WO2019148419A1 - 一种添加酪氨酸提高食药用真菌胞外多糖产量的方法 - Google Patents

一种添加酪氨酸提高食药用真菌胞外多糖产量的方法 Download PDF

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WO2019148419A1
WO2019148419A1 PCT/CN2018/074936 CN2018074936W WO2019148419A1 WO 2019148419 A1 WO2019148419 A1 WO 2019148419A1 CN 2018074936 W CN2018074936 W CN 2018074936W WO 2019148419 A1 WO2019148419 A1 WO 2019148419A1
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fermentation
tyrosine
yield
medium
medicinal
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PCT/CN2018/074936
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French (fr)
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丁重阳
马忠宝
徐萌萌
王琼
许正宏
赵丽婷
刘高强
顾正华
石贵阳
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江南大学
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Priority to PCT/CN2018/074936 priority Critical patent/WO2019148419A1/zh
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

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  • the invention relates to a method for adding tyrosine to increase the yield of extracellular polysaccharides of edible medicinal fungi, and belongs to the field of microbial fermentation.
  • Edible and medicinal fungi belonging to the Basidiomycetes or Ascomycetes, are widely distributed in the world because of their good nutritional functions and medicinal value. Dietetic fungi are not only delicious, but also rich in digestible proteins, carbohydrates, fiber, vitamins, minerals and antioxidants that contribute to good health. Bioactive substances rich in edible and medicinal fungi, including polysaccharides, lectins, lactones, terpenoids and alkaloids, have been widely studied and applied. As its main bioactive substance, polysaccharide (PS) has anti-tumor, immune regulation, anti-inflammatory, analgesic, anti-viral, anti-oxidation, blood sugar lowering and liver protection effects, and can prevent chronic radiation.
  • PS polysaccharide
  • Ganoderma lucidum is a typical representative of edible and medicinal fungi.
  • Polysaccharide is one of the main biological active substances of Ganoderma lucidum. It has the functions of immune regulation, anti-tumor, anti-oxidation, blood sugar lowering and liver protection. These biological activities also make Ganoderma lucidum polysaccharides have great application potential in the fields of food, medicine, cosmetics and the like.
  • a first object of the present invention is to provide a method for increasing the yield of extracellular polysaccharides of edible medicinal fungi by adding tyrosine at the beginning of the fungal fermentation or during the fermentation.
  • the fungus is a medicinal fungus.
  • the medicinal and medicinal fungi include, but are not limited to, Ganoderma lucidum, fungus, shiitake mushroom, Swine fever, white fungus, ash tree flower, medlar, Yunzhi, Hericium erinaceus, Cordyceps sinensis and common organisms thereof.
  • the fungus includes, but is not limited to, Ganoderma lucidum, Pleurotus ferulae, Pleurotusgeesteranus, Grilfolafrondosa.
  • the method is the addition of tyrosine at 0 to 96 h of fermentation.
  • the final concentration of the tyrosine is from 0.1 to 3 g/L.
  • the method is to inoculate the fungus into a fermentation medium and add tyrosine to the fermentation medium.
  • the fermentation medium is a liquid fermentation medium commonly used for medicinal fungi.
  • each L contains 30 g of glucose, 1 g of tryptone, 5 g of yeast powder, 1 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate heptahydrate, 0.1 g of vitamin B 1 , and a natural pH.
  • a second object of the present invention is to provide a method for producing a fungal extracellular polysaccharide by inoculating a fungus into a fermentation medium, and adding tyrosine to the fermentation medium at the beginning of the fungal fermentation or during the fermentation. Fermentation at 25-33 ° C, 150 ⁇ 200r ⁇ min -1 for 5 ⁇ 7d.
  • inoculation is carried out in an inoculum amount of 3 to 6 g of wet weight mycelium/L medium.
  • the invention also provides for the use of the method in the preparation of a product comprising an exopolysaccharide.
  • the invention increases the yield of extracellular polysaccharides of the edible and medicinal fungi by adding tyrosine in the fermentation process without increasing the original fermentation cycle, and the maximum increase rate is more than 25%, which greatly reduces the food.
  • the production cost of medicinal fungal polysaccharides is beneficial to industrial production and product application.
