A fermentation process for Cordyceps militaris mycelium containing golden flowers

Abstract

The invention discloses a fermentation process for Cordyceps militaris mycelia containing golden flowers. The fermentation process comprises the following steps: uniformly mixing fish meal, glucose, potassium dihydrogen phosphate, magnesium sulfate and water, sterilizing, naturally cooling to room temperature, inoculating Cordyceps militaris strains under aseptic conditions, and culturing at a constant temperature; uniformly mixing peanut powder, glucose, linolenic acid, potassium dihydrogen phosphate, magnesium sulfate and water, sterilizing, naturally cooling to room temperature, inoculating strains obtained by the culture under aseptic conditions, and culturing at a constant temperature; uniformly mixing Fucha powder, corn starch, skim milk powder, nutrient solution and water, sterilizing, naturally cooling to room temperature, inoculating strains obtained by the culture under aseptic conditions, culturing at a constant temperature until reducing sugar is less than 0.1%, and harvesting by tanking; and vacuum concentrating the obtained product at 30-40°C, cooling to 40-50°C, keeping the temperature for 10-30 minutes, then heating to 30-40°C, keeping the temperature for 1-2 hours, freeze-drying, and crushing and sieving to obtain the Cordyceps militaris mycelia containing golden flowers.

Description

Technical Field

[0001] The invention relates to the technical field of Cordyceps gunnii fermentation, in particular to a fermentation process of Cordyceps gunnii mycelium containing golden flowers. Background Art

[0002] Gunni Cordyceps and Cordyceps sinensis belong to the same genus Cordyceps. Gunni Cordyceps, also known as Sub-sweet Cordyceps and Hawks Cordyceps, parasitizes the larvae of soil-dwelling insects belonging to the family Bat Moth. Its fruiting bodies emerge from the host's head, with a white stalk and a gray to gray-black apex. They are oblong to columnar, solitary, dichotomously branched, or borne in clusters, measuring 8-22 mm x 5-8 mm. At maturity, they are clearly demarcated from the stalk and lack a sterile apex. The ascocarp is pseudo-ovate or ampoule-shaped, buried, with the orifice exposed at maturity. Gunni Cordyceps is widely distributed in provinces such as Hunan and Guizhou.

[0003] At present, a lot of research has been done on the isolation of Cordyceps militaris strains, determination of asexual forms, nutritional components, and pharmacological experiments. Modern scientific research shows that Cordyceps militaris mycelium is rich in functional ingredients such as cordycepic acid and cordycepin, and their content is basically the same as that of Cordyceps sinensis. Cordyceps militaris mycelium also has many functions that promote human health, such as improving human immunity, resisting fatigue, preventing cardiovascular and cerebrovascular diseases, and anti-tumor. The product has significant health benefits and is an ideal substitute for Cordyceps sinensis, with broad development prospects.

[0004] Cordycepic acid, one of the main active ingredients in the mycelium of Cordyceps guniana, is a weak acid that is temperature-sensitive and easily volatile when heated. Cordycepin, another main active ingredient in this mycelium, is an adenosine analogue and has poor thermal stability. With the continuous improvement of people's living standards, the demand for Cordyceps guniana is increasing. Therefore, improving the fermentation process of Cordyceps guniana mycelium and increasing the content of its active ingredients, cordycepic acid and cordycepin, holds great research potential. Summary of the Invention

[0005] The invention aims to solve the shortcomings in the prior art and proposes a fermentation process for Cordyceps militaris mycelium containing golden flower.

[0006] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0007] S1. Mix fish meal, glucose, potassium dihydrogen phosphate, magnesium sulfate, and water evenly, sterilize, and naturally cool to room temperature. Inoculate Cordyceps militaris under aseptic conditions and culture at a constant temperature of 20-25°C for 4-10 days.

[0008] S2. Mix peanut powder, glucose, linolenic acid, potassium dihydrogen phosphate, magnesium sulfate, and water, sterilize, and naturally cool to room temperature. Aseptically inoculate the bacteria obtained by culture in S1, and culture at a constant temperature of 20-24°C for 1-2 days. Maintain a stirring speed of 140-200 r / min during the culture process.

