WO2001005938A1 - A method for producing mycelia of ganoderma lucidum containing selenium and other trace elements - Google Patents

A method for producing mycelia of ganoderma lucidum containing selenium and other trace elements Download PDF

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WO2001005938A1
WO2001005938A1 PCT/CN2000/000201 CN0000201W WO0105938A1 WO 2001005938 A1 WO2001005938 A1 WO 2001005938A1 CN 0000201 W CN0000201 W CN 0000201W WO 0105938 A1 WO0105938 A1 WO 0105938A1
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ganoderma lucidum
selenium
ganoderma
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Quancai Li
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Quancai Li
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
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    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution

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  • the present invention relates to a method for producing ganoderma, and in particular, the present invention relates to a method for producing ganoderma mycelium containing selenium and other various elements. current technology
  • Ganoderma G an0 dermaLucidum The taste is mild and the temperature is warm, which can nourish the deficiency of the cold, and replenish the solid.
  • Selenium is one of the essential trace elements in the human body. Selenium is found in many functional proteases in the body. Compared with inorganic selenium, organic selenium is more conducive to human body absorption and has good safety. Selenium compounds can scavenge lipid free radicals and hydroxyl radicals. Selenium can affect the expression and regulation of cancer cell genes, and selenium intervention has been used to obtain good anti-cancer effects in cancer-prone areas. China has a low-selenium zone covering 15 provinces from northeast to southwest. The average daily selenium for adults in China is 26.42ug, and the normal daily demand for adults is 50-200ug.
  • iron, zinc, copper, manganese, selenium, chromium, fluorine, and iodine are essential elements of the human body.
  • the human body's requirements for these elements are: 10-20 mg of iron, 15 mg of zinc, 2-3 mg of copper, 3 ⁇ 4 50-200 g, 2.5-5 mg of manganese, 100ug of cobalt, 50-120ug of chromium, and 2 of fluorine 1-3mg, iodine 1.5mg.
  • These elements each have their special functions for the synthesis of certain adenine and hormones in the human body, energy conversion, reproduction, growth, development, brain thinking and memory, information transmission, and transmission of the genetic code.
  • Elements are the only basic nutrients that cannot be synthesized by the body itself. When the elements coexist in the human body, there is not only a quantitative relationship, but more importantly, a proportional relationship and a complex interaction, which are specifically manifested in synergy and antagonism, or both.
  • the existence of trace elements in Chinese medicine is mainly to form complexes with organic matter.
  • the amino acids, peptides, proteins, and nucleic acids contained in the mycelium of Ganoderma lucidum can be used as excellent complexes of chromium compounds.
  • Ganoderma lucidum mycelium synthesizes complexes with selenium and a series of trace elements, which not only enriches traditional Chinese medicine, but also provides a new approach for the study of trace elements in traditional Chinese medicine.
  • the purpose of the present invention is to adopt an advanced liquid deep fermentation process, use a special liquid culture solution adapted to the process, and add selenium and a series of trace elements to prepare a multi-linguine ganoderma selenium mycelium dry product and a water agent, and make it It can supplement biological selenium for selenium-deficient people, and has curative effects on liver, cardiovascular diseases, tumors, and AIDS prevention.
  • the invention provides a method for producing ganoderma mycelium, including:
  • red ganoderma mycelium was cultured in a medium of 50-200ppm sodium selenite;
  • the medium for cultivating red ganoderma mycelium of the present invention preferably further contains ferrous sulfate 200-1000 ppm, zinc sulfate 500-3000 ppm, copper sulfate 50-200 ppm, and manganese sulfate 100-500 ppm.
  • ferrous sulfate 200-1000 ppm preferably further contains ferrous sulfate 200-1000 ppm, zinc sulfate 500-3000 ppm, copper sulfate 50-200 ppm, and manganese sulfate 100-500 ppm.
  • the biological selenium dry product selenium is> 1000 ppm
  • the aqueous agent selenium is> 50 ppm. Its toxicity is extremely low, and its maximum tolerance to mice is 1080 times higher than the maximum safe intake of human diet. Examples
  • the primary seed tank is 50 liters, the fixed volume is 35 liters, and the inoculation volume is 1500-2000 ml (5% -7%).
  • 500 liters of secondary seed tank 350 liters of fixed volume, 35 liters (10%) of primary seed liquid.