  • Polysaccharide extraction take 100mL fermentation broth filtrate, add 4 times of 95% alcohol, stir for 20min, centrifuge at 4000r ⁇ min -1 for 5min, remove protein, add 2.25 times 95% alcohol, stir for 20min, then let stand at 4 °C overnight. The solution was centrifuged at 10,000 r ⁇ min -1 for 5 min, the supernatant was removed, and 30 mL of distilled water was added to the precipitate to dissolve and dissolve. The solution was centrifuged at 10,000 r ⁇ min -1 for 10 min, and the clear solution was a solution of water-soluble polysaccharide.
  • the content of polysaccharide was determined by using a phenol sulfuric acid method, and the measurement system was 2 mL of a sample solution, 1 mL of 6% phenol, and 5 mL of concentrated sulfuric acid. After cooling, the OD value was measured at a wavelength of 490 nm.
  • Ganoderma lucidum seed culture Take 0.5cm 2 size of the bacteria, inoculate the seed culture medium with 80mL/250mL flask, and incubate at 150r ⁇ min -1 and 30°C for 7d.
  • Ganoderma lucidum fermentation culture 150 mL of flask was added to 150 mL of fermentation medium, and sterilized at 115 ° C for 20 minutes. The inoculum was 0.5 g wet ganoderma mycelium, cultured at 150 r ⁇ min -1 and 30 ° C for 7 days.
  • the seed and the fermentation medium contained 30 g of glucose, 1 g of tryptone, 5 g of yeast powder, 1 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate heptahydrate, 0.1 g of vitamin B 1 , and a natural pH per L of the fermentation medium.
  • the medium and the culture method were the same as in Example 1, except that tyrosine was added to the fermentation medium at 0 h to a final concentration of 0.1 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.330 g/L, which was 10% higher than that of Example 1.
  • the medium and the culture method were the same as in Example 1, except that tyrosine was added to the fermentation medium at 0 h to a final concentration of 3 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.345 g/L, which was 15% higher than that of Example 1.
  • the medium and the culture method were the same as in Example 1, except that tyrosine was added to the fermentation medium at 0 h to a final concentration of 1 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.366 g/L, which was 22% higher than that of Example 1.
  • the medium and culture method were the same as in Example 1, except that tyrosine was added at 48 h of fermentation. Its final concentration was 0.5 g/L.
  • tyrosine was added at 48 h of fermentation. Its final concentration was 0.5 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.375 g/L, which was 25% higher than that of Example 1.
  • the medium and the culture method were the same as in Example 1, except that tyrosine was added at 96 h of fermentation. The final concentration was 2.5 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.351 g/L, which was 17% higher than that of Example 1.
  • Ganoderma lucidum seed culture Take 0.5cm 2 size of the bacteria, inoculate the seed culture medium with 80mL/250mL flask, and incubate at 150r ⁇ min -1 and 30°C for 7d.
  • Ganoderma lucidum fermentation culture 150 mL of flask was added to 150 mL of fermentation medium, and sterilized at 115 ° C for 20 minutes. The inoculum was 0.5 g wet ganoderma mycelium, cultured at 150 r ⁇ min -1 and 30 ° C for 7 days.
  • the seed and fermentation medium contained 20 g of glucose, 10 g of corn flour, 10 g of bran, 3 g of potassium dihydrogen phosphate, 2 g of magnesium sulfate heptahydrate, and natural pH.
  • the yield of extracellular polysaccharide was measured during the fermentation. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.325 g/L.
  • the medium and the culture method were the same as in Example 7, except that tyrosine was added to the fermentation medium at 0 h to a final concentration of 1 g/L.
  • the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.387 g/L, which was 19% higher than that of Example 7.
  • Ganoderma lucidum seed culture Take 0.5cm 2 size of the bacteria, inoculate the seed culture medium with 80mL/250mL flask, and incubate at 150r ⁇ min -1 and 30°C for 7d.
  • Ganoderma lucidum fermentation culture 3 L fermentation medium was added to a 5 L fermentor, and sterilized at 115 ° C for 20 minutes.
