TW200835791A - Method for artificially propagating fruit body stroma of Antrodia Camphorate - Google Patents

Abstract

The present invention provides a method for artificially propagating fruit body of Antrodia Camphorate. The invention also provides formulas of liquid fermentation culture mediums to be used for growing most types of fungi. The method also implants cultures of an Antrodia Camphorate into wood filled within the flask to perform mycelium cultivation , and then propagates fruit bodies in a flask containing air. The flask is sealed by porous rubber and the pores are filled with cotton. Relative humidity in the flask can be kept to propagate Antrodia Camphorate while preventing other types of fungi or bacteria from intruding the flask and exhausting gases generated by the fermentation metabolism of fungi. The invention can increase the productivity of the fungi, and stimulates fungi to generate metabolite, especially for the metabolite containing medical active ingredients.

Description

200835791 九、發明說明（1) <發明背景> 1·發明的範圍： 本發明—r牛樟芝-子實體j之繁育方法；本發明同^»1液相發 酵靜置培養基處方，可用於大部分真菌之生長，時該方法包括將一樟芝菌種植入 以瓶裝盛之木料上進行菌絲體培養，並續於含有空氣之瓶中進行子實體培育。該瓶 係以有孔洞之橡膠塞封口，並裝讎花入膠雛洞中，一方面阻隔雜菌入侵並可確 保培養瓶中樟芝菌成長所需之相醒度，另一方面可排放眞菌因發酵代謝所生氣 體。不僅提高真菌之產量，同時刺激其衍生代謝物，特別是含有醫藥活性成分的代 謝物办 2·有關技籠域之敍述： 牛樟芝（Antrodia camphorate) 子菌 basidiomycete 多孔菌科 Polyporaceae，生長季200835791 IX. Inventive Description (1) <Invention Background> 1 Scope of the Invention: The present invention - a method for breeding r antarctica - fruiting body j; the present invention is the same as the liquid phase fermentation static medium prescribed for the liquid phase, which can be used for large For the growth of part of the fungus, the method comprises planting a sphagnum sinensis into the bottled wood for mycelium culture, and continuing the cultivation of the fruit body in a bottle containing air. The bottle is sealed with a rubber plug with a hole, and the flower is inserted into the rubber hole. On the one hand, it blocks the invasion of bacteria and ensures the awakening degree required for the growth of the Phytophthora in the culture bottle. On the other hand, the bottle can be discharged. The bacteria produce a gas by fermentation. It not only increases the yield of fungi, but also stimulates its metabolites, especially the metabolites containing medicinal active ingredients. 2. Description of the technology cage: Antrodia camphorate, basidiomycete Polyporaceae, growing season

節；春、夏、秋，棲息於中、低海拔之台灣國寶級保育樹種1"牛樟樹 Cinnamomumcomphora』上，在腐朽之牛樟木材上，引起木材褐腐朽，是導致牛樟木 腐朽中空的元兒^牛樟芝是台灣特有之樟薄孔菌Antrodia dnnamomea，享有森林中「台 灣紅寶石」之美譽。其子實體一年生至多年生，呈鮮血紅色、橘紅色至淡肉桂色， 味呈黃樟香氣、性苦寒。產期在每年六月至十一月，十一月以後受温度之影響其成 長緩慢。早期原住民上山工作，在口中含一小片就可整天不用喝水、而且防止宿醉， 民間傳說其具殺蟲、保肝、解毒功能。現代硏究證實；牛樟芝富含三鞀類、固醇類、 #醣成份 > 可增强肝功能、抗氧化，晚疫力腳制麵之效用。 由於牛樟芝野外數量不多，而民間需求有增無減，稱得上是全世界最昂貴的眞菌， 也因此導致「山老鼠（盜林者)」濫砍»的目標。目前生物科技已能人工培養樟芝 菌絲體（市面上·，分有液態深層發酵繁殖菌絲體及固體培育牛樟菌絲體兩類）以菌 絲體商品當作代用品。但各種培養牛樟菌絲體之製程，均有其操控極限和困難，不 能也無法去完整臨摹野生子實體成長，導致產出 之品質效用有其極限，難達野生子實體相同標準，致價碼高漲難以滿足需求。 3·先前技術： 野生牛樟芝因受牛樟樹保育已明令禁採，雖有人利用颱風過境擇取牛樟漂流木，鋸 成段木並灑上牛樟菌種，經4-6個月削下牛樟芝子實體薄層販售，但生長和培育條件 管控困難，爲數仍有限無法滿足需求。一般樟芝菌種之保存^氏零下8〇度液態氮 中。而繁麵麥芽抽出物（ΜΕΑ)、甚至酵母抽出物培養基（YEA例：2%«^糖0·5% 酵母抽出物）或馬鈴薯培養基（PDA)等培養，並可於處方中添加0.1%磷酸氫二鉀 及0.05%硫酸鎂，借以促進菌絲之成長。該等培養基注入平版皿中呈固相並温^氏 15-32度，均瓶靜置培養。 通常液態深讎酵繁殖牛樟雜絲體方法；先以特定培養基調製之固相平顧靜置 培養2周，再將菌絲置於三角瓶中並以經調製液體培養基，以攝氏25-26度進行震盪 培養7-10天之後移入相似條件之種子罐攪拌通氣培養，然後轉人相似條件、轉速 9_、通氣量爲0.5wm之小型（500公升）發丽培養，再轉入中型（5公噸）發 酵槽培養，再升級入更大型發酵槽中培養，其各級之攪拌通氣培養均約7-10天。 200835791 九、發明說明（2) 一般培養條件爲1%之接種量，轉速90_ι、通氣量爲0.5™温度爲攝氏28度（例； 以五十噸槽進行三十五噸培養，八天後可得紅色菌絲，再行固液分離，或有發酵全 液以温度攝氏55度減壓濃縮後進行冷凍乾燥，收率可達1.8%其多醣體含量則在8% 以上），且多醣體含量與菌絲多寡成正比〇 惟一般深層液態發酵方法；均需進行固液分離，有廢液排放污染環境之問題。且因 該發酵廢液之排放常導致溶解於水中的小分子活性成分流失，液態發酵常僅能取得 菌絲細胞壁內所含之物質成分。 固體培養牛樟菌方法；先齡樟芝菌種以太空行菌絲體培養，並續於空氣中進 行子實體纖，且利用該方法培觀之樟芝固體培養物（專利公告號00565430) 〇 該專利係一種培養固體樟芝之方法，·以BCRC 35398爲菌種來源植在太空包內，且該 太空包內含之培養基係由60-70%選自蕈類、植物之莖、梗、果實及木屑之纖維物、 20·30%馬鈴薯、5-15%雑、1-10%葡萄糖、052%磷酸二氫鉀及0.1-1%硫組成， 其溼度爲6仏80%之間，並調整其pH値爲中性；根據該專利之方法，太空包之培養 基係由65%草本植物之莖、梗、果實或木屑、20%馬鈴薯、10%米糠、3.5%葡萄糖、 1%磷酸二氫鉀及0.5%硫酸鎂所組成。且其中子實體栽培期間之日間溫度係介於20 至30°C之間，而夜間麵係介於8-14°C之間，且控制其日夜間溫差於12至16°C之 間。其溫度爲5至32°C間，含有0.24%之二氧化碳之空氣下進行菌絲體培養（培養 進行50-80天），隨之移除太空包，使之暴露於二氧化碳含量低於1%且空氣淫度介 於90^95%之間之空氣流通條件下，進行子實體栽培一段時間（子實體培養須20^50 天）至圓錐形子實體形成。 但此固體培養牛樟菌之方法，受限於太空包內所含培養基材，此基材在提供營養成 份後易軟化且此時已移除太空包，一方面易受雜菌汚染無法久置。故需在特定期內 行採收，致其圓錐形子實體形成時麵，所形成之表層膜甚薄。 類均具極佳吸附金屬之能力，例;2006年日本麒麟啤酒之關係生技產品『姬 松茸-巴西蘑菇』就發生過高重金屬含量而下市，而我國也發現國產生鮮1r巴西蘑 菇』中亦隱藏有相同問題-究其因不外乎巴西蘑菇、洋菇覆土性需求所造成❹在現 今工業發達所造成空氣和環境土壤污染，導致野生之 牛樟芝所含銅、錳、鋁、硼、銻明顯高於人工培育牛樟芝菌絲體。 牛樟芝菌固、液態培養首見文獻於1997年師大簡狄源等之硏究，其中添加0.1^0.3% 樟腦結晶可助於菌絲體生長。其後1998年瘳英明指出添加0.02%之 樟木萃出液（10%之樟木屑水）可有縦的成長，2000年許勝傑等發現樟樹 精油中常見如teipineol、safrolr、camphor成分皆有助於樟芝菌絲體生長。 爲防止環境污染導致野生牛樟芝商品具高重金屬含量，以及盜砍牛樟木、盜採牛樟 芝，唯有人工培育牛樟链體（子實體）一途。如比一來·，除了能供應市場之需求， 還能爲台灣國寶r牛樟木j保育樹及天然牛樟芝留下生路。Festival; spring, summer, autumn, inhabited in the middle and low altitude Taiwan national treasure-level conservation tree species 1 " burdock Cinnamomumcomphora, on the decaying burdock wood, causing brown decay of the wood, is the cause of the decay of the burdock wood ^ Antrodia is the endemic Antrodia dnnamomea, which is unique to Taiwan. It enjoys the reputation of “Taiwan Ruby” in the forest. Its fruiting body is born from perennial to perennial. It is blood red, orange-red to pale cinnamon, and its taste is yellow scent and bitter cold. The production period is from June to November every year, and after November, the growth is slow due to the temperature. Early aborigines went to work in the mountains, and in the mouth, they could drink water without drinking water and prevent hangovers. Folklore has the functions of killing insects, protecting liver and detoxifying. It is confirmed by modern research; Antrodia camphorata is rich in triterpenoids, sterols, and #saccharide ingredients> It can enhance liver function, anti-oxidation, and effect of late-effect foot-making. Because there are not many wild animals in the wild, and the demand for the private sector has increased, it is the most expensive sputum in the world, which has led to the goal of "Mountain Mouse (Pirates)". At present, biotechnology has been able to artificially culture the mycelium of Antrodia camphorata (on the market, there are two types of liquid deep-fermentation and propagation mycelium and solid cultivation of Burdock mycelium), and the mycelium products are used as substitutes. However, all kinds of processes for cultivating burdock mycelium have their control limits and difficulties. They cannot and cannot complete the growth of wild fruiting bodies in Linyi, which leads to the limit of the quality utility of output, and it is difficult to reach the same standard of wild fruiting bodies. It is difficult to meet the demand. 3. Previous technology: Wild burdock has been banned for burdock conservation. Although some people use typhoon to choose burdock driftwood, saw it into segments and sprinkle with burdock strain, and cut burdock for 4-6 months. The physical thin layer is sold, but the growth and cultivation conditions are difficult to control, and the number is still limited to meet the demand. In general, the storage of Antrodia strains is in the liquid nitrogen of minus 8 degrees. The flour is extracted from the malt extract (ΜΕΑ), or even the yeast extract medium (YEA case: 2% «^ sugar 0. 5% yeast extract) or potato medium (PDA), and 0.1% can be added to the prescription. Dipotassium hydrogen phosphate and 0.05% magnesium sulfate to promote the growth of hyphae. The medium was injected into a lithographic dish in a solid phase and the temperature was 15-32 degrees, and the flasks were statically cultured. Usually, the liquid deep-fermentation method is used to propagate the burdock filament method; the solid phase prepared by the specific medium is firstly incubated for 2 weeks, and the hyphae are placed in a triangular flask and the prepared liquid medium is used at 25-26 Celsius. After 7-10 days of shaking culture, the seed tanks of similar conditions were transferred to agitated aeration culture, and then transferred to a small (500 liter) larval culture with similar conditions, rotation speed of 9 _, and aeration of 0.5 wm, and then transferred to medium (5 metric tons). The fermentation tank is cultured and then upgraded to a larger fermentation tank for cultivation, and the agitation culture of each stage is about 7-10 days. 200835791 IX. Description of invention (2) The general culture condition is 1% inoculation, the rotation speed is 90_ι, the ventilation is 0.5TM, the temperature is 28 degrees Celsius (for example; 35 tons of cultivation is carried out in 50 tons of tank, after 8 days Red hyphae are obtained, and then solid-liquid separation is carried out, or the whole fermentation liquid is concentrated under a reduced pressure of 55 degrees Celsius, and then freeze-dried, the yield is up to 1.8%, the polysaccharide content is above 8%, and the polysaccharide content is obtained. It is directly proportional to the number of hyphae, but the general deep liquid fermentation method; both need solid-liquid separation, and there is a problem that the waste liquid discharges the environment. Moreover, since the discharge of the fermentation waste liquid often causes the loss of the small molecule active ingredient dissolved in the water, the liquid fermentation often only obtains the substance components contained in the mycelial cell wall. Method for solid-culture of Burdock bacteria; Agrobacterium sinensis strains are cultured in space mycelium, and continue to be in the air for fruiting body fibers, and the solid culture of Antrodia camphorata cultivated by this method (Patent Bulletin No. 00565430) The patent is a method for cultivating solid Antrodia camphorata, which is planted in a space bag with BCRC 35398 as a strain source, and the medium contained in the space pack is 60-70% selected from the group consisting of mites, stems and stems of plants. Fruit and wood fiber, 20.30% potato, 5-15% strontium, 1-10% glucose, 052% potassium dihydrogen phosphate and 0.1-1% sulfur, the humidity is between 6 and 80%, and The pH is adjusted to be neutral; according to the method of the patent, the medium of the space package is 65% of herbaceous stems, stems, fruits or sawdust, 20% potato, 10% rice bran, 3.5% glucose, 1% dihydrogen phosphate Potassium and 0.5% magnesium sulfate. The daytime temperature during the fruiting body cultivation period is between 20 and 30 °C, and the nighttime facial system is between 8-14 °C, and the day and night temperature difference is controlled between 12 and 16 °C. The mycelial culture (culture for 50-80 days) is carried out at a temperature of 5 to 32 ° C, containing 0.24% of carbon dioxide, and the space capsule is removed to expose it to a carbon dioxide content of less than 1%. Under air circulation conditions of between 90 and 95%, the fruiting body is cultivated for a period of time (the body culture is required to be 20^50 days) to form a conical body. However, the solid culture method of Burdock is limited to the culture medium contained in the space package. The substrate is softened after providing nutrients and the space bag has been removed at this time. On the one hand, it is susceptible to contamination by bacteria. Set. Therefore, it is necessary to harvest in a certain period of time, so that the conical sub-solids form the surface, and the surface film formed is very thin. The class has excellent ability to adsorb metal. For example, in 2006, the related biotechnology product of Japanese Kirin beer, Agaricus blazei-Brazilian mushroom, was produced under high heavy metal content, and China also found that the country produced fresh 1r Brazilian mushroom. It also hides the same problem - it is caused by the soil and environmental soil pollution caused by the current industrial development of Brazil's mushroom and mushroom soil, resulting in the copper, manganese, aluminum, boron and strontium contained in the wild Antrodia camphorata. Significantly higher than artificially cultivated mycelium of Antrodia camphorata. The first and foresight of the culture and liquid culture of Antrodia camphorata was studied in 1997. The addition of 0.1^0.3% camphor crystals can help mycelium growth. Later, in 1998, Yan Yingming pointed out that the addition of 0.02% eucalyptus extract (10% sassafras water) can grow awkwardly. In 2000, Xu Shengjie discovered that common eucalyptus oils such as teipineol, safrolr and camphor are helpful. Mycelium growth of Antrodia camphorata. In order to prevent environmental pollution, the wild Antrodia camphorata products have high heavy metal content, as well as stealing burdock wood and stealing burdock, only artificially cultivating the burdock chain (fruit body). For example, in addition to being able to supply the market, it can also leave a path for Taiwan's national treasure r burdock j conservation tree and natural burdock.

