KR102631606B1 - Monochamus alternatus solid medium composition for producing fruitbodies of cordyceps and its manufacturing method - Google Patents
Monochamus alternatus solid medium composition for producing fruitbodies of cordyceps and its manufacturing method Download PDFInfo
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- FGRBYDKOBBBPOI-UHFFFAOYSA-N 10,10-dioxo-2-[4-(N-phenylanilino)phenyl]thioxanthen-9-one Chemical compound O=C1c2ccccc2S(=O)(=O)c2ccc(cc12)-c1ccc(cc1)N(c1ccccc1)c1ccccc1 FGRBYDKOBBBPOI-UHFFFAOYSA-N 0.000 description 1
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- AAWZDTNXLSGCEK-ZHQZDSKASA-N Quinic acid Natural products O[C@H]1CC(O)(C(O)=O)C[C@H](O)C1O AAWZDTNXLSGCEK-ZHQZDSKASA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/50—Inoculation of spawn
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
본 발명은 동충하초 자실체를 생산하기 위한 솔수염하늘소 고체배지 조성물 및 이의 제조방법에 관한 것으로, 보다 상세하게는 솔수염하늘소 또는 열수 추출된 솔수염하늘소로 제조된 고체배지에 배양된 동충하초 자실체는 기존의 동충하초 자실체를 생산하기 위한 고체배지들에 비해서 발이기간이 짧아서 동충하초 자실체 생장이 효율적이며, 가장 높은 자실체 길이 및 중량을 나타냈으며, 동충하초 자실체 생장의 효율성을 평가를 위해 일반적으로 사용되는 척도인 biological efficiency의 경우, 솔수염하늘소 고체배지를 사용했을 때 높은 것으로 확인됨에 따라, 동충하초 자실체 생장량 증대의 효과가 확인됨에 따라, 상기 고체배지 제조방법은 동충하초 자실체를 효율적으로 생산하기 위한 고체배지용 조성물로 제공될 수 있다.The present invention relates to a solid medium composition for producing Cordyceps sinensis fruiting bodies and a method for producing the same. More specifically, the Cordyceps sinensis fruiting bodies cultured on a solid medium prepared from Cordyceps sinensis or hot water extracted Cordyceps sinensis are prepared from the existing Cordyceps sinensis fruiting bodies. Compared to solid media for producing Cordyceps sinensis fruiting bodies, the germination period is shorter, so the growth of Cordyceps sinensis fruiting bodies is efficient, and it showed the highest fruiting body length and weight, as well as biological efficiency, a measure commonly used to evaluate the efficiency of Cordyceps sinensis fruiting body growth. In this case, as it was confirmed that the effect of increasing the growth amount of Cordyceps sinensis fruiting bodies was confirmed to be high when using a solid medium of Cordyceps sinensis, the solid medium manufacturing method can be provided as a composition for a solid medium for efficiently producing Cordyceps sinensis fruiting bodies. there is.
Description
본 발명은 동충하초 자실체 생산을 위한 솔수염하늘소 고체배지 조성물 및 이의 제조방법에 관한 것이다.The present invention relates to a solid medium composition for the production of Cordyceps sinensis fruiting bodies and a method for producing the same.
동충하초(冬蟲夏草)는 겨울에는 벌레, 여름에는 버섯으로 나온다는 뜻에서 이름이 붙여졌다. 대부분이 곤충병원성진균(entomopathogenic fungi)으로 약 800여종이 알려져 있으며, 기주곤충에 감염되어 내부에서 균사 생장을 하여 자실체를 생성하는 것이 큰 특징이다. 이러한 동충하초는 아시아(중국, 한국, 일본 등)에서 예전부터 약초로 쓰여 왔다. 그 효능으로는 면역력 증강, 성인병 예방, 천식, 빈혈에 좋은 약재로 사용되어져 왔다.Cordyceps sinensis (冬蟲夏草) got its name from the meaning that it comes out as bugs in the winter and as mushrooms in the summer. About 800 species are known, most of which are entomopathogenic fungi, and their main characteristic is that they infect host insects and grow internally to produce fruiting bodies. Cordyceps sinensis has long been used as a medicinal herb in Asia (China, Korea, Japan, etc.). It has been used as a good herbal medicine for enhancing immunity, preventing adult diseases, asthma, and anemia.
동충하초는 동의보감을 통해서 오랫동안 많은 효능이 인정되어 각종 성인병 질환의 치료 및 건강유지를 위하여 사용되어 왔다. 중국에서는 육상선수들이 동충하초를 복용하고 기적의 기록들을 나타낸다고 하여 동충하초를 3대 천연식품 건강명약으로 인정하고 있다.Cordyceps sinensis has long been recognized for its many efficacy through Donguibogam and has been used to treat various adult diseases and maintain health. In China, Cordyceps sinensis is recognized as one of the top three natural food health medicines, as track and field athletes take Cordyceps sinensis and show miraculous records.
