배양근의 제조방법은 산삼주근의 무균화 및 산삼 부정근을 유도하는 산삼 부정근 유도단계(S1단계)와; 상기 단계에서 유도시킨 산삼부정근으로부터 산삼배양근을 대량생산하기 위하여 액체배양하는 액체배양단계(S2단계)와; 상기 단계에서 생장이 완료된 산삼배양과 별도로 산삼액체 배양시 배양시작시 또는 배양중 산삼배양근의 생물배양기 내로 균근성버섯 또는 부후성 버섯의 균사체 접종시 멸균된 배양근의 배양액으로 균질기(Homogenizer)를 이용하여 균사체를 분쇄한 후 감염원을 준비하는 접종원 준비단계(S3단계)와; 상기 단계에서 생물반응기를 이용하여 산삼배양근의 접종 및 외생균근성 버섯 균사체를 동시접종 또는 배양종료 0 ∼ 205일전 접종구를 통해 접종 및 목재 부후성버섯 균사체를 버섯수확 전 0 ∼ 15일전 접종구를 통해 버섯균사체를 산삼배양근에 접종하는 접종단계(S4단계)로 이루어진다.1 is a flow chart for executing the method for mass production of wild ginseng cultured root infected with mushroom bacteria of the present invention, FIG. 2 is a photograph showing an irregular root induced from the main root tissue of wild ginseng root, 4 is a photograph of wild ginseng cultured co-cultured by infection with matsutake mushroom mycelium, FIG. 5 is an enlarged photograph of wild ginseng cultured co-cultured after infection with matsutake mushroom mycelium. The method for preparing wild ginseng cultured root co-infected with useful mushroom mycelium by co-culture of epigenetic medicinal mushroom mycelium and wild ginseng culture root is a wild ginseng root induction step (step S1); a liquid culturing step (step S2) of liquid culturing in order to mass-produce wild ginseng root from the wild ginseng root induced in the above step; In the case of culturing wild ginseng liquid separately from the wild ginseng culture that has been grown in the above step, use a homogenizer as the culture medium of the sterilized cultured root when inoculating the mycelium of mycorrhizal fungi or decaying mushroom into the biological incubator of the wild ginseng cultured root during culture. an inoculum preparation step (step S3) of preparing an infectious agent after crushing the mycelium; In the above step, using a bioreactor, inoculate wild ginseng cultured root and ectopic mycelium simultaneously or inoculate through the inoculation port 0 to 205 days before the end of culture and inoculate the wood-rotting mushroom mycelium 0 to 15 days before mushroom harvest It consists of an inoculation step (step S4) of inoculating the mushroom mycelium into the wild ginseng cultured root.
이하, 본 발명의 구체적인 사항을 도면과 실시예를 들어 상세히 설명하지만 본 발명의 권리범위가 이들 실시예에만 한정하지 아니함은 통사의 기술자에게 당연하다 할 것이다.Hereinafter, the specific details of the present invention will be described in detail with reference to drawings and examples, but it will be natural for those skilled in the art that the scope of the present invention is not limited only to these examples.
[실시예 1][Example 1]
산삼의 소독 및 치상후 주근으로부터 부정근 유도Disinfection of wild ginseng and induction of irregular roots from post-dental main roots
소백산(경북 영주)에서 채취한 40년생 자연산 산삼의 뿌리로부터 주근만 남기고 부착 세근을 모두 제거하였다. 주근의 뿌리에 붙은 흙을 포함한 여러 종류의 이물질을 멸균수로 세척하고 70% 에탄올로 30초∼ 2분, 3% NaOCl에 20분 정도 표면살균한 후 멸균된 증류수로 3회 수세하였다. 표면살균된 산삼의 주근을 중심으로 밑부분의 직경이 4mm 이하 인 것은 제거하고 제거된 윗부분의 직경이 4mm 이상의 것으로 세로방향으로 두께 4mm의 간격으로 절단 한 후 부정근을 유도하기 위하여 WPM(Woody plant media, Duchefa사), MS 배지를 바람직하게는 WPM배지를 기본으로 하는 배지에 2,4D 0.1mg/L∼0.5mg/L과 IBA를 5mg/L 혼용첨가된 배지에 유도하였다(도 1).From the roots of 40-year-old wild ginseng harvested from Sobaeksan (Yeongju, Gyeongbuk), all the attached fine roots were removed, leaving only the main root. Various kinds of foreign substances including soil attached to the root of the main root were washed with sterile water, surface sterilized with 70% ethanol for 30 seconds to 2 minutes and 3% NaOCl for 20 minutes, and then washed with sterile distilled water 3 times. Centering on the main root of surface-sterilized wild ginseng, remove those with a diameter of less than 4mm at the bottom, cut those with a diameter of 4mm or more at the removed top, and cut them at intervals of 4mm in thickness in the vertical direction to induce irregular roots. , Duchefa), MS medium was preferably induced in a medium containing 0.1 mg/L to 0.5 mg/L of 2,4D and 5 mg/L of IBA to a medium based on WPM medium (FIG. 1).