Claims (1)

200835791 十、申請專利範圍（1): 1·根據本發明說明的第1項範圍之方法，該培養基係由牛樟木料65%、馬鈴薯20%、 米糠10%、蔗糖3.5%、磷隨二鉀1%及硫瞧0.5%所組成。 1根據發明說明的第1項範圍之方法，該培養基係由牛樟木料65%、豆漿30% (該黄豆漿瓶和賴1:7 -1:14比率硏製而成）蔗糖3.5%、磷隨二鉀1%及硫賺 0.5%所組成。 3·根據本發明說明的第1項範圍之方法，該培養基係由牛樟木料65%、營養液3〇%(該 營養液係以酵母抽出物0.5-3%)、蔗糖3·5%、磷隨二鉀1%及硫麵0.5%所組成。 4·根據本發明說明的第1項範圍之方法，該培養基係由牛樟木料65%、營養液3〇% (該 營養液舰麥雜出物0.5-3% )、蔗糖3.5%、磷酸氫二鉀1%及硫麵〇·5%所組成。200835791 X. Patent application scope (1): 1. According to the method of the first aspect of the present invention, the culture medium is composed of burdock wood 65%, potato 20%, rice bran 10%, sucrose 3.5%, phosphorus with dipotassium 1% and sulphur 瞧 0.5%. 1 according to the method of the first aspect of the invention, the culture medium is 65% of burdock wood, 30% of soybean milk (the soymilk bottle and Lai 1:7 -1:14 ratio), sucrose 3.5%, phosphorus It consists of 1% dipotassium and 0.5% sulfur. 3. The method according to the first aspect of the present invention, wherein the culture medium is 65% of burdock wood, 3% by weight of nutrient solution (the nutrient solution is 0.5-3% of yeast extract), and 3.5% of sucrose, Phosphorus consists of 1% dipotassium and 0.5% sulfur surface. 4. The method according to the first aspect of the present invention, wherein the culture medium is 65% of burdock wood, 3% by weight of nutrient solution (0.5-3% of the nutrient solution, sucrose, 3.5%, hydrogen phosphate) Dipotassium 1% and sulphur surface 〇·5%. 5·根據本發明說明的第1項範圍之方法，該培養基係由牛樟木料65%、營養液3〇% (該 營養液係以酵母抽出物0.5-2%、麥芽抽出物0.5-2%)、蔗糖15%、磷酸氫二鉀1% 及硫麟0.5%所組成。 6·根據本發明說明的第i項範圍之方法，該培養基係由樟木科（本棒、油棒、 芳樟、陰陽樟）木料65%、馬鈴薯20%、米糠10%、蔗糖3.5%、磷酸氫二鉀1%及硫 麵〇·5%所組成。 7·根據本發明說明的第1項範圍之方法，該培養基係由滕香杉芝生長及金針菇繁育 的杉科（如香杉等）梢65%、馬鈴薯20%、雜10%、蔗糖3.5%、磷酸氫二鉀1% 及硫難0.5%所組成。 8·根據本發明說明的第1項範圍之方法，該培養基係由適於各類香菇、木耳等薛菌繁 育的雜;^ (如相思樹、油桐樹等）65%、馬鈴薯20%、 10%、蔗糖3.5%、磷 驗二鉀1%及硫麵0.5%所組成。 9.根據本發明的第}項範圍之方法，該培養基係將馬鈴薯、米糠和水萃出之液，再加 入蔗糖3.5%、磷酸氫二鉀1%及硫麵〇·5%所組成營養液。 1〇· _本發明說明的第1項範圍之方法，牛樟等_4木料需經含水量、濕度管控，先 ^水【以烘箱控温65 C乾燥8小時，駐水ΐ〇·3-14·5% (Ave.12.5%)】，乾燥後賺 g乾淨水【浸泡3-8天（5天爲最佳），使吸附含水量約35%】，再浸塗經調配並已 均質攪碎之讎狀牛讎菌賺面。該培養總將馬鈴薯20%、雜腦以水萃出, ^加入蔗糖15%、讎氫二鉀1%及硫_ 〇·5%和甲素〇.5%-2%所組成之稠 200835791 十、申請專利範園（2): 11.根據本發明說明的第1項範圍之方法，牛樟等樟科木料含水量、濕係以自 然風乾之木料去浸泡乾淨水3-8天（5天爲最佳）使吸附含水量約35%，苒塗浸經 調配並已均質攪碎之翻狀牛_菌纖面。該培養麵將馬鈴^ 2〇% 以水萃出，再加入蔗糖3.5%、磷醜二鉀1%及硫纖α5%和甲基纖維素〇偏％ 所組成之稠狀營養液。 ΐ2·本發明，月的第1項範圍之方法，各a#科（香杉）、雜梅等，需經含水量、 濕度管控’先祛水【以烘箱控温65 C乾燥8小時，去水ι〇·3%44·5%( Ave.12.5%)】， 後再獅S調配之培養基面，該培養^^將馬鈴薯2〇%、ι〇%以 水萃出，再加入蔗糖3·5%、磷酸氫二鉀1%及硫酸鎂〇·5%所組成之營養液（浸泡3_8 天，5天爲最佳使吸附含水量約35%)，再經以攝氏121度殺菌60分鐘，備供^5. The method according to the first aspect of the present invention, wherein the culture medium is 65% of burdock wood and 3% by weight of nutrient solution (the nutrient solution is 0.5-2% of yeast extract, and malt extract 0.5-2). %), 15% sucrose, 1% dipotassium hydrogen phosphate and 0.5% sulphur. 6. The method according to the scope of item i of the present invention, wherein the medium is composed of 65% wood of the genus Aphididae, the oil bar, the fragrant barium, the yin and yang, the potato 20%, the rice bran 10%, and the sucrose 3.5%. It consists of dipotassium hydrogen phosphate 1% and sulphur surface 〇·5%. 7. The method according to the first aspect of the present invention, wherein the medium is 65% of the cedar family (such as cedar, etc.), 20% of the potato, 10% of the miscellaneous, and 3.5% of the sucrose, which are grown by T. sylvestris and Flammulina velutipes. It consists of dipotassium hydrogen phosphate 1% and sulfur difficulty 0.5%. 8. The method according to the first aspect of the present invention, wherein the culture medium is made of a variety suitable for various types of mushrooms, fungus, and the like; 6 (e.g., acacia, tung tree, etc.) 65%, potato 20%, 10%, sucrose 3.5%, phosphorus test potassium 1% and sulfur surface 0.5%. 9. The method according to the scope of the invention, wherein the medium is a solution of potato, rice bran and water, and a nutrient solution comprising sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulphate 5%. . 1〇· _ The method of the first item of the present invention, the burdock, etc. _4 wood material needs to be controlled by water content and humidity, first water [drying in oven at 65 C for 8 hours, stationed in water ΐ〇·3- 14·5% (Ave.12.5%)】, after drying, earn g clean water [soaked for 3-8 days (5 days is the best), so that the adsorption water content is about 35%], then dip-coated and blended and homogenized The smashed burdock burdock earns noodles. The culture always takes 20% of the potato and the miscellaneous brain is extracted with water. ^ Adds sucrose 15%, bismuth hydrogen dipotassium 1%, sulfur _ 〇 · 5% and A 〇 5% 5% - 2% thick 200835791 Patent application garden (2): 11. According to the method of the first item of the present invention, the water content of the burdock and the like, and the moisture of the wet system are soaked in clean water for 3-8 days (5 days). It is optimal to make the adsorbed water content about 35%, and the mash is immersed in the turned-over bovine porridge. The culture surface was extracted with water, and then a thick nutrient solution composed of sucrose 3.5%, phosphorus ugly potassium 1%, sulfur fiber α 5% and methyl cellulose oxime % was added. Ϊ́2· The invention, the method of the first item of the month, each a# section (香杉), miscellaneous plum, etc., need to be controlled by water content and humidity, 'first simmering water' [drying in oven at 65 C for 8 hours, dewatering 〇 〇 3% · · · 3% 3% 3% 3% 3% 3% 3% 3% 3% 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 调 S S S S S S S S S S S S Nutrient solution consisting of %, dipotassium hydrogen phosphate 1% and magnesium sulfate 5% 5% (soaked for 3_8 days, 5 days is the best to make the adsorbed water content about 35%), and then sterilized at 121 degrees Celsius for 60 minutes. For ^ 13. 根據本發明說明的第1項範圍之方法，各種杉科（香杉）、雜木料等，含水量、濕 度管控，廳自然賺之賴去浸讎調配之培養基纖面，該培養麵將馬鈴薯 20%、雜10%以水萃出，再加入蔗糖3.5%、磷酸氫二鉀1%及硫_ 〇·5%所組成 之營養液（浸泡3-8天，5天爲最佳使吸附含水量約35%)，再經以攝氏121度殺菌 60分鐘，備供菌。 14. 根據本發明的第1項範圍之方法，其中菌絲體培養之溫度爲28°0 15. 根據本發明說明的第1項範圍之方法，其瓶中空氣溼度爲80% 〇 16. 根據本發明說明的第1項範圍之方法，其中該子實體栽培時之日間溫度爲24^261 ’ 而夜間溫度爲1042°C 〇 17. 根據本發明說明的第1項範圍之方法，其中其日夜間溫差爲15°C 〇 18·讎本發明說明的第1項範圍之方法，成長光線以七分陰三分陽爲佳，其菌絲體培 養進行4045天，而子實體培養須90420天（含以上）。 19.根據本發明說明的第1項範圍之方法，其於菌絲體培養階段前，可先處以一牛_ 種大量培養，步驟包含(1)自液態氮保存活化的菌種取一小塊洋雜絲塊，移入新鮮 培養基中顏溫下培養；_菌種生長至旺盛時，將該菌纖種至殺過菌之五穀雜 糧（例如；米粒、麥粒或麥片）之中續於怪溫下培養；（3府該五榖雜糧中之菌種長 猶絲時，取出雜菌絲塊，然後置入經調配之_狀營養液中，再以均質讎碎 成樟芝種菌液，即可提供大量接賴源。 200835791 十、申請專利範圍（3): 20·據本發明說明的第1項範圍之方法，菌絲體培養階段，包含一牛樟菌種大量培養步 驟，包含以液體培養發酵以大量培養菌種，斜面試管培養後，將大量繁殖好的菌種 接種之5 _瞒養液中，於24»261下以160 rpm旋剛隱培養約u天，續以90rpm 往返振盪培養約14天，又MM 20升液體發酵液中^^養天；其中發酵培養 液之配方爲0·1-1%酵母抽出物、0.1-1%麥芽抽獅、1-3%蔗糖、0·05»0·5%磷酸氫二鉀、 0.01-0.1% 硫酸鎂。 21.根據本發明說明的第1項範圍之方法’牛樟等形狀；不限於塊狀、條狀或 片狀 &gt; 其尺寸大小 '長度 '鎌可視容嚣大小麵高决定。 22·根據本發明說明的第1項範圍之方法，容罷材質；不限於玻璃或塑膠。 23.根據本發明說明的第1項範圍之方法，容器形式;不限於瓶、罐或桶。 24·根據本發明說明的第1項範圍之方法，瓶塞材質；不限於橡膠或砂膠❶ 25·讎本發明說明的第1項範圍之方法，碳源;不酿蔗糖、黑糖、麵糖或果糖。 26·根據本發明說明的第1項範圍之方法，氮源；不限於米糠、黄豆、酵母抽出物、麥 芽抽出物。 27. 根據本發明說明的第1項範圍之方法，元素；不酸氯υ酸二氨· 28. 根據本申請專利範圍第20條之方法，其液體發酵培養，培養液之配方爲α5%酵母 抽出物、0.5%麥芽抽出物、2%蔗糖、0·1%ϋ酸氫二鉀及0 Q5%硫〇 29·根據本申請專利範圍第10條、第11條、第19條之方法，其粘稠狀營養液係將馬 鈴薯20%、雜10%以水萃出，再加入蔗糖3·5%、磷酸氫二鉀1%及硫_ 〇·5%和 甲基纖維素1%所組成之稠狀營養液。本粘稠狀營養液氮源；不限於米糠、黃豆、 酵母抽出物、麥芽抽娜❶碳源；不瞻薄糖、黑糖、麵糖或果糖。 30.根據本申請專利範圍第10條、第11條之方法，牛棒等各種木料_乾淨水5 天後，其自然釋出之樟樵出液（樟木層水），因含te^ed、safrolr、c卿hor成 分，可當替代水用於浸泡香杉、相思樹等木料。 200835791 七、指定代表圖： (一) 本案指定代表圖為：第（1)圖。 (二) 本代表圖之元件符號簡單說明：13. According to the method of the first aspect of the present invention, various species of cedar (scented cedar), miscellaneous wood, etc., water content and humidity control, and the hall naturally relies on the medium surface of the dip-mixed medium, and the culture surface will be Potato 20%, miscellaneous 10% is extracted with water, and then added nutrient solution consisting of sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur _ 〇 · 5% (soaking for 3-8 days, 5 days is the best for adsorption) The water content is about 35%), and then sterilized for 60 minutes at 121 degrees Celsius. 14. The method according to the first aspect of the present invention, wherein the mycelial culture temperature is 28°. 15. The method according to the first aspect of the present invention, wherein the air humidity in the bottle is 80% 〇16. The method of the first aspect of the invention, wherein the fruiting temperature of the fruiting body is 24^261' during the cultivation, and the nighttime temperature is 1042 °C. 17. The method according to the first item of the present invention, wherein the day The nighttime temperature difference is 15 °C 〇18·雠 The method of the first item of the present invention, the growth light is preferably seven-point yin and three-point yang, the mycelial culture is carried out for 4045 days, and the fruit body culture is required to be 90,420 days ( Including the above). 19. A method according to the scope of the first aspect of the present invention, which may be preceded by a large amount of culture before the mycelial culture stage, the step comprising: (1) taking a small piece from the liquid nitrogen-activated strain. The foreign silk is transferred to a fresh medium and cultured at a temperature of aging; when the strain grows to a strong state, the strain is fermented into the sterilized whole grains (for example, rice grains, wheat grains or cereals). Under the cultivation; (3) When the strain of the five amaranth grains is long, the hyphae are taken out, and then placed in the formulated _-shaped nutrient solution, and then homogenized and chopped into the aglythous inoculum. A large number of sources are provided. 200835791 X. Patent application scope (3): 20. According to the method of the first aspect of the present invention, the mycelium culture stage comprises a large number of culture steps of a burdock strain, including liquid culture Fermentation is carried out in a large number of cultured strains, and after culturing in a slanted test tube, a large number of well-preserved strains are inoculated into the 5 _ broth, and cultured at 24 rpm at 160 rpm for about 5 days, followed by shaking culture at 90 rpm. About 14 days, and MM 20 liters of liquid fermentation broth The formula of the nutrient solution is 0.1-1% yeast extract, 0.1-1% malt lion, 1-3% sucrose, 0.05»0.5% dipotassium hydrogen phosphate, 0.01-0.1% magnesium sulfate. 