동충하초는 작물들 중 단백질 함량이 가장 높은 (건조함량의 28%이상) 군에 속하고, 면역력을 강화시키는 물질이 다량 함유되어 있다. 동충하초의 면역활성물질 중에는 키닉산(quinic acid)의 이성체로 밝혀진 '코디세핀 (Cordycepin :3'-deoxy-adenosine)' 이 포함되어 있으며, 동충하초 중에서도 밀리타리스 동충하초(Cordyceps militaris)가 다른 동충하초에 비하여 코디세핀의 함량이 월등히 높다. 코디세핀은 핵산 물질로서 세포의 유전정보에 관여하면서 저하된 면역기능을 활성화하여 정상세포가 암세포로 전환되는 것을 방지하는 작용을 한다. 동충하초를 투입한 임상 실험에서 암세포를 죽이는 면역세포인 NK세포(natural killer cell/자연살해세포)와 면역세포에서 분비되는 사이토카인의 함량이 18%~25% 증가되는 것으로 확인되어 혈액암 치료용으로 활용되고, 건강기능식품으로서도 많은 관심을 받고 있다. 이러한 결과를 토대로 국내에서는 코디세핀이 함유된 동충하초가 버섯으로는 유일하게 건강 기능식품으로 식약처 인증을 받았다. 또한 동충하초는 혈압과 당뇨를 조절하는 효능과 폐와 간 기능 등에도 많은 도움을 주는 것으로 임상연구결과가 발표되고 있다.Cordyceps sinensis belongs to the group with the highest protein content among crops (more than 28% of dry content) and contains a large amount of substances that strengthen immunity. Among the immune-activating substances of Cordyceps sinensis, 'Cordycepin (3'-deoxy-adenosine)', which has been found to be an isomer of quinic acid, is included. Among Cordyceps sinensis, Cordyceps militaris is more effective than other Cordyceps sinensis. The content of cordycepin is significantly higher. Cordycepin is a nucleic acid substance that participates in the genetic information of cells and acts to prevent normal cells from converting into cancer cells by activating decreased immune function. In clinical trials using Cordyceps sinensis, it was confirmed that the content of NK cells (natural killer cells), which are immune cells that kill cancer cells, and the content of cytokines secreted by immune cells increased by 18% to 25%, making it suitable for the treatment of blood cancer. It is used and is receiving a lot of attention as a health functional food. Based on these results, Cordyceps sinensis, which contains cordycepin, is the only mushroom in Korea to be certified by the Ministry of Food and Drug Safety as a health functional food. In addition, clinical research results show that Cordyceps sinensis is effective in controlling blood pressure and diabetes, and is helpful for lung and liver function.
다만, 동충하초는 버섯류 중에서도 많은 소비가 이루어지고 있지만 천연에서 채집되는 양은 매우 적어서 수요를 충당할 수 없으므로 전 세계의 많은 농가들이 인공재배법으로 재배사를 만들어서 대량 재배하고 있으나 동충하초는 성장속도가 느리고, 외부환경에 민감하여 농약을 살포할 수 없는 식물이어서 재배가 매우 까다로워 오랫동안 재배를 해 온 농가에서도 동충하초 재배의 수확률이 저조한 문제점이 있어서 동충하초 자실체 생장량 증대를 위한 고체 배지 제조 공정을 위한 연구가 지속적으로 이루어지고 있는 실정이다.However, although Cordyceps sinensis is widely consumed among mushrooms, the amount collected from nature is very small and cannot meet the demand. Therefore, many farmers around the world are cultivating large quantities by creating cultivation boxes using artificial cultivation methods. However, Cordyceps sinensis has a slow growth rate and is subject to environmental conditions It is a plant that cannot be sprayed with pesticides because it is sensitive to it, so its cultivation is very difficult. Even farms that have been cultivating Cordyceps sinensis for a long time have problems with low yields when cultivating Cordyceps sinensis. Therefore, research is continuously being conducted on the solid medium manufacturing process to increase the growth of Cordyceps sinensis fruiting bodies. There is a situation.
이에, 본 발명은 동충하초 자실체 생장량 증대를 위해서 솔수염하늘소 또는 열수 추출된 솔수염하늘소를 이용한 고체배지의 제조방법을 제공하고자 한다.Accordingly, the present invention seeks to provide a method for producing a solid medium using Cordyceps sinensis fruiting body growth or hot water-extracted Cordyceps sinensis.
또한, 본 발명의 다른 목적은 상기 제조방법으로 얻어진 동충하초 자실체 생산 고체배지용 조성물을 제공하는데 있다.In addition, another object of the present invention is to provide a composition for a solid medium for producing Cordyceps sinensis fruiting bodies obtained by the above production method.
또한, 본 발명의 다른 목적은 상기 제조방법으로 얻어진 동충하초 자실체 생산 고체배지용 조성물을 이용하여 동충하초 자실체 생산방법을 제공하는데 있다.In addition, another object of the present invention is to provide a method for producing Cordyceps sinensis fruiting bodies using the composition for a solid medium for producing Cordyceps sinensis fruiting bodies obtained by the above production method.
본 발명은 1) 곡류에 솔수염하늘소 또는 열수 추출된 솔수염하늘소를 첨가하는 단계; 2) 상기 1)단계에서 제조된 혼합물에 물을 첨가하는 단계; 및 3) 상기 2)단계에서 제조된 혼합물을 고온 멸균하는 단계를 포함하여 이루어지는 것을 특징으로 하는 솔수염하늘소 고체배지의 제조방법을 제공한다.The present invention includes the steps of 1) adding pine needles or hot water extracted pine needles to grains; 2) adding water to the mixture prepared in step 1); and 3) sterilizing the mixture prepared in step 2) at high temperature.
본 발명은 곡류 30 내지 40 중량%, 열수 추출된 솔수염하늘소 8 내지 17 중량% 및 잔량의 물을 포함하는, 동충하초 자실체 생산용 고체배지용 조성물을 제공한다.The present invention provides a composition for a solid medium for the production of Cordyceps sinensis fruiting bodies, comprising 30 to 40% by weight of grains, 8 to 17% by weight of hot water-extracted seaweed, and the remaining amount of water.