  • sterilize the seed tank (135'C-140 ° C, 2.5-2.8 kg / cm 2 ) for 1 hour, fill the fermentation broth and then sterilize the fermentation broth: preheat the interlayer to 90 'C, 10 After a few minutes, the steam was raised to 115 V, 1.2 kg / cm 2 , and the temperature was kept constant for 8 minutes. Then the temperature was lowered and the temperature was reduced to 28 ° C.
  • Inoculation must use a dedicated inoculation tube and inoculation bottle. Fermentation temperature is 25'C—28 ° C, PH5.0-5.5, ventilation volume 1: 0.5—1.0, stirring speed is 180 rpm. Quality control sampling during the whole fermentation: at 0, check for miscellaneous bacteria, 0D value, initial sugar: 12, 24, 36, 48 hours, check for 0D, residual sugar, PH value. 48 hours normal value: PH5, 0, 0D net length of 0.5, one unit of sugar consumption.
  • the 5 ton fermentation tank has a fixed capacity of 3500 liters and an inoculation volume of 350 liters.
  • the high-temperature boiled seed oil is added as a defoamer at 2% of the fermentation broth.
  • the fermentation temperature is 25'C-26'C.
  • the initial ventilation is 1: 0.5, and it can be increased to 1 in the middle. : 1.
  • Stirring rate is 180 rpm. Measure sugar, nitrogen, and pH every 12 hours after 72 hours of culture. Measure sugar, nitrogen, pH, and sterility every 12 hours after 72 hours of incubation.
  • the standard of irrigation is : PH5, residual sugar 2.5, ammonia nitrogen ⁇ 30 ⁇ g / ml.
  • the qualified fermentation liquid in the fermentor was centrifuged, rinsed with sterile water, filtered with 100 mesh, and freeze-dried to obtain dried Ganoderma lucidum selenium. Add twice the sterile water to the washed mycelium, and heat it to 50 ° C. After 35-50 minutes, the mycelium will dissolve automatically. After aseptic filling, the mycelium solution will be obtained.
  • Duolian Ganoderma selenium dry product contains selenium> 1000ppm.
  • the liquid preparation contains selenium> 50 ⁇ ! 11, and the contents of iron, zinc, copper, manganese, iodine, chromium, fluorine, and boron all exceed the content of conventional ganoderma mycelium fermentation products.

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Abstract

A method for producing mycelia of Ganoderma lucidum. It comprises: a, culturing the seed of Ganoderma lucidum CGMCC 0475 in the medium containing glucose 1.5-2.5 %(w/w), corn steep powder 2-3 %(w/w), peptone 0.5-1 %(w/w), beef extract 1.5-2.5 % (w/w), KH2PO4 0.1-0.2 %(w/w), Na2SeO3 30-50ppm, pH 5.0-5.5; b, inoculating above seed culture into the medium containing glucose 1-2 %(w/w), corn steep powder 3-4 %(w/w), peptone 0.5-1 %(w/w), beef extract 2-3 %(w/w), Na2SeO3 30-50ppm, pH 5.0-5.5 so as to obtain the mycelia of Ganoderma lucidum; c, purifying and preparing mycelia. The mycelia of the invention contain > 1000ppm selenium (dry weight), or > 50ppm selenium (wet weight).

Description

含有硒和其它多种元素的灵芝的生产方法 技术领域  Method for producing ganoderma containing selenium and various other elements Technical field
本发明涉及一种灵芝的生产方法， 具体地说， 本发明涉及含有硒和其它多种元 素的灵芝菌丝体的生产方法。 现有技术  The present invention relates to a method for producing ganoderma, and in particular, the present invention relates to a method for producing ganoderma mycelium containing selenium and other various elements. current technology
灵芝 G an0dermaLucidum。 味淡、 性温， 能补润虚寒， 扶正固本。 《神农本草经》 记载： "主耳聋、 利关节、 保神、 益精气、 坚筋骨， 好颜色"。 《本草纲目》 "久服、 轻身不老延年"。 现代多指标综合研究表明： 灵芝深层发酵液与灵芝子实体含有相似 的化学成分和药理作用， 自然灵芝含硒元素 0. 5-2. 0ppm。 Ganoderma G an0 dermaLucidum. The taste is mild and the temperature is warm, which can nourish the deficiency of the cold, and replenish the solid. "Shen Nong's Materia Medica" records: "The main deafness, sharp joints, protects the soul, replenishes vital energy, strengthens bones and bones, good color". "Compendium of Materia Medica""Jiufu, light weight, not old and prolonged". Modern multi-indicator comprehensive research shows that: Ganoderma lucidum deep fermentation broth and Ganoderma lucidum fruit body contain similar chemical components and pharmacological effects, and natural Ganoderma lucidum contains selenium element 0.5 to 2.0 ppm.