[0009] S3. Mix the Fucha powder, corn starch, skim milk powder, nutrient solution, and water evenly, sterilize at 130-140°C for 1-2 minutes, cool naturally to room temperature, inoculate the strain obtained by culture in S2 under sterile conditions, and culture at a constant temperature of 20-24°C until the reducing sugar content in the system is less than 0.1%, then harvest the mixture; during the culture process, maintain the ventilation ratio at 1:0.4-0.6 and the stirring speed at 140-200 r / min;

[0010] S4. Add the product obtained in S3 to a vacuum concentrator and concentrate under vacuum at 30-40°C until the mycelium concentration is 1-1.2 g / cm 3 , cool to -40~-50℃, keep warm for 10-30min, then heat to 30-40℃, keep warm for 1-2h, freeze-dry, crush and sieve to obtain Cordyceps militaris mycelium containing golden flower.

[0011] Preferably, in S1, the mass ratio of fish meal, glucose, potassium dihydrogen phosphate and magnesium sulfate is 1-5:1-2:0.01-0.1:0.01-0.1.

[0012] Preferably, in S2, the mass ratio of peanut powder, glucose, linolenic acid, potassium dihydrogen phosphate, and magnesium sulfate is 10-20:1-5:1-5:0.01-0.1:0.01-0.1.

[0013] Preferably, in S2, the inoculation amount of the strain obtained by culture in S1 is 10-20%.

[0014] Preferably, ventilation is maintained during the culture of S2, with a ventilation rate of 1:0.4-0.6.

[0015] Preferably, in S3, the nutrient solution comprises, by mass percentage, 15-25% potato flour, 0.5-1.5% sucrose, 0.1-0.3% sodium nitrate, 0.01-0.03% magnesium sulfate, 0.1-0.3% potassium dihydrogen phosphate, 0.001-0.003% VB1, 0.002-0.006% VB6, 0.002-0.006% niacin, and the balance is water.

[0016] Preferably, in S3, the mass ratio of Fucha powder, corn starch, skimmed milk powder and nutrient solution is 5-10:1-5:1-5:1-2.

[0017] Preferably, in S3, the inoculation amount is 5-10%.

[0018] Preferably, ventilation is maintained during the culture of S3, with a ventilation rate of 1:0.4-0.6.

[0019] Preferably, during the vacuum concentration process of S4, the vacuum degree is 0.04-0.08 MPa.

[0020] Preferably, in S4, after vacuum concentration, the temperature is lowered to -40 to -50°C at a rate of 2-5°C / min.

[0021] Preferably, in S4, the heating rate from -40 to -50°C to 30-40°C is 0.1-1°C / min.

[0022] The technical effects of the present invention are as follows:

[0023] (1) The Pu'er tea powder used in the present invention contains golden flowers, which can secrete amylase and oxidase, effectively promoting the conversion of starch and other substances in the culture medium into monosaccharides and catalyzing the oxidation of polyphenol compounds. In addition, even after high-temperature sterilization at 100-110°C, the golden flowers are not easily inactivated. The strains obtained by S2 culture are inoculated into the system, which can not only provide a large amount of easily absorbed nutrients for the mycelium of Cordyceps militaris, but also induce the production of more mycelium. It has high safety, high bioavailability and high nutritional value.

[0024] (2) The present invention adopts skimmed milk powder as a nitrogen source, which contains a large amount of high-quality protein and is an animal protein. In combination with Fu'er tea powder, Fu'er tea powder is a fully fermented tea, and its internal nutrients are thoroughly fermented. Fu'er tea powder and skimmed milk powder can provide sufficient amino acids for the mycelium of Cordyceps sinensis. The golden flower in Fu'er tea powder can decompose corn starch and skimmed milk powder. The nutrients in the culture medium are extremely diverse and sufficient, providing the best nitrogen source and various mineral elements for the Cordyceps sinensis mycelium at this stage, which can significantly promote the growth of the Cordyceps sinensis mycelium in the fermentation tank and make the mycelium grow faster and fuller.