  • the inoculation amount was 10 g wet weight ganoderma lucidum mycelium, 150 r ⁇ min -1 , the aeration was 1.5 L ⁇ min -1 , and cultured at 30 ° C for 7 d.
  • the seed and fermentation medium contained 20 g of glucose, 10 g of corn flour, 10 g of bran, 3 g of potassium dihydrogen phosphate, 2 g of magnesium sulfate heptahydrate, and natural pH.
  • Tyrosine was continuously fed during the fermentation period of 24 to 96 hours to a final concentration of 1 g/L. During the fermentation, the yield of Ganoderma lucidum extracellular polysaccharide was determined. The results showed that the yield of Ganoderma lucidum polysaccharide was 0.406 g/L, which was 25% higher than that of Example 7.
  • Seed culture of Pleurotus ostreatus Take 0.5cm 2 size of the bacteria, inoculate the seed culture medium with 80mL/250mL flask, and incubate at 150r ⁇ min -1 and 28°C for 7d.
  • Fermentation culture of Pleurotus ostreatus 150 mL of fermentation flask was added to a 500 mL flask and sterilized at 115 ° C for 20 minutes.
  • the inoculum amount was 0.5 g wet weight mycelium, cultured at 150 r ⁇ min -1 and 28 ° C for 7 days.
  • the seed and the fermentation medium contained 30 g of glucose, 1 g of tryptone, 5 g of yeast powder, 1 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate heptahydrate, 0.1 g of vitamin B 1 , and a natural pH per L of the fermentation medium.
  • the medium and the culture method were the same as those in Example 10. The difference was that the experimental strain was selected from the mushroom, and tyrosine was added to the fermentation medium at 0 h to a final concentration of 1 g/L. During the fermentation, the yield of extracellular polysaccharides of Pleurotus ostreatus was measured, and the results showed that the yield of exopolysaccharide of Pleurotus ostreatus was 0.280 g/L, which was 12% higher than that of Example 10.
  • the medium and the culture method were the same as those in Example 10, except that the experimental strain was selected from the genus Mushroom. During the fermentation process, the yield of extracellular polysaccharides of Pleurotus ostreatus was determined, and the results showed that the yield of exopolysaccharide of Pleurotus ostreatus was 0.270 g/L.
  • the medium and the culture method were the same as those in Example 10. The difference was that the experimental strain was selected from the oyster mushroom, and tyrosine was added to the fermentation medium at 0 h to a final concentration of 2 g/L. During the fermentation, the yield of the exopolysaccharide of Pleurotus ostreatus was measured, and the results showed that the yield of exopolysaccharide of Pleurotus ostreatus was 0.319 g/L, which was 18% higher than that of Example 12.
  • the medium and the culture method were the same as those in Example 10 except that the experimental strain was selected from Grifola frondosa. During the fermentation, the yield of extracellular polysaccharide of Grifola frondosa was determined, and the yield of the extracellular polysaccharide of Grifola frondosa was 0.180 g/L.
  • the medium and the culture method were the same as those in Example 10, except that the experimental strain was selected from Grifola frondosa, and tyrosine was added to the fermentation medium at a concentration of 2.5 g/L at 0 h.
  • the yield of the extracellular polysaccharide of Grifola frondosa was measured, and the results showed that the yield of the extracellular polysaccharide of Grifola frondosa was 0.198 g/L, which was 10% higher than that of Example 14.
  • phenylalanine was added to the fermentation process of edible fungi such as fungus, shiitake mushroom, hog, white fungus, alfalfa, Yunzhi, Hericium erinaceus, and Cordyceps sinensis, and the results showed that The yield of extracellular polysaccharides has increased to varying degrees.
  • Example 2 In the same manner as in Example 1, except that an amino acid having a final concentration of 1 g/L was added to the medium, and the yield of extracellular polysaccharide was measured, and the results are shown in the following table.
  • Table 1 shows changes in the production of extracellular polysaccharides after adding different amino acids
  • the medicinal and medicinal fungi referred to in the present application is not limited to fungi which are known to be medicinal and medicinal to produce extracellular polysaccharides at the present stage, and fungi of known or unknown species having similarities with known medicinal and medicinal fungi are suitable.
  • the method of the present application is capable of achieving an approximate effect of an increase in the production of extracellular polysaccharides.