21. The method according to the first aspect of the present invention, the shape of the burdock or the like; not limited to a block shape, a strip shape or a sheet shape&gt; the size 'length' is determined by the height of the visible volume. 22 The method of the first aspect of the invention, the material of the invention, is not limited to glass or plastic. 23. The method according to the scope of the first aspect of the invention, the container form; not limited to a bottle, a can or a barrel. The method of the first item of the description, the material of the stopper; not limited to the rubber or the mortar ❶ 25 · The method of the first item of the invention, the carbon source; no sucrose, brown sugar, facial sugar or fructose. The method according to the first aspect of the present invention, the nitrogen source; not limited to rice bran, soybean, yeast extract, and malt extract. 27. The method according to the first aspect of the present invention, the element; the acid chloride Acid diamine. 28. According to the method of Article 20 of the patent application, the liquid fermentation culture, the culture liquid The formula is α5% yeast extract, 0.5% malt extract, 2% sucrose, 0.1% dipotassium hydrogen citrate and 0% 5% thiopurine 29. According to the scope of the patent, Article 10, Article 11, In the method of 19, the viscous nutrient solution extracts 20% of the potato and 10% of the miscellaneous water, and then adds 3% of sucrose, 1% of dipotassium hydrogen phosphate, and sulphur _ 〇·5% and methyl fiber. A thick nutrient solution consisting of 1%. This viscous nutrient solution nitrogen source; not limited to rice bran, soybean, yeast extract, malt extracting carbon source; not thin sugar, brown sugar, facial sugar or fructose. 30. According to the method of Articles 10 and 11 of the patent scope of the present application, various kinds of wood materials such as beef rods and _ clean water for 5 days, the natural release of the sputum liquid (the eucalyptus layer water), because te^ed , safrolr, c qing ingredients, can be used as a substitute for water to soak the cedar, acacia and other wood. 200835791 VII. Designated representative map: (1) The representative representative of the case is: (1). (2) A brief description of the symbol of the representative figure: 八、本案若有化學式時，請揭示最能顯示發明特徵的化學式： 200835791 發明專利說明書8. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention: 200835791 Patent Description (本說明書格式、順序及粗體字，請勿任意更動，※記號部分請勿填寫） ※申請案號：096106426 ※申請日期：2007/02/26 焱1^0：分類： 一、 發明名稱： 一^&gt;^纖F牛樟芝-子實體j之繁育方法 METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate 0 二、 申請人：（共1人） 姓名或名稱：李培榮/Li, Pei-Jung 代表人：李培榮/Li, Pei-Jung 住居所或營業所地址： 苗栗縣三灣鄕銅鏡村一鄰十四號 No.l4,Alleyl, Timg Ching Village Sanwan hsiang, Miaoli, 352 Taiwan (R.O.C·) 國籍：中華民國/R.O.C _三、發明人：（共1人) 姓名：李培榮/Li，Pei-Jung 國籍：中華民國/R.ac 200835791 發明專利說明書 卜年6月 --&quot; ——一 (本說明書格式、順序及粗體字，請勿任意更動，※記號部分請勿填寫） ※申請案號：0961⑽426 ※申請日期：2007/02/26 兹1卩€分類：&quot; 、 ^Olh Wo^O (2006.01)&quot; 一、發明名稱： 一歡工纖&quot;牛樟芝孑實體』之繁育方法 METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF ⑩ .、申請人：（共1人） 姓名或名稱··李培榮/Li, Pei-Jung 代表人：李培榮/Li，Pei-Jung 住居所或營業所地址： 苗栗縣三灣鄉銅鏡村一鄰十四號 No. 14, Alley 1, Tung Ching Village Sanwan hsiang, Miaoli, 352 Taiwan (R.O.C.) 國籍：中華民國/R.O.C φ 三、發明人：（共1人） 姓名：李培榮/U, Pei-Jung 國籍：中華民國/R.O.C 200835791 四、聲明事項： 口主事=7期二為，二，/工-款㈣^ 口 = 前已向下列國家(地區)申曰請專利： 门^依x理國豕（地區）、巾請日、申請案號順序註記】 0料槪第二十七條第-卵際優先權： 籲 □無主張專利法第二十七條第-項國際優先權： □=張專利法第二十九條第一項國内優先權： 【式請依··申請曰、申請案SI順序註記】 0主張專利法第三十條生物材料： D須寄存生物材料者： 國内生物材料【格式請依：寄存機構、日期、號碼順序註記】 % S外生物材料【格式請依：寄存國家、機構、曰期、號碼順序註記】 0不須寄存生物材料者： 所屬技術倾中具有通常知識者綠獲㈣，不須寄存。 200835791 四、聲明事項： □主張專利法第二十二條第二項□第一款或□第二款規定之事實，其 事實發生曰期為：年月曰。 □申請前已向下列國家（地區）申請專利： 【格式請依：受理國家（地區）、申請曰、申讀案號順序註記】 口有主張專利法第二十七條第一項國際優先權： □無主張專利法第二十七條第一項國際優先權：(The format, order, and bold type of this manual should not be changed at all. ※Please do not fill in the mark. ※Application number: 096106426 ※Application date: 2007/02/26 焱1^0: Classification: 1. Name of the invention: ^^&gt;^ Fiber F Antrodia camphorata - fruiting body j breeding method METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate 0 II. Applicant: (1 in total) Name: Li Peirong/Li, Pei-Jung Representative :Li Peirong/Li, Pei-Jung Residence or Office Address: No.14, No.14, Bronze, Sanlitun, Bronze, Sanlitun, Miaoli, 352 Taiwan (ROC·) Nationality: Zhonghua Republic of China/ROC _3. Inventor: (1 in total) Name: Li Peirong/Li, Pei-Jung Nationality: Republic of China/R.ac 200835791 Invention Patent Specification June--&quot; ——1 (This manual format , order, and bold characters, please do not change any more. ※Please do not fill in the ※ part. ※Application number: 0961(10)426 ※Application date: 2007/02/26 1卩€ Classification: &quot; , ^Olh Wo^O (2006.01 )&quot; First, the name of the invention METHOD OF GENERATION OF YING YING TEXTILES &quot; 樟 樟 孑 MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET MET Li, Pei-Jung Address of residence or establishment: No. 14, No. 14, Bianjing Village, Sanwan Township, Miaoli County No. 14, Alley 1, Tung Ching Village Sanwan hsiang, Miaoli, 352 Taiwan (ROC) Nationality: Republic of China/ROC φ III. Inventor: (1 in total) Name: Li Peirong/U, Pei-Jung Nationality: Republic of China/ROC 200835791 IV. Statement: Speaking of the main event = 7 issues, 2, / work - section (4) ^ mouth = Before applying for patents to the following countries (regions): Doors ^ According to the order of the country (region), the date of the towel, and the order of the application number] 0 Item 槪 Article 27 - The egg's priority: □ No claim for patent law Article 27 - International priority: □ = Article 29 of the Patent Law Article 1 Domestic Priority: [Please apply according to the application, the application sequence SI note] 0 claims patent law Article 30 Biological materials: D must deposit biological materials : Domestic biomaterials [format: please note: registration agency, date, number order] % S external biological materials [format please follow: registration country, organization, period, number order note] 0 no need to deposit biomaterials: The technology belongs to the general knowledge of the green (4), no need to register. 200835791 IV. Declarative matters: □ Proposal for the facts as stipulated in the second paragraph or the second paragraph of Article 22 of the Patent Law, the fact that the period of occurrence is: year and month. □ Before applying, apply for patents from the following countries (regions): [Format please: Accept the country (region), application 曰, order of application case number] The patent claim law Article 27 The first international priority : □ No claim for patent law Article 27, first international priority: □主張專利法第二十九條第一項國内優先權： 【格式請依：申請日、申請案號順序註記】 3主張專利法第三十條生物材料： [□須寄存生物材料者： 國内生物材料【格式請依：寄存機構、日期、號碼順序註記】□ Propose the first domestic priority of Article 29 of the Patent Law: [Please follow the format: application date, application case number] 3 Claim Patent Law Article 30 Biological Materials: [On the need to deposit biological materials: Domestic biomaterials [format please note: order of depository, date, number order] 國外生物材料【格式請依：寄存國家、機構、日期、號碼順序註記】 @不須寄存生物材料者： 所屬技術領域中具有通常知識者易於獲得時，不須寄存。 2 200835791 九、發明說明（1) 1·發明所羼之技麵域： 牛樟芝（Antrodia camphorate) 子菌 basidiomycete 多孔菌科 Myporaceae，生長季 節；春、夏、秋，棲息於中、低海拔之台灣國寶級保育樹種f牛樟樹Cinnamomum comphora』上，在腐朽之牛樟木材上，引起木材褐腐朽，是導致牛樟木腐朽中空的 元兒°牛樟芝是台之樟薄孔菌Antrodia dimamomea，享有森林中Γ台灣紅寶石j 之美譽。其子實年生至多年生，雖衄色，橘紅輕淡肉赌，味呈黃樟香 氣、性苦寒。產期在每年六月至十一月，十一月以後受温度之影響其成長緩慢。早 期原住民上山工作，在口中含一小片就可整天不用喝水、而且防止宿醉，民間傳說 、保肝、解毒功能。現代硏究證實；牛樟芝富含三蔽類、固醇類、多醣成 份，可增强肝功能、抗氧化、咖免疫力及抑制麵之效用。 由於牛樟芝野外數量不多，而民間需求有增無減，稱得上是全世界最昂貴的眞菌， 也因此導致「山老鼠（盜林者)」濫砍盜採的目標。目前生物繼已能人工培養樟芝 菌絲體（市面上；分有液態深臟酵繁麵絲體及固體培育牛樟菌絲體兩類）以菌 絲體商品當作代用品。但各種培養牛樟菌絲體之製程，均有其操控極限和困難，不 能也無法去完整臨摹野生子實體成長，導致產出之品質效用有麵限，難達野生子 實體相同標準 &gt; 致價碼高漲難以滿足需求。 2·先前技術： 野生牛樟芝因受牛機保育已明令禁採，雖有人利用颱風過境擇取牛樟漂流木，鋸 成段木並灑上牛樟麵，經4-6個月削下牛樟芝子實體薄層販售，但生長和培育條件 管控困難，爲數仍有限無法滿足需求樟芝麵之保存紐氏零下80度液態氮 中。而繁殖以麥芽抽出物（ΜΕΑ)、甚至酵母抽出物培養基（YEA例:2%««糖〇.5% 酵母抽出物）或馬鈴薯培養基(PDA)等培養，並可於處方中添加0.1%磷酸氫二鉀 及0.05%硫酸鎂，借以促進菌絲之成長。該等培養基注入平版皿中呈固相並温控攝氏 15-32度，均適以靜置培養。 通常液態深臟酵繁殖牛樟芝菌絲體方法；先以特定培養基調製之固相平版皿靜置 培養2周，再將菌絲置於三角瓶中並以經調製液體培養基，以攝氏25-26度進行震盪 培養7-10天之後移入相似條件之種子罐鰣通氣培養，然後轉入相似條件、轉速 90rpm、通氣量爲(X5wm之小型（500公升）發酵槽培養，再轉入中型（5公噸）發 酵槽培養，再升級入更大型發酵槽中培養，其各級之攪拌通氣培養均約740天。 一般培養條件爲1%之接種量，轉速90rpm、通氣量爲0.5wm温度爲攝氏28度（例； 以五十噸槽進行三十五_培養，八天後可得紅色菌絲，再行固液分離，或有發酵全 液^温®攝氏55度減壓濃縮後進行冷凍乾燥，收率可達1.8%其多_含量則在8% 以上），且多醣體含量與菌絲多寡成正比❶ 惟一^深層液態發酵方法；均需進行固液分離，有廢液排放污染環境之問題。且因 該發酵廢液之排放常導_解於水中的小奸活性成分流失，麵發酵常僅能取得 菌絲細胞壁內所含之物質成分。 200835791 九、發明說明（1) 1·發明所羼之技術領域： 牛樟芝（Antrodia camj)hoiate&gt;^8f basidiomycete 多孔菌科 Myporaceae ,生長季 節；春、夏、秋，棲息於中、低海拔之台灣國寶級保育Μ **牛樟樹Cimamomm amphoraj上，之牛樟树上,引起辅褐網，是導致牛樟木腐朽中空的 元兇。牛樟芝是台灣特有之樟薄孔菌AntaxMa dmamomea，享有森林中Γ台灣紅寶石」 之美譽。其子實體一年生至多年生，騰雌色，紅色至淡肉馳 &gt; 味呈黃樟香 氣、性苦寒。產期在每年六月至十一月，十一月以後受温度之影響其成長緩慢◎早 期原住民上山工作，在口中含一小片就可整天不用喝水、而且防止宿醉，民間讎 類麵、保肝、解毒功能，現代硏究證實；牛樟芝富含三_、固_、多醣成 份，可增强肝功能、抗氧化、增加免疫力及抑制臟之效用。Foreign biomaterials [format: please note: country, organization, date, number order note] @Do not need to deposit biomaterials: When there is general knowledge in the technical field, it is not easy to obtain. 2 200835791 IX. Description of invention (1) 1. The technical domain of the invention: Antrodia camphorate, basidiomycete, Myporaceae, growth season; spring, summer, autumn, Taiwan national treasure inhabiting medium and low altitude The graded tree species, Cinnamomum comphora, is on the decaying burdock wood, causing the wood to be brown and decaying. It is the genus of the burdock decaying hollow. The burdock is the Antrodia dimamomea, which enjoys the forest and the Taiwanese ruby. The reputation of j. Its son is born to perennial, although the twilight, orange red light meat gambling, the taste is yellow scent, bitter cold. The production period is from June to November every year, and after November, it is slow to grow due to temperature. Early Aboriginal work in the mountains, including a small piece in the mouth, can not drink water all day, and prevent hangovers, folklore, liver protection, detoxification. It has been confirmed by modern research; Antrodia camphorata is rich in tri-console, sterols and polysaccharides, which can enhance liver function, anti-oxidation, coffee immunity and inhibit the effect of noodles. Because there are not many wild animals in the wild, and the demand for the private sector has increased, it is the most expensive sputum in the world, which has led to the goal of “Mountain Mouse (Pirates)”. At present, the organism has been able to artificially culture the mycelium of Antrodia camphorata (on the market; there are two types of liquid deep-soiled fermented silk and solid burdock mycelium), which are used as substitutes for mycelium products. However, all kinds of processes for cultivating burdock mycelium have their control limits and difficulties. They cannot and cannot complete the growth of wild fruiting bodies in Linyi, resulting in a limit on the quality of output and the difficulty of reaching the same standard for wild fruiting bodies. The price tag is so high that it is difficult to meet the demand. 