[실시예 2][Example 2]
유도된 부정근의 액체배양 및 산삼배양근의 공기부양 생물반응기 배양Liquid culture of induced irregular root and air-floating bioreactor culture of wild ginseng cultured root
절단되어 치상된 산삼 주근조직으로부터 캘러스가 1차 유도되고 이 캘러스로부터 20일 후 정도부터 부정근이 유도 되었으며, 유도되어 생장하는 부정근의 길이가 3cm이상 생장후 캘러스에서 생장되는 부정근을 적출하고 다시 SH배지를 기본으로 하는 배지에 5mg/L IBA를 첨가한 고체 배지에 유도된 부정근을 1.5cm∼ 2cm 길이로 절단 한 후 치상하였다. 치상된 산삼배정근 단편은 측 세근이 유도되고 35일 후 무균적으로 수거된 부정근을 기본배지로써 WPM(woody plant media), mB5(modified Gamberg), MS (Murashige and Skoog), 1/2 MS, B5(Gamberg), SH(Schenk and Hildebrandt), LP(Quoirin and Lepoivre), Whit , GD(Gresshoff and Doy), DKW(Driver and Kuniyuki) 또는 DCR(Gupfa and Durzan)을 사용하였다. 필요시 상기배지에 2.0 ∼ 8.0mg/L IBA를 첨가하여 배양할 수 있으며, 바람직하게는 WPM(Woody plant media)을 기본배지로써 가장 바람직 하기는 5.0 ∼ 7.0mg/L IBA를 첨가한 액체배지를 이용하고 2L 공기부양 생물반응기를 이용하여 산삼 배양근을 생산하였으며(도 3), 배양된 산삼 배양근 5 ∼ 100g을 상기 액체배지 조성으로 20L 생물 반응기에 적정 운용량(working volume) 15L 배양배지를 제조한 후 산삼 배양근을 접종하고 증식하였다.The callus was first induced from the cut and dented wild ginseng main root tissue, and the irregular root was induced from about 20 days after the callus. The irregular root induced in a solid medium supplemented with 5 mg/L IBA in a medium based on . For the dentate ginseng dorsal root fragments, WPM (woody plant media), mB5 (modified Gamberg), MS (Murashige and Skoog), 1/2 MS, B5, aseptically harvested irregular roots 35 days after lateral fine root induction, were used as a basal medium. (Gamberg), SH (Schenk and Hildebrandt), LP (Quoirin and Lepoivre), Whit , GD (Gresshoff and Doy), DKW (Driver and Kuniyuki) or DCR (Gupfa and Durzan) were used. If necessary, it can be cultured by adding 2.0 to 8.0 mg/L IBA to the medium, preferably WPM (Wooody plant media) as a basic medium, and most preferably 5.0 to 7.0 mg/L IBA is added to the liquid medium. Wild ginseng cultured roots were produced using a 2L air flotation bioreactor (FIG. 3), and 5 to 100 g of cultured wild ginseng cultured roots were prepared with the liquid medium composition in a 20L bioreactor with an appropriate working volume (working volume) 15L culture medium. Then, wild ginseng cultured roots were inoculated and proliferated.
[실시예 3][Example 3]
생물반응기를 이용한 송이버섯등 버섯 균사체의 공동 감염배양Co-infectious culture of mushroom mycelium such as matsutake using a bioreactor
상기 실시예 2에 기술한 20L 생물반응기를 이용하여 35일 ∼ 40일간 배양한 후 산삼 배양근 생체량이 반응기를 2/3정도를 체울 정도로 생장이 되면 배양된 송이버섯을 포함하는 식용 또는 약용 버섯 균사체를 균질기로 분쇄하여 접종원을 준비한 다음 생물 반응기의 접종구를 통해 분쇄된 균사체를 수확 전 20일전에 접종하고 20일 동안 송이버섯등 버섯균사체에 감염이 충분히 된 산삼 배양근의 생체를 제조하였다(도 4).After culturing for 35 to 40 days using the 20L bioreactor described in Example 2, when the biomass of wild ginseng cultured roots grows enough to fill about 2/3 of the reactor, edible or medicinal mushroom mycelium containing the cultured matsutake mushroom is prepared. After preparing the inoculum by grinding with a homogenizer, the mycelium crushed through the inoculation port of the bioreactor was inoculated 20 days before harvest, and the living body of wild ginseng cultured roots sufficiently infected with mushroom mycelium such as matsutake mushroom for 20 days was prepared (FIG. 4) .