또한, 본 발명은 가) 액체배양된 동충하초 균사체를 상기 제조된 솔수염하늘소 고체배지용 조성물에 투입하고 암배양 하는 단계; 및 나) 상기 가)단계에 의해 암배양 후 형광등 조건의 배양기로 이동하여 자실체를 배양하는 단계를 포함하는 것을 특징으로 하는 동충하초 자실체의 생산방법을 제공한다. In addition, the present invention includes the following steps: a) adding liquid-cultured Cordyceps sinensis mycelium to the prepared composition for a solid medium for Solsulsulyeomseo and culturing it in the dark; and b) providing a method for producing Cordyceps sinensis fruiting bodies, which includes the step of cultivating the fruiting bodies by moving them to an incubator under fluorescent lighting conditions after dark culturing in step a) above.
본 발명에 따른 솔수염하늘소 고체배지로 동충하초 자실체를 생산하는 경우, 기존의 동충하초 자실체를 생산하기 위한 고체배지들에 비해서 발이기간이 짧아서 동충하초 자실체 생장이 효율적이며, 가장 높은 자실체 길이 및 중량을 나타냈으며, 동충하초 자실체 생장의 효율성을 평가를 위해 일반적으로 사용되는 척도인 자실체 생산량의 경우, 솔수염하늘소 고체배지를 사용했을 때 높은 것으로 확인됨에 따라, 동충하초 자실체 생장량 증대의 효과를 이룰 수 있다.When producing Cordyceps sinensis fruiting bodies with the solid medium of Cordyceps sinensis according to the present invention, the germination period is shorter than the existing solid media for producing Cordyceps sinensis fruiting bodies, so the growth of Cordyceps sinensis fruiting bodies is efficient, and the fruiting body length and weight are the highest. , fruiting body production, which is a measure commonly used to evaluate the efficiency of Cordyceps sinensis fruiting body growth, was confirmed to be high when using the Solseumbibae Hansol solid medium, which can achieve the effect of increasing the amount of Cordyceps sinensis fruiting body growth.
도 1은 본 발명에 따른 열수 추출된 솔수염하늘소를 이용한 고체배지 제조방법 및 이를 이용한 동충하초 자실체의 생산방법을 나타낸 모식도이다.
도 2는 동충하초가 성숙기가 되었음을 확인할 수 있는 동충하초 자실체 끝부분의 헤드를 나타낸다.
도 3는 고체배지 제조 공정에 따른 동충하초 자실체 생장 상태로, (가)는 열수 추출하지 않은 솔수염하늘소 배지로 생산한 동충하초 자실체, (나)는 열수 추출된 솔수염하늘소 배지로 생산한 동충하초 자실체를 나타낸다.
도 4은 RSM(Response surface methodology) 분석법을 통해 최적 고체배지 제조 조건에 대한 예측모델을 나타낸 것으로, X1은 열수 추출된 솔수염하늘소 투입량(g), X2은 수분함량(%), X3은 배양기간(days)을 나타낸다.
도 5는 RSM 분석법을 통해 최적 고체배지 제조 조건에 대한 예측모델의 최적화를 한 것을 확인한 결과이다.Figure 1 is a schematic diagram showing a method for producing a solid medium using hot water-extracted pine needles and a method for producing Cordyceps sinensis fruiting bodies using the same according to the present invention.
Figure 2 shows the head of the tip of the fruiting body of Cordyceps sinensis, which confirms that the Cordyceps sinensis has reached maturity.
Figure 3 shows the growth state of Cordyceps sinensis fruiting bodies according to the solid medium manufacturing process, (a) is Cordyceps sinensis fruiting bodies produced with a hot-water extracted medium, and (b) shows Cordyceps sinensis fruiting bodies produced with hot-water extracted medium. indicates.
Figure 4 shows a prediction model for optimal solid medium manufacturing conditions through RSM (Response surface methodology) analysis, where X 1 is Input amount of hot water extracted pine beard sky cow (g), Moisture content ( % ), Indicates the culture period (days).
Figure 5 shows the results confirming the optimization of the prediction model for optimal solid medium manufacturing conditions through RSM analysis.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
동충하초는 기주의 대상이 되는 곤충의 몸에 침입하여 기주의 양분을 통해 자실체를 형성시키는 약용버섯으로, 동물성 단백질을 영양성분으로 하기 때문에 일반 버섯류에서는 찾아볼 수 없는 유용한 물질, 즉 인간의 각종 질병치료에 이용되어질 수 있는 유용한 성분을 포함하여 기능성 식품으로서의 고부가가치가 있으나, 대량 재배가 어렵고 외부환경에 민감하여 재배가 매우 까다로운 문제점이 있는 바, 상기 문제점을 해결하기 위하여 본 발명자들은 열수 추출된 솔수염하늘소를 이용한 고체배지를 개발하여 본 발명을 완성하였다.Cordyceps sinensis is a medicinal mushroom that invades the body of a host insect and forms a fruiting body through nutrients from the host. Since it uses animal protein as a nutritional ingredient, it is a useful substance that cannot be found in ordinary mushrooms, that is, the treatment of various human diseases. Although it has high added value as a functional food, including useful ingredients that can be used in food, it has the problem of being difficult to cultivate in large quantities and being sensitive to the external environment, making cultivation very difficult. In order to solve the above problems, the present inventors developed hot water extracted pine beard. The present invention was completed by developing a solid medium using Sky Cow.
본 발명은 1) 곡류에 솔수염하늘소 또는 열수 추출된 솔수염하늘소를 첨가하는 단계; 2) 상기 1)단계에서 제조된 혼합물에 물을 첨가하는 단계; 및 3) 상기 2)단계에서 제조된 혼합물을 고온 멸균하는 단계를 포함하여 이루어지는 것을 특징으로 하는 솔수염하늘소 고체배지의 제조방법을 제공한다.The present invention includes the steps of 1) adding pine needles or hot water extracted pine needles to grains; 2) adding water to the mixture prepared in step 1); and 3) high-temperature sterilization of the mixture prepared in step 2) above.
