Rapid separation and purification method of cordyceps militaris fruit body water-soluble peptide polysaccharide
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A peptidoglycan, water-soluble technology, applied in the field of extraction of biologically active substances, can solve the problems of cumbersome process, low benefit, pollution, etc., and achieve the effect of process simplification, cost saving and simple process
Inactive Publication Date: 2010-11-03
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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[0004] The purpose of the present invention is to aim at the defects of complex process, high cost, pollution, low benefit and difficulty in large-scale production in the methods for separating and purifying Cordyceps sporocarp polysaccharides at home and abroad; to propose a method with simple process, fast speed, A rapid separation and purification method of water-soluble peptidoglycan from fruiting bodies of Cordyceps militaris with high efficiency, no pollution, high product quality and low cost
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Embodiment 1
[0026] Follow these steps:
[0027] 1) drying the fruiting body of Cordyceps militaris, pulverizing, passing through a 200-mesh sieve, and setting aside;
[0028]2) Ultrapure water extraction: According to the ratio of Cordyceps militaris fruiting body powder: ultrapure water = 1:20, extract in three times, the specific ratio of adding water each time is 1:10, 1:5, 1:5 respectively, and the temperature is controlled at Keep at about 100°C for 4 hours, centrifuge at 5,000 rpm for 20 minutes, and collect the supernatant; combine the supernatants extracted three times, and filter through a 0.22 μm membrane to obtain the filtrate component CF1;
[0029] 3) Molecular sieve separation: Put the filtrate CF1 on the column of Superose 6 as the chromatographic gel, the chromatographic instrument is AKTAPurifier 100 equipped with Shimadzu RID-10A differential detector, the mobile phase is ultrapure water, and the flow rate is 1.0ml / min. Three bands 280, 260, 215λm ultraviolet detection ...
Embodiment 2
[0033] In this example, the fruiting body powder of Cordyceps militaris: ultrapure water is 1:30, add water and extract three times at 1:10, 1:10, and 1:10, and the temperature is controlled at about 90°C; the molecular sieve is Sephadex G-50 column The ion exchange column is outside the DEAE Sepharose CL-6B column, and all the other processes, conditions, steps, and materials are the same as in Example 1.
Embodiment 3
[0035] In this example, the fruiting body powder of Cordyceps militaris: ultrapure water is 1:40, add water and extract three times at 1:15, 1:15, and 1:10, and the temperature is controlled at about 80°C; the molecular sieve is Sephadex G-100 column The ion-exchange column is outside the column of DEAE cellulose D-32, and all the other processes, conditions, steps, and materials are the same as in Example 1.
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Abstract
The invention discloses a rapid separation and purification method of the water soluble peptide polysaccharide from Cordyceps Militaris and belongs to the extraction technology field of the bioactive substances. The method comprises drying and smashing the Cordyceps Militaris, obtaining the component CF1 by extracting the ultrapure water, obtaining the component CF1-1 by the separation from the molecular sieve, obtaining the component CF1-1-1 and CF1-1-2 by separation and purification from the ionic exchange column, and obtaining the CF1-1-1-1, CF1-1-2-1 and CF1-1-2-2 by separation from the affinity chromatography column. By adopting the purification method of smashing the Cordyceps Militaris and direct introduction of the water extract on column, the invention greatly promotes the efficiency and decreases the cost, in addition, the entire separation and purification process does not need the organic solvent, thereby really achieving the purpose of green, rapidness and high efficiencyof the separation of the polysaccharide. The method can be widely applied in the separation and purification technology of the water soluble peptide polysaccharide from the Cordyceps Militaris in thefactory and the laboratory.
Description
technical field [0001] The invention relates to the technical field of extraction of bioactive substances, in particular to a method for rapidly extracting and purifying water-soluble peptidoglycan from fruiting bodies of Cordyceps militaris. Background technique [0002] Cordyceps (Cordyceps) fungus infects host insects to make them zombified, and grows sporozoites from infected zombies under suitable conditions, thus forming fruiting bodies with a specific form of insect (zombie) grass (fungus sporozoite) complex structure. my country's unique Cordyceps sinensis is a species of the genus Cordyceps. At present, there have been detailed reports on the asexual forms of many famous Chinese caterpillar fungus such as Cordyceps militaris, cicadae and Gurney. Pharmacological studies on Cordyceps sinensis and its anamorphs have shown that Cordyceps sinensis and its anamorphs have various pharmacological effects. Cordyceps contains a variety of physiologically active substances, ...
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