[0025] (3) Cordyceps acid, one of the main active ingredients of Cordyceps mycelium, is a weak acid that is sensitive to temperature and easily volatile when heated. Cordycepin, one of the main active ingredients of Cordyceps mycelium, is an adenosine analogue and has poor thermal stability. SOD enzyme, one of the main active ingredients of Cordyceps mycelium, is a biological enzyme that easily denatures and loses its biological activity when exposed to high temperatures. The present invention achieves the concentration and drying process of Cordyceps mycelium under lower processing temperature conditions, and compared with the existing technology, better preserves the activity of cordyceps acid and cordycepin in the Cordyceps mycelium.

[0026] (4) Through extensive experiments, the present invention has shown that the addition of Fucha powder significantly increases the content of cordycepic acid and cordycepin in Cordyceps mycelium. Testing has shown that compared with the prior art, the cordycepic acid content of the Cordyceps mycelium of the present invention is increased by 20-40%, the cordycepin content is increased to over 60%, and the SOD enzyme activity is also increased by 20-30%, significantly improving the product's efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 1-2 is a comparison chart of the mycelia obtained in Example 5 and Comparative Examples 1-2 and their quality. DETAILED DESCRIPTION

[0028] The present invention will be further explained below with reference to specific embodiments.

[0029] Example 1

[0030] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0031] S1. Mix 1 kg of fish meal, 1 kg of glucose, 0.01 kg of potassium dihydrogen phosphate, 0.01 kg of magnesium sulfate, and 90 kg of water, sterilize at 110°C for 10 min, cool naturally to room temperature, inoculate Cordyceps militaris under aseptic conditions, and culture at a constant temperature of 20°C for 4 days;

[0032] S2. Mix 10 kg peanut powder, 1 kg glucose, 1 kg linolenic acid, 0.01 kg potassium dihydrogen phosphate, 0.01 kg magnesium sulfate, and 70 kg water, sterilize at 110°C for 10 min, cool naturally to room temperature, and aseptically inoculate the bacteria obtained in S1 at an inoculum size of 10%. Cultivate at a constant temperature of 20°C for 1 day; maintain the ventilation rate at 1:0.4 during the cultivation process, and maintain the stirring speed at 140 r / min.

[0033] S3, 5kg of Fucha powder, 1kg of corn starch, 1kg of skim milk powder, 1kg of nutrient solution, and 70kg of water were mixed evenly, sterilized at 130℃ for 1min, cooled naturally to room temperature, and inoculated with the strain obtained from S2 culture under sterile conditions at an inoculum size of 5%. The culture was cultured at a constant temperature of 20℃ until the reducing sugar content in the system was less than 0.1%, and then the culture was harvested; the ventilation rate was maintained at 1:0.4 during the culture process, and the stirring speed was maintained at 140r / min during the culture process;

[0034] The nutrient solution comprises, by mass percentage, 15% potato starch, 0.5% sucrose, 0.1% sodium nitrate, 0.01% magnesium sulfate, 0.1% potassium dihydrogen phosphate, 0.001% VB1, 0.002% VB6, 0.002% niacin, and the balance being water;

[0035] S4: Add the product obtained in S3 to a vacuum concentration device and concentrate at a temperature of 30°C and a vacuum degree of 0.04 MPa until the mycelium concentration is 1 g / cm 3 , cool to -40℃ at a rate of 2℃ / min, keep warm for 10min, heat to 30℃ at a rate of 0.1℃ / min, keep warm for 1h, freeze-dry, add to a grinder and grind, pass through a 100-mesh sieve to obtain Cordyceps militaris mycelium containing golden flowers.

