Grifola frondosa strain, culture method and application thereof
A cultivation method, the technology of Grifola frondosa, applied in the field of microorganisms, can solve the problems of difficulty in obtaining strains, unsolved problems of extraction and purification of Grifola frondosa polysaccharides, and high production costs
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Embodiment 1
[0037] Embodiment 1 bacterial cell culture
[0038]The subculture method adopts slant subculture, and the slant medium adopts potato agar enriched medium, and the ingredients are as follows: 200g potato; 20g glucose; 1g potassium dihydrogen phosphate; 0.5g magnesium sulfate; 20g agar; 1000mL water; 1 Appropriate amount, natural pH.
[0039] The slant culture medium is prepared according to the conventional method; the inoculation culture, according to the requirements of aseptic operation, is inoculated on the ultra-clean bench in the aseptic room, and inoculated by the double-slope inoculation method. After inoculation, culture at 27±1°C for 1 day, you can see creeping hyphae and basal hyphae grow from around the grafted tissue block, and then the creeping hyphae elongate into aerial hyphae, and there are also directly elongate into aerial hyphae. After 4 to 7 days, the young mycelium covered the slope, and the mycelium was white and fluffy, thick and developed, and felt li...
Embodiment 2
[0040] Embodiment 2 Shake flask culture
[0041] Medium for liquid culture: 1% potato, 1% bran, 2% glucose, 0.7% corn flour, 0.3% peptone, 0.14% yeast powder, 0.05% potassium dihydrogen phosphate, and 0.1% magnesium sulfate.
[0042] Prepare the culture medium and put it into the Erlenmeyer flask with a filling coefficient of 0.2-0.4, seal it with a cotton plug, and sterilize it. Grifola frondosa strains are inoculated according to 5-10% (V / V) of the loading amount, and cultured on a shaking table. The temperature is 25-30°C, and the rotation speed is 90-150r / min. After 2 days of culture, a large number of new mycelium balls can be seen, and after 3 to 5 days, a large number of mycelia are formed in the liquid medium, and the mycelium grows in suspension in the liquid medium, and the forms include branched mycelium, flocculated mycelium, and granular fungus Silk.
[0043] Cultivate for 3-5 days according to the above-mentioned method, and the yield of mycelium of Grifola fr...
Embodiment 3
[0044] Embodiment 3 fermentation culture
[0045] For three-stage fermentation, the scale-up ratio of the fermenter is 1:10. The filling factor of the seed tank and fermenter is 50-80%, and the liquid seed inoculation amount is 5-10% (V / V).
[0046] Culture conditions: culture medium C / N is 20-30:1, pH value is 4-7, temperature is 24-29°C, tank pressure is 0.5-1.5Mp, stirring speed is 100-600r / min, dissolved oxygen content is 10-100% .
[0047] The fermenter is gradually enlarged. In the synthetic or semi-synthetic medium containing sufficient nitrogen and carbon sources, the mycelia of Grifola frondosa can grow and reproduce in large quantities under aerobic conditions, so that the amount of mycelia can be continuously accumulated. The growth cycle of the seed tank is 24-48 hours, and the growth cycle of the fermenter is 36-72 hours. The biomass can be increased by hundreds of times within a few days by means of continuous amplification, and a growth cycle can be completed...
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