상기 곡류는 귀리, 현미, 쌀 및 보리로 이루어진 군으로부터 선택된 어느 하나 이상인 것으로 이에 한정되는 것은 아니다. The grains are not limited to one or more selected from the group consisting of oats, brown rice, rice, and barley.
상기 귀리는 다른 곡류에 비하여 단백질, 필수 아미노산, 수용성 섬유질이 풍부하기 때문에 동충하초 자실체 생산을 위한 영양분 역할을 수행할 수 있다.Since the oats are rich in protein, essential amino acids, and soluble fiber compared to other grains, they can serve as nutrients for Cordyceps sinensis fruiting body production.
상기 열수 추출된 솔수염하늘소는 a) 솔수염하늘소와 증류수를 혼합한 후 가열하는 단계; b) 상기 a)단계에서 제조된 혼합물을 감압 여과하는 단계; 및 c) 상기 b)단계에 의해 생성된 잔사를 건조하는 단계를 포함하여 제조될 수 있다.The hot water-extracted pine needles include the steps of a) mixing pine needles and distilled water and then heating them; b) filtering the mixture prepared in step a) under reduced pressure; and c) drying the residue generated in step b).
상기 a)단계는 증류수 100 중량부에 대하여 솔수염하늘소를 2.5 내지 7.5 중량부로 혼합한 후 50 내지 70℃에서 2 내지 4시간 동안 가열할 수 있다. 상기 솔수염하늘소의 함량 범위 내에 포함되면 동충하초 자실체의 최초 발이기간이 4.7 내지 6.0일, 원기 수가 16.7 내지 22.7개, 자실체 성숙 기간이 31 내지 39일이 소요될 수 있으나, 상기 솔수염하늘소의 함량 범위인 2.5 중량부보다 낮을 경우, 자실체의 최초 발이기간이 8.3일 내지 10.0일, 원기 수가 10.0 내지 15.0개, 자실체 성숙 기간이 40 내지 50일 소요되고, 상기 솔수염하늘소의 함량 범위인 7.5 중량부보다 높을 경우 자실체의 최초 발이기간이 6.5 내지 9.5일, 원기 수가 12.5 내지 16.0개, 자실체 성숙 기간이 40 내지 50일 소요되어 발이기간 및 자실체 성숙 기간이 길어지고 원기수가 감소하는 문제가 야기될 수 있다.In step a), 2.5 to 7.5 parts by weight of pine beetroot can be mixed with 100 parts by weight of distilled water and then heated at 50 to 70°C for 2 to 4 hours. If it is included within the content range of the above-mentioned pine needles, the first germination period of the Cordyceps sinensis fruiting body may be 4.7 to 6.0 days, the number of primordia may be 16.7 to 22.7, and the fruiting body maturation period may take 31 to 39 days, but the content range of the pine needles rays above is If it is lower than 2.5 parts by weight, the first germination period of the fruiting body takes 8.3 to 10.0 days, the number of primordia is 10.0 to 15.0, and the fruiting body maturation period takes 40 to 50 days, and the content is higher than 7.5 parts by weight, which is the content range of the above. In this case, the initial germination period of the fruiting body takes 6.5 to 9.5 days, the number of primordia is 12.5 to 16.0, and the maturation period of the fruiting body takes 40 to 50 days, which may cause the problem of the germination period and fruiting body maturation period being prolonged and the number of primordia decreasing.
상기 c)단계는 상기 b)단계에 의해 생성된 잔사를 15 내지 25℃에서 40 내지 56시간 동안 건조할 수 있다.In step c), the residue produced in step b) may be dried at 15 to 25°C for 40 to 56 hours.
상기 1)단계는 곡류 100 중량부에 대하여 열수 추출된 솔수염하늘소 20 내지 50 중량부로 첨가할 수 있다. 상기 열수 추출된 솔수염하늘소의 함량 내에 포함되면 동충하초 자실체의 중량이 26.4 내지 27.5 g, 자실체 길이가 54.3 내지 55.1 mm, biological efficiency(=fruit body weight/medium weight × 100)가 26.2 내지 29.5 %로 나타냈지만, 상기 열수 추출된 솔수염하늘소 함량인 20 중량부 이하에서는 자실체의 중량이 7.2 내지 20.0 g, 자실체 길이가 15.1 내지 30.0 mm, biological efficiency가 6.3 내지 10.5 %이고, 상기 열수 추출된 솔수염하늘소 함량인 50 중량부 이상에서는 자실체의 중량이 9.8 내지 21.0 g, 자실체 길이가 34.7 내지 50.0 mm, biological efficiency가 9.4 내지 20.0 %로 자실체의 중량 및 길이 생산량에 문제가 야기될 수 있다.In step 1), 20 to 50 parts by weight of hot water-extracted seaweed can be added to 100 parts by weight of grain. When included within the content of the hot water extracted Cordyceps sinensis, the weight of the Cordyceps sinensis fruiting body is 26.4 to 27.5 g, the fruiting body length is 54.3 to 55.1 mm, and the biological efficiency (=fruit body weight/medium weight × 100) is 26.2 to 29.5%. However, at 20 parts by weight or less, which is the content of the hot water-extracted pine needles, the weight of the fruiting body is 7.2 to 20.0 g, the length of the fruiting body is 15.1 to 30.0 mm, and the biological efficiency is 6.3 to 10.5%, and the content of the hot water extracted pine needles is Above 50 parts by weight, the weight of the fruiting body is 9.8 to 21.0 g, the length of the fruiting body is 34.7 to 50.0 mm, and the biological efficiency is 9.4 to 20.0%, which may cause problems with the weight and length production of the fruiting body.