[0036] Example 2

[0037] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0038] S1. Mix 5 kg fish meal, 2 kg glucose, 0.1 kg potassium dihydrogen phosphate, 0.1 kg magnesium sulfate, and 100 kg water, sterilize at 130°C for 20 min, cool naturally to room temperature, inoculate Cordyceps militaris under aseptic conditions, and culture at a constant temperature of 25°C for 10 days;

[0039] S2. Mix 20 kg peanut powder, 5 kg glucose, 5 kg linolenic acid, 0.1 kg potassium dihydrogen phosphate, 0.1 kg magnesium sulfate, and 80 kg water, sterilize at 130 ° C for 20 min, cool naturally to room temperature, and inoculate the bacteria obtained by culture in S1 under sterile conditions with an inoculum size of 20%. Culture at a constant temperature of 24 ° C for 2 days; maintain the ventilation rate at 1:0.6 during the culture process, and maintain the stirring speed at 200 r / min;

[0040] S3, 10kg Fucha powder, 5kg corn starch, 5kg skim milk powder, 2kg nutrient solution, and 80kg water were mixed evenly, sterilized at 140℃ for 2min, cooled naturally to room temperature, and inoculated with the strain obtained by culture in S2 under sterile conditions at an inoculum size of 10%. The mixture was cultured at a constant temperature of 24℃ until the reducing sugar content in the system was less than 0.1%, and then harvested. During the culture process, the ventilation volume was maintained at 1:0.6, and the stirring speed was maintained at 200r / min.

[0041] The nutrient solution comprises, by mass percentage, 25% potato starch, 1.5% sucrose, 0.3% sodium nitrate, 0.03% magnesium sulfate, 0.3% potassium dihydrogen phosphate, 0.003% VB1, 0.006% VB6, 0.006% niacin, and the balance is water;

[0042] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 40°C and a vacuum degree of 0.08 MPa until the mycelium concentration reaches 1.2 g / cm 3 , cool to -50℃ at a rate of 5℃ / min, keep warm for 30min, heat to 40℃ at a rate of 1℃ / min, keep warm for 2h, freeze-dry, add to a grinder and grind, pass through a 120-mesh sieve to obtain Cordyceps militaris mycelium containing golden flowers.

[0043] Example 3

[0044] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0045] S1. Mix 2 kg of fish meal, 1.7 kg of glucose, 0.02 kg of potassium dihydrogen phosphate, 0.07 kg of magnesium sulfate, and 92 kg of water, sterilize at 128°C for 13 min, cool naturally to room temperature, inoculate Cordyceps militaris under aseptic conditions, and culture at a constant temperature of 24°C for 6 days;

[0046] S2. Mix 18 kg peanut powder, 2 kg glucose, 4 kg linolenic acid, 0.02 kg potassium dihydrogen phosphate, 0.07 kg magnesium sulfate, and 73 kg water, sterilize at 128 ° C for 12 min, cool naturally to room temperature, and inoculate the bacteria obtained in S1 culture under sterile conditions with an inoculum size of 17%. Culture at a constant temperature of 21 ° C for 1.5 days; maintain the ventilation rate at 1:0.55 during the culture process, and maintain the stirring speed at 160 r / min;

[0047] S3, 8kg of Fucha powder, 2kg of corn starch, 4kg of skim milk powder, 1.3kg of nutrient solution, and 77kg of water were mixed evenly, sterilized at 132℃ for 1.5min, cooled naturally to room temperature, and inoculated with the strain obtained from S2 culture under sterile conditions at an inoculum size of 8%. The culture was cultured at a constant temperature of 21℃ until the reducing sugar content in the system was <0.1%, and then the culture was harvested; the ventilation volume was maintained at 1:0.55 during the culture process, and the stirring speed was maintained at 140r / min during the culture process;

[0048] The nutrient solution comprises, by mass percentage, 24% potato starch, 0.8% sucrose, 0.25% sodium nitrate, 0.015% magnesium sulfate, 0.25% potassium dihydrogen phosphate, 0.0015% VB1, 0.005% VB6, 0.003% niacin, and the balance being water;

[0049] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 37°C and a vacuum degree of 0.05 MPa until the mycelium concentration reaches 1.15 g / cm 3 , cooled to -48°C at a rate of 3°C / min, kept warm for 15 minutes, heated to 33°C at a rate of 0.8°C / min, kept warm for 1.7 hours, freeze-dried, added to a grinder and crushed, passed through a 105-mesh sieve, and obtained Cordyceps militaris mycelium containing golden flowers.