이때, 배양온도는 23 ∼ 25℃, pH 5.6~5.8, 공기 주입량은 0 ∼0.99vvm(volume/volume/min)이며, 접종시기는 버섯 균사체의 생장 정도에 따라, 송로버섯, 능이버섯, 큰갓버섯 등의 생장이 느린 균사체는 초기 산삼 배양근 접종 20일 후 균사체를 접종하고, 외생균근성 버섯 균사체중 생장이 빠른 균사체는 수확 20일 전, 바람직하게는 수확 15일 전에 접종구를 통해 접종할 수 있다. 한조건, 목재부후성 버섯 균사체는 수확 전 15일전 바람직하게는 버섯 균사체의 수확 10일 전 접종하여 산삼 배양근에 감염을 유도하였다. 생물 반응기내로 공기 주입량은 버섯종에 따라 균사체 생장의 차이와 산삼 배양근 표피조직으로 감염정도가 다르므로 공동 감염배양중 공기 주입량 증감조절에 따라 감염수율을 최대로 하였다(도 5).At this time, the culture temperature is 23 ~ 25 ℃, pH 5.6 ~ 5.8, the air injection amount is 0 ~ 0.99vvm (volume/volume/min), and the inoculation time depends on the growth degree of the mushroom mycelium, truffle, neungi mushroom, champignon mushroom Mycelium with slow growth on the back is inoculated 20 days after the initial wild ginseng cultured root inoculation, and mycelium with fast growth among the mycelium of ectopic mycorrhizal mushrooms can be inoculated through the inoculation port 20 days before harvest, preferably 15 days before harvest. . Under one condition, wood-rotting mushroom mycelium was inoculated 15 days before harvest, preferably 10 days before harvest of mushroom mycelium, to induce infection in wild ginseng cultured roots. Since the amount of air injected into the bioreactor differs depending on the mushroom species, the mycelial growth and the degree of infection with wild ginseng cultured root epidermal tissue, the infection yield was maximized by adjusting the air injection amount during co-infection and culture (FIG. 5).
[실시예 4][Example 4]
식품조성물의 용도 Use of food composition
공동 감염 배양되는 버섯균사체는 식품에 사용이 가능한 원료로서 사용가능하며 예컨대 외생균근성 버섯 균사체로는 갓버섯(큰갓버섯, Parasol mushroom Macrolepiota procera), 검은서양송로(Black Truffles, Tuber melanosporum), 기와버섯(청버섯,청갈버섯, Russula virescens), 까치버섯(먹버섯, Polyozellus multiplex (Underw.) Murrill), 꾀꼬리버섯(Chanterelle, Cantharellus cibarius Fr.), 노루털버섯(Sarcodon imbricatus (L.) P. Karst), 능이(향)버섯(Sarcodon asparatus), 다박잎새버섯(Grifola albicans Imaz.), 다발방패버섯(다발구멍장이버섯, Albatrellus confluens (Alb. & Schwein.) Kotl. & Pouzar / Polyporus confluens (Alb. & Schwein.) Fr.), 달걀버섯(Amantita caesarea), 땅송이(Tricholoma terreum), 먹물버섯(Coprinus comatus), 송로버섯(Truffle, Tuber aestivum (Wulfen) Vitt. / Tuber melanosporum Vittad.), 송이버섯(송이, Pine mushroom, Tricholoma matsutake (S.Ito & S. Imai)Singer), 블랙모렐즈(Black Morels, Morchella elata), 흰서양송로(송로버섯, White Truffles, Tuber magnatum Pico), 흰송이(White-flower lousewort, Tricholoma japonicum A.Kawam.), 흰꾀꼬리버섯(Cantharellus pallidus Lloyd), 흰조각광대버섯(Amanita esculenta Hongo et Matsuda), 흰주름버섯(Agaricus arvensis Schaeff ex Fr.), 젖버섯류(Lactarius spp.), 그물버섯류(Suillus spp.), 무당버섯류(Russula spp.)의 자실체로부터 분리된 균사체와, 목재 부후성 버섯으로 개암버섯(Naematoloma sublateritium), 구름버섯(운지버섯, Turkey tail, Trametes versicolor / Coriolus versicolor / Polystictus versicolor), 꽃송이버섯(Sparassis crispa), 꽃흰목이(Tremella foliacea Pers.), 나도팽나무버섯(맛버섯, Pholiota nameko), 노랑느타리(Gold oyster mushroom, Pleurotus cornucopiae var. citrinopileatus), 노랑먹물버섯(Coprinus radians (Desm.) Fr.), 노루궁뎅이버섯(Hericium erinacium), 느타리버섯(느타리, Oyster mushroom, Pleurotus ostreatus), 느티만가닥버섯(Beech mushroom, Hypsizygus marmoreus (Peck) H.E. Bigelow), 