硒是人体必需的微量元素之一， 硒存在于肌体的多种功能蛋白酶。 有机硒较之 于无机硒更有利于人体的吸收和具有很好的安全性。 硒化合物能清除脂质过氧自由 基和羟自由基。 硒可以影响癌细胞基因的表达和调控， 在癌症高发区采用硒干预获 得良好的防癌效果。 我国从东北到西南有一覆盖 15个省区的低硒地带。 我国成人日 硒平均 26. 42ug, 成人正常日需量为 50— 200ug。  Selenium is one of the essential trace elements in the human body. Selenium is found in many functional proteases in the body. Compared with inorganic selenium, organic selenium is more conducive to human body absorption and has good safety. Selenium compounds can scavenge lipid free radicals and hydroxyl radicals. Selenium can affect the expression and regulation of cancer cell genes, and selenium intervention has been used to obtain good anti-cancer effects in cancer-prone areas. China has a low-selenium zone covering 15 provinces from northeast to southwest. The average daily selenium for adults in China is 26.42ug, and the normal daily demand for adults is 50-200ug.
占人体总量为 0. 05%的 41 种微量元素中， 铁、 锌、 铜、 锰、 硒、 铬、 氟、 碘 等是人体必需元素。 人体对这些元素的需要量（ 日需量)是： 铁 10— 20mg， 锌 15mg， 铜 2— 3mg， ¾ 50-200 g, 锰 2. 5 — 5mg， 钴 100ug，铬 50— 120ug， 氟 2. 1— 2. 3mg， 碘 1. 5mg。 这些元素对人体的某些腺素和激素的合成， 能量转换， 生殖、 生长、 发 育， 大脑的思维与记忆， 信息的传导， 遗传密码的传递等各有其特殊功能。 元素是 唯一不能在人体内自行合成的最基本的营养素。 元素在人体共存时不但有数量的关 系， 更重要的是有比例关系和复杂的相互作用， 具体表现在协同作用和拮抗作用或 二者兼而有之。  Of the 41 trace elements that account for 0.05% of the human body, iron, zinc, copper, manganese, selenium, chromium, fluorine, and iodine are essential elements of the human body. The human body's requirements for these elements (daily demand) are: 10-20 mg of iron, 15 mg of zinc, 2-3 mg of copper, ¾ 50-200 g, 2.5-5 mg of manganese, 100ug of cobalt, 50-120ug of chromium, and 2 of fluorine 1-3mg, iodine 1.5mg. These elements each have their special functions for the synthesis of certain adenine and hormones in the human body, energy conversion, reproduction, growth, development, brain thinking and memory, information transmission, and transmission of the genetic code. Elements are the only basic nutrients that cannot be synthesized by the body itself. When the elements coexist in the human body, there is not only a quantitative relationship, but more importantly, a proportional relationship and a complex interaction, which are specifically manifested in synergy and antagonism, or both.
微量元素在中药的存在形式主要是和有机物形成络合物， 灵芝菌丝体所含氨基 酸、 肽、 蛋白质、 核酸等可作为铬合物的优良配合体。 灵芝菌丝体与硒元素及系列 微量元素合成络合物， 不仅丰富了中医中药， 还为中药微量元素的研究提供了新的 途径。  The existence of trace elements in Chinese medicine is mainly to form complexes with organic matter. The amino acids, peptides, proteins, and nucleic acids contained in the mycelium of Ganoderma lucidum can be used as excellent complexes of chromium compounds. Ganoderma lucidum mycelium synthesizes complexes with selenium and a series of trace elements, which not only enriches traditional Chinese medicine, but also provides a new approach for the study of trace elements in traditional Chinese medicine.