2. Previous technology: Wild burdock has been banned for cattle breeding. Although some people use typhoon to choose the burdock driftwood, saw it into segments and sprinkle with burdock, and cut the burdock fruit body after 4-6 months. Thin layers are sold, but the growth and cultivation conditions are difficult to control, and the number is still limited. The demand for 樟 面 之 之 保存 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽 纽The breeding is carried out by using malt extract (ΜΕΑ) or even yeast extract medium (YEA case: 2% «« glycoside. 5% yeast extract) or potato medium (PDA), and 0.1% can be added to the prescription. Dipotassium hydrogen phosphate and 0.05% magnesium sulfate to promote the growth of hyphae. The medium was injected into a lithographic dish in a solid phase and temperature-controlled at 15-32 degrees Celsius, and all were suitable for static culture. Usually, the liquid deep-fermentation method is used to propagate the mycelium of Antrodia camphorata; the solid phase platter prepared by the specific medium is firstly cultured for 2 weeks, and the hyphae are placed in a triangular flask and adjusted to a liquid medium at 25-26 degrees Celsius. After 7-10 days of shaking culture, transfer to a similar condition of the seed tank, aeration culture, and then transfer to similar conditions, speed of 90 rpm, ventilation (X5wm small (500 liter) fermentation tank culture, and then transferred to medium (5 metric tons) The fermentation tank is cultured, and then upgraded to a larger fermentation tank for cultivation. The aeration and aeration cultures of each stage are about 740 days. The general culture condition is 1% inoculum, the rotation speed is 90 rpm, and the aeration amount is 0.5 wm, and the temperature is 28 degrees Celsius. For example, thirty-five _ cultures are carried out in a 50-ton trough, red hyphae can be obtained after eight days, and then solid-liquid separation is carried out, or the whole liquid is heated at a temperature of 55 ° C and concentrated under reduced pressure, followed by lyophilization. Up to 1.8% of its _ content is above 8%), and the polysaccharide content is proportional to the number of mycelium ❶ unique ^ deep liquid fermentation method; both need to be solid-liquid separation, there is a problem of waste liquid discharge pollution. Due to the fermentation waste liquid The introduction of the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Camj)hoiate&gt;^8f basidiomycete Polyporaceae Myporaceae, growing season; spring, summer, autumn, inhabiting medium- and low-altitude Taiwan national treasure-level conservation Μ 樟 burdock Cimamomm amphoraj, on the burdock tree, causing auxiliary brown net It is the culprit leading to the decaying hollow of the burdock. The burdock is the unique AntanMa dmamomea of Taiwan, which enjoys the reputation of “Taiwan ruby in the forest.” Its fruit body is annual to perennial, and it is tanned, red to light. It has a yellow scent and a bitter cold. The production period is from June to November every year. After November, it is slow to grow due to the temperature. ◎ Early aborigines went up the mountain and had a small piece in the mouth to drink water all day. And to prevent hangovers, folk scorpion noodles, liver protection, detoxification function, modern research confirmed; burdock is rich in three _, solid _, polysaccharide components, can enhance liver function, Oxidation, and increase immunity to suppress the dirty utility. 由於牛樟芝野外數量不多，而民間需求有增無減，稱得上是全世界最昂貴的眞菌， 也因此導致「山老鼠（盜林者)j濫砍盜採的目標。目前生物科技已能人工培養樟芝 菌絲體（市面上；分有液態深臟酵繁麵絲體及固體培育牛樟菌絲體兩類）以菌 絲體商品當作代用品。但各種培養牛樟菌絲體之製程，均有其操控極限和困難，不 能也無法去完整臨摹野生子實體成長，導致產出之品質效用有難限，生子 實體相同標準，致價碼高漲難以滿足需求。 2·先前技術： 野生牛樟芝因受牛樟樹保育已明令禁採，雖有人利用颱風過境擇取牛樟漂流木，鋸 成段木並灑上牛樟菌種，經4-6個月削下牛樟芝子實體薄層販售，但生長和培育條件 管控困難，爲數仍有限無法滿足需求。-Μ樟芝菌種之保存麵氏零下80度液態氮 中。而繁雜麥芽抽出物（ΜΕΑ )、甚至酵母抽出物培養基（YEA例：2%麵糖0.5% 酵母抽出物）或馬鈴薯培養基（PDA)等培養，並可於處方中添加0.1%磷酸氫二鉀 及0.05%硫_，借以讎菌絲之成長。該等培養基注入平顧中呈固相並温控攝氏 15-32度，均適以靜置培養。 通常麵深讎酵繁殖牛樟芝菌絲體方法；先以特定培養基調製之固相平臟靜置 培養2周，再將菌絲置於三角瓶中並以經調製液體培養基，以攝氏25-26度進行震盪 培養740天之後移入相似條件之種子罐攪拌通_養，然後轉入相似條件、轉速 90rpm、通氣量爲0.5vvm之小型（500公升）發酵槽培養，再轉入中型（5公噸）發 酵槽培養，再升級入更大型發酵槽中培養，其各級之攪持通氣培養均約7-10天。 一^養條件爲1%之接種量，轉速90rpm、通氣量爲a5wm温度爲攝氏28度（例； 以五十噸槽進行三十五噸培養，八天後可得紅色菌絲，再行固液分離，或有發酵全 液以温度攝氏55度減壓濃縮後進行冷凍乾燥，收率可達1挪其多醣體含量則在8% 以上），且多醒含量與菌絲多寡成正比。 惟一^深層液態發酵方法；均需進行固液分離，有廢液排放污染環境之問題。且因 該發酵廢液之排放常導致溶解於水中的小肝活性成分流失，液態發酵常僅能取得 菌絲細胞壁內所含之物質成分◊ 5 200835791 九、發明說明（2) 固體培養牛樟菌方法·，先將牛樟芝菌種以太空包進行菌絲體培養，並續於空氣中進 行子實體栽培，且利用該方法培養得之樟芝固體培養物（專利公告號⑻565430) 〇 該專利係一種培養固體樟芝之方法；以BCRC 35398爲菌種來源植在太空包內，且該 太空包內含之培養基係由你70%選自蕈類、植物之莖、梗、果實及木屑之纖維物、 2〜30%馬鈴薯、545%米糠、140%«^、0.5-2鑛酸二氫鉀及0.1-1%硫^1^組成， 其溼度爲胳80%之間，並調整其pH値爲中性；_該專利之方法’太空包之培養 基係由65%草本植物之莖、梗、果實或木屑、20%馬鈴薯、10%米糠、3·5%葡萄糖、 酸二氫鉀及0.5%硫組成。且其中子實體栽培期間之日間溫度係介於2〇 至30°C之間，而夜間溫度係介於844t：之間，且控制其日夜鬮溫差於12至16°C之 間。其溫度爲5至32°C間，含有0.24%之二氧化碳之空氣下進行菌絲體培養（培養 進行50*80天），隨之移除太空包，使之暴露於二氧化碳含量低於1%且空氣溼度介 於90*95%之間之空氣流通條件下，進行子實體栽培一段時間（子實體培養須2化50 天）至圓錐形子實體形成❹ 讎固體培養牛樟菌之方法，受限於太空包內所含培養基材，此謝在提供營養成 份後易軟化且此時已移除太空包，一方面易受雜菌汚染無法久置。故需在特定期內 行採收，致其圓錐形子實體形成時麵，形成之表層膜甚薄。 一^猫菌類均具極佳吸附金屬之能力，例;2006年日本麒麟啤酒之關係生技產品姬 松茸-巴西蘑菇j就發生過高重金屬含量而下市，而我國也發現國產雜r巴麵 菇』中亦隱藏有相同問題。究其因不外乎巴西蘑链、洋菇覆土性需求所造成。在現 今工業發達所造成空氣麵魅壤污染，導致野生之牛樟芝、錳、鋁、硼、 銻明顯高於人工培育牛樟芝菌絲體。 牛樟芝菌固、液態培養首見文獻於1997年師大MC源等之硏究，其中添加0.1^0.3% 樟腦結晶可助於菌絲體生長。其後1998年廖英明指出添加0娜之樟木萃出液（10% 之樟木屑水）可有最好的成長，2000年許勝傑等發現樟樹精油中常見如terpined、 safrole、camphor成分皆有助於樟芝菌絲體生長6爲防止環境污染導致野生牛樟芝商 品具高重金屬含量 &gt; 以及撤牛穌 '纖牛樟芝 &gt; 唯有ΛΧ培育牛樟薛體（子實 體）一途。如比一來；除了能供應市場之需求，還能爲台灣國寶Γ牛樟木j保育樹 及天然牛樟芝留下娜。 3.發明的內容： 本發明係提供一種人工栽培r牛樟芝-子實體j之繁育方法；本發明同時提供液相 發酵靜置培養基處方，可用於大部分真菌之生長。同時該方法包括將一樟芝菌麵 入以瓶容器裝盛且經調理減菌後之濕木料上進行菌絲體培養（第1圖），並續於含 有空氣之瓶中進行子實體培育。麵讎有孔洞之橡膠塞封口，並裝塡棉花入膠塞 孔洞中，一方面阻隔雜菌入侵並可確保培養瓶中樟芝菌成長所需之相對濕度，另一 方面可排放眞菌因發酵代謝所生氣體。不僅提高真菌之產量，同時刺激其衍生代謝 物，特別是含有醫藥活性成分的代謝物。 200835791 九、發明說明（2) 固體培養牛樟菌方法:先齡樟^^^太空儀行菌絲體培養，並續於空氣中進 行子實體栽培，且利用該方法培養得之樟芝固體培養物（專利公告號00565430)。 該專利係一種培養固體樟芝之方法；以BCRC 35398爲菌種來源植在太空包內，且該 太空包內含之培養基係由6(K70%選自蕈類、植物之莖、梗、果實及木肩之纖維物、 20·30%馬鈴薯、545%雜、140%葡纖、0.5-2%磷酸二氫鉀及0.1-1%硫組成， 其淫度爲60*80%之間，並調整其pH値爲中性；讎該專利之方法，太空包之培養 基係由65%草本植物之莖、梗、果實或木屑、20%馬鈴薯、10%米糠、3.5%葡萄糖、 1%磷酸二氫鉀及0.5%硫組成。且其中子實體栽培期間之日間溫度係介於20 至30°C之間，而夜間溫度係介於844°C之間，且控制其日夜間溫差於12至16°C之 間。其溫度爲5：至32°C間，含有0.24%之二氧化碳之空氣下進行菌絲體培養（培養 進行50观天），隨之移除太空包，使之暴露於二氧化碳含量低於1%且空氣溼度介 於90^95%之間之空氣流通條件下，進行子實體栽培一段時間（子實體培養須瓜50 天）至圓錐形子實體形成众 雛固體培酙樟菌之方法，受酸太空包內養謝，此紐營養成 份後易軟化且此時已移除太空包，一方面易受雜菌汚染無法久置。故需在特定期內 行採收 &gt; 致其圓錐形子實體形成時雖，形成之表層膜甚薄。 一般琏菌類均麵佳吸附金屬之能力，例；2006年曰本麒麟啤酒之關係生技產品 Γ姬松茸-巴西蘑菇j就發生過高重金屬含量而下市，而我國也發現國產生鮮 Γ巴西蘑菇《I中亦隱藏有相同問題。究其因不外乎巴西蘑菇、洋链覆土性需求所 造成。在現今工業發達所造成空氣和環社壤污染，致野生之牛樟芝發 鋁、硼、銻明顯高於人工培育牛樟芝菌絲體。 牛樟芝菌固、液態培養首見文獻於1997年師大簡秋源等之硏究，其中添加0.1-0.3% 樟腦結晶可助於菌絲體生長。其後1998年廖英明指出添加_%之樟辟出液(臟 之樟木屑水）可有最好的成長，2000年許勝傑等麵樟樹精油中常見如terpined、 safrole ~ camphor成分皆有助於樟芝菌絲體生長。爲防止環牺染導嚮生牛樟芝商 品具高重金屬含量，以及盜砍牛樟木、盜採牛樟芝，唯有人工培育牛樟_ (子實 體）一途❶如比一來；除了能供應市場之需求，麗爲台灣國寶p牛樟木j保育樹 及天然牛樟芝留下娜。 3.發明的內容： 本發明係提供一種人工f牛樟芝-子實體』之繁育方法；本發明同時提供液相發 _黯養基處方，可用於大部分真菌之生長❶同時該方贿括將一樟芝菌麵入 以瓶容嚣裝盛且經調理減菌後之濕木料上進行菌絲體培養（如附件），並續於含有空 氣之瓶中進行子實體培育。該瓶係嫌孔洞之橡膠塞封口，並裝塡棉花入膠塞孔洞 中，一方面阻隔雜菌入侵並可確保培養瓶中樟芝菌成長所需之相對濕度，另一方面 可排放眞菌因發酵代謝所生氣體。不僅提高真菌之產量，同時刺激其衍生代謝物， 特別是含有醫藥活性成分的代謝物。 6 200835791 九、發明說明（3): 4·實施方式： 本發明係將繁芽抽ffiti MEA，母抽出物培養基皿或馬鈴薯舰培養基 等·養樟芝菌絲》移ΛΜ調配處方之麥片培養基中擴大培養（一種繁殖真菌之固 態發酵培養方法，中華民國發明第1231825號專利），再將生長旺盛滿是樟芝菌絲之 麥片和無菌粘稠狀營養觀均質觀碎成樟芝種菌液。然後；再將稠狀樟芝菌液注 入或浸塗讎調理且減菌後容器內之濕木材上，借以羅菌絲在木材上之成長並温 控於攝氏15-32度，直至容器（玻璃）內之木材有牛樟芝子實體形成於表面。 當參照較佳實施所描述之有關發明細節後，則刪熟知相技術之xm進行之各種 ^^^讎，麵不脫離本發明申請專利範圍所述之精神及範圍。 5·【圖式簡單說明】：一種人工栽培『牛樟芝-子實體』之繁育方法製程圖。Because there are not many wild animals in the wild, and the demand for the private sector has increased, it is the most expensive sputum in the world, which has led to the goal of “snakes and stalkers”. It can artificially cultivate the mycelium of Antrodia camphorata (on the market; there are two types of liquid deep-soiled fermented silk and solid burdock mycelium), which are used as substitutes for mycelium products. The process of the body has its control limits and difficulties, and it cannot and cannot complete the growth of wild fruiting bodies in the whole Linyi, which leads to the difficulty in the quality effect of the output. The birth of the child entity is the same standard, and the price increase is difficult to meet the demand. 2. Pre-technical: Wild Niu Zhizhi has been banned for burdock conservation. Although some people use the typhoon to choose the burdock driftwood, saw it into segments and sprinkle with the burdock strain, and cut the thin layer of the burdock fruit body for 4-6 months. However, the control of growth and cultivation conditions is difficult, and the number is still limited, which cannot meet the demand. - The preservation of A. sinensis strains is in the liquid nitrogen of minus 80 degrees, while the mixed malt extract (ΜΕΑ) and even the yeast extract medium YEA case: 2% facial sugar 0.5% yeast extract) or potato medium (PDA), etc., and 0.1% potassium dihydrogen phosphate and 0.05% sulfur _ can be added to the prescription, whereby the growth of sputum hyphae. Injecting into the solid phase and temperature control of 15-32 degrees Celsius, both are suitable for static culture. Usually deep-faced fermentation of the Astragalus membranaceus mycelium method; first fixed solid medium prepared by a specific medium for 2 weeks of static culture Then, the hyphae were placed in a triangular flask and incubated in a conditioned liquid medium at a concentration of 25-26 degrees Celsius for 740 days, then transferred to a similar condition of the seed tank, and then transferred to a similar condition, rotating at 90 rpm, and ventilated. The small (500 liter) fermentation tank with a volume of 0.5vvm is cultured, transferred to a medium-sized (5 metric ton) fermentation tank, and then upgraded to a larger fermentation tank. The aeration and aeration cultures at each level are about 7-10 days. The condition of inoculation is 1% inoculation, the rotation speed is 90 rpm, the aeration is a5wm, the temperature is 28 degrees Celsius (for example; 35 tons of culture is carried out in 50 tons of tank, red hyphae can be obtained after 8 days, and then Solid-liquid separation, or fermentation of whole liquid at a temperature of 55 degrees Celsius After shrinking, freeze-drying, the yield can reach 1%, the polysaccharide content is above 8%), and the amount of wake-up is proportional to the amount of hyphae. The only deep liquid fermentation method; both need solid-liquid separation, waste The liquid discharge pollutes the environment. And because the discharge of the fermentation waste liquid often leads to the loss of the active components of the small liver dissolved in the water, the liquid fermentation often only obtains the material components contained in the cell wall of the mycelium. 