[0050] Example 4

[0051] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0052] S1. 4 kg fish meal, 1.3 kg glucose, 0.08 kg potassium dihydrogen phosphate, 0.03 kg magnesium sulfate, and 97 kg water were mixed evenly, sterilized at 120°C for 17 min, cooled naturally to room temperature, inoculated with Cordyceps militaris under aseptic conditions, and cultured at a constant temperature of 22°C for 8 days;

[0053] S2. Mix 12 kg peanut powder, 4 kg glucose, 2 kg linolenic acid, 0.08 kg potassium dihydrogen phosphate, 0.03 kg magnesium sulfate, and 77 kg water, sterilize at 120 ° C for 18 min, cool naturally to room temperature, and inoculate the bacteria obtained by culture in S1 under sterile conditions with an inoculum size of 13%. Culture at a constant temperature of 23 ° C for 1.5 days; maintain the ventilation rate at 1:0.45 during the culture process, and maintain the stirring speed at 180 r / min;

[0054] S3, 6kg of Fucha powder, 4kg of corn starch, 2kg of skim milk powder, 1.7kg of nutrient solution, and 72kg of water were mixed evenly, sterilized at 137℃ for 1.5min, cooled naturally to room temperature, and inoculated with the strain obtained from S2 culture under sterile conditions at an inoculum size of 6%. The culture was cultured at a constant temperature of 23℃ until the reducing sugar content in the system was less than 0.1%, and then the mixture was harvested; the ventilation volume was maintained at 1:0.45 during the culture process, and the stirring speed was maintained at 200r / min during the culture process;

[0055] The nutrient solution comprises, by mass percentage, 16% potato starch, 1.2% sucrose, 0.15% sodium nitrate, 0.025% magnesium sulfate, 0.15% potassium dihydrogen phosphate, 0.0025% VB1, 0.003% VB6, 0.005% niacin, and the balance being water;

[0056] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 33°C and a vacuum degree of 0.07 MPa until the mycelium concentration reaches 1.05 g / cm 3 , cooled to -44°C at a rate of 4°C / min, kept warm for 25 minutes, heated to 37°C at a rate of 0.2°C / min, kept warm for 1.3 hours, freeze-dried, added to a grinder and crushed, passed through a 115-mesh sieve, and obtained Cordyceps militaris mycelium containing golden flowers.

[0057] Example 5

[0058] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0059] S1. Mix 3 kg of fish meal, 1.5 kg of glucose, 0.05 kg of potassium dihydrogen phosphate, 0.05 kg of magnesium sulfate, and 95 kg of water, sterilize at 124 ° C for 15 min, cool naturally to room temperature, inoculate Cordyceps militaris under sterile conditions, and culture at a constant temperature of 23 ° C for 7 days;

[0060] S2. Mix 15 kg peanut powder, 3 kg glucose, 3 kg linolenic acid, 0.05 kg potassium dihydrogen phosphate, 0.05 kg magnesium sulfate, and 75 kg water, sterilize at 124 ° C for 15 min, cool naturally to room temperature, and inoculate the bacteria obtained by culture in S1 under sterile conditions with an inoculum size of 15%. Culture at a constant temperature of 22 ° C for 1.5 days; maintain the ventilation rate at 1:0.5 during the culture process, and maintain the stirring speed at 170 r / min;

[0061] S3, 7kg of Fucha powder, 3kg of corn starch, 3kg of skim milk powder, 1.5kg of nutrient solution, and 75kg of water were mixed evenly, sterilized at 135℃ for 1.5min, cooled naturally to room temperature, and inoculated with the strain obtained from S2 culture under sterile conditions at an inoculum size of 7%. The culture was cultured at a constant temperature of 22℃ until the reducing sugar content in the system was less than 0.1%, and then the culture was harvested; the ventilation rate was maintained at 1:0.5 during the culture process, and the stirring speed was maintained at 170r / min during the culture process;

[0062] The nutrient solution comprises, by mass percentage, 20% potato starch, 1% sucrose, 0.2% sodium nitrate, 0.02% magnesium sulfate, 0.2% potassium dihydrogen phosphate, 0.002% VB1, 0.004% VB6, 0.004% niacin, and the balance is water;

[0063] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 35°C and a vacuum degree of 0.06 MPa until the mycelium concentration reaches 1.1 g / cm 3 , cool to -46°C at a rate of 3.5°C / min, keep warm for 20 minutes, heat to 35°C at a rate of 0.5°C / min, keep warm for 1.5 hours, freeze-dry, add to a grinder and grind, pass through a 110-mesh sieve to obtain Cordyceps militaris mycelium containing golden flowers.