在公知的技术中， 中国专利申请号 94115520. X使用微生物制备有机硒提取剂的 方法， 中国专利申请号 95105605. 0 微量元素的生物转化及酶化提取方法， 中国 专利申请号 96119695. 5深层发酵法生产灵芝保健饮料， 中国专利申请号 96107080. 3 富含多种微量元素的灵芝菌丝体及其生产方法， 中国专利申请号 98113198. 5灵芝 10 吨发酵罐工艺等技术。 其不足之处是： 未充分发挥硒元素与灵芝菌丝体的主导作用， 定量偏低， 配套的系列微量元素未科学地定性定量， 品种不明， 用量不准； 经深层 发酵的菌线体含无机元素未被有效清除； 深层发酵只有机械， 物理指标而无具体综 合效应的生物指标； 缺乏将灵芝菌丝体溶解的有效方法。 发明详细描述 In the well-known technology, Chinese Patent Application No. 94115520. X Method for preparing organic selenium extractant using microorganisms, Chinese Patent Application No. 95105605. 0 Method for biological transformation and enzymatic extraction of trace elements, China Patent application No. 96119695. 5 deep fermentation method for the production of Ganoderma lucidum health drink, Chinese patent application No. 96107080. 3 Ganoderma lucidum mycelium rich in various trace elements and its production method, China Patent Application No. 98113198. 5 Ganoderma 10 tons fermentation tank process And other technologies. Its shortcomings are: the leading role of selenium and ganoderma mycelium is not fully exerted, and the quantitative is low, and the supporting series of trace elements have not been scientifically qualitatively and quantified, the variety is unknown, and the amount is inaccurate; the mycelium content after deep fermentation Inorganic elements have not been effectively removed; deep fermentation only has mechanical and physical indicators without specific comprehensive biological indicators; lack of effective methods to dissolve ganoderma mycelium. Detailed description of the invention
本发明的目的是： 采用先进的液体深层发酵工艺， 使用适应于本工艺的专用液 体培养液， 并添加硒元素及系列微量元素， 制取多元灵芝硒菌丝体干品和水剂、 使 之能为缺硒人群补充生物硒， 并在肝、 心血管疾病、 肿瘤、 艾滋病防治等方面具有 疗效。  The purpose of the present invention is to adopt an advanced liquid deep fermentation process, use a special liquid culture solution adapted to the process, and add selenium and a series of trace elements to prepare a multi-linguine ganoderma selenium mycelium dry product and a water agent, and make it It can supplement biological selenium for selenium-deficient people, and has curative effects on liver, cardiovascular diseases, tumors, and AIDS prevention.
本发明提供了一种灵芝菌丝体的生产方法， 包括：  The invention provides a method for producing ganoderma mycelium, including:
1、 在含有葡萄糖 1. 5— 2. 5% (重量）， 玉米粉 2— 3% (重量）， 蛋白胨 0. 5- 1 % (重量）， 牛肉膏 1. 5— 2. 5% (重量），磷酸二氢钾 0. 1— 0. 2 % (重量），亚硒酸钠 30 -50ppm, PH5. 0- 5. 5 的培养基中培养赤灵芝 （保藏单位： 中国菌种保藏管理委员 会普通微生物中心， 简称 CGMCC; 保藏单位地址： 中国， 北京， 中关村； 保藏日期： 2000年 7月 13日； 保藏号： CGMCC 0475 ) 种子；  1. In the case of containing glucose 1.5-2.5% (weight), corn flour 2-3% (weight), peptone 0.5- 1% (weight), beef paste 1.5-2.5% (weight) ), Potassium dihydrogen phosphate 0.1-0.2% (weight), sodium selenite 30 -50ppm, PH5. 0-5. 5 culture medium (preserved by: Chinese strains collection management committee Common Microbial Center, abbreviated as CGMCC; depository address: Zhongguancun, Beijing, China; date of deposit: July 13, 2000; deposit number: CGMCC 0475) seed;
2、将培养出的赤灵芝种子接种在含有葡萄糖 1一 2% (重量）， 玉米粉 3— 4% (重 量）， 蛋白胨 0. 5— 1 % (重量）， 牛肉膏 2— 3% (重量）， PH5. 0-5. 5, 亚硒酸纳 50— 200ppm的培养基中培养出赤灵芝菌丝体；  2. Inoculate the cultivated red ganoderma seeds in glucose containing 1-2% by weight, corn flour 3-4% by weight, peptone 0.5-1% by weight, beef extract 2-3% by weight ), PH5. 0-5. 5, red ganoderma mycelium was cultured in a medium of 50-200ppm sodium selenite;
3、 纯化和制取培养出的赤灵芝菌丝体。  3. Purify and prepare the cultured red ganoderma mycelium.