2008 5 200835791 IX. 2) Solid culture of Burdock method · First, the A. sinensis strain is cultured in a space capsule for mycelium culture, and continues to be cultivated in the air for fruit body cultivation, and the solid culture of A. camphorata cultured by the method is obtained (Patent Announcement No.) (8) 565430) 〇 This patent is a method for cultivating solid Antrodia camphorata; it is planted in a space bag with BCRC 35398 as the source of the strain, and the medium contained in the space capsule is 70% selected from the group consisting of scorpions, stems and stems of plants. , fruit and wood fiber, 2~30% potato, 5455% rice bran, 140% «^, 0.5-2 potassium dihydrogenate and 0.1-1% sulfur ^1^, the humidity is between 80% And adjust its pH to neutral; The method of 'culture space bag from 65% of the herb-based group of stems, stalks, fruits or wood chips, 20% potato, 10% rice bran, 3.5% glucose, 0.5% potassium dihydrogen and sulfur. The daytime temperature during the fruiting body cultivation period is between 2〇 and 30°C, and the nighttime temperature is between 844t:, and the temperature difference between day and night is controlled between 12 and 16 °C. Mycelium culture (cultured for 50*80 days) at a temperature of 5 to 32 ° C containing 0.24% carbon dioxide, followed by removal of the space capsule to expose to a carbon dioxide content of less than 1% Under the air circulation condition of air humidity between 90*95%, the fruiting body is cultivated for a period of time (the fruiting body culture must be 2 to 50 days) to the conical fruit body to form the ❹ solid solid culture of the burdock, limited The culture medium contained in the space package is easy to soften after providing nutrients and the space bag has been removed at this time. On the one hand, it is susceptible to contamination by bacteria. Therefore, it is necessary to harvest in a certain period of time, so that the conical sub-solids form the surface, and the surface film formed is very thin. A cat fungus has excellent ability to adsorb metal. For example, in 2006, the Japanese biochemical product of Kirin beer, Agaricus blazei-Brazilian mushroom j, was produced with high heavy metal content, and China has also found domestically produced r-bar surface. The same problem is hidden in the mushroom. This is due to the fact that the Brazilian mushroom chain and the mushroom cover soil demand. In the current industrial development, the pollution of the air surface is caused by the pollution of the air, resulting in the wild cattle Antrodia, manganese, aluminum, boron, and strontium are significantly higher than the artificial cultivation of A. sinensis mycelium. The original and literature of the culture and liquid culture of Antrodia camphorata was studied in 1997. The addition of 0.1^0.3% camphor crystals can help mycelium growth. Later, in 1998, Liao Yingming pointed out that the addition of 0 Na's eucalyptus extract (10% sassafras water) can have the best growth. In 2000, Xu Shengjie discovered that common eucalyptus oils such as terpined, safrole and camphor are helpful. In the growth of mycelium of A. camphor. 6 to prevent environmental pollution, the content of high-heavy metal in wild A. angustifolia &gt; and the removal of the cow's fiber, and the only way to cultivate the calf (small body). In addition to being able to supply the market, it can also be reserved for Taiwan's national treasure yak yak j jani tree and natural yak. 3. Disclosure of the Invention: The present invention provides a method for cultivating artificially cultivated r. chinensis-fruit body j; the present invention also provides a liquid phase fermentation static medium formulation which can be used for the growth of most fungi. At the same time, the method comprises the step of inoculating a scutellaria sinensis into a wet container of a bottle container and conditioning the sterilized fungus (Fig. 1), and continuing the cultivation of the fruit body in a bottle containing air. The rubber plug is sealed with a hole in the face, and the cotton is inserted into the hole of the rubber plug. On the one hand, the invasion of the bacteria is blocked, and the relative humidity required for the growth of the Phytophthora in the culture bottle can be ensured. On the other hand, the fermentation of the bacterium can be discharged. Metabolize the gas produced. It not only increases the yield of fungi, but also stimulates its derived metabolites, especially metabolites containing pharmaceutically active ingredients. 200835791 IX. Description of invention (2) Method for solid-culture of Burdock: The mycelium cultured by the ancestor 樟^^^ spacecraft, and continued to be cultivated in the air for fruit body cultivation, and the solid culture of Antrodia camphorata cultivated by the method (patent bulletin number 05546430). The patent is a method for cultivating solid Antrodia camphorata; it is planted in a space bag with BCRC 35398 as a strain source, and the medium contained in the space bag is 6 (K70% is selected from the group consisting of a scorpion, a stem, a stem, and a fruit of a plant). And the fiber of the wooden shoulder, 20.30% potato, 5455% miscellaneous, 140% Portuguese fiber, 0.5-2% potassium dihydrogen phosphate and 0.1-1% sulfur, the degree of lust is between 60*80%, and Adjusting its pH to neutral; 雠 the method of this patent, the space bag culture medium is 65% of herbaceous stems, stems, fruits or sawdust, 20% potato, 10% rice bran, 3.5% glucose, 1% dihydrogen phosphate Potassium and 0.5% sulfur, and the temperature during the fruiting period is between 20 and 30 °C, while the nighttime temperature is between 844 °C, and the temperature difference between day and night is controlled at 12 to 16 °. Between C. Mycelium culture (50 days of culture) was carried out in air containing 0.24% carbon dioxide at a temperature of 5: to 32 ° C, and the space capsule was removed to expose it to low carbon dioxide. Under the air circulation condition of 1% and air humidity between 90 and 95%, the fruiting body is cultivated for a period of time (the fruit body culture must be 50) The method of forming a solid solid cultured bacteria into a conical fruit body, which is cultivated by the acid space package, is easy to soften after the nutrient composition, and the space bag has been removed at this time, and is susceptible to contamination by bacteria on the one hand. Therefore, it is necessary to harvest within a certain period of time. Although the surface film formed by the conical body is very thin, the general surface of the fungus is good for adsorbing metals. For example, the relationship between Sakamoto Kirin Beer in 2006 Biotech products Γ 松 松 - 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西 巴西Caused by sexual demand. In the current industrial development caused by air and environmental pollution, the wild Astragalus sinensis hair, aluminum, boron, strontium is significantly higher than the artificial cultivation of A. sinensis mycelium. Astragalus bacillus fungus, liquid culture first seen in 1997 The addition of 0.1-0.3% camphor crystals can help the growth of mycelium. In 1998, Liao Yingming pointed out that adding _% 樟 樟 樟 (dirty swarf sawdust water) can have the best Growth In 2000, Xu Shengjie and other common eucalyptus essential oils, such as terpined and safrole ~ camphor, all contribute to the growth of A. sinensis mycelium. In order to prevent the ring-staining, the raw material of the A. sinensis has high heavy metal content, as well as the slashing of the burdock and the argus Only artificially cultivated burdock _ (sub-entity) is a one-way approach; in addition to the market demand, Li is Taiwan's national treasure p burdock j conservation tree and natural burdock leaves Na. 3. Summary of the invention: The present invention The invention provides a method for breeding an artificial f. Antrodia camphorata-fruit body; the invention also provides a liquid phase hair-feeding base prescription, which can be used for the growth of most fungi, and the bribe includes a group of Zhizhi bacteria into a bottle Mycelium culture (such as attachment) is carried out on wet wood after conditioning and sterilizing, and continues to be cultured in a bottle containing air. The bottle is sealed with a rubber plug of the hole, and the cotton is inserted into the hole of the rubber plug to block the invasion of the bacteria and ensure the relative humidity required for the growth of the Phytophthora in the culture bottle. On the other hand, the bacterium can be discharged. The gas produced by fermentation metabolism. It not only increases the yield of fungi, but also stimulates its derived metabolites, especially metabolites containing pharmaceutically active ingredients. 