Strain of rhizopus stolonifer FL-3 and application thereof in extracting pachymaran
A technology of Rhizopus stolonifera and FL-3 is applied in the application field of microbial fermentation technology in the extraction of tuckahoe polysaccharide, which can solve the problems such as large difference in polysaccharide yield, and achieve the effects of improving extraction yield and promoting dissociation.
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Embodiment 1
[0025] 1. Screening of strain FL-3
[0026] (1) Add about 10 g of Poria cocos powder that has been crushed through a 60-mesh sieve into a 250-mL Erlenmeyer flask, then add 20 mL of sterile water to moisten it, and incubate at a constant temperature of 28°C for 4 days. Dilute the enriched culture covered with mold with sterile water 1 × 10 6 After doubling, spread it on the PDA plate medium, culture at 28°C for 2 days, pick out mold colonies with different colors and shapes, transfer them to fresh PDA medium, place them in 28°C for 3 days, and obtain 6 purely cultured strains, respectively Numbered FL-1~FL-6. Add 5 mL of sterile water to the fresh plate cultures of the 6 strains, stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spores with sterile water. The concentration is 1×10 8 individual / mL.
[0027] (2) 250-mL triangular flask is sterilized by dry heat at 160°C for 2 hours, add 10 g of Poria cocos pow...
Embodiment 2
[0040] Rhizopus stoloniferus FL-3 is applied to the extraction of polysaccharides, and can be operated according to the following steps:
[0041] (1) Preparation of spore liquid of Rhizopus stoloniferus FL-3: cryopreserved Rhizopus stoloniferus FL-3 (freeze-dried tube or plate colony) was inoculated on fresh PDA plate medium, cultured at 28°C for 3 days, and added 5 mL Put sterile water in a petri dish, stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 1×10 with sterile water. 8 individual / mL. Described PDA plate medium composition and preparation method are the same as embodiment 1.
[0042] (2) 500-mL Erlenmeyer flask was sterilized by dry heat at 160°C for 2 hours, added 20 g of Poria cocos powder crushed through a 60-mesh sieve, added 60 mL of sterile water, and then inoculated with 2 mL of Rhizopus stoloniferus FL- 3 spore liquid, stir well. The mouth of the triangular flask was ...
Embodiment 3
[0047] Rhizopus stoloniferus FL-3 is applied to the extraction of polysaccharides, and can be operated according to the following steps:
[0048] (1) Preparation of spore liquid of Rhizopus stoloniferus FL-3: cryopreserved Rhizopus stoloniferus FL-3 (freeze-dried tube or plate colony) was inoculated on fresh PDA plate medium, cultured at 28°C for 3 days, and added 5 mL Put sterile water in a petri dish, stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 2×10 with sterile water 8 individual / mL. Described PDA plate medium composition and preparation method are the same as embodiment 1.
[0049] (2) 2-L Erlenmeyer flask was sterilized by dry heat at 160°C for 2 hours, added 50 g of Poria cocos powder crushed through a 60-mesh sieve, added 150 mL of sterile water, and then inoculated with 5 mL of the spore liquid prepared in step (1) of FL-3 , stir well. The mouth of the triangular flask w...
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