[0064] Comparative Example 1

[0065] A fermentation process of Acanthosporium spp. containing golden flowers comprises the following steps:

[0066] S1, 3kg fish meal, 1.5kg glucose, 0.05kg potassium dihydrogen phosphate, 0.05kg magnesium sulfate, and 95kg water were mixed evenly, sterilized at 124°C for 15min, cooled naturally to room temperature, inoculated with Pseudomonas aeruginosa under sterile conditions, and cultured at a constant temperature of 23°C for 7 days;

[0067] S2, 15kg peanut powder, 3kg glucose, 3kg linolenic acid, 0.05kg potassium dihydrogen phosphate, 0.05kg magnesium sulfate, 75kg water were mixed evenly, sterilized at 124℃ for 15min, cooled naturally to room temperature, inoculated with the powdery spores obtained by culture in S1 under sterile conditions, with an inoculum size of 15%, and cultured at a constant temperature of 22℃ for 1.5 days; during the culture process, the ventilation rate was maintained at 1:0.5, and the stirring speed was maintained at 170r / min;

[0068] S3, 7kg of Fucha powder, 3kg of corn starch, 3kg of skim milk powder, 1.5kg of nutrient solution, and 75kg of water were mixed evenly, sterilized at 135℃ for 1.5min, cooled naturally to room temperature, and inoculated with the powdery spores obtained by culture in S2 under sterile conditions, with an inoculum size of 7%, and cultured at a constant temperature of 22℃ until the reducing sugar content in the system was <0.1%, and then harvested; the ventilation rate was maintained at 1:0.5 during the culture process, and the stirring speed was maintained at 170r / min during the culture process;

[0069] The nutrient solution comprises, by mass percentage, 20% potato starch, 1% sucrose, 0.2% sodium nitrate, 0.02% magnesium sulfate, 0.2% potassium dihydrogen phosphate, 0.002% VB1, 0.004% VB6, 0.004% niacin, and the balance is water;

[0070] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 35°C and a vacuum degree of 0.06 MPa until the mycelium concentration reaches 1.1 g / cm 3 , cool to -46°C at a rate of 3.5°C / min, keep warm for 20 minutes, heat to 35°C at a rate of 0.5°C / min, keep warm for 1.5 hours, freeze-dry, add to a grinder and grind, pass through a 110-mesh sieve to obtain powdery spores containing golden flowers.

[0071] Comparative Example 2

[0072] A fermentation process for Cordyceps militaris mycelium containing golden flowers comprises the following steps:

[0073] S1. Mix 3 kg of fish meal, 1.5 kg of glucose, 0.05 kg of potassium dihydrogen phosphate, 0.05 kg of magnesium sulfate, and 95 kg of water, sterilize at 124 ° C for 15 min, cool naturally to room temperature, inoculate Cordyceps militaris under sterile conditions, and culture at a constant temperature of 23 ° C for 7 days;

[0074] S2. Mix 15 kg peanut powder, 3 kg glucose, 3 kg linolenic acid, 0.05 kg potassium dihydrogen phosphate, 0.05 kg magnesium sulfate, and 75 kg water, sterilize at 124 ° C for 15 min, cool naturally to room temperature, and inoculate the bacteria obtained by culture in S1 under sterile conditions with an inoculum size of 15%. Culture at a constant temperature of 22 ° C for 1.5 days; maintain the ventilation rate at 1:0.5 during the culture process, and maintain the stirring speed at 170 r / min;

[0075] S3. Evenly mix 7 kg of green tea powder, 3 kg of corn starch, 3 kg of skim milk powder, 1.5 kg of nutrient solution, and 75 kg of water. Sterilize at 135°C for 1.5 min, cool naturally to room temperature, and aseptically inoculate the strain obtained in S2 at an inoculum size of 7%. Cultivate at a constant temperature of 22°C until the reducing sugar content in the system is less than 0.1%, then place in a tank for harvesting; maintain an aeration ratio of 1:0.5 during the cultivation process, and maintain a stirring speed of 170 r / min during the cultivation process;