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Abstract

一种提高食药用真菌胞外多糖产量的方法以及该方法在制备含有胞外多糖的产品方面的应用，该方法为将真菌接种至发酵培养基中，在真菌发酵初始时或发酵过程中加入酪氨酸，酪氨酸的终浓度为0.1-3g/L。

Description

一种添加酪氨酸提高食药用真菌胞外多糖产量的方法 技术领域
本发明涉及一种添加酪氨酸提高食药用真菌胞外多糖产量的方法，属于微生物发酵领域。
背景技术
食药用真菌，隶属担子菌纲或子囊菌纲，在世界分布广泛，因其具有很好的营养功能和药用价值。食药用真菌不仅味道鲜美，而且富含大量的易消化的蛋白质、碳水化合物、纤维、维生素、矿物质和抗氧化剂，有助于身体健康。食药用真菌所富含的生物活性物质，包括多糖，凝集素，内酯，萜类化合物和生物碱等，已被广泛研究与应用。作为其主要生物活性物质，多糖(PS)具有抗肿瘤、免疫调节、抗炎、止痛、抗病毒、抗氧化、降血糖和护肝作用，而且可以防止慢性辐射。其中灵芝在食药用真菌中作为一个典型的代表，多糖是灵芝主要的生物活性物质之一，具有免疫调节、抗肿瘤、抗氧化、降血糖、保护肝脏等活性。这些生物活性也使得灵芝多糖在食品、医药、化妆品等领域具有巨大的应用潜力。
目前，通过液体深层发酵技术培养食药用真菌已经成为获取食药用真菌多糖的主要发酵方式，相较于固态发酵，其发酵周期短，提取成本降低，但是，其多糖产量相对较低，如何大幅提高食药用真菌多糖的发酵产量，以进一步降低生产成本，以实现在上述领域的广泛应用已成为食药用真菌液体发酵急需解决的问题。
发明内容
本发明的第一个目的是提供一种提高食药用真菌胞外多糖产量的方法，是在真菌发酵初始时或发酵过程中加入酪氨酸。
在本发明的一种实施方式中，所述真菌为食药用真菌。
在本发明的一种实施方式中，所述食药用真菌包括但不限于灵芝、木耳、香菇、猪苓、银耳、灰树花、茯苓、云芝、猴头菇、冬虫夏草及其常见的生物分类学上亲缘关系相近或性状相近的食药用真菌子实体或菌丝体。
在本发明的一种实施方式中，所述真菌包括但不限于灵芝(Ganoderma lucidum)、阿魏蘑(Pleurotusferulae)、秀珍菇(Pleurotusgeesteranus)、灰树花(Griflolafrondosa)。
在本发明的一种实施方式中，所述方法是在发酵0～96h添加酪氨酸。
在本发明的一种实施方式中，所述酪氨酸的终浓度为0.1～3g/L。
在本发明的一种实施方式中，所述方法是将真菌接种至发酵培养基中，向发酵培养基中添加酪氨酸。
在本发明的一种实施方式中，所述发酵培养基为食药用真菌常用液体发酵培养基。例如，每L含有葡萄糖30g，胰蛋白胨1g，酵母粉5g，磷酸二氢钾1g，七水硫酸镁1g，维生素B 10.1g，自然pH。
本发明的第二个目的是提供一种真菌胞外多糖的生产方法，所述方法是将真菌接种至发酵培养基中，在真菌发酵初始时或发酵过程向发酵培养基中加入酪氨酸，于25～33℃，150～200r·min -1发酵5～7d。
在本发明的一种实施方式中，按3～6g湿重菌丝体/L培养基的接种量进行接种。
本发明还提供所述方法在制备含有胞外多糖的产品方面的应用。
有益效果：本发明在发酵过程中通过添加酪氨酸，在不增加原有发酵周期的基础上，显著提高了食药用真菌胞外多糖产量，最大提高幅度达25％以上，大大降低了食药用真菌多糖的生产成本，有利于工业化生产和产品应用。
具体实施方式
多糖提取：取100mL发酵液滤液，加4倍的95％酒精，搅拌20min，4000r·min -1离心5min，去除蛋白，上清液加2.25倍95％酒精，搅拌20min后，于4℃静置过夜。溶液经10000r·min -1离心5min，去上清，沉淀加30mL蒸馏水振荡溶解，10000r·min -1离心10min，清液即为水溶性多糖的溶液。
多糖测定：测定多糖的含量，采用苯酚硫酸法，测定体系为2mL的样品溶液，1mL的6％的苯酚，5mL的浓硫酸。冷却后在490nm的波长下测定OD值。