6 200835791 IX. Inventive Note (3): 4. Implementation Mode: The present invention is to transfer the ffiti MEA, the mother extract culture medium dish or the potato ship culture medium, etc. Enlarged culture (a solid-state fermentation culture method for breeding fungi, Patent No. 1231825 of the Republic of China), and then homogenizes the growth of the oatmeal and the aseptic viscous nutrient view into a medicinal solution of Antrodia camphorata. Then; inject or dip the thick Zhizhi bacteria solution onto the wet wood in the container after sterilizing and reducing the bacteria, so that the mycelium grows on the wood and is temperature controlled at 15-32 degrees Celsius until the container (glass The wood inside is formed on the surface of the body of the burdock. </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; 5·[Simple description of the schema]: A process map for the artificial cultivation of the breeding method of "Bag Beans - Fruit Body". —J f 5000ml往返Μ»培 成長變化 並取得菌 絲體及代 謝勸烘乾 燥供比對 供存·查d in - 通氣攪ί雜養 抬操作接種 取樣以HPLC定性定1 比對 ! 上架觀察並監視成長 Λ #得最诖之成分含董與 培育條沣 成品加工 培養室殺菌 消毒 —-- |類菌台操作 !接種 200835791 九、發明說明（3): 4·實施方式： 本發明係將繁芽抽出物MEA '酵母抽出物培養基YEA或馬鈴薯PDA培養基 等所培養樟芝菌絲’移入經調配處方之麥片培養基中擴大培養（一觀殖真菌之固 態發酵培養方法，中華民國發明第1231825號專利），再將生長旺盛滿是樟芝菌絲之 麥片和無菌翻狀營養雜均質麵碎成樟魏菌液。然後；再將稠狀樟芝菌液注 入或浸塗於經調理且減菌後容器內之濕木材上，借以促進菌絲在木材上之成長並温 控於攝氏15·32度，直至容嚣（玻璃等）內之木材有牛樟芝子實體形成於表面。 當參照_實施所插脱有關發明細節後，則獅熟知本項技術之人所進行之各種 修改及替換，將仍不脫離本發明申請專利範圍所述之精神及範圍。 • 5·【圖式簡單說明】： 第1圖：一種人工栽培r牛樺芝-子實體』之繁育方法製程圖❶ 200835791 五、中文發明摘要： 潑明之名稱：一默工纖f牛樟芝·子實體』之繁育方法） 本發明係提供一種人工11牛樟芝-子實體j之繁育方法；本發明同時提供液相 發酵靜置培養基處方，可用於大部分真菌之生長。同時該方^^將一樟魏觀 入以瓶容嚣裝盛且經調理減菌後之濕木料上進行菌絲體培養（第1圖）， 有空氣之瓶中進行子實體培育。觀讎有孔洞之橡膠塞封口，並_道^^二口 孔洞中一方面阻隔雜菌入侵並職保培養瓶中樟魏成長所需之入膠塞 方面可排放眞菌因發酵代謝所生氣體❶不讎高真菌之產裊，同時锻,另〜 物，特別是含有醫藥活性成分的代謝物。 、顯聰生代謝—J f 5000ml round-trip 培» Peer growth change and obtain mycelium and metabolism to dry and dry for comparison and storage. Check d in - ventilating ί 养 抬 操作 操作 操作 操作 以 以 以 以 以 以 以 以 以 以 以 以 以 HPLC HPLC HPLC HPLC HPLC HPLC HPLC HPLC HPLC Monitoring growth Λ # The most acclaimed ingredients including Dong and cultivating strips 沣 沣 沣 沣 — — — — — — — — — — 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 Bud extract MEA 'Yeast extract medium YEA or potato PDA medium and other cultivated Antrodia sinensis 'transplanted into the blended prescription of the oatmeal medium to expand the culture (a solid fermentation culture method of the colony fungus, the Republic of China invention No. 1231825 patent ), and then grow the flour that is full of A. sinensis and the aseptically turned nutrient-like homogenized surface into the sputum. Then; inject or dip the thick Antrodia camphorata liquid onto the wet wood in the conditioned and reduced bacteria container, thereby promoting the growth of the hyphae on the wood and controlling the temperature at 15·32 degrees Celsius until the volume is allowed The wood in the (glass, etc.) is formed on the surface of the body of the burdock. </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; • 5·[Simple description of the schema]: Figure 1: Process diagram for the breeding method of artificially cultivated r-Betula chinensis-fruit body ❶ 200835791 V. Abstract of Chinese invention: The name of the priming: one murder fiber f burdock The invention provides a method for breeding artificial 11 Antrodia camphorata-fruiting body j; the invention also provides a liquid phase fermentation static medium prescription, which can be used for the growth of most fungi. At the same time, the party ^^ will be immersed in a bottle of oysters and cultured on the wet wood after conditioning and sterilizing (Fig. 1), and the fruit body is cultivated in an air bottle. Guanlan has a rubber plug seal for the hole, and _dao^^ two holes in the hole on the one hand to block the invasion of bacteria and the protective rubber bottle in the nursery bottle can be discharged into the rubber plug. It is not high in the production of fungi, but also in the forging, another thing, especially metabolites containing pharmaceutically active ingredients. Xiancong metabolism 六、英文發明摘要： METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate ° 200835791 五、中文發明摘要： 潑明之名稱：-®AX^ r牛樟芝孑實體j之繁育方法） 本發明係提供一種人工^ r牛樟芝-子實體j之繁育方法；本發明同時提供液相 發酵靜觀養基處方，可用於大部分真菌之生長。同時該方鮑括將一樟Μ讎 入以瓶容器麵且經麵減菌後之濕·上進行菌絲體培養（如附件），並續舱 有空氣之瓶中進行子實體培育。該瓶係似有孔洞之橡膠塞封口，並裝塡棉花入膠塞 孔洞中，一方面闘雜菌入侵並可確保培養瓶中樟芝菌成長所需之相對濕度，另-方面可排放眞菌因發酵代謝所生氣體。不讎高真菌之產量，同時刺激其衍生代謝 物，特別是含有醫藥活性成分的代謝物。 六、英文發明摘要： METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate ° 200835791 十、申請專利範圍（1): L 本發B月說B月的第3項內容之方法，該培養基係由牛棒木料65%、馬鈴馨2〇%、 米糠跳、蔗糖3.5%、磷醜二鉀1%及硫隨0.5%所組成。 2·根據發明，明的第3項內容之方法，該培養基係由牛樟木料65%、豆漿3〇% (該黃職觀和賴1:7 · 1:14比率硏製而成）蔗糖3.5%、磷賴二鉀1%及硫麵 〇·5%所組成。 3· _本發B月說明的第3項內容之方法，該培養基係由牛棒木料65%、營養液3〇% (該 營養液係以酵母抽出物〇·5·3%)、蔗糖3·5%、磷酸氫二鉀1%及硫麵0.5%所組成。 4·麵本發明說明的第3項內容之方法，該培養麵由牛樟木料65%、營養液3〇% (該 • 營養液観麥雜出物0.5-3%)、蔗糖3.5%、磷酸氫二鉀1%及硫隱0.5%所組成❶ 5·根據本發明說明的第3項內容之方法，該培養基係由牛樟木料65%、營養液3〇% (該 營養液係以酵母抽_ 〇·5-2%、麥芽抽052% )、蔗糖3·5%、磷酸氫二郵1% 及硫麵0.5%所組成。 6·根據本發明說明的第3項內容之方法，該培養^^由樟_($#、油樟、芳樟、 陰陽樟）木料65%、馬鈴薯20%、米糠鹏、蔗糖3·5%、鱗醜二鉀1%及硫麵 0.5%所組成。 7·根據本發明說明的第3項內容之方法，該培養基係由適於香杉芝生長及金針薛繁育 的杉科（如香杉等）槲65%、馬鈴薯20%、米糠10%、蔗糖3.5%、磷酸氫二鉀1% 及硫麵0.5%所組成。 馨 8·根據本發明說明的第3項內容之方法，該培養基係由逾於各類香菇、木耳等顏繁 育的雜桃（如相思樹、油桐«#) 65%、馬鈴薯20%、_ 10%、蔗糖3.5%、磷 酸氫二鉀1%及硫麵0·5%所組成。 λ根據本發明的第3項內容之方法，該培養基係將馬鈴薯、米糠和水萃出之液，再加 入蔗糖3.5%、磷酸氫二鉀1%及硫麵0.5%所組成營養液。 10·麵本發明說明的第4項實施方式之方法，牛樟等_祕需經含水量、濕度管 控，先祛水【以烘箱控温65 75 C乾燥8小時，祛水10.3445% (Ave.12.5%)】，乾 燥後再吸附乾淨水【浸泡3-8天（5天爲最佳），使吸附含水量約35%】。再浸塗經 調配並已均質攪碎之粘稠狀牛擁菌於表面。該培養基係將馬鈴薯20%、米糠10% 以水萃出，再加入蔗糖3.5%、磷酸氫二鉀1%及硫酸鎂〇·5%和甲基纖維素〇·5%-2% 所組成之稠狀營養液。 200835791 十、申請專利範園（1): 1.麵本發明說明的第3項內容之方法，該培養基係由牛樟木料65%、馬鈴薯20%、 米糠10%、蔗糖3.5%、磷酸氫二鉀1%及硫麵〇·5%所組成。 1根據發明^月的第3項內容之方法，該培養基係由牛樟# 65%、_ 30% (該黄豆漿瓶和水以1:7 -1:14比率硏製而成）蔗糖3.5%、磷醜二鉀1%及硫_ 0.5%所組成众 3·根據本發明說明的第3項內容之方法，該培養基係由牛樟木料65%、營養液3〇%(該 營養液讎酵母抽出物0.5-3% )、蔗糖3‘5%、磷酸氫二鉀1%及硫麵〇·5%所組成。 4·讎本翻說明的第3項內容之方法，該培養基係由牛樟木料65%、營養液3〇% (該 Φ 營養液備麥芽抽0.5-3% )、蔗糖15%、磷酸氫二鉀1%及硫隱0.5%所組成。 5·根據本發明說明的第3項內容之方法，該培養基係由牛樟木料65%、營養液30% (該 營養液係以酵母抽出物0·5·2%、麥芽抽出物0.5-2% )、蔗糖3.5%、磷醜二鉀1% 及硫麵0.5蕭組成。 6·根據本發明說明的第3項內容之方法，該培養基係由樟木科(香樟、芳樟)木料65%、 馬鈴薯20%、雜10%、蔗糖15%、磷醜二鉀1%及硫麵0·5%所組成。 7·根據本發明說明的第3項內容之方法，該培養基係由適於金針薛、香链等繁育的杉 科（如紅杉等）木料65%、馬鈴薯20%、米糠10%、蔗糖3.5%、磷酸氫二鉀1%及硫 瞧0·5%所組成❶ 8. 根據本發明說明的第3項內容之方法，該培養基係由適於各類香薛、木耳等蔽菌繁 • 育的雜木料（如相思樹、木棉花樹等)65%、馬鈴薯20%、米糠10%、蔗糖3.5%、 磷酸氫二鉀1%及硫纖0.5%所組成。 9. 根據本發明的第3項內容之方法，該培養基係將馬鈴薯、米糠和水萃出之液，再加 入蔗糖3.5%、磷酸氫二鉀1%及硫隱0·5%所組成營養液。 10·根據本發明說明的第4項實施方式之方法，牛樟等賴需經含水量、濕度管 控，先驗水【以_控温65 - 75 C乾燥8小時，餘水10.344.5% ( Aml2.5% )】，乾 燥後再吸附乾淨水【浸泡3-8天（5天爲最佳），使吸附含水量約35%】。再浸塗經 調配並已均質攪碎之棚狀牛麵菌織面。該培養麵將馬鈴薯20%、雜10% 以水萃出，再加入蔗糖3.5%、磷酸氣二鉀1%及硫瞧〇·5%和甲基纖維素〇.5%-2% 所組成之稠狀營養液。 8 200835791 十、申請專利範圍（2): 11·根據本發明說明的第4項實施方式之方法，牛樟等樟科木料含水量、触等，探 均if牛Sf⑦以培養讎將馬鈴薯継、雜 0.5:2S成温 顧麵0.5%和甲咖素 12.根據本發明說明的第4項實施方式之方法，各種杉科（香杉）、雜^^菩，霱經含 水量、濕度管控，先駐水【以烘箱控温65-75C乾燥8小時駐 Ave.12.5%)】，乾燥後再吸附浸讎調配之培養基於表面，該培養基係馬 20%、雑10%以水萃出，再加入蔗糖3·5%、磷酸氫二鉀1%及硫“ ％VI. Abstracts of English invention: METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate ° 200835791 V. Abstract of Chinese invention: The name of Pluron: -®AX^ r Breeding method of burdock 孑 孑 entity j) The present invention provides a manual ^ r The method for breeding burdock-fruit body j; the invention also provides a liquid phase fermentation nucleus prescription, which can be used for the growth of most fungi. At the same time, the party will enclose the mycelium culture (such as the attachment) on the wet surface of the bottle container and the surface of the bottle, and renew the air bottle to carry out the fruit body cultivation. The bottle is similar to a rubber plug seal with a hole, and is fitted with cotton into the hole of the rubber plug. On the one hand, the inoculating bacteria can ensure the relative humidity required for the growth of the Phytophthora in the culture bottle, and the other can discharge the bacterium. Gas produced by fermentation metabolism. It does not increase the yield of fungi, but also stimulates its derived metabolites, especially metabolites containing pharmaceutically active ingredients. Sixth, English invention summary: METHOD FOR PROPAGATING FRUIT BODY (STROMA) OF Antrodia camphorate ° 200835791 X. Patent application scope (1): L This is the method of the third item of B month, which is made of beef rod Wood 65%, Ma Lingxin 2%, rice bran, sucrose 3.5%, phosphorus ugly potassium 1% and sulfur with 0.5%. 2. According to the invention, the method of the third item, the culture medium is made of burdock wood 65%, soy milk 3〇% (the ratio of Huang Shiguan and Lai 1:7 · 1:14) sucrose 3.5 %, phosphorus lysine 1% and sulphur surface 〇 · 5%. 3· _ The method of the third item described in the month of this month, the culture medium is 65% of bovine stick wood, 3〇% of nutrient solution (the nutrient solution is yeast extract 〇·5·3%), and sucrose 3 · 5%, dipotassium hydrogen phosphate 1% and sulfur surface 0.5%. 4. The method of the third aspect of the present invention, wherein the culture surface is 65% of burdock wood, 3〇% of nutrient solution (the nutrient solution buckwheat miscellaneous material 0.5-3%), sucrose 3.5%, phosphoric acid Dihydrogen potassium 1% and sulfur cryptiness 0.5% composition 5. According to the method of the third aspect of the present invention, the culture medium is 65% of burdock wood and 3% by weight of nutrient solution (the nutrient solution is extracted by yeast) _ 〇·5-2%, malt extract 052%), sucrose 3.5%, hydrogen phosphate 2% and sulfur surface 0.5%. 6. According to the method of the third aspect of the present invention, the culture is composed of 樟_($#, oil 樟, 樟, 阴阳阴) wood 65%, potato 20%, rice 糠鹏, sucrose 3.5% It consists of 1% phobia, 1% potassium and 0.5% sulfur surface. 7. The method according to the third aspect of the present invention, wherein the culture medium is composed of a cedar family (such as cedar, etc.) suitable for the growth of the cedar and the golden needle, 65% of the potato, 20% of the potato, 10% of the rice bran, and sucrose. It consists of 3.5%, dipotassium hydrogen phosphate 1% and sulfur surface 0.5%. Xin 8· According to the method of the third aspect of the present invention, the culture medium is composed of more than a variety of mushrooms, fungus and other mixed peaches (such as acacia, tung oil «#) 65%, potato 20%, _ 10%, sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur surface 0.5%. λ According to the method of the third aspect of the present invention, the culture medium is a solution of potato, rice bran and water, and a nutrient solution comprising sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur surface 0.5%. 