在本发明的用于培养赤灵芝菌丝体的培养基中最好还含有硫酸亚铁 200— lOOOppm, 硫酸锌 500— 3000ppm， 硫酸铜 50— 200ppm， 硫酸锰 100— 500ppm。 此外 还可以再添加碘化锌 10 - lOOppm, 铬酸钾 10—100ppm， 氯化钴 10— lOOppm, 氟 硼酸钠 20 — 200pptn。  The medium for cultivating red ganoderma mycelium of the present invention preferably further contains ferrous sulfate 200-1000 ppm, zinc sulfate 500-3000 ppm, copper sulfate 50-200 ppm, and manganese sulfate 100-500 ppm. In addition, you can also add zinc iodide 10-100 ppm, potassium chromate 10-100 ppm, cobalt chloride 10-100 ppm, sodium fluoroborate 20-200 pptn.
上述配方中还可以使用含有硒或其它微量元素的化合物， 只要它们易溶于水（或 热水）， 无毒、 无腐蚀作用。 在本发明的方法所使用的培养基中硒元素含量相当于人 体日需量 80— 100% , 其它微量元素添加量相当于人体日需量的 10— 30% , 元素添 加总量 <发酵液的千分之二。 本发明的方法可以在任何一种公知的用于灵芝培养的条件下进行。 在本发明中未被转化的无机元素被有效地清除， 保证了制剂的优质和低毒。 本 发明中生物硒干品硒〉1000ppm， 水剂硒〉50ppm。 其毒性极低， 对小鼠的最大耐受 量大于人体膳食最高安全摄入量的 1080倍。 实施例 Compounds containing selenium or other trace elements can also be used in the above formula, as long as they are easily soluble in water (or hot water), non-toxic and non-corrosive. The content of selenium in the medium used in the method of the present invention is equivalent to 80-100% of the human daily demand, and the amount of other trace elements is equivalent to 10-30% of the daily human demand. Two thousandths. The method of the present invention can be performed under any of the well-known conditions for culturing ganoderma. In the present invention, the unconverted inorganic elements are effectively removed, which ensures the high quality and low toxicity of the preparation. In the present invention, the biological selenium dry product selenium is> 1000 ppm, and the aqueous agent selenium is> 50 ppm. Its toxicity is extremely low, and its maximum tolerance to mice is 1080 times higher than the maximum safe intake of human diet. Examples
本发明中各种组分的含量均是以重量为基础， 除非特别说明。  The contents of the various components in the present invention are based on weight unless otherwise specified.
灵芝斜面母种： 赤灵芝 （保藏号： CGMCC0475)  Ganoderma Lucidum Inclined Female Species: Red Ganoderma (Deposit No .: CGMCC0475)
将培养液装入 500毫升三角瓶内， 每瓶装量为 100毫升， 用牛皮纸和棉花封口。 在 1.5Kg I cm2压力下灭菌 30分钟， 冷却至 30Ό时接种。 每瓶接入 2cm2的种子， 首先在 24Ό — 26'C恒温条件下培养 24小时， 再置于旋转式摇床上培养。 摇床偏心 距为 4cm， 振荡频率为 150— 180转 /分。 室温（或摇床自动控温） 24'C— 28°C， 培养 72-96 小时， 停止培养。 合格的摇瓶种子培养液是： 清澈透明， 镜检菌丝球直径 1 -10 mm, 并按标准镜检后方可做种子液。 Fill the culture medium into a 500 ml Erlenmeyer flask, each containing 100 ml, and seal it with kraft paper and cotton. Sterilize under 1.5Kg I cm 2 pressure for 30 minutes and inoculate when cooled to 30 ° C. Seeds of 2 cm 2 are inserted into each bottle, and first cultivated at a constant temperature of 24Ό-26'C for 24 hours, and then cultivated on a rotary shaker. The eccentricity of the shaker is 4 cm, and the oscillation frequency is 150-180 rpm. Room temperature (or automatic temperature control of the shaker) 24'C—28 ° C, incubate for 72-96 hours, and stop cultivation. Qualified shake flask seed culture solution is: clear and transparent, microscopic examination of mycelium ball diameter 1-10 mm, and according to standard microscopy can be used as seed solution.
