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

Inactive Publication Date: 2018-11-06
JIANGSU UNIV
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  • Application Information

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Problems solved by technology

[0004] After searching, there is no report on the selective separation of fungal exopolysaccharides using a system composed of tert-butanol and inorganic salt ammonium sulfate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a method for separating fungus extracellular polysaccharides by utilizing a three-phase system, belongs to the technical field of bioactive substance separation and purification, and particularly relates to selective separation of fungus extracellular polysaccharides by utilizing a system consisting of tert-butanol and an inorganic salt that is ammonium sulfate. The methodmainly includes steps of subjecting a fungus fermentation solution to treatment with ultrasonic waves having a sound energy density of 15 W / cm<3> for 10 min; adding the ammonium sulfate and the tert-butanol; performing vibration in a water bath at a constant temperature of 35 DEG C for 30 min; performing centrifugal separation; subjecting a lower inorganic salt phase to dialysis, concentration andspray drying to obtain fungus extracellular polysaccharide powder; and subjecting an upper organic phase to vacuum rotary evaporation to recover the tert-butanol. The three-phase system is suitable for industrial production of the fungus extracellular polysaccharides, and has rapid, efficient and environmentally friendly effects. The organic phase tert-butanol can be recycled and reutilized so that the method is safe, convenient and low in pollution, the production cost is reduced. The method can be widely applied for extracellular polysaccharide separation of various fungus fermentation solutions.

Description

technical field [0001] The invention relates to a method for separating fungal extracellular polysaccharides using a three-phase system, in particular to a system composed of tert-butanol and inorganic salt ammonium sulfate for the selective separation of fungal extracellular polysaccharides, and belongs to the technical field of separation and purification of biologically active substances. Background technique [0002] At present, the method of obtaining exopolysaccharide from fungal liquid fermentation broth at home and abroad is mainly ethanol precipitation method. However, the alcohol precipitation process requires a large amount of ethanol, which causes a great waste of ethanol, and the crude polysaccharide needs separation processes such as protein removal and decolorization, and the operation process is cumbersome. The commonly used protein removal method-Sevag method requires the use of chloroform, which will react with oxygen in the air when exposed to light, and g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 闫景坤王瑶瑶仇雯漪丁志超武利霞蔡吴丹
Owner JIANGSU UNIV
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