10. The method of the fourth embodiment of the present invention, the burdock, etc. _ secretly controlled by water content, humidity, first simmering water [drying at an oven temperature of 65 75 C for 8 hours, drowning water 10.3445% (Ave. 12.5%)], after drying, then absorb the clean water [soaking for 3-8 days (5 days is the best), so that the adsorption water content is about 35%]. Re-dip coating the viscous bovine congee that has been formulated and homogenized to the surface. The medium is prepared by extracting 20% potato and 10% rice bran with water, and then adding sucrose 3.5%, dipotassium hydrogen phosphate 1%, magnesium sulfate 5%, and methyl cellulose 〇·5%-2%. Thick nutrient solution. 200835791 X. Application for Patent Park (1): 1. The method of the third item of the present invention, which is composed of burdock wood 65%, potato 20%, rice bran 10%, sucrose 3.5%, hydrogen phosphate II. It consists of 1% potassium and 5% sulphur surface. 1 According to the method of the third item of the invention, the culture medium is made up of burdock #65%, _ 30% (the soymilk bottle and water are made at a ratio of 1:7 to 1:14), sucrose 3.5%. a method according to the third aspect of the present invention, which comprises 65% of burdock wood and 3% by weight of nutrient solution (the nutrient solution 雠 yeast) It is composed of 0.5-3% of extract, 3'5% of sucrose, 1% of dipotassium hydrogen phosphate and 5% of sulphur. 4. The method of the third item of the description, the culture medium is 65% of burdock wood, 3〇% of nutrient solution (the Φ nutrient solution is prepared by extracting 0.5-3% of malt), sucrose 15%, hydrogen phosphate Dipotassium 1% and sulfur cryptiness 0.5%. 5. The method according to item 3 of the present invention, wherein the culture medium is 65% of burdock wood and 30% of nutrient solution (the nutrient solution is 0. 5·2% of yeast extract, and 0.5 of malt extract). 2%), sucrose 3.5%, phosphorus ugly potassium 1% and sulfur surface 0.5 Xiao. 6. The method according to the third aspect of the present invention, wherein the culture medium is 65% of eucalyptus (Taro, fragrant), 20% of potatoes, 10% of impurities, 15% of sucrose, 1% of phosphorus ugly potassium. And sulfur surface composed of 0. 5%. 7. The method according to the third aspect of the present invention, wherein the culture medium is composed of 65% wood, 20% potato, 10% rice bran, and sucrose 3.5, which are suitable for the growth of the golden needle, the fragrant chain, and the like. %, the composition of dipotassium hydrogen phosphate 1% and thiopurine 0.5% 8. According to the method of the third aspect of the present invention, the medium is suitable for various types of sclerophylla, fungus, etc. The wood (such as acacia, kapok tree, etc.) 65%, potato 20%, rice bran 10%, sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur fiber 0.5%. 9. The method according to item 3 of the present invention, wherein the medium is a solution of potato, rice bran and water, and then a nutrient solution comprising 3.5% of sucrose, 1% of dipotassium hydrogen phosphate and 5% of sulfur. . 10. According to the method of the fourth embodiment of the present invention, the burdock and the like need to be controlled by water content and humidity, and the a priori water is dried for 8 hours at a temperature of 65-75 C, and the remaining water is 10.344.5% ( Aml 2.5%)], after drying, then absorb the clean water [soaking for 3-8 days (5 days is the best), so that the adsorption water content is about 35%]. Re-dip coating the shed-like beef noodles that have been blended and homogenized. The culture surface extracts 20% of the potato and 10% of the water by water, and then adds sucrose 3.5%, dipotassium phosphate 1%, thiopurine·5% and methylcellulose 〇.5%-2%. Thick nutrient solution. 8 200835791 X. Patent application scope (2): 11. According to the method of the fourth embodiment described in the present invention, the water content, the touch, etc. of the burdock and the like, and the average of iff Sf7 to cultivate the mash, Miscellaneous 0.5: 2S into a temperature of 0.5% and caffeine 12. According to the method of the fourth embodiment described in the present invention, various species of cedar (scented cedar), miscellaneous sage, sputum, water content, humidity control, First stationed in water [drying at 65-75C in oven temperature for 8 hours in Ave.12.5%)], after drying, adsorb the medium prepared on the surface of the dipping medium. The medium is 20% horse and 10% water is extracted with water. Add sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur "% 讎耐動35%)，麵贼121麟菌 13.讎本發明說明的第4項實施方式之方法，各種杉科(香杉）、雜雜等，含水量、 濕度管控’係以自然風乾之木料去浸泡經調配之培養基於表面3·8天(6天爲最佳）, 該培養基係將馬鈴薯20%、米糠10%以水萃出，再加入蔗糖3·5%、磷醜二鉀1% 及硫酸鎂0.5%所組成之營養液，再經以攝氏121度殺菌60分鐘，備供移接牛樟種 菌。 、 14.根據本發明的第4項實施方式之方法，其中菌絲體培養之溫度爲28它。 15·讎本發明說明的第4項實施方式之方法，其瓶中空氣^爲80% 〇 16·根據本發明說明的第4項實施方式之方法，其中該子實體栽培時之日間溫度爲 24-26〇C，而夜間雛爲 10-12〇C ◊ 17·根據本發明說明的第4項實施方式之方法，其中其日夜間溫差爲15°C。 18· _本發明說明的第4項實施方式之方法，成長光線以七分陰三分陽爲佳，其菌絲 體培養進行4045天，而子實體培養須9(M20天（含以上）。 19·讎本發明說明的第4項實施方式之方法，其於菌絲體培養階段前，可先顔一牛 樟菌歡難養，步媒包含⑴自灘氮保存活化的麵取一小塊洋菜菌_，移入 新鮮培養基中臟溫下培養；(2梅離生長至旺盛時，將該菌種接魅殺過菌之五 榖雜糧（例如；米粒、麥粒或麥片）之中續於恆溫下培養；⑶#該五榖雜糧中之菌 種長麵絲時，取出粒狀菌絲塊，然後置入經調配之粘稠狀營養液中，再以均質機 攪碎成樟芝種菌液，即可提供大量接麵源。 9 200835791 十、申請專利範圍（2): 11·根據本發明說明的第4項實施方式之方法，牛樟等樟科木料含水量、濕度管控，係 以自然雌;去浸泡乾淨水孓8天（6瑶最佳）使酣含水量約35%。再塗 浸經調配並已均質攪碎之粘稠狀牛籠菌赚面❶該培養麵將馬鈴薯鶴、雜 10%以水萃出，再加入蔗糖3·5%、磷酸氫二鉀1%及硫麵0.5%和甲基纖ϋ素 0.5%_2%所組成之稠狀營養液。 12·麵本發明說明的第4項實施方式之方法，各種杉科（紅杉）、雜木料（如相思樹、 木棉花樹）等，需經含水量、濕度管控，_水【以烘箱控温65-75C_ Μ傭 駐水10·3%-14·5% Ave.115%)】，_後再吸隱讎調配之培養基纖面，該培養 麵將馬鈴薯廳、雑10%财萃出，再加入蔗糖3·5%、磷_二鉀1%及硫酸 鎂〇·5%所組成之營養液（浸泡从天，5天爲最佳使吸附含水量約35%)，再經以攝 氏121度殺菌60分鐘，讎移接牛。 13·讎本發明說明的第4項實施方式之方法，各艱科（紅杉）、雜桃（如相思樹、 木棉花樹）等，含水量、濕度管控，係以自然雌之木料去浸泡經調配之培養基於 表面3-8天（6天爲最佳），該培養麵將馬鈴薯20%、米糠10%以水萃出，再加入 蔗糖3.5%、磷醜二鉀1%及硫讎0.5%所組成之營養液，再經以攝氏121度殺菌 60分鐘，備供移接牛樟種菌。 14·根據本發明的第4項實施方式之方法，其中菌絲體培養之溫度爲2ΤΟ 15·根據本發明說明的第4項實施方式之方法，其瓶中空氣溼度爲80%。 16·根據本發明說明的第4項實施方式之方法，其中該子實體栽培時之日間溫度爲 2冬26。(：，而夜間濉爲1042〇0 17·根據本發明說明的第4項實施方式之方法，其中其日夜間溫_1〇45乞。 18·根據本發明說明的第4項實施方式之方法，成長光線以七分陰三分陽爲佳，其菌絲 體培養進行4045天，而子實體培養須90420天（含以上）〇 19.讎本發明說明的第4項實施方式之方法，其於菌絲體培養階段前，可先處以一牛 樟Μλ量培養，步驟包含⑴自纖氮保存活化的菌種取一小塊洋菜菌絲塊，移入 新鮮培養基中於怪溫下培養；(2府菌種生長至旺盛時，將該菌麵種至殺過菌之五 g糧（例如；米粒、麥粒或麥片）之中續於怪溫下培養；(3府該五穀雜糧中之菌 種長漏絲時，取出雛菌絲塊,然後置ΛΙ莖調配之_狀營養液中，再以均質機 攪碎成樟雜菌液，即可麟大量接 9 200835791 十、申請專利範圍（3): 20· _：$：#明說B月的第3項內容之方法，菌絲體培養階段，包含一牛棒菌種大量培養步 驟，包含以液體培養發酵以大量培養麵，斜面試管培養後，將大量繁薪的菌種 接種515升發_養液中，於24-26°C下以1«)聊旋轉腿培養約14天，續以90ipm 往培養給14天，又接種至20升液體發酵液中養14天；其中發酵培養 液之配方爲0·Η爾母抽出物、〇·μ1%麥芽抽_、1-3%蔗糖、α〇5·0·5%磷隨二鉀、 0.01-0.1% 硫酸鎂。 21·根據本發明說明的第3項內容之方法，牛樟等形狀；不限於塊狀、條狀或片狀， 其尺寸大小、長度、趙可視容默小麵高决定。 22·根據本發明說明的第3項內容之方法，容嚣材質；不限於玻璃或塑膠。 23·根據本發明說明的第3項內容之方法，容器形式·，不限於瓶、罐或桶。 24·根據本發明說明的第3項內容之方法，容器封口不限於瓶塞或瓶蓋且其材質；不限 橡膠或麵或PU海綿亦或塑膠和鐵蓋。 25.根據本發明說明的第3項內容之方法，碳源·,不暇於蔗糖、黑糖、葡萄糖或果糖❶ 26·娜本發明說明的第3項內容之方法，氮源；不限於米糠、黄豆、酵母抽出物、 27·根據本發明說明的第3項內容之方法，微量元素；不限於磷酸氫二鉀、磷酸二氫鉀。 28.樹i本申請專利範圍第20條之方法，其液體發酵培養，培養液之配方爲0.5%酵母 抽出物、0.5%麥芽抽出物、2%蔗糖、0.1%磷醜二鉀及〇娜硫瞧。 29·根據本申請專利範圍第10條、第11條、第19條之方法，其粘稠狀營養液係將馬 鈴薯20%、雑10%以水萃出，再加入蔗糖3.5%、磷酸氫二鉀1%及硫隱〇·5%和 甲素1%所組成之稠狀營養液。本粘稠狀營養液氮源；不限於米糠、黄豆、 酵母抽出物、麥芽抽出物。碳源；不限於蔗糖、黑糖、麵糖或果糖。 30.根據本申請專利範圍第10條、第11條之方法，牛樟等各種樟科木料浸泡乾淨水5 天後’其自然釋出之樟木溶出液（樟木屑水），因含terpineol、safrole、camphor成 分，可當替代水用於浸泡香杉、相思樹等木料。 31·根據本申請專利範圍第1〇條、第lljp之方法，該培養基係由牛樟等各種樟科木料 65%、水30%、蔗糖3·5%、磷酸氫二鉀1%及硫薩0 5%所組成。 10 200835791 十、申請專利範圍（3): 20·據本發明說明的第3項內容之方法，菌絲體培養階段，包含一牛樟菌種大量培養步 驟，包含以液體培養發酵以大量培養麵，斜面試管培養後，將大量繁殖好的麵 接種之5 養液中，於24«26°C下以160 rpm旋轉培養約14天，續以9〇jpm 往養約14天，又接種至20升液體發酵液中^培養14天；其中發酵培養 液之配方爲0.14%酵母抽出物、0.14%麥芽抽出物、1-3%蔗糖、0·05-0·5%磷酸氫二鉀、 0则·赚酸鎂。 21.根據本發明說明的第3項內容之方法，牛樟等形狀；不限於塊狀、條狀或片狀， 其尺寸大小、長度、離可灘器大小纖高决定❶ 22·根據本發明說明的第3項內容之方法，容器材質；不限於玻璃或塑膠。 23·根據本發明說明的第3項內容之方法，容器形式•，不限於瓶、罐、桶或RP袋^ 24. 根據本發明說明的第3項內容之方法,容器封口不限於瓶塞、瓶蓋或繩，且其材質； 不限橡膠或砂膠或PU讎亦或塑膠和鐵蓋❶ 25. 根據本發明說明的第3項內容之方法，碳源;不_蔗糖、黑糖、葡萄糖或果糖。 26·根據本發明說明的第3項內容之方法，氮源；不限於米糠、黄豆、酵母抽出物、 麥芽抽出物。 27·根據本發明說明的第3項內容之方法，微量元素;不限於磷酸氫二鉀、磷酸二氫鉀。 28. 根據本申請專利範圍第20條之方法，其液體發酵培養，培養液之配方爲酵母 抽出物、0.5%麥芽抽出物、2%蔗糖、0.1%磷醜二鉀及〇·〇5%硫薩。 29. 根據本申請專利範圍第10條、第11條、第19條之方法，其粘稠狀營養液係將馬 鈴薯20%、雑10%以水萃出，再加入蔗糖3·5%、二鉀1%及硫麵〇·5%和 甲基纖維素1%所組成之稠狀營養液。本粘稠狀營養液氮源；不限於米糠、黄豆、 酵母抽出物、麥芽抽出物。碳源·，不限於蔗糖、黑糖、麵糖或果糖。 30. 根據本申請專利範圍第10條 '第11條之方法，牛樟等各種樟科木料浸泡乾淨水5 天後，其自然釋出之樟木溶出液（樟木屑水），因含t_eol、safrole、camphor成 分，可當替代水用於浸泡香杉、相思樹等木料。 31·根據本申請專利範圍第10條、第11條之方法，該培養基係由牛樟等各種樟科木料 65%、水30%、蔗糖3.5%、磷酸氫二鉀1%及硫麵0.5%所組成。 10雠 雠 3 3 3 3 , 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 121 The medium to be immersed was immersed on the surface for 3·8 days (the best for 6 days). The medium was extracted with 20% potato and 10% rice bran with water, and then added with sucrose 3.5% and phosphorus ugly potassium 1%. The nutrient solution consisting of 0.5% magnesium sulfate is sterilized for 60 minutes at 121 degrees Celsius for the transfer of the burdock. 14. The method according to the fourth embodiment of the present invention, wherein the mycelium culture temperature is 28%. The method of the fourth embodiment of the present invention, wherein the air temperature in the bottle is 80% 〇16. The method according to the fourth embodiment of the present invention, wherein the fruit temperature during the cultivation of the fruit body is 24 -26〇C, and the night chick is 10-12〇C ◊ 17. The method of the fourth embodiment according to the present invention, wherein the day and night temperature difference is 15 °C. 18. The method according to the fourth embodiment of the present invention, wherein the growth light is preferably seven minutes and three cents, and the mycelial culture is carried out for 4045 days, and the fruit body culture is carried out for 9 days (M20 days (including above). 19. The method according to the fourth embodiment of the present invention, wherein before the mycelial culture stage, the bacterium can be raised with a bacterium, and the bacterium comprises (1) taking a small piece from the surface of the beach nitrogen storage activation. Acacia _, migrating into fresh medium and cultivating under dirty temperature; (2) When the plum grows to a strong state, the cultivar is succulent in the glutinous glutinous rice (for example, rice, wheat or cereal) (3)# When the long-term silk of the strain in the five-cropped grains is taken out, the granular hyphae are taken out, and then placed into the viscous nutrient solution prepared, and then pulverized into a bryophyte liquid by a homogenizer. A large number of junction sources can be provided. 