一级种子罐 50升， 定容 35升， 接种量为 1500— 2000毫升（5%— 7%)。 二级种 子罐 500升， 定容 350升， 接入一级种子液 35升（10%)。 接种前， 先将种子罐灭菌 (135'C— 140°C， 2.5-2.8 公斤 / cm2) 1 小时， 装入发酵液再对发酵液灭菌： 先预 热夹层至 90 'C, 10分钟后， 再放蒸汽升至 115 V， 1.2公斤 /cm2 ， 恒温 8分钟， 然后降温， 降至发酵液为 28'C时接种。 接种必须使用专用接种管、 接种瓶。 发酵 温度 25'C— 28°C， PH5.0-5.5, 通风量 1: 0.5— 1.0， 搅拌速率 180转 /分。 发酵 全过程抽样质监： 0时， 查杂菌， 0D值、 初糖： 12、 24、 36、 48小时， 查 0D， 残 糖， PH值。 48小时正常值： PH5、 0、 0D净长值 0.5， 耗糖一个单位。 The primary seed tank is 50 liters, the fixed volume is 35 liters, and the inoculation volume is 1500-2000 ml (5% -7%). 500 liters of secondary seed tank, 350 liters of fixed volume, 35 liters (10%) of primary seed liquid. Before inoculation, sterilize the seed tank (135'C-140 ° C, 2.5-2.8 kg / cm 2 ) for 1 hour, fill the fermentation broth and then sterilize the fermentation broth: preheat the interlayer to 90 'C, 10 After a few minutes, the steam was raised to 115 V, 1.2 kg / cm 2 , and the temperature was kept constant for 8 minutes. Then the temperature was lowered and the temperature was reduced to 28 ° C. Inoculation must use a dedicated inoculation tube and inoculation bottle. Fermentation temperature is 25'C—28 ° C, PH5.0-5.5, ventilation volume 1: 0.5—1.0, stirring speed is 180 rpm. Quality control sampling during the whole fermentation: at 0, check for miscellaneous bacteria, 0D value, initial sugar: 12, 24, 36, 48 hours, check for 0D, residual sugar, PH value. 48 hours normal value: PH5, 0, 0D net length of 0.5, one unit of sugar consumption.
5吨发酵罐定容 3500升， 接种量 350升。 接种前， 先将发酵罐灭菌（14(TC, 2.8 公斤 /cm2)l 小时， 加入发酵液再对发酵液灭菌（115'C— 118'C)50— 60分钟后， 降 温至 25'C。 在接种时， 将高温煮沸后的莱籽油按发酵液的 2%加入作消泡剂。 发酵 温度 25'C— 26'C。 前期通风量 1:0.5, 中期可加大至 1:1。 搅拌速率为 180转 /分。 培养 72小时后每隔 12小时测糖、 氮、 PH值。 培养 72小明后每隔 12小时测糖、 氮、 PH、 无菌度。 放灌标准是： PH5， 残糖 2.5， 氨态氮 <30亳克 /毫升。 The 5 ton fermentation tank has a fixed capacity of 3500 liters and an inoculation volume of 350 liters. Before inoculation, first sterilize the fermentation tank (14 (TC, 2.8 kg / cm 2 ) for 1 hour, add the fermentation broth, and then sterilize the fermentation broth (115'C— 118'C) for 50-60 minutes, then cool down to 25 'C. At the time of inoculation, the high-temperature boiled seed oil is added as a defoamer at 2% of the fermentation broth. The fermentation temperature is 25'C-26'C. The initial ventilation is 1: 0.5, and it can be increased to 1 in the middle. : 1. Stirring rate is 180 rpm. Measure sugar, nitrogen, and pH every 12 hours after 72 hours of culture. Measure sugar, nitrogen, pH, and sterility every 12 hours after 72 hours of incubation. The standard of irrigation is : PH5, residual sugar 2.5, ammonia nitrogen <30 亳 g / ml.
发酵罐合格发酵液经离心分离， 再加无菌水冲洗， 100 目过滤， 冷冻干燥后即 得多元灵芝硒干品。 将清洗后的菌丝体添加 2倍的无菌水， 升温至 50°C， 35-50 分钟后菌丝体自 溶， 无菌灌装后即得菌丝体水剂。 The qualified fermentation liquid in the fermentor was centrifuged, rinsed with sterile water, filtered with 100 mesh, and freeze-dried to obtain dried Ganoderma lucidum selenium. Add twice the sterile water to the washed mycelium, and heat it to 50 ° C. After 35-50 minutes, the mycelium will dissolve automatically. After aseptic filling, the mycelium solution will be obtained.
每 5吨发酵罐可产出干菌丝体 70公斤， 即产出率〉 2%，菌丝体水剂 1400公斤， 即产出率为 40%。 多元灵芝硒干品含硒〉1000ppm。 水剂含硒>50卯!11， 铁、 锌、 铜、 锰、 碘、 铬、 氟、 硼含量均超过常规灵芝菌丝体发酵品含量。  For every 5 ton fermenter, 70 kg of dried mycelium can be produced, that is, the yield is> 2%, and 1,400 kg of mycelium water preparation, which is 40%. Duolian Ganoderma selenium dry product contains selenium> 1000ppm. The liquid preparation contains selenium> 50 卯! 11, and the contents of iron, zinc, copper, manganese, iodine, chromium, fluorine, and boron all exceed the content of conventional ganoderma mycelium fermentation products.
