Radial flow chromatography for polysaccharides of mushroom hyphae

A technology for mycelial polysaccharide and chromatographic separation is applied in the field of radial flow chromatographic separation of Lentinus edodes mycelium polysaccharide, which can solve the problem that radial chromatography has not been applied, and achieves improved comprehensive utilization rate and added value, high flow rate, and improved The effect of biological activity

Active Publication Date: 2011-12-21
杭州雪域生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, radial chromatography has not been applied in the separation of polysaccharide products

Method used

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  • Radial flow chromatography for polysaccharides of mushroom hyphae
  • Radial flow chromatography for polysaccharides of mushroom hyphae
  • Radial flow chromatography for polysaccharides of mushroom hyphae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Degreasing: Take dried shiitake mushroom mycelia, crush them, pass through a 50-mesh sieve, put the 50-mesh sieve powder into a 1000ml beaker, add absolute ethanol, stir until the powder is completely submerged, and degrease at room temperature for 24 hours. Remove the yellow-brown supernatant by filtration, add absolute ethanol to the filter residue, repeat the degreasing at room temperature for 24 hours, remove the yellow-brown supernatant by filtration, and obtain the filter residue, which is the defatted mushroom mycelium powder.

[0045] 2. Ultrasonic-assisted water extraction: add distilled water to the degreased mushroom mycelium powder obtained in step 1 to form a feed liquid, add according to the material-liquid ratio of 1g degreased mushroom mycelium powder plus 15mL distilled water, and use 8KHz ultrasonic wave at 50°C Extract for 2.0 h, centrifuge at 3000 rpm for 15 min, evaporate the supernatant in a rotary evaporator at 45°C under reduced pressure, and c...

Embodiment 2

[0055] 1. Degreasing: take dried shiitake mushroom mycelia, crush them, pass through a 120-mesh sieve, put the powder through a 120-mesh sieve into a 1000ml beaker, add absolute ethanol, stir until the powder is completely submerged, and degrease at room temperature for 24 hours. Remove the yellow-brown supernatant by filtration, add absolute ethanol to the filter residue, repeat the degreasing at room temperature for 24 hours, remove the yellow-brown supernatant by filtration, and obtain the filter residue, which is the defatted mushroom mycelium powder.

[0056] 2. Ultrasonic-assisted water extraction: Add distilled water to the degreased shiitake mycelium powder obtained in step 1 to form a feed liquid, add according to the ratio of 1g of degreased shiitake mycelium powder to 5mL distilled water, and use 20KHz ultrasonic wave at 80°C Extract for 0.5h, centrifuge at a speed of 5000rpm for 30min, evaporate the supernatant in a rotary evaporator at 45°C under reduced pressure, ...

Embodiment 3

[0066] 1. Degreasing: Take the dried shiitake mushroom mycelium, crush it, pass it through an 80-mesh sieve, put the powder passed through the 80-mesh sieve into a 1000ml beaker, add absolute ethanol, stir until the powder is completely submerged, and degrease at room temperature for 24 hours. Remove the yellow-brown supernatant by filtration, add absolute ethanol to the filter residue, repeat the degreasing at room temperature for 24 hours, remove the yellow-brown supernatant by filtration, and obtain the filter residue, which is the defatted mushroom mycelium powder.

[0067] 2. Ultrasonic-assisted water extraction: add distilled water to the degreased shiitake mycelium powder obtained in step 1 to form a feed liquid, add according to the ratio of 1g of degreased shiitake mycelium powder to 10mL distilled water, and use 15KHz ultrasonic wave at 65°C Extract for 1.0 h, centrifuge at 4000 rpm for 20 min, evaporate the supernatant in a rotary evaporator at 45°C under reduced pre...

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Abstract

The invention discloses a radial flow chromatography separating method of mushroom mycelia polysaccharides, comprising the following steps of: crushing mushroom mycelia; degreasing and extracting biological active polysaccharide with water at room temperature; removing protein; dialyzing to remove salt; purifying the radial flow chromatography; and freeze drying, wherein eluent used in the purification of the radial flow chromatography is one of a Tris-hydrochloric acid buffer solution, a phosphate buffer solution and a barbital-hydrochloric acid buffer solution; and anion exchange filler is agarose gel with an aglycone group of diethyl aminoethyl. The separating method does not influence a natural structure and the separating effect of the mushroom mycelia polysaccharides; the obtained mushroom mycelia polysaccharides has high content, retains the bioactivity and can be used as a pharmaceutical raw material (such as injection and the like); and the method has short process, low energy consumption and low cost, thereby ensuring the stable and quick separation of polysaccharides and being convenient for indusial production.

Description

technical field [0001] The invention relates to the technical field of chromatographic separation of polysaccharides, in particular to a radial flow chromatographic separation method of mushroom mycelia polysaccharides. Background technique [0002] Lentinula edodes is a Basidiomycetes white mushroom family fungus, is one of the world's famous edible fungi and medicinal fungi. Lentinus edodes is rich in active polysaccharides. Modern scientific research has proved that lentinan is a non-specific immune stimulant, which has many active functions such as inhibiting tumors, anti-virus, anti-oxidation and enhancing immunity, without any toxic side effects. [0003] The polysaccharide is extracted from the shiitake mushroom mycelium obtained by submerged fermentation, and the yield is large, the cycle is short, easy to control, and convenient for industrialized production. At present, the fractional purification of polysaccharides adopts the chromatography method of adsorption a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00B01D15/10A61P31/12A61P35/00A61P37/04A61P39/06
Inventor 贺亮吴学谦程俊文李卫旗吴庆其李海波魏海龙
Owner 杭州雪域生物技术有限公司
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