Extraction method of active peptides of scapharca subcrenata

A technology of active peptide and extraction method, which is applied in the field of extraction of clam active peptide

Inactive Publication Date: 2017-05-10
尤世元
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The above patent documents describe in detail the extraction process and pharmacological activity of active substances in cockles, but more

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  • Extraction method of active peptides of scapharca subcrenata

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[0011] The present invention will be further described below in conjunction with the examples.

[0012] Shell the cockles, take the cockle meat, wash with water at 4°C, homogenate, add distilled water at a low temperature (4°C) for extraction for 2 hours at a weight ratio of 1:1, flocculate, centrifuge to remove slag, take the supernatant, and After the supernatant was lyophilized, it was subjected to gel column chromatography with Sephadex G25 (0.03mol / 1NH 4 HCO 3 elution buffer), after collecting the peaks respectively, take the peaks with biological activity and then use anionic Sephadex DEAE-Sephadex A25 ion exchange column chromatography (PH8.5 Tris-HC1 buffer, 0 ~ 0.03mol / NaCl Gradient elution), the active peaks were collected, and after freeze-drying, desalted with Sephadex G15 gel column to obtain natural small molecule active peptides with high purity. The active peptide is a naturally extracted substance without any chemical modification, and the extraction process...

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Abstract

The invention provides an extraction method of active peptides of scapharca subcrenata. The method comprises the steps as follows: scapharca subcrenata is shelled, cleaned, homogenized, extracted at a low temperature with distilled water, flocculated and centrifuged for slag removal; a supernate is taken, lyophilized and subjected to gel column chromatography, peaks are collected, ion exchange column chromatography is performed for separation and purification, active peaks are collected, lyophilized and desalted with a gel column, natural active peptides with higher purity are obtained, obtained small-molecular peptides are purified by a C18 reverse phase column, and the active peptides with purity being 95% or higher are obtained. The molecular weight of the scapharca subcrenata small-molecular peptides obtained with the method is 500-3000 Da, and the biological activity of the small-molecular peptides is effectively protected in the extraction process. The active peptides have significant anti-tumor activity, in-vitro anti-tumor experiments show that the tumor inhibition rate reaches 52%, and the active peptides can be used for the development and application of novel anti-tumor drugs.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of biological extraction, and relates to a method for separating and extracting active peptides from cockles by using column chromatography technology, in particular to a method for extracting active peptides in cockles. [0003] Background technique: [0004] Cockles belong to the phylum Molluscs, Class Phyllobranches, Order Dendontia, and the family Clamidae. It is one of the rich marine organisms in my country and has a long history of medicinal use. Its shell is named corrugated fruit, which has been recorded in past materia medica. It has the functions of reducing phlegm, softening hardness, reducing acid and relieving pain. The cockle meat is a soft part, which is mostly contained in dietotherapy herbal medicines. The clam as a whole is used as medicine, which has the function of shell and meat. It can nourish the blood and warm the middle, strengthen the spleen and stomach, and is suitable for ...

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Application Information

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IPC IPC(8): C07K1/20C07K1/18
Inventor 张玉孙亮黄绪凤
Owner 尤世元
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