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A method for purification and identification of disulfide bond polypeptides in Astragalus membranaceus

A technology of disulfide bonds and astragalus, which is applied in the field of enrichment and identification of disulfide bond polypeptides, can solve the problems of cumbersome operation and many post-processing processes, and achieves the effect of simple operation and few steps.

Active Publication Date: 2021-11-23
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the separation and purification methods of peptides and proteins still need to rely on special treatment, such as: urea extraction, enzymatic preparation, etc., the operation is relatively cumbersome, and there are many post-processing processes

Method used

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  • A method for purification and identification of disulfide bond polypeptides in Astragalus membranaceus
  • A method for purification and identification of disulfide bond polypeptides in Astragalus membranaceus
  • A method for purification and identification of disulfide bond polypeptides in Astragalus membranaceus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] 1. Enrichment of polypeptide compounds in Radix Astragali

[0059] Crush and dry 0.5kg of Astragalus slices, add 4L of 70% ethanol aqueous solution, ultrasonically extract twice at 40°C for 1 hour each time, and filter once to obtain Astragalus ethanol extract and Astragalus filter residue; Concentrate Astragalus extract to alcohol-free volume, Add deionized water to suspend, add ethyl acetate to the suspension at a volume ratio of 1:1 to obtain an extract containing ethyl acetate and an extract not containing ethyl acetate; take the extract not containing ethyl acetate , add n-butanol according to the volume ratio of 1:1 to obtain an extract containing n-butanol and an extract not containing n-butanol; the extract containing n-butanol is concentrated to dryness, and the obtained dry product Ia; The substance Ia was suspended with deionized water, extracted twice with an organic reagent (n-butanol: dichloromethane = 4:1) according to the volume ratio of 3:1, left to sta...

Embodiment 2

[0063] 1. Enrichment of polypeptide compounds in Radix Astragali

[0064] Crush and dry 0.5kg of Astragalus slices, add 4L of 70% ethanol aqueous solution, ultrasonically extract twice at 40°C for 1 hour each time, and filter once to obtain Astragalus ethanol extract and Astragalus filter residue; Concentrate Astragalus extract to alcohol-free volume, Add deionized water to suspend, add acetone to the suspension at a volume ratio of 1:1 to obtain acetone-containing extracts and acetone-free extracts; take the acetone-free extracts and add them at a volume ratio of 1:1 n-butanol, to obtain the extract containing n-butanol and the extract containing n-butanol; concentrating the extract containing n-butanol to dryness to obtain a dry product Ib; suspending the dry product Ib with deionized water According to the volume ratio of 3:1, the organic reagent (n-butanol: dichloromethane = 4:1) was extracted twice, left to stand, and the middle white precipitate was taken, concentrated t...

Embodiment 3

[0068] 1. Enrichment of polypeptide compounds in Radix Astragali

[0069] Crush and dry 0.5kg of Astragalus slices, add 4L of 70% ethanol aqueous solution, ultrasonically extract twice at 40°C for 1 hour each time, and filter once to obtain Astragalus ethanol extract and Astragalus filter residue; Concentrate Astragalus extract to alcohol-free volume, Add deionized water to suspend, add dichloromethane to the suspension according to the volume ratio of 1:1 to obtain an extract containing dichloromethane and an extract without dichloromethane; take the extract without ethyl acetate , add n-butanol according to the volume ratio of 1:1 to obtain an extract containing n-butanol and an extract not containing n-butanol; the extract containing n-butanol is concentrated to dryness, and the obtained dry product Ic; The substance Ic was suspended with deionized water, extracted twice with an organic reagent (n-butanol: dichloromethane = 5:1) according to the volume ratio of 3:1, left to...

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Abstract

The invention relates to a method for enriching and identifying polypeptides from Astragalus membranaceus. The identified dried Astragalus membranaceus slices are ultrasonically extracted with