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Preparation method for separating and purifying water-soluble protein of ginseng

A water-soluble protein, separation and purification technology, applied in the field of separation, purification and preparation of ginseng water-soluble protein, can solve the problems of no antifungal, antiviral and anti-proliferation activities, and achieve a short cycle, quick effect and simple purification process. Effect

Inactive Publication Date: 2007-12-26
CHANGCHUN UNIV OF CHINESE MEDICINE
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Problems solved by technology

Then they isolated a ribonuclease with a molecular weight of 23kDa from ginseng flower buds, but it had no antifungal, antiviral and antiproliferative activities (Protein Expression and Purification.2004, 33:195~199)

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  • Preparation method for separating and purifying water-soluble protein of ginseng

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Embodiment 1

[0020] Embodiment 1: Separation and purification of 5 kinds of ginseng proteins

[0021] 1. Extraction of total ginseng protein: Weigh 100g of fresh ginseng and cut it into small pieces, add 300ml of 0.01mol / L Tris-HCl buffer solution containing 0.15mol / L NaCl and pH=7.4 for homogenization, and add 3000ml of 0.15 0.01mol / L Tris-HCl buffer solution with mol / L NaCl pH=7.4 was extracted at 4°C for 24 hours, centrifuged at 4°C and 6000r / min for 30min to obtain 3100ml of supernatant; Then add 3000ml 0.01mol / L Tris-HCl buffer solution containing 0.15mol / L NaCl and pH=7.4 to the ginseng sediment, extract at 4°C for 24 hours, centrifuge at 4°C and 6000r / min 30min, to obtain 3000ml of supernatant; combine the supernatant obtained by 2 extractions to obtain 6100ml of ginseng total protein crude extract;

[0022] 2. Ammonium sulfate precipitation: Add 3148 grams of solid ammonium sulfate (80% saturation) to the crude extract of 6100ml ginseng total protein obtained in the above steps, l...

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Abstract

This invention relates to a method for preparing ginseng proteins by buffer extraction, ammonium sulfate precipitation and affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration chromatography. The method comprises: extracting ginseng juice, adding solid ammonium sulfate to 80% saturation, performing equilibrium dialysis with Tris-HCl buffer, centrifuging at a high speed, performing ultrafiltration, and purifying by different kinds of chromatography to obtain 5 kinds of inseng proteins GP1-GP5. GP1 is RNA-like enzyme. GP3 is chitinase-like protein. GP4 is ginsengribonuclease. GP2 and GP5 are reported for the first time. The method has such advantages as good purification effect, large loading volume and high sample yield, and can be scaled up for mass production. The samples, except for GP4, show a single band in SDS-PAGE polyacrylamide gel electrophoresis, which confirms that the samples reach electrophoretic pure.

Description

technical field [0001] The invention belongs to the technical field of protein separation and purification, and in particular relates to a preparation method for separation and purification of ginseng water-soluble protein by combining buffer extraction, ammonium sulfate precipitation and various column chromatography techniques. Background technique [0002] The research on ginseng protein and peptides began in the 1960s, mainly focusing on the extraction method of ginseng peptides. In 1963, F.Gstirnor et al. of Germany detected a variety of amino acids from ginseng root for the first time: glutamic acid, cysteine, tyrosine, etc. In 1966, they (Arch Pharm.1966, 299(11): 934-937) used electrophoresis to obtain 5 small peptides from the methanol water extract of Korean white ginseng, but the primary structure was not determined. In 1980, Japanese Ando and Okada (Planta Medica. 1980, 38: 18-21) found that there is a substance in ginseng water extract that strongly inhibits ad...

Claims

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Application Information

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IPC IPC(8): C07K1/36C07K4/10C07K14/415C07K1/34C07K1/30
Inventor 赵雨张巍李红艳惠歌李银清姜先刚唐任能幺宝金
Owner CHANGCHUN UNIV OF CHINESE MEDICINE
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