땅찌만가닥버섯(Lyophyllum shimeji (Kawam.) Hongo), 만가닥버섯(Lyopyllum ulmarium), 망태버섯(Dictyophora indusiata), 목이버섯(목이, Tree ear, Auricularia auricula-judae (Bull.) Wettst), 버들송이(Agrocybe cylindracea (DC. ex Fr.) Maire), 분홍느타리(Pink oyster mushroom, Pleurotus salmoneostramineus), 붉은덕다리버섯(Laetiporus sulphureus Murr var. miniatus), , 뽕나무버섯(Armillariella mellea), 사철느타리(Oyster mushroom, Pleurotus florida), 산느타리(Oyster mushroom, Pleurotus pulmonarius Qu'el), 석이버섯(Umbilicaria esculenta (Miyoshi) Minks), 신령버섯(흰들버섯, 아가리쿠스Agaricus blazei Murrill), 아까시재목버섯(장수버섯, Fomitella fraxinea (Fr.) Imaz.), 아위버섯(Pleurotus ferulae), 여름느타리(Oyster mushroom, Pleurotus sajor-caju), 영지버섯(Ganoderma lucidum), 왕송이(Wangsongi, Tricholoma giganteum), 잎새버섯(Grifola frondosa), 잿빛만가닥버섯(Lyophyllum decastes (Fr. ex Fr.) Sing / Tricholoma aggregatum Gill.), 전복느타리버섯(Oyster mushroom, Pleurotus abalonus), 털목이버섯(털목이, Auricularia polytticha Sacc.), 팽이버섯(팽나무버섯, Winter mushroom, Flammulina velutipes Sing.), 표고버섯(마고, 표고, 향담, Oak Mushroom, Lentinus edodes (Berk.) Sing. / Cortinellus edodes (Berk.) S. Ito & S. Imai), 황금목이(Tremella mesenterica Retz.), 흰목이버섯(White jelly fungi, Tremella fuciformis Beck.), 흰우단버섯(대형흰우단버섯, Leucopaxillus giganteus (Sow.:ex.Fr.) Sing), 식품의 제한적 원료로써 곰보버섯(Morchella esculenta Pers.), 말굽버섯(Fomes fomentarius (L.) Gill.), 모첼라코니카(검은곰보버섯, Conic Moral, Himalayan Morel Mushroom, Black Moral, Morchella conica Pers.), 진흙버섯(목질진흙버섯, 상황버섯, Phellinus linteus / Phellinus baumii / Sanghuangporus sanghuang), 차가버섯(검은자작나무버섯, Chaga Mushroom, Inonotus obliquus (persoon) pilat / Fuscoporia obliqua Persoon ex fries)의 자실체에서 분리된 균사체를 포함하며, 국내 식품공전에 등재는 되지 않았으나, 면역증강 효능에 의한 건강기능성 식품으로 인증된 번데기동충하초(Cordyceps militaris), 눈꽃동충하초(Paecilomyces japonica), 박쥐나방 동충하초(Cordyceps sinensis)의 자실체에서 분리된 균사체가 포함되는 것은 자명하다.Mushroom mycelium that is co-infected and cultured can be used as a raw material that can be used in food, for example, as an exogenous mycorrhizal mushroom mycelium (Parasol mushroom Macrolepiota procera), Black Truffles, Tuber melanosporum , tile mushroom (Blue mushroom, Blue brown mushroom, Russula virescens ), Black mushroom (Polyozellus multiplex (Underw.) Murrill), Oriole (Chanterelle, Cantharellus cibarius Fr.), Sarcodon imbricatus (L.) P. Karst ), Neungyi (scented) mushroom ( Sarcodon asparatus ) , Grifola albicans Imaz. & Schwein.) Fr.), Egg Mushroom ( Amantita caesarea ), Ground Mushroom ( Tricholoma terreum ), Ink Mushroom ( Coprinus comatus ), Truffle, Tuber aestivum (Wulfen) Vitt. / Tuber melanosporum Vittad. (Pine mushroom, Tricholoma matsutake (S.Ito & S. Imai)Singer), Black Morels, Morchella elata , White Truffles, Tuber magnatum Pico White-flower lousewort, Tricholoma japonicum A.Kawam.), White oriole ( Cantharellus pallidus Lloyd), White-flowered clown mushroom ( Amanita esculenta Hongo et Matsuda), White wrinkled mushroom ( Agaricus arvensis ) Schaeff ex Fr.), mycelium isolated from the fruiting body of Lactarius spp., Suillus spp., and radish ( Russula spp.) Mushroom (Turkey tail, Trametes versicolor / Coriolus versicolor / Polystictus versicolor ), oyster mushroom ( Sparassis crispa ), white oyster mushroom ( Tremella foliacea Pers. ), oyster mushroom ( Pholiota nameko ), Gold oyster mushroom, Pleurotus cornucopiae var. citrinopileatus ), Coprinus radians (Desm. Fr.), Hericium erinacium, Oyster mushroom, Pleurotus ostreatus , Beech mushroom, Hypsizygus marmoreus (Peck) HE Bigelow), Lyophyllum shimeji (Kawam.) Hongo), Lyophyllum ulmarium , Dictyophora indusiata , Wood ear mushroom (Tree ear, Auricularia auricula-judae (Bull.) Wettst ), Agrocybe cylindracea (DC. ex Fr.) Maire), Pink oyster mushroom, Pleurotus salmoneostramineus , Laetiporus sulphureus Murr var. miniatus, , Armillariella mellea (Oyster mushroom, Pleurotus florida ), Oyster mushroom ( Pleurotus pulmonarius Qu'el ), Umbilicaria esculenta (Miyoshi) Minks) , Fomitella fraxinea (Fr.) Imaz.), Pleurotus ferulae , Oyster mushroom, Pleurotus sajor-caju , Ganoderma lucidum , Wangsongi, Tricholoma giganteum , Grifola frondosa ), Lyophyllum decastes (Fr. ex Fr.) Sing / Tricholoma aggregatum Gill.), Oyster mushroom, Pleurotus abalonus , Auricularia polytticha Sacc.), Enoki mushroom (Winter mushroom, Flammulina velutipes Sing.), Shiitake (Margo, Shiitake, Hyangdam, Oak Mushroom, Lentinus edodes (Berk.) Sing. / Cortinellus edodes (Berk.) S. Ito & S. Imai), Tremella mesenterica Retz., White jelly fungi, Tremella fuciformis Beck. (Sow.:ex.Fr.) Sing), Morchella esculenta Pers., Fomes fomentarius (L.) Gill. Himalayan Morel Mushroom, Black Moral, Morchella conica Pers .), Chaga (Chaga Mushroom, Inonotus obliquat (persoon) pilin) / Contains mycelium isolated from the fruiting body of Fuscoporia obliqua Persoon ex fries, which has not been registered in the National Food Code, but has been certified as a health functional food due to its immune enhancing effect ( Cordyceps militaris ), Paecilomyces japonica ) , It is self-evident that the mycelium isolated from the fruiting body of Cordyceps sinensis is included.
이하, 본 발명의 식품조성물로서 다양한 제형의 제조예를 들어 설명하지만, 이밖에 식품학상 다른 제형에도 본 발명에 속하는 것은 물론이다.Hereinafter, examples of the preparation of various formulations as the food composition of the present invention will be described, but it is of course also included in the present invention to other formulations in terms of food science.
[제조예 1][Production Example 1]
산제의 제조Preparation of powders
이하에서 본 발명의 상기 실시예 3의 버섯균사체에 감염된 산삼 배양근의 생체, 건체 또는 그 추출물을 유효성분으로 하는 각종 제형의 예를 기재하지만, 본 발명의 제제가 이에 국한되는 것은 아니다. 버섯균사체에 감염된 산삼 배양근 추출물 분말 20mg, 유당 100mg 및 탈크 10mg 의 성분들을 혼합하고 기밀포에 충진하여 본 발명 일 실시에의 제형으로써 산제를 제조하였다.Hereinafter, examples of various formulations using the living body, dried body or extract thereof of wild ginseng cultured root infected with the mushroom mycelium of Example 3 of the present invention as an active ingredient are described, but the preparation of the present invention is not limited thereto. A powder was prepared as a formulation according to an embodiment of the present invention by mixing ingredients of 20 mg of wild ginseng cultured root extract powder infected with mushroom mycelium, 100 mg of lactose and 10 mg of talc, and filling in an airtight bag.