본 발명은 곡류 30 내지 40 중량%, 열수 추출된 솔수염하늘소 8 내지 17 중량% 및 잔량의 물을 포함하는, 동충하초 자실체 생산용 고체배지용 조성물을 제공한다.The present invention provides a composition for a solid medium for the production of Cordyceps sinensis fruiting bodies, comprising 30 to 40% by weight of grains, 8 to 17% by weight of hot water-extracted seaweed, and the remaining amount of water.
이때, 상기 곡류의 함량인 30 내지 40 중량%에서는 자실체의 중량 26.4 내지 27.5 g, 자실체 길이 54.3 내지 55.1 mm, biological efficiency가 26.2 내지 29.5%로 나타냈지만, 상기 곡류의 함량인 30 중량%보다 낮을 경우, 자실체의 중량 9.8 내지 17.5 g, 자실체 길이 15.1 내지 31.5 mm, biological efficiency가 9.4 내지 16.3%이고, 상기 곡류의 함량인 40 중량비보다 높을 경우, 자실체의 중량 17.5 내지 26.4 g, 자실체 길이 31.5 내지 54.3 mm, biological efficiency가 16.3 내지 26.2%로 자실체의 중량 및 길이 생산량에 문제가 야기될 수 있다.At this time, at the content of 30 to 40% by weight of the grain, the weight of the fruiting body was 26.4 to 27.5 g, the length of the fruiting body was 54.3 to 55.1 mm, and the biological efficiency was 26.2 to 29.5%, but when the content was lower than the content of 30% by weight of the grain, , the weight of the fruiting body is 9.8 to 17.5 g, the length of the fruiting body is 15.1 to 31.5 mm, the biological efficiency is 9.4 to 16.3%, and if it is higher than the weight ratio of 40, which is the content of the grain, the weight of the fruiting body is 17.5 to 26.4 g, and the length of the fruiting body is 31.5 to 54.3 mm. , the biological efficiency is 16.3 to 26.2%, which may cause problems with the weight and length production of fruiting bodies.
또한, 상기 열수 추출된 솔수염하늘소는 솔수염하늘소 내부의 탄수화물이 감소함에 따라 상대적으로 단백질과 무기물 함량이 증가한 상태이므로 동충하초 자실체의 생장을 가속화시키는 역할을 수행한다. 상기 열수 추출된 솔수염하늘소의 함량인 8 내지 17 중량%에서는 자실체의 중량 26.4 내지 27.5 g, 자실체 길이 54.3 내지 55.1 mm, biological efficiency가 26.2 내지 29.5 %로 나타냈지만, 상기 열수 추출된 솔수염하늘소의 함량인 8 중량%보다 낮을 경우, 자실체의 중량 17.5 내지 26.4 g, 자실체 길이 31.5 내지 54.3 mm, biological efficiency가 16.3 내지 26.2%이고, 상기 열수 추출된 솔수염하늘소의 함량인 17 중량%보다 높을 경우, 자실체의 중량 9.8 내지 17.5 g, 자실체 길이 15.1 내지 31.5 mm, biological efficiency가 9.4 내지 16.3%로 자실체의 중량 및 길이 생산량에 문제가 야기 될 수 있다. In addition, the hot water-extracted seaweed porridge plays a role in accelerating the growth of Cordyceps sinensis fruiting bodies because the protein and mineral contents relatively increase as the carbohydrates inside the sea urchin are reduced. At 8 to 17% by weight, which is the content of the hot water-extracted pine needles, the weight of the fruiting body was 26.4 to 27.5 g, the length of the fruiting body was 54.3 to 55.1 mm, and the biological efficiency was 26.2 to 29.5%, but the If the content is lower than 8% by weight, the weight of the fruiting body is 17.5 to 26.4 g, the length of the fruiting body is 31.5 to 54.3 mm, the biological efficiency is 16.3 to 26.2%, and if it is higher than the content of 17% by weight of the hydrothermally extracted pine cone, The weight of the fruiting body is 9.8 to 17.5 g, the length of the fruiting body is 15.1 to 31.5 mm, and the biological efficiency is 9.4 to 16.3%, which may cause problems with the weight and length production of the fruiting body.
상기 열수 추출된 솔수염하늘소는 증류수 100 중량부에 대하여 솔수염하늘소를 2.5 내지 7.5 중량부로 혼합한 후 50 내지 70℃에서 2 내지 4시간 동안 가열하고 여과한 잔사를 건조하여 수득할 수 있다.The hot water-extracted seaweed can be obtained by mixing 2.5 to 7.5 parts by weight of seaweed seaweed with 100 parts by weight of distilled water, heating at 50 to 70°C for 2 to 4 hours, and drying the filtered residue.
본 발명은 가) 액체배양된 동충하초 균사체를 상기 제조된 솔수염하늘소 고체배지용 조성물에 투입하고 암배양 하는 단계; 및 나) 상기 가)단계에 의해 암배양 후 형광등 조건의 배양기로 이동하여 자실체를 배양하는 단계를 포함하는 것을 특징으로 하는 동충하초 자실체의 생산방법을 제공한다.The present invention includes the following steps: a) adding liquid-cultured Cordyceps sinensis mycelium to the prepared composition for a solid medium and culturing it in the dark; and b) providing a method for producing Cordyceps sinensis fruiting bodies, which includes the step of cultivating the fruiting bodies by moving them to an incubator under fluorescent lighting conditions after dark culturing in step a) above.
상기 동충하초는 밀리타리스 동충하초(Cordyceps militaris)일 수 있지만, 이에 한정되는 것은 아니다.The Cordyceps sinensis is Militaris Cordyceps ( Cordyceps) militaris ), but is not limited to this.