实施例1
灵芝种子培养：取0.5cm 2大小的菌块，接种到装液量80mL/250mL三角瓶的种子培养基中，150r·min -1、30℃培养7d。
灵芝发酵培养：500mL三角瓶中加入150mL发酵培养基，115℃灭菌20分钟。接种量为0.5g湿重灵芝菌丝体，150r·min -1、30℃培养7d。
所述种子与发酵培养基每L含有葡萄糖30g，胰蛋白胨1g，酵母粉5g，磷酸二氢钾1g，七水硫酸镁1g，维生素B 10.1g，自然pH。
发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.300g/L。
实施例2
培养基和培养方法同实施例1，区别在于，0h在发酵培养基中添加酪氨酸，使其终浓度为0.1g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.330g/L，相对于实施例1提高了10％。
实施例3
培养基和培养方法同实施例1，区别在于，0h在发酵培养基中添加酪氨酸，使其终浓度为3g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.345g/L，相对于实施例1提高了15％。
实施例4
培养基和培养方法同实施例1，区别在于，0h在发酵培养基中添加酪氨酸，使其终浓度为1g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.366g/L，相对于实施例1提高了22％。
实施例5
培养基和培养方法同实施例1，区别在于，在发酵48h时添加酪氨酸。使其终浓度为0.5g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.375g/L，相对于实施例1提高了25％。
实施例6
培养基和培养方法同实施例1，区别在于，在发酵96h时添加酪氨酸。使其终浓度为2.5g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.351g/L，相对于实施例1提高了17％。
实施例7
灵芝种子培养：取0.5cm 2大小的菌块，接种到装液量80mL/250mL三角瓶的种子培养基中，150r·min -1、30℃培养7d。
灵芝发酵培养：500mL三角瓶中加入150mL发酵培养基，115℃灭菌20分钟。接种量为0.5g湿重灵芝菌丝体，150r·min -1、30℃培养7d。
所述种子与发酵培养基每L含有葡萄糖20g，玉米粉10g，麸皮10g，磷酸二氢钾3g，七水硫酸镁2g，自然pH。
发酵过程中，测定胞外多糖的产量。结果显示，灵芝多糖的产量为0.325g/L。
实施例8
培养基和培养方法同实施例7，区别在于，0h在发酵培养基中添加酪氨酸，使其终浓度为1g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.387g/L，相对于实施例7提高了19％。
实施例9
灵芝种子培养：取0.5cm 2大小的菌块，接种到装液量80mL/250mL三角瓶的种子培养 基中，150r·min -1、30℃培养7d。
灵芝发酵培养：5L发酵罐中加入3L发酵培养基，115℃灭菌20分钟。接种量为10g湿重灵芝菌丝体，150r·min -1，通气量为1.5L·min -1，30℃培养7d。
所述种子与发酵培养基每L含有葡萄糖20g，玉米粉10g，麸皮10g，磷酸二氢钾3g，七水硫酸镁2g，自然pH。
在24～96小时的发酵过程中连续流加酪氨酸，使其终浓度为1g/L。发酵过程中，测定灵芝胞外多糖的产量。结果显示，灵芝多糖的产量为0.406g/L，相对于实施例7提高了25％。
实施例10
阿魏蘑种子培养：取0.5cm 2大小的菌块，接种到装液量80mL/250mL三角瓶的种子培养基中，150r·min -1、28℃培养7d。
阿魏蘑发酵培养：500mL三角瓶中加入150mL发酵培养基，115℃灭菌20分钟。接种量为0.5g湿重菌丝体，150r·min -1、28℃培养7d。