9 200835791 X. Patent application scope (2): 11. According to the method of the fourth embodiment of the present invention, the moisture content and humidity control of the burdock and the like are natural. Female; to soak clean water for 8 days (6 Yao is the best) to make the water content of the sputum about 35%. The viscous bovine bacterium containing the homogenized granules is used to make the noodles. The culture surface is extracted with 10% of the potato crane and the miscellaneous water, and then added with sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur surface 0.5%. And a thick nutrient solution composed of methyl fibrin 0.5%_2%. 12. The method of the fourth embodiment of the present invention, various cedar (redwood), miscellaneous wood (such as acacia, kapok) Tree), etc., need to be controlled by water content and humidity, _ water [with oven temperature control 65-75C_ Μ 驻 驻 10 10 10 10 10 10 10 10 10 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 115 The culture medium has a 10% excretion of the potato bowl and the glutinous rice, and then a nutrient solution composed of sucrose 3.5%, phosphorus-dipotassium 1% and magnesium sulfate 5% 5% (soaked from the day, 5 The best time is to make the adsorption water content about 35%), and then sterilize for 60 minutes at 121 degrees Celsius, and then transfer the cattle. 13· The method of the fourth embodiment of the present invention, each of the hardships (Sequoia) , peach (such as acacia, kapok tree), water content, humidity control, the natural female wood to soak the prepared medium on the surface for 3-8 days (6 days is the best), the culture surface will Ma Ling 20%, rice bran 10% is extracted with water, and then added nutrient solution consisting of sucrose 3.5%, phosphorus ugly potassium 1% and glucosin 0.5%, and then sterilized for 60 minutes at 121 degrees Celsius for preparation of transfer burdock The method according to the fourth embodiment of the present invention, wherein the temperature of the mycelium culture is 2 ΤΟ 15. The method according to the fourth embodiment of the present invention has a humidity of 80% in the bottle. The method according to the fourth embodiment of the present invention, wherein the fruit temperature during the cultivation of the fruit body is 2 winter 26. (:, and the nighttime is 1042 〇 0 17) The fourth embodiment according to the present invention The method, in which day and night temperature is 〇 〇 45 乞. 18. According to the method of the fourth embodiment of the present invention, the growth light is preferably seven-point yin and three-point yang, the mycelial culture is carried out for 4045 days, and the fruit body culture is required to be 90420 days (including above) 〇 19. The method according to the fourth embodiment of the present invention, which can be cultured in the amount of burdock λ before the mycelial culture stage, and the step comprises: (1) taking a small piece of the genus of the genus The silk is transferred to a fresh medium and cultured at a strange temperature; (when the 2 species grows to a strong level, the surface of the fungus is planted into the 5 g of the sterilized food (for example, rice, wheat or cereal). It is strange to cultivate under temperature; (3) When the strain of the grain in the grain is long, the young hyphae are taken out, and then the stalk nutrient solution is placed in the stalk, and then mixed with a homogenizer to form a mixed liquid. It can be connected to a large number of 9 200835791 X. Patent application scope (3): 20· _:$:# The method of Ming said the third item of B month, the mycelium culture stage contains a large number of culture steps of a species of Bovine Including a large amount of culture surface fermented by liquid culture, and a large number of well-grown bacteria after being cultured in a beveled test tube Inoculate 515 liters of _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ The formula of the fermentation broth is 0·Mer's mother extract, 〇·μ1% malt pumping _, 1-3% sucrose, α〇5·0·5% phosphorus with dipotassium, 0.01-0.1% magnesium sulfate 21. The method according to the third aspect of the present invention, the shape of the burdock or the like; not limited to a block shape, a strip shape or a sheet shape, the size, the length, and the height of the Zhao visible capacitance are determined. The method of the third aspect of the invention is a material, not limited to glass or plastic. 23. The method according to the third aspect of the present invention, the container form, is not limited to a bottle, a can or a barrel. According to the method of the third aspect of the invention, the container seal is not limited to the stopper or the cap and the material thereof; and the rubber or the surface or the PU sponge or the plastic and the iron cover are not limited. 25. The third item according to the description of the present invention Method, carbon source, not sucrose, brown sugar, glucose or fructose ❶ 26· Na, the method of the third item of the invention, nitrogen source; It is not limited to rice bran, soybean, yeast extract, 27. The method according to item 3 of the present invention, trace elements; not limited to dipotassium hydrogen phosphate, potassium dihydrogen phosphate. The method is liquid fermentation culture, and the formulation of the culture solution is 0.5% yeast extract, 0.5% malt extract, 2% sucrose, 0.1% phosphorus bismuth potassium and sulphate. 29· According to the scope of patent application In the method of 10, 11 and 19, the viscous nutrient solution extracts 20% of potato and 10% of water by water, and then adds sucrose 3.5%, dipotassium hydrogen phosphate 1% and sulfur concealment. A thick nutrient solution consisting of 5% and 1% of A. The viscous nutrient solution nitrogen source; not limited to rice bran, soybean, yeast extract, malt extract. Carbon source; not limited to sucrose, brown sugar, facial sugar or fructose. 30. According to the method of Articles 10 and 11 of the patent scope of the present application, the eucalyptus wood and the like are immersed in clean water for 5 days, and then the naturally released eucalyptus solution (the sassafras water) contains terpineol. The safrole and camphor ingredients can be used as a substitute for water to soak wood such as cedar and acacia. 31. According to the method of the first paragraph and the lljp of the patent scope of the present application, the culture medium is composed of various carp, such as burdock, 65%, 30% water, 3.5% sucrose, 1% potassium dihydrogen phosphate, and thiosalate. Composition of 0 5%. 10 200835791 X. Patent application scope (3): 20. According to the method of the third aspect of the present invention, the mycelium culture stage comprises a large number of culture steps of a burdock strain, including a large number of culture surfaces by liquid culture fermentation. After culturing the slanted test tube, inoculate the 5 nutrient solution inoculated with a large number of seeds, rotate at 160 rpm at 160 rpm for about 14 days, continue to feed for 9 days at 9 〇 jpm, and inoculate to 20 The liquid fermentation broth is cultured for 14 days; wherein the fermentation broth is formulated as 0.14% yeast extract, 0.14% malt extract, 1-3% sucrose, 0.05-0.5% dipotassium hydrogen phosphate, 0 Then earn magnesium. 21. The method according to the third aspect of the present invention, the shape of a calf or the like; not limited to a block shape, a strip shape or a sheet shape, the size, the length, and the size of the fiber from the beach can be determined according to the present invention. The method of item 3 of the description, the material of the container; not limited to glass or plastic. 23. The method according to item 3 of the present invention, the container form, not limited to a bottle, a can, a bucket or an RP bag. 24. According to the method of claim 3 of the present invention, the container closure is not limited to a stopper, Bottle cap or rope, and its material; not limited to rubber or sand glue or PU 雠 or plastic and iron cover ❶ 25. Method according to item 3 of the present invention, carbon source; not sucrose, brown sugar, glucose or fructose. 26. A method according to the third aspect of the present invention, the nitrogen source; not limited to rice bran, soybean, yeast extract, malt extract. 27. A method according to the third aspect of the present invention, the trace element; not limited to dipotassium hydrogen phosphate or potassium dihydrogen phosphate. 28. The liquid fermentation culture according to the method of Article 20 of the patent application of the present application, wherein the formulation of the culture solution is yeast extract, 0.5% malt extract, 2% sucrose, 0.1% phosphorus ugly potassium and 〇·〇 5% Thiesa. 29. According to the method of Articles 10, 11, and 19 of the scope of the patent application, the viscous nutrient solution extracts 20% of the potato and 10% of the mash, and then adds sucrose to 3.5% and two. A thick nutrient solution consisting of 1% potassium, thiophene quinone·5% and 1% methylcellulose. The viscous nutrient solution nitrogen source; not limited to rice bran, soybean, yeast extract, malt extract. The carbon source is not limited to sucrose, brown sugar, facial sugar or fructose. 30. According to the method of Article 10 of the patent scope of the present application, the eucalyptus wood, such as burdock, is soaked in clean water for 5 days, and the eucalyptus solution (woodworm water) naturally released is contained in t_eol. The safrole and camphor ingredients can be used as a substitute for water to soak wood such as cedar and acacia. 31. According to the method of Articles 10 and 11 of the patent scope of the present application, the culture medium is composed of various carp, such as burdock, 65%, 30% water, 3.5% sucrose, 1% potassium hydrogen phosphate, and 0.5% sulfur surface. Composed of. 10