Method for producing feed additives rich in heat-resistant protease and cordycepin by solid-state fermentation of Cordyceps militaris
A feed additive and solid-state fermentation technology, which is applied in the field of solid-state fermentation of Cordyceps militaris to produce feed additives rich in heat-resistant protease and cordycepin, achieving the effects of no three wastes, stable process, and less land occupation
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Embodiment 1
[0030] Embodiment 1, the screening of Cordyceps militaris bacterial strain
[0031] Isolation and screening of the above-mentioned Cordyceps militaris (Cordyceps militaris) bacterial classification:
[0032] 1. Isolation and screening of bacterial strains
[0033] In August 2015, wild Cordyceps militaris was collected in Yimeng Mountain, Yinan County, Shandong Province. The collection site was in a forest with loose soil and rich humus. The ambient temperature was 20-28°C and the air humidity was 70-80%. A total of 9 wild Cordyceps strains were collected, and the Cordyceps was slowly peeled off from the layer, and the surface of the Cordyceps was rinsed repeatedly with clean water until it was cleaned. Blot the surface moisture of Cordyceps dry with sterile filter paper. Then disinfect the surface with 75% ethanol or 0.1 L mercury, and rinse with sterile water again. Then, carefully cut out a small piece from the Cordyceps militaris subunit, put it on the activated solid me...
Embodiment 2
[0057] Embodiment 2: compound mutagenesis and breeding method of Cordyceps militaris bacterial strain
[0058] (1) Activation culture of strains The wild strain XZCM6 isolated in Example 1 was transferred to PDA solid slant medium for activation culture, and cultured at a constant temperature of 25-28°C for 4-5 days. The strains were activated twice as above.
[0059] (2) Preparation of strain mycelium Take 1 cm from the activated mycelium solid plate medium 2 The mycelium block was inoculated into a 250mL Erlenmeyer flask filled with 40mL PDA mycelium medium, and cultured with constant temperature shaking at 24-28°C and 160-200 r / min for 3-7d. Mycelia liquid medium formula: glucose 20-30g, peptone 5-8g, 20% potato extract 200ml, (NH 4 ) 2 S0 4 3~5g, KH 2 P0 4 1~2g, MgS0 4 ·7H 2 0 1~2g, yeast powder 4~6g, add water to make up to 1L, pH 6.5~7.0, sterilize at 121℃ for 20min.
[0060] (4) Protoplast preparation MgS0 4 ·7H 20. Mannitol was dissolved in double-distille...
Embodiment 3
[0064] Embodiment 3 Preparation of solid-state fermentation strain
[0065] This embodiment 3 illustrates the method for preparing a heat-resistant protease-rich and cordycepin feed additive by using Cordyceps militaris HYCM12 as a strain solid-state fermentation method, including the following steps:
[0066] Step 1, strain activation: first, inoculate the original seed of the preserved Cordyceps militaris HYCM12 test tube strain on the activation medium of a petri dish, and incubate in an incubator at 20-26° C. in the dark for 5-7 days to obtain first-grade seeds.
[0067] Activated slant medium: 200 g of potatoes, washed and chopped, add 1000 mL of water and boil until boiling for 20 min, filter with gauze, add 20 g of glucose to the filtrate, 1 mL of inorganic salt solution, 15 g of agar and continue to boil, fully dissolve and then pack into test tubes , each test tube is about 4 mL, and the culture medium is sterilized at 121°C for 20 min. After the sterilization is comp...
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Abstract
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