所述种子与发酵培养基每L含有葡萄糖30g，胰蛋白胨1g，酵母粉5g，磷酸二氢钾1g，七水硫酸镁1g，维生素B 10.1g，自然pH。
发酵过程中，测定阿魏蘑胞外多糖的产量，结果显示，阿魏蘑胞外多糖的产量为0.250g/L。
实施例11
培养基和培养方法同实施例10，区别在于，实验菌株选用阿魏蘑，而且0h在发酵培养基中添加酪氨酸，使其终浓度为1g/L。发酵过程中，测定阿魏蘑胞外多糖的产量，结果显示，阿魏蘑胞外多糖的产量为0.280g/L，相对于实施例10提高了12％。
实施例12
培养基和培养方法同实施例10，区别在于，实验菌株选用秀珍菇。发酵过程中，测定秀珍菇胞外多糖的产量，结果显示，秀珍菇胞外多糖的产量为0.270g/L。
实施例13
培养基和培养方法同实施例10，区别在于，实验菌株选用秀珍菇，而且0h在发酵培养基中添加酪氨酸，使其终浓度为2g/L。发酵过程中，测定秀珍菇胞外多糖的产量，结果显示，秀珍菇胞外多糖的产量为0.319g/L，相对于实施例12提高了18％。
实施例14
培养基和培养方法同实施例10，区别在于，实验菌株选用灰树花。发酵过程中，测定灰树花胞外多糖的产量，结果显示，灰树花胞外多糖的产量为0.180g/L。
实施例15
培养基和培养方法同实施例10，区别在于，实验菌株选用灰树花，而且0h在发酵培养基中添加酪氨酸，使其终浓度为2.5g/L。发酵过程中，测定灰树花胞外多糖的产量，结果显示，灰树花胞外多糖的产量为0.198g/L，相对于实施例14提高了10％。
实施例16
采用与实施例1～15相同的策略，分别在木耳、香菇、猪苓、银耳、茯苓、云芝、猴头菇、冬虫夏草等食药用真菌的发酵过程中添加苯丙氨酸，结果显示，胞外多糖的产量具有不同程度的提高。
对照例1
按照实施例1相同的方式，区别在于，向培养基中加入终浓度1g/L的氨基酸，测定胞外多糖产量，结果如下表所示。
表1添加不同氨基酸后的胞外多糖产量变化
Figure PCTCN2018074936-appb-000001
注：“-”表示产量降低。
本申请所指的食药用真菌不局限于现阶段为人们所知食药用产胞外多糖的真菌，与已知的食药用真菌存在相似性的已知或未知种属的真菌皆适用于本申请的方法，并能够达到胞外多糖产量提高的近似效果。
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做各种的改动与修饰，因此本发明的保护范围应该以权利要求书所界定的为准。

Claims (10)

  1. 一种提高食药用真菌胞外多糖产量的方法，其特征在于，在真菌发酵初始时或发酵过程中加入酪氨酸。
  2. 根据权利要求1所述的方法，其特征在于，所述真菌为食药用真菌。
  3. 根据权利要求1或2所述的方法，其特征在于，所述真菌包括灵芝(Ganoderma lucidum)、阿魏蘑(Pleurotus ferulae)、秀珍菇(Pleurotus geesteranus)或灰树花(Griflola frondosa)。
  4. 根据权利要求1～3任一所述的方法，其特征在于，在发酵0～96h添加酪氨酸。
  5. 根据权利要求4任一所述的方法，其特征在于，酪氨酸的终浓度为0.1～3g/L。
  6. 一种真菌胞外多糖的生产方法，其特征在于，将真菌接种至发酵培养基中，在真菌发酵初始时或发酵过程向发酵培养基中加入酪氨酸。
  7. 根据权利要求6所述的方法，其特征在于，所述酪氨酸通过分批补加或连续流加的方式加入。
  8. 根据权利要求7所述的方法，其特征在于，所述方法是在发酵0～96h添加终浓度为0.1～3g/L的酪氨酸，于25～33℃，150～200r·min -1发酵5～7d。
  9. 根据权利要求6～8任一所述的方法，其特征在于，按3～6g湿重菌丝体/L培养基的接种量进行接种。
  10. 权利要求1～9任一所述方法在制备含有胞外多糖的产品方面的应用。
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