상기 암배양은 22~26℃의 배양기에서 12 내지 16일 동안 암배양할 수 있다. 상기 암배양 조건에서는 자실체의 최초 발이기간이 평균 4.7일, 원기 수가 평균 25개로 나타냈지만, 상기 암배양 조건을 벗어나면 최초 발이기간이 6.3 내지 8.3일, 원기 수가 16 내지 22개로 생장하여 발이기간이 길어지고 원기수가 감소하는 문제가 야기될 수 있다.The cancer culture can be cultured in an incubator at 22 to 26°C for 12 to 16 days. Under the above dark culture conditions, the first germination period of the fruiting body was 4.7 days on average and the average number of primordia was 25. However, if the above dark culture conditions were exceeded, the first germination period was 6.3 to 8.3 days and the number of primordia was 16 to 22, resulting in a short germination period. This may cause problems such as lengthening and reduction in original number.
상기 자실체 배양은 최초 발이가 발생한 시점을 기준으로 40 내지 60일간 배양할 수 있다. 상기 배양기간에서는 자실체의 중량 26.4 내지 27.5 g, 자실체 길이 54.3 내지 55.1 mm, biological efficiency가 26.2 내지 29.5 %로 나타냈지만, 상기 배양기간에서 40일 이하로 배양할 경우, 자실체 중량 9.8 내지 17.5 g, 자실체 길이 15.1 내지 31.5 mm, biological efficiency가 9.4 내지 16.3 %이며, 상기 배양기간에서 60일 이상으로 배양할 경우, 자실체 중량 19.5 내지 26.5 g, 자실체 길이 30.5 내지 53.5 mm, biological efficiency가 20 내지 25 %로 자실체의 중량 및 길이 생산량에 문제가 야기될 수 있다.The fruiting body culture can be cultured for 40 to 60 days based on the time when the first germination occurs. In the above culture period, the weight of the fruiting body was 26.4 to 27.5 g, the length of the fruiting body was 54.3 to 55.1 mm, and the biological efficiency was 26.2 to 29.5%. However, when cultured for less than 40 days in the above culture period, the weight of the fruiting body was 9.8 to 17.5 g, the fruiting body was 54.3 to 55.1 mm in length, and the biological efficiency was 26.2 to 29.5%. The fruiting body has a length of 15.1 to 31.5 mm, a biological efficiency of 9.4 to 16.3%, and when cultured for more than 60 days in the above culture period, a fruiting body weight of 19.5 to 26.5 g, a fruiting body length of 30.5 to 53.5 mm, and a biological efficiency of 20 to 25%. Problems may arise in weight and length production.
상기 동충하초 자실체의 최대 생장량을 위한 최적 배양 조건은 열수 추출된 솔수염하늘소의 투입량, 수분함량 및 배양기간과 하기의 수학식 1과 같은 상관관계를 가지는 것일 수 있다.The optimal culture conditions for the maximum growth amount of the Cordyceps sinensis fruiting body may have a correlation with the input amount, moisture content, and culture period of the hot water-extracted pine needles, as shown in Equation 1 below.
[수학식 1][Equation 1]
Y1 = -244.5075+1.8595X1+4.30810X2+2.21867X3-0.0018X1X2-0.002X1X3-0.000267X2X3-0.1487X1 2-0.021388X2 2-0.023522X3 2 Y 1 = -244.5075 + 1.8595X 1 + 4.30810X 2 + 2.21867X 3 -0.0018X 1 2
상기 수학식 1에서 Y1은 밀리타리스 동충하초 자실체의 생장량 (g/bottle), X1은 열수 추출된 솔수염하늘소의 투입량 (g), X2는 수분함량 (%) 및 X3는 배양기간 (days)을 나타낸다.In the above equation 1, Y 1 is the growth amount of the fruiting body of Militaris Cordyceps sinensis (g/bottle), (days).
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, the present invention will be described in detail through examples to aid understanding. However, the following examples only illustrate the content of the present invention and the scope of the present invention is not limited to the following examples. Examples of the present invention are provided to more completely explain the present invention to those skilled in the art.
<< 실시예Example 1> 동충하초 자실체 생산량을 고효율로 증대시키기 위한 1> To increase Cordyceps sinensis fruiting body production with high efficiency 고체배지solid medium 제조 공정 Manufacture process
도 1은 본 발명의 실시예에 따른 열수 추출한 솔수염하늘소를 이용한 고체배지 제조방법부터 동충하초 자실체 생산방법까지 나타낸 모식도이다.Figure 1 is a schematic diagram showing the method for producing a solid medium using hot water-extracted seaweed porridge according to an embodiment of the present invention to the method for producing Cordyceps sinensis fruiting bodies.
1. 솔수염하늘소를 이용한 열수 추출방법1. Thermal water extraction method using pine needles.
솔수염하늘소의 열수 추출은 둥근바닥플라스크에서 솔수염하늘소 20 g에 증류수 400g을 투입한 후 냉각관을 연결하여 60 ℃에서 3시간 가열하였다 .For hot water extraction of pine beetle, 400 g of distilled water was added to 20 g of pine beetle in a round bottom flask, then a cooling tube was connected and heated at 60°C for 3 hours.
열수 추출 후, Whatman filter paper (No. 2)를 이용하여 감압 여과하고 잔사를 48시간 실온에서 건조시켰다.After hot water extraction, it was filtered under reduced pressure using Whatman filter paper (No. 2), and the residue was dried at room temperature for 48 hours.
이렇게 열수 추출된 솔수염하늘소를 획득하여 고체배지의 원료로 사용하였다.In this way, we obtained the hot-water-extracted pine cone and used it as a raw material for the solid medium.
2. 솔수염하늘소를 이용한 고체배지 제조공정2. Solid medium manufacturing process using pine needles.
