A method for separating fungus extracellular polysaccharides by utilizing a three-phase system
An extracellular polysaccharide and three-phase system technology is applied in the field of separation and purification of biologically active substances to achieve the effects of reducing production costs, selective separation, and optimizing separation conditions
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Embodiment 1
[0017] Embodiment one: take Phellinus fermented liquid as research object
[0018] Step 1, ultrasonic pretreatment: take 10mL of fungal fermentation liquid in a 50mL centrifuge tube, place it in an ultrasonic washing tank, and use 15w / cm 3 Ultrasonic treatment for 10 minutes;
[0019] Step 2: Use a three-phase system to separate fungal exopolysaccharides: add 1.0 g of ammonium sulfate to the fermentation broth pretreated by ultrasonic waves in step 1, shake and dissolve, then add 8.0 g of tert-butanol, ammonium sulfate, tert-butanol and fermentation broth The mass ratio is 0.1:0.8:1.0. The mixture was shaken in a water bath at a constant temperature of 35°C for 30 minutes, and centrifuged at 4500 rpm for 10 minutes. The extraction rate of polysaccharide in the lower inorganic salt phase was measured to be 56.84%, and the residual rate of protein was 4.54%.
[0020] Step 3: Take out the lower inorganic salt phase obtained in Step 2, dialyze with distilled water for 48 hours,...
Embodiment 2
[0022] Embodiment two: take Cordyceps fermented liquid as research object
[0023] The test treatment process was the same as in Example 1, except that the mass ratio of ammonium sulfate 3.0 g, tert-butanol 16.0 g ammonium sulfate, tert-butanol and fermentation broth was 0.3:1.6:1.0. The extraction rate of polysaccharide in the lower inorganic salt phase was measured to be 48.19%, and the residual rate of protein was 3.92%.
Embodiment 3
[0024] Embodiment 3: Taking Ganoderma lucidum fermented liquid as research object
[0025] The test treatment process was the same as that in Example 1, except that the mass ratio of ammonium sulfate 2.0 g, tert-butanol 12.0 g ammonium sulfate, tert-butanol and fermentation broth was 0.2:1.2:1.0. The extraction rate of polysaccharide in the lower inorganic salt phase was measured to be 52.94%, and the residual rate of protein was 3.42%.
[0026] As can be seen from the above examples, the system composed of tert-butanol and inorganic salt ammonium sulfate can be used for the selective separation of exopolysaccharides of Phellinus phellinus, Ganoderma lucidum, and Cordyceps sinensis, with green, fast and efficient effects, and the Free protein is mainly enriched in the mesophase.
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