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Method for separating polysaccharide and protein by using choline ionic liquid two-phase aqueous system

A two-phase aqueous system, choline ion technology, applied in the field of separation of polysaccharides and proteins, to achieve the effect of reducing production costs, selective separation, and optimization of separation conditions

Active Publication Date: 2015-08-26
东台海滨科技创业园管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[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

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment one: take Phellinus mycelium as research object

[0018] Step 1, degreasing: Phellinus mycelium was crushed, wrapped in absorbent cotton cloth, placed in a 500 mL Soxhlet extractor, added 200-300 mL petroleum ether (boiling range: 30-60°C), refluxed at 35°C for 6 h, Such degreasing treatment was performed twice, and then dried naturally to obtain degreasing Phellinus mycelium.

[0019] Step 2, prepare the Phellinus Phellinus aqueous extract: Weigh 100 g of Phellinus Phellinus mycelia after degreasing in Step 1, add 1000 mL of distilled water at a solid-liquid ratio of 1:10g / mL, place in a 2000 mL round bottom flask, and stir thoroughly After uniformity, reflux extraction 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 and evaporated in a 45°C rotary evaporator under reduced pressure, concentrated to 1.5 times the original volume, dialyzed in distilled water for 48 h, and va...

Embodiment 2

[0024] Embodiment two: take mushroom fruiting body as research object

[0025] The test treatment process is the same as in Example 1, except 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] Embodiment three: take Cordyceps sinensis mycelium as research object

[0027] The test treatment process is the same as in Example 1, except 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 polysaccharide from Cordyceps sinensis in the lower inorganic salt phase was 71.9%.

[0028] As can be seen from the above examples, [Ch]Cl / K 3 PO 4 The composed ionic liquid two-phase aqueous system can be used for the selective separation of polysaccharides and proteins of Phellinus lentinus, shiitake mushrooms, and 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 using a choline ionic liquid two-phase system, in particular to ionic liquid choline chloride [Ch]Cl and inorganic phosphate K 3 PO 4 The composed ionic liquid two-phase aqueous 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 separation and purification methods for 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 in order to obtain uniform active ingredients. Among them, ethanol fractional precipitation is a simple and effective method, but the protein is also precipitated when th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00C07K1/14
Inventor 闫景坤马海乐王振斌裴娟娟姜宇
Owner 东台海滨科技创业园管理有限公司
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