Phellinus linteus mycelia active glucoprotein and use thereof and preparation
A technology of Phellinus mycelium and glycoprotein, which is applied in the direction of fungi, antibacterial drugs, and medical raw materials derived from fungi, and can solve the problems of separation and purification of antibacterial active glycoproteins, identification of physical and chemical properties, and related biological functions. Report and other issues, to achieve the effect of low production cost, high yield of glycoprotein, and broad application prospects
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Embodiment 1
[0044] Phellinus AS3.637 was purchased from China Microorganism Culture Collection Center. The strains were first cultured on slant media. The test tubes and tampons used for culture need to be sterilized at 121°C for 30 minutes, then put into the culture medium and sterilized at 121°C for 30 minutes, and placed on an inclined plane to cool to solidify into a slope. Then the ultra-clean bench was inoculated. The slant culture was carried out at a constant temperature of about 25°C for a period of 7 days. Cut the slant bacteria into small pieces and inoculate them into the seed culture medium. After the medium is divided, put a cotton plug on the Erlenmeyer flask to prevent external microorganisms from entering the medium and cause pollution, and ensure good ventilation performance. . The composition of the seed culture solution is: glucose 20g, yeast extract 5g, corn steep liquor 10g, bran 50g, MgSO 4·7H 2 O 0.5g, KH 2 PO 4 1g, the rest is water, and the pH is natural;...
Embodiment 2
[0046] 100g of Phellinus mycelium was added to 1 times the volume of water, and the homogenate was broken, extracted at 4°C, centrifuged, discarded insoluble matter, and the extract was collected. After the extract was concentrated, ammonium sulfate was added to a concentration of 80% (m / V). Stand still for 24 hours, high-speed centrifuge to take the precipitate, add a small amount of water to redissolve the above precipitate, put the precipitate reconstituted solution into an 8000 Dalton dialysis bag, dialyze with deionized water, collect the dialysate, concentrate, and load the sample on DEAE-Sepharose Fast Flow The chromatographic column uses distilled water and NaCl as eluents for stepwise elution, collects the glycoprotein complex components eluted by the NaCl solution, and freezes and dries each collected solution.
[0047] Take each tube and measure the UV absorption at 280nm, and get 4 peaks. After the antibacterial activity test, B and C are the active ingredients, wh...
Embodiment 3
[0049] After adding 3 times the volume of water to 100g Phellinus mycelium, the homogenate was broken, leached at 15°C, centrifuged, discarded the insoluble matter, collected the extract, concentrated the extract and added ammonium sulfate to a concentration of 85% (m / V), Stand still for 36 hours, high-speed centrifuge to take the precipitate, add a small amount of water to redissolve the precipitate, put the precipitate reconstituted solution into an 8000 Dalton dialysis bag, dialyze with deionized water, collect the dialysate, concentrate, and load the sample on the DEAE-Sepharose Fast Flow layer The column was analyzed, and distilled water and NaCl were used as eluents for stepwise elution, and the glycoprotein complex components eluted by the NaCl solution were collected, and each collected solution was freeze-dried.
[0050] Take each tube and measure the UV absorption at 280nm, and get 4 peaks. After the antibacterial activity test, B and C are the active ingredients, wh...
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