[제조예 2] [Production Example 2]
정제의 제조 manufacture of tablets
버섯균사체에 감염된 산삼 배양근의 생체, 건체 및 추출물 분말 10mg, 옥수수전분 100mg, 유당 100mg, 스테아린산 마그네슘 2mg의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 본 발명 실시예의 제형으로서 정제를 제조하였다.After mixing the components of the living body, dried and extract powder 10mg, cornstarch 100mg, lactose 100mg, and magnesium stearate 2mg of wild ginseng cultured root infected with mushroom mycelium, it is compressed according to a conventional tablet manufacturing method to prepare a tablet as a formulation of the present invention did
[제조예 3] [Production Example 3]
캡슐제의 제조 manufacture of capsules
버섯균사체에 감염된 산삼 배양근 생체, 건체 및 추출물 분말 10mg, 결정성 셀룰로오스 3mg, 락토오스 14.8mg와 마그네슘 스테아레이트 0.2mg를 통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충진하였다.Cultured wild ginseng root infected with mushroom mycelium, dried and extract powder 10 mg, crystalline cellulose 3 mg, lactose 14.8 mg, and magnesium stearate 0.2 mg were mixed with the above ingredients according to a conventional capsule preparation method and filled in a gelatin capsule.
[제조예 4] [Production Example 4]
액상파우치의 제조Production of liquid pouch
버섯균사체에 감염된 산삼배양근 생체, 건체 및 추출물 2ml, 홍삼농축액 1g, 식물혼합농축액 2g과 석류농축액 3g, 프락토올리고당10g 등을 통상의 파우치제조방법에 따라 상기의 성분을 혼합하고 파우치에 충진하여 제조하였다.Cultured wild ginseng root infected with mushroom mycelium, dried and extract 2ml, red ginseng concentrate 1g, plant mixed concentrate 2g, pomegranate concentrate 3g, fructooligosaccharide 10g, etc. are prepared by mixing the above ingredients according to the usual pouch manufacturing method and filling the pouch did
[제조예 5] [Production Example 5]
스틱젤리의 제조Production of Stick Jelly
버섯균사체에 감염된 산삼배양근 생체, 건체 및 추출물 2ml, 과일농축액 3g, 프락토올리고당 5g 비타민 및 무기질, 겔화제 등을 통상의 스틱젤리 제조방법에 따라 상기의 성분을 혼합하고 스틱파우치에 충진하여 제조하였다.Cultured wild ginseng root infected with mushroom mycelium, dried and extract 2ml, fruit concentrate 3g, fructooligosaccharide 5g vitamins and minerals, gelling agent, etc. were prepared by mixing the above ingredients according to a conventional stick jelly manufacturing method and filling a stick pouch. .
[제조예 6][Production Example 6]
소환의 제조crafting of summons
버섯균사체에 감염된 산삼배양근 생체, 건체 및 추출물 분말 20mg, 현미분말 1g, 찹쌀분말 0.8g 등을 통상의 환 제조방법에 따라 상기의 성분을 혼합, 제환하고 포장, 제조하였다.Wild ginseng cultured living body infected with mushroom mycelium, dried body and extract powder 20 mg, brown rice powder 1 g, glutinous rice powder 0.8 g, etc. were mixed, pilled, packaged, and manufactured with the above ingredients according to a conventional pill manufacturing method.
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| A study on the immune-enhancing efficacy of the mycelium extract of the Sangha mushroom mycelium by confirming the secretion-producing effect of Cytokine, a physiologically active substance that mediates between cells secreted by leukocytes (Song Ho-yeon, 2012) |
| Bioactive substances isolated from mushroom mycelium (Moradali, 2007) |
| Effect of pine mushroom liquid cultured polysaccharide on immune system cytokine production, ergone promotion from old mycelium, and inhibition of growth of cancer cells of ergon. Liquid culture of mushroom mycelium and production of useful substances on the increase of content through liquid culture of Ergothionine (Lee Wi-Young, 2008) |
| Research data describing the synergistic effects such as antioxidant and anticancer effects of fermented enoki mushroom, oyster mushroom, and cordyceps mushroom mycelium (Miye Son, 2007) |
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