[0076] The nutrient solution comprises, by mass percentage, 20% potato starch, 1% sucrose, 0.2% sodium nitrate, 0.02% magnesium sulfate, 0.2% potassium dihydrogen phosphate, 0.002% VB1, 0.004% VB6, 0.004% niacin, and the balance is water;

[0077] S4: Add the product obtained in S3 to a vacuum concentrator and concentrate at a temperature of 35°C and a vacuum degree of 0.06 MPa until the mycelium concentration reaches 1.1 g / cm 3 , cool to -46°C at a rate of 3.5°C / min, keep warm for 20 minutes, heat to 35°C at a rate of 0.5°C / min, keep warm for 1.5 hours, freeze-dry, add to a grinder and grind, pass through a 110-mesh sieve to obtain Cordyceps militaris mycelium containing golden flowers.

[0078] The mycelium obtained was weighed using the method of Example 5 and Comparative Examples 1-2. Figure 1 shown.

[0079] The obtained mycelium was subjected to high performance liquid chromatography to determine the contents of cordycepic acid and cordycepin, as follows:

[0080] 1. Determination of cordycepic acid content

[0081] 1.1 Plotting the Standard Curve: Accurately weigh 0.64 g of cordycepic acid standard into a 50 mL volumetric flask. Dose 1, 2, 3, 4, 5, and 10 mL of the stock solution, dilute to 50 mL with 20% methanol solution. Filter through a 0.45 μm filter and plot a standard curve based on peak area and cordycepic acid content.

[0082] 1.2 Chromatographic conditions were as follows: the chromatographic column was an Intersil NH2 column (4.6 mm × 250 mm), the mobile phase was acetonitrile-water (72:28, v / v), the flow rate was 1.0 mL / min, the column temperature and the heating temperature in the differential detector were both 35°C, and the injection volume was 20 μL;

[0083] 1.3 Accurately weigh the sample again, add 20% methanol aqueous solution, ultrasonicate at room temperature for 30 minutes, let it stand, take the supernatant and filter it through a 0.22μm filter membrane, and inject it for detection.

[0084] 2. Determination of cordycepin content

[0085] 2.1 Accurately weigh the cordycepin standard and dilute to 100 mL with 50% methanol as a stock solution. Pipette 0.1, 0.5, 1, 2, 3, 4, and 5 mL of the stock solution to 10 mL, respectively. Detect through a 0.22 μm membrane and draw a standard curve based on the peak area and content.

[0086] 2.2 Chromatographic conditions: The chromatographic column was an ODS column (4.6 mm × 250 mm) from Unimicro Technologies, the mobile phase was methanol-water (15%-85%, v / v), the flow rate was 1 mL / min, the column temperature was 35°C, and the injection volume was 10 μL.

[0087] 2.3 Accurately weigh the sample again, add 20% methanol aqueous solution, ultrasonicate at room temperature for 30 minutes, let it stand, take the supernatant and filter it through a 0.22μm filter membrane, and inject it for detection.

[0088] The results are as follows Figure 1 As shown by Figure 1 It can be seen that the method of Example 5 can effectively induce the production of more Cordyceps gunnii mycelia and promote the growth of Cordyceps gunnii mycelia, so that the mycelia content far exceeds that of the comparative example; and the content of cordycepic acid and cordycepin also exceeds that of the comparative example, indicating that the present invention can significantly increase the content of cordycepic acid and cordycepin in Cordyceps sinensis mycelia by adding Fu'er tea powder.

[0089] The mycelia obtained in Example 5 and Comparative Examples 1-2 were tested for SOD enzyme activity, with commercially available Cordyceps sinensis mycelia as a control group. The results are as follows:

[0090] Change rate compared with the control group, % Example 5 27.39 Comparative Example 1 8.35 Comparative Example 2 7.26

[0091] It can be seen from the above table that the present invention not only increases the content of cordycepic acid and cordycepin in the product, but also increases the activity of SOD enzyme, thereby significantly improving the efficacy of the product.

