Method for extracting atractylodes macrocephala polysaccharide by utilizing microbial fermentation method
A microbial fermentation method and atractylodes polysaccharide technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high enzyme cost, long time consumption, low polysaccharide extraction rate, etc., to achieve easy dissolution and promote solution The effect of improving the separation and extraction yield
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Embodiment 1
[0024] Screening and identification of embodiment 1 fermentation strain
[0025] 1. Screening of Rhizopus oryzae CH5 strain
[0026] (1) Add 20g of Zizhi powder crushed through a 40-mesh sieve into a 250-mL Erlenmeyer flask, then add 30mL of sterile water to moisten it, and incubate at 28°C for 4 days. Dilute the enriched culture covered with mold with sterile water 1 × 10 6 After doubling, it was spread on the PDA plate medium, and cultured at 28°C for 3 days, and fungal colonies with different colors and shapes were picked and transferred to fresh PDA medium, and cultured at 28°C for 3 days to obtain 7 purely cultured strains (No. CH1-CH7 respectively), inoculated with fresh PDA medium, and cultured at 28°C for 3 days. Add 5 mL of sterile water to each of the fresh plate cultures of the 7 strains, and stir with an inoculating loop to suspend the spores. The spore liquid was transferred to a sterile test tube, and the spore concentration was adjusted to 2×10 with sterile w...
Embodiment 2
[0043] Embodiment 2 Rhizopus oryzae CH5 is applied to the extraction of Atractylodes macrocephala polysaccharide
[0044] (1) Preparation of Rhizopus oryzae CH5 spore liquid: Plate colonies of Rhizopus oryzae CH5 preserved at 4°C were inoculated on fresh PDA plate medium, cultured at a constant temperature of 28°C for 3 days, added 5 mL of sterile water to the plate culture, and used Agitate the inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 3×10 with sterile water. 8 individual / mL. Described PDA plate medium composition and preparation method are the same as embodiment 2.
[0045] (2) 500-mL triangular flask was sterilized by dry heat at 160°C for 2 hours, added 20 g of Atractylodes macrocephala powder crushed through a 60-mesh sieve, added 50 mL of sterile water, and then inoculated with 2 mL of CH5 spore liquid prepared in step (1), and stirred uniform. The mouth of the triangular flask was ...
Embodiment 3
[0050] Embodiment 3 Rhizopus oryzae CH5 is applied to the extraction of Atractylodes macrocephala polysaccharide:
[0051] (1) Preparation of Rhizopus oryzae CH5 spore liquid: Plate colonies of Rhizopus oryzae CH5 preserved at 4°C were inoculated on fresh PDA plate medium, cultured at a constant temperature of 28°C for 3 days, added 5 mL of sterile water to the plate culture, and used Agitate the inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 4×10 with sterile water. 8 individual / mL. Described PDA plate medium composition and preparation method are the same as embodiment 1.
[0052] (2) 2-L triangular flask was sterilized by dry heat at 160°C for 2 hours, added 50 g of Atractylodes macrocephala powder crushed through a 60-mesh sieve, added 150 mL of sterile water, and then inoculated with 5 mL of Rhizopus oryzae CH5 spore liquid prepared in step (1) , stir well. The mouth of the triangular flas...
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