多元灵芝硒急性毒性试验报告 (最大耐受量试验)结论： 经最大浓度， 最大容量， 每日灌胃给药三次， 连续观察七天， 除少数动物有大便拉稀的现象外， 未出现任何 中毒症状， 饮食活动良好， 无一例死亡， 灵芝硒对小鼠的最大耐受量〉24g / kg, 亦 即〉 7200ug / kg。 此剂量大于人 (60kg)膳食每日硒 400ug最高安全摄入量 （6.67ug / kg) 的 1080倍。  Multiple Ganoderma Lucidum Selenium Acute Toxicity Test Report (Maximum Tolerance Test) Conclusion: After maximum concentration, maximum volume, three times a day intragastric administration, continuous observation for seven days, except for a few animals with thin stool, no symptoms of poisoning The dietary activity was good without any death. The maximum tolerance of Ganoderma selenium to mice was> 24g / kg, that is,> 7200ug / kg. This dose is 1080 times the maximum safe intake (6.67ug / kg) of 400ug daily selenium in a human (60kg) diet.

Claims

1. 一种灵芝菌丝体的生产方法， 包括- a. 在含有葡萄糖 1.5— 2.5% (重量）， 玉米粉 2— 3% (重量）， 蛋白胨 0. 5— 1% (重量）， 牛肉膏 1.5— 2.5% (重量）， 磷酸二氢钾 0.1— 0.2% (重量）， 亚硒酸钠 30 -50ppm, PH5.0—5.5的培养基中培养赤灵芝 CGMCC0475种子； 1. A method for the production of ganoderma mycelium, comprising-a. In a diet containing 1.5-2.5% by weight of glucose, 2-3% by weight of corn flour, 0.5-1% by weight of peptone, beef extract 1.5-2.5% (weight), potassium dihydrogen phosphate 0.1-0.2% (weight), sodium selenite 30-50ppm, pH 5.0-5.5 cultured Ganoderma lucidum CGMCC0475 seeds;
b. 将培养出的赤灵芝种子接种在含有葡萄糖 1一 2% (重量）， 玉米粉 3— 4% (重 量）， 蛋白胨 0.5— 1% (重量）， 牛肉膏 2— 3% (重量）， PH5.0-5.5, 亚硒酸纳 50— 200ppm的培养基中培养出赤灵芝菌丝体；  b. Inoculate the cultivated red ganoderma seeds with glucose 1-2% (weight), corn flour 3-4% (weight), peptone 0.5-1% (weight), beef extract 2-3% (weight), Red ganoderma mycelium was cultured in a medium with a pH of 5.0-5.5 and sodium selenite at 50-200 ppm;
c. 纯化和制取培养出的赤灵芝菌丝体。  c. Purifying and preparing the cultured Ganoderma lucidum mycelium.
2. 按照权利要求 1所述的方法， 其中， 用于培养赤灵芝菌丝体的培养基中还含 有硫酸亚铁 200— lOOOppm, 硫酸锌 500— 3000ppm， 硫酸铜 50— 200ppm， 硫酸猛 100 — 500ppm。  2. The method according to claim 1, wherein the medium for cultivating the mycelia of Ganoderma lucidum further contains ferrous sulfate 200-1000 ppm, zinc sulfate 500-3000 ppm, copper sulfate 50-200 ppm, and fibrous sulfate 100- 500ppm.
3. 按照权利要求 2所述的方法， 其中， 用于培养赤灵芝菌丝体的培养基中还含 有碘化锌 10 -lOOppm, 铬酸钾 10— lOOppm, 氯化钴 10— lOOppm, 氟硼酸钠 20 — 200ppm。  3. The method according to claim 2, wherein the medium for cultivating the mycelia of Ganoderma lucidum further contains zinc iodide 10-100 ppm, potassium chromate 10-100 ppm, cobalt chloride 10-100 ppm, fluoboric acid Sodium 20 — 200 ppm.