[0092] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A fermentation process for Cordyceps militaris mycelium containing golden flowers, characterized in that: The steps include: S1. Mix fish meal, glucose, potassium dihydrogen phosphate, magnesium sulfate, and water evenly, sterilize, and naturally cool to room temperature. Inoculate Cordyceps militaris under aseptic conditions and culture at a constant temperature of 20-25°C for 4-10 days. S2. Mix peanut powder, glucose, linolenic acid, potassium dihydrogen phosphate, magnesium sulfate, and water, sterilize, and naturally cool to room temperature. Aseptically inoculate the bacteria obtained by culture in S1, and culture at a constant temperature of 20-24°C for 1-2 days. Maintain a stirring speed of 140-200 r / min during the culture process. S3. Mix the Fucha powder, corn starch, skim milk powder, nutrient solution, and water evenly, sterilize at 130-140°C for 1-2 minutes, cool naturally to room temperature, inoculate the strain obtained by culture in S2 under sterile conditions, and culture at a constant temperature of 20-24°C until the reducing sugar content in the system is less than 0.1%, then harvest the mixture; during the culture process, maintain the ventilation ratio at 1:0.4-0.6 and the stirring speed at 140-200 r / min; S4. Add the product obtained in S3 to a vacuum concentrator and concentrate under vacuum at 30-40°C until the mycelium concentration is 1-1.2 g / cm 3 , cool to -40~-50℃, keep warm for 10-30min, then heat to 30-40℃, keep warm for 1-2h, freeze-dry, crush and sieve to obtain Cordyceps militaris mycelium containing golden flower.

2. The fermentation process of the Cordyceps militaris mycelium containing golden flower according to claim 1, characterized in that: In S1, the mass ratio of fish meal, glucose, potassium dihydrogen phosphate, and magnesium sulfate is 1-5:1-2:0.01-0.1:0.01-0.

1.

3. The fermentation process of Cordyceps militaris mycelia containing golden flower according to claim 1, characterized in that: In S2, the mass ratio of peanut powder, glucose, linolenic acid, potassium dihydrogen phosphate, and magnesium sulfate is 10-20:1-5:1-5:0.01-0.1:0.01-0.

1.

4. The fermentation process of Cordyceps militaris mycelia containing golden flower according to claim 1, characterized in that: In S2, the inoculum size of the strain obtained from S1 culture is 10-20%.

5. The fermentation process of the Cordyceps militaris mycelium containing golden flower according to claim 1, characterized in that: During the culture process of S2, ventilation was maintained at a rate of 1:0.4-0.

6.

6. The fermentation process of the Cordyceps militaris mycelium containing golden flower according to claim 1, characterized in that: In S3, the nutrient solution includes, by mass percentage, the following: potato starch 15-25%, sucrose 0.5-1.5%, sodium nitrate 0.1-0.3%, magnesium sulfate 0.01-0.03%, potassium dihydrogen phosphate 0.1-0.3%, VB1 0.001-0.003%, VB6 0.002-0.006%, niacin 0.002-0.006%, and the balance is water.

7. The fermentation process of Cordyceps militaris mycelia containing golden flower according to claim 1, characterized in that: In S3, the mass ratio of Fucha powder, corn starch, skimmed milk powder and nutrient solution is 5-10:1-5:1-5:1-2.

8. The fermentation process of the mycelium of Cordyceps militaris containing golden flower according to claim 1, characterized in that: In S3, the inoculum amount of the bacteria obtained from S2 culture is 5-10%.

9. The fermentation process of Cordyceps militaris mycelia containing golden flower according to claim 1, characterized in that: During the vacuum concentration process of S4, the vacuum degree is 0.04-0.08 MPa.

10. The fermentation process of Cordyceps militaris mycelia containing golden flowers according to claim 1, characterized in that: In S4, after vacuum concentration, the temperature is lowered to -40 to -50°C at a rate of 2-5°C / min; The heating rate from -40 to -50°C to 30-40°C is 0.1-1°C / min.

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