A method for separating polysaccharides and proteins using choline ionic liquid two-phase system
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A two-phase aqueous system and choline ion technology, applied in the field of separation of polysaccharides and proteins, to achieve the effects of reducing production costs, optimizing separation conditions, and selective separation
Active Publication Date: 2017-02-01
东台海滨科技创业园管理有限公司
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[0004] After retrieval, there is no use of choline chloride [Ch]Cl and inorganic phosphate K 3 PO 4 Report on the Selective Separation of Polysaccharides and Proteins Using Constructed Ionic Liquid Two-Phase System
Method used
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Examples
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Embodiment 1
[0017] Example 1: Taking Phellinus igniarius mycelium as the research object
[0018] Step 1. Degreasing: pulverize the mycelium of Phellinus igniarius, wrap it in absorbent cotton cloth, put it in a 500 mL Soxhlet extractor, add 200~300 mL petroleum ether (boiling range: 30~60℃), reflux at 35℃ for 6 hours, The defatted treatment was carried out twice, and dried naturally to obtain defatted Phellinus igniarius mycelium.
[0019] Step two, prepare the water extract of Phellinus igniarius: Weigh 100 g of Phellinus igniarius mycelium after defatting in step 1, add 1000 mL of distilled water at a material-to-liquid ratio of 1:10g / mL, place it in a 2000 mL round bottom flask, and stir thoroughly After homogenization, reflux and extract at 95°C for 8 h. The mixture was centrifuged in a 300 mL plastic centrifuge tube at 5000 rpm for 10 min. The supernatant was combined with a 45°C rotary evaporator and evaporated under reduced pressure, concentrated to 1.5 times the original volume, dial...
Embodiment 2
[0024] Example 2: Taking Shiitake Mushroom Fruit Body as Research Object
[0025] The experimental treatment process is the same as in Example 1, the difference is that the ionic liquid [Ch]Cl 2.0 g, K 3 PO 4 4.0 g and 4.0 mL of distilled water, the mass ratio of the three was 0.5:1.0:1.0. The extraction rate of protein in the upper ionic liquid phase was 86.3%, and the extraction rate of lentinan in the lower inorganic salt phase was 55.5%.
Embodiment 3
[0026] Example 3: Taking Cordyceps mycelium as the research object
[0027] The experimental treatment process is the same as in Example 1, the difference is that the ionic liquid [Ch]Cl 1.2 g, K 3 PO 4 3.2 g and 4.0 mL of distilled water, the mass ratio of the three was 0.3:0.8:1.0. The extraction rate of protein in the upper ionic liquid phase was 85.7%, and the extraction rate of Cordyceps polysaccharide in the lower inorganic salt phase was 71.9%.
[0028] It can be seen from the above examples that [Ch]Cl / K 3 PO 4 The composed ionic liquid water two-phase system can be used for the selective separation of polysaccharides and proteins of Phellinus igniarius, Shiitake, Cordyceps sinensis, and has green, fast and efficient effects.
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Abstract
The invention relates to a method for separating polysaccharide and protein by using a choline ionic liquid two-phase aqueous system, belongs to the technical field of bioactive substance separation and purification, and particularly relates to application of the ionic liquid two-phase aqueous system composed of choline chloride [Ch]Cl and inorganic phosphate K3PO4 in selective separation of fungal polysaccharide and protein. The method is mainly characterized by comprising the following steps: after degreasing crushed fungi mycelium or sporocarp with petroleum ether, using hot water with the temperature of 95 DEG C for reflux extraction for 8 hours, centrifuging, concentrating, dialyzing, and freeze-drying, so as to obtain a fungus aqueous extract; carrying out extraction separation by using the ionic liquid two-phase aqueous system composed of choline chloride[Ch]C1 and inorganic phosphate K3P04; carrying out lower-layer inorganic salt phase dialysis, ethanol precipitation and freeze-drying so as to obtain fungal polysaccharide; carrying out extraction on an upper-layer ionic liquid phase by using dichloromethane, concentrating and recycling an ionic liquid. The ionic liquid two-phase aqueous system provided by the invention is suitable for industrial production of active polysaccharides, has the effects of being green, quick and efficient, the ionic liquid can be recycled for use, the production cost is lowered, and the ionic liquid two-phase aqueous system can be commonly used for the selective separation of various fungal polysaccharide and protein.
Description
Technical field [0001] The invention relates to a method for separating polysaccharides and proteins by using a two-phase aqueous two-phase system of choline ionic liquid, particularly ionic liquid choline chloride [Ch]Cl and inorganic phosphate K 3 PO 4 The composed ionic liquid aqueous two-phase system is used for the selective separation of fungal polysaccharides and proteins, and belongs to the technical field of separation and purification of biologically active substances. Background technique [0002] There are many methods for separation and purification of fungal polysaccharides, such as ethanol fractional precipitation, ion exchange chromatography, gel filtration chromatography, and membrane filtration. Researchers must choose the correct separation and purification method according to the properties of polysaccharides to obtain uniform active ingredients. Among them, ethanol fractionation precipitation is a simple and effective method, but the protein is also precipitat...
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