PCT/CN2000/000201 1999-07-15 2000-07-14 A method for producing mycelia of ganoderma lucidum containing selenium and other trace elements WO2001005938A1 (en)

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AT500435A1 (en) * 2004-05-18 2005-12-15 G N V Gesundheits Und Naturpro Aphrodisiac food supplement contains erytroxylum catuaba, avena satavia and urtica dioca
SG129276A1 (en) * 2003-04-10 2007-02-26 Auric Pacific Nutritech Pte Lt Large-scale production method for cordyceps and ganoderma
CN104620853A (en) * 2015-02-04 2015-05-20 深圳福山生物科技有限公司 Culture method of ganoderma lucidum spore powder or sporocarp rich in selenium
CN105009930A (en) * 2014-04-25 2015-11-04 江苏中祥高科技实业有限公司 Production method for selenium-enriched ganoderma mycelium
CN105176832A (en) * 2015-07-23 2015-12-23 广西仙草堂制药有限责任公司 Preparation method of selenium-enriched lucid ganoderma strain
US20170280629A1 (en) * 2016-04-01 2017-10-05 Ami Cosmetic Co., Ltd. Methods of Producing Ganoderma Lucidum Mycelia Having Enhanced Ganodermanondiol Content
CN110257261A (en) * 2019-07-04 2019-09-20 硒利技术研发有限公司 A kind of production method of selenium-rich antrodia mycelia
CN112353719A (en) * 2020-11-02 2021-02-12 郭光波 Anti-aging lucid ganoderma face cream and preparation method thereof
CN113767812A (en) * 2021-09-23 2021-12-10 山西农业大学山西功能食品研究院 Ganoderma strain and culture method of ganoderma lucidum fruiting body
CN114214208A (en) * 2021-12-14 2022-03-22 江南大学 High-selenium-resistance ganoderma lucidum mutant strain and application thereof
CN114250153A (en) * 2021-12-15 2022-03-29 江南大学 High-selenium-resistance Ganoderma lucidum JNUSE-200 and selenium-rich fermentation strategy thereof
CN115053752A (en) * 2022-06-30 2022-09-16 安徽金寨仙芝灵生物科技有限公司 Novel ganoderma lucidum strain cultivation method

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CN1170758A (en) * 1996-07-17 1998-01-21 谭新国 Glossy ganoderma mycelium rich in micro elements and its producing method

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SG129276A1 (en) * 2003-04-10 2007-02-26 Auric Pacific Nutritech Pte Lt Large-scale production method for cordyceps and ganoderma
AT500435A1 (en) * 2004-05-18 2005-12-15 G N V Gesundheits Und Naturpro Aphrodisiac food supplement contains erytroxylum catuaba, avena satavia and urtica dioca
AT500435B1 (en) * 2004-05-18 2006-10-15 G N V Gesundheits Und Naturpro Aphrodisiac food supplement contains erytroxylum catuaba, avena satavia and urtica dioca
CN105009930A (en) * 2014-04-25 2015-11-04 江苏中祥高科技实业有限公司 Production method for selenium-enriched ganoderma mycelium
CN105009930B (en) * 2014-04-25 2018-03-09 江苏中祥高科技实业有限公司 A kind of Se-rich lucid ganoderma mycelium production method
CN104620853A (en) * 2015-02-04 2015-05-20 深圳福山生物科技有限公司 Culture method of ganoderma lucidum spore powder or sporocarp rich in selenium
CN105176832A (en) * 2015-07-23 2015-12-23 广西仙草堂制药有限责任公司 Preparation method of selenium-enriched lucid ganoderma strain
US20170280629A1 (en) * 2016-04-01 2017-10-05 Ami Cosmetic Co., Ltd. Methods of Producing Ganoderma Lucidum Mycelia Having Enhanced Ganodermanondiol Content
CN110257261A (en) * 2019-07-04 2019-09-20 硒利技术研发有限公司 A kind of production method of selenium-rich antrodia mycelia
CN112353719A (en) * 2020-11-02 2021-02-12 郭光波 Anti-aging lucid ganoderma face cream and preparation method thereof
CN113767812A (en) * 2021-09-23 2021-12-10 山西农业大学山西功能食品研究院 Ganoderma strain and culture method of ganoderma lucidum fruiting body
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CN114214208A (en) * 2021-12-14 2022-03-22 江南大学 High-selenium-resistance ganoderma lucidum mutant strain and application thereof
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CN114250153B (en) * 2021-12-15 2023-07-25 江南大学 High selenium-resistant ganoderma Ganoderma lucidum JNUSE-200 and selenium-rich fermentation strategy thereof
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