귀리 14.7 g, 상기 열수 추출된 솔수염하늘소 5.3 g 및 물 20 g을 첨가하여 혼합한 후에 오토클레이브에서 121 ℃, 30분 동안 고온 멸균하여 배지를 제조하였다.A medium was prepared by adding and mixing 14.7 g of oats, 5.3 g of the hot water-extracted seaweed and 20 g of water, and then sterilizing the mixture in an autoclave at 121°C for 30 minutes.
3. 상기 고체배지에 의한 동충하초 자실체의 생산방법3. Method for producing Cordyceps sinensis fruiting bodies using the above solid medium
사부로오드 덱스트로즈 배지(Sabouraud dextrose broth; 펩톤 10 g, 덱스트로즈 20 g, 증류수 1 L; pH 5.6)에 7일간 액체배양된 동충하초 균사체 현탁액 5 mL를 앞서 준비된 고체배지에 투입 후 24 ℃ 배양기에서 14일간 암배양하였다. 상기 동충하초 균사체는 밀리타리스 동충하초(Cordyceps militaris)를 이용하였다.5 mL of Cordyceps sinensis mycelium suspension cultured in Sabouraud dextrose broth (10 g of peptone, 20 g of dextrose, 1 L of distilled water; pH 5.6) for 7 days was added to the previously prepared solid medium and placed in an incubator at 24°C. Cancer culture was performed for 14 days. The Cordyceps sinensis mycelium is Militaris Cordyceps ( Cordyceps militaris ) was used.
암배양 종료 후, 온도 20 ℃, 습도 60 %, 24시간 형광등 조건의 배양기로 이동한 다음 60일간 자실체를 배양하였다.After completion of dark culture, the cells were moved to an incubator with a temperature of 20°C, humidity of 60%, and fluorescent lighting for 24 hours, and the fruiting bodies were cultured for 60 days.
4. 4. 고체배지solid medium 제조 공정에 따른 동충하초 자실체의 Cordyceps sinensis fruiting body according to the manufacturing process 발이기간Eating period , 원기 수 및 자실체 성숙기, primordia number and fruiting body maturity stage
발이는 자실체가 형성되는 과정 중 형태적, 기능적으로 성숙하기 이전의 단계를 의미하며, 처음 원기가 발생한 날을 기준으로 발이기간을 기록하게 된다. 성숙기는 도 2와 같이 동충하초 자실체의 끝부분에 헤드가 발생되었을 때를 의미하며, 헤드는 일반적으로 두께가 두꺼워지고 돌기가 발생된 부분으로 나타낸다. Germination refers to the stage before morphological and functional maturity in the process of fruiting body formation, and the germination period is recorded based on the date when the first primordia occurs. The maturity stage refers to when a head is generated at the end of the fruiting body of Cordyceps sinensis as shown in Figure 2, and the head is generally indicated as a part that is thick and has protrusions.
표 1은 고체배지 제조 공정에 따른 동충하초의 자실체의 발이기간, 원기 수 및 자실체 성숙기를 나타낸다. 각 값은 평균 ± SE (n = 5)로 표시된다. Table 1 shows the germination period, number of primordia, and fruiting body maturity period of the fruiting body of Cordyceps sinensis according to the solid medium manufacturing process. Each value is expressed as mean ± SE (n = 5).
그 결과 동충하초 자실체의 원기 수는 솔수염하늘소 또는 열수 추출된 솔수염하늘소로 고체배지를 제조했을 때 유의성있게 높은 것으로 확인되었으며, 자실체 성숙기까지의 소요기간이 짧은 것으로 확인되었다. 따라서, 솔수염하늘소 고체배지를 제조하는 것이 동충하초 자실체 생산에 효율적일 것으로 판단되며, 바람직하게는 열수 추출된 솔수염하늘소 배지를 제조하는 것이 더 효율적이었다.As a result, it was confirmed that the number of primordia of Cordyceps sinensis fruiting bodies was significantly higher when a solid medium was prepared with Sulwhasoo or hot-water-extracted Sulwhasin asparagus, and the time required for fruiting bodies to reach maturity was confirmed to be short. Therefore, it is believed that producing a solid medium for Cordyceps sinensis is efficient for producing Cordyceps sinensis fruiting bodies, and it is preferable to produce a hot water extracted medium for Cordyceps sinensis.
5.5. 고체배지solid medium 제조 공정에 따른 동충하초 자실체의 배, 길이, 자실체 중량 및 biological efficiency Doubling, length, fruiting body weight, and biological efficiency of Cordyceps sinensis fruiting bodies according to the manufacturing process
표 2는 동충하초 자실체 생장에 대한 고체배지 공정의 영향을 나타냈으며 배지 중량을 포함한 자실체의 중량, 자실체 길이 및 BE(biological efficiency (%) = fruit body weight/medium weight × 100)를 나타낸다. 각 값은 평균 ± SE (n = 5)로 표시된다.Table 2 shows the effect of the solid medium process on the growth of Cordyceps sinensis fruiting bodies and shows the weight of the fruiting body including the medium weight, fruiting body length, and BE (biological efficiency (%) = fruit body weight/medium weight × 100). Each value is expressed as mean ± SE (n = 5).
그 결과, 표 2와 도 3에 따르면, 솔수염하늘소 또는 열수 추출한 솔수염하늘소를 이용한 고체배지를 이용했을 때, 높은 자실체 길이 및 중량을 나타내는 것으로 확인되었다.As a result, according to Table 2 and Figure 3, it was confirmed that high fruiting body length and weight were observed when using a solid medium using pine needles or hot water extracted pine needles.
또한, 동충하초 자실체 생장의 효율성을 평가를 위해 일반적으로 사용되는 척도인 biological efficiency의 경우, 솔수염하늘소 고체배지를 사용했을 때 높은 것으로 확인되었으며, 바람직하게는 열수 추출된 솔수염하늘소 배지를 제조하는 것이 더 높은 것을 확인할 수 있었다.In addition, biological efficiency, a measure commonly used to evaluate the efficiency of fruiting body growth of Cordyceps sinensis, was found to be high when solid medium was used, and it is preferable to produce hot water extracted medium. It was confirmed that it was higher.
<< 실시예Example 2> 동충하초 자실체 생산량 증대를 위한 2> To increase Cordyceps sinensis fruiting body production 고체배지solid medium 제조공정의 최적화 Optimization of manufacturing processes
열수 추출된 솔수염하늘소 투입량, 수분함량 및 배양 기간을 변수로 하고, Design-expert version 12를 이용하여 표 3과 같이 동충하초 자실체 생산량을 위한 최적의 코드 및 실측값을 얻었으며, 하기 수학식 1을 이용하여 최적 고체배지 제조 조건을 도출하였다.Using the input amount, moisture content, and culture period extracted with hot water as variables, the optimal code and actual measured value for Cordyceps sinensis fruiting body production were obtained as shown in Table 3 using Design-expert version 12, and Equation 1 below was used. The optimal solid medium manufacturing conditions were derived using this method.
[수학식 1][Equation 1]
Y1 = -244.5075+1.8595X1+4.30810X2+2.21867X3-0.0018X1X2-0.002X1X3-0.000267X2X3-0.1487X1 2-0.021388X2 2-0.023522X3 2 Y 1 = -244.5075 + 1.8595X 1 + 4.30810X 2 + 2.21867X 3 -0.0018X 1 2
상기 수학식 1에서 Y1은 밀리타리스 동충하초 자실체의 생장량 (g/bottle), X1은 열수 추출된 솔수염하늘소의 투입량 (g), X2는 수분함량 (%) 및 X3는 배양기간 (days)을 나타낸다.In the above equation 1, Y 1 is the growth amount of the fruiting body of Militaris Cordyceps sinensis (g/bottle), (days).
그 결과, 도출된 회귀식으로부터 동충하초 자실체 생산량을 위한 최적 고체배지 제조 조건을 도 4 및 도 5와 같이 도출할 수 있었으며 귀리 14.7 g, 열수 추출된 솔수염하늘소(전건중량기준) 5.3 g, 증류수 20.0 g 및 배양기간 46.4 일의 조건으로 배양할 때, 자실체 생산량이 최대인 것으로 확인되었다.As a result, from the derived regression equation, the optimal solid medium manufacturing conditions for Cordyceps sinensis fruiting body production could be derived as shown in Figures 4 and 5, including 14.7 g of oats, 5.3 g of hot water-extracted pine needles (based on total dry weight), and 20.0 g of distilled water. It was confirmed that the fruiting body production was maximum when cultured under conditions of 46.4 g and culture period of 46.4 days.
또한, 표 4에 의한 ANOVA 분석으로 동충하초 자실체 생산을 위한 최적 고체배지 제조 조건에 대한 예측모델의 신뢰성 테스트 결과로 도출된 동충하초 자실체 생산을 위한 최적 고체배지는 자실체 생산량을 최대 27.7225 g/bottle로 예상하였으며, 실측한 결과 28.1 g/bottle의 유사한 자실체 생산량을 나타내었다(R2=0.8737).In addition, as a result of the reliability test of the prediction model for the optimal solid medium production conditions for Cordyceps sinensis fruiting body production through ANOVA analysis according to Table 4, the optimal solid medium for Cordyceps sinensis fruiting body production was expected to produce a maximum fruiting body production of 27.7225 g/bottle. , the actual measurement results showed a similar fruiting body production of 28.1 g/bottle (R 2 =0.8737).
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (12)
2) 상기 1)단계에서 제조된 혼합물에 물을 첨가하는 단계; 및
3) 상기 2)단계에서 제조된 혼합물을 고온 멸균하는 단계
를 포함하여 이루어지는 것을 특징으로 하는 솔수염하늘소 고체배지의 제조방법으로써,
상기 열수 추출된 솔수염하늘소는
a) 증류수 100 중량부에 대하여 솔수염하늘소를 2.5 내지 7.5 중량부로 혼합한 후 50 내지 70℃에서 2 내지 4시간 동안 가열하는 단계;
b) 상기 a)단계에서 제조된 혼합물을 감압 여과하는 단계; 및
c) 상기 b)단계에 의해 생성된 잔사를 건조하는 단계
를 포함하여 제조되는 것을 특징으로 하는 솔수염하늘소 고체배지의 제조방법.1) Adding hot water-extracted pine needles to oats;
2) adding water to the mixture prepared in step 1); and
3) High-temperature sterilization of the mixture prepared in step 2) above.
As a method for producing a solid medium for pine beetles, characterized in that it comprises,
The thermal water extracted pine beard sky cow is
a) mixing 2.5 to 7.5 parts by weight of pine needles per 100 parts by weight of distilled water and then heating at 50 to 70°C for 2 to 4 hours;
b) filtering the mixture prepared in step a) under reduced pressure; and
c) drying the residue produced in step b).
A method for producing a solid medium for pine beetles, characterized in that they are manufactured including.
나) 상기 가)단계에 의해 암배양 후 형광등 조건의 배양기로 이동하여 자실체를 배양하는 단계
를 포함하는 것을 특징으로 하는 동충하초 자실체의 생산방법. A) Injecting the liquid-cultured Cordyceps sinensis mycelium into the composition for a solid medium prepared according to claim 1 and culturing it in the dark;
b) After culturing in the dark in step a) above, moving to an incubator under fluorescent lighting conditions and cultivating the fruiting body.
A method of producing Cordyceps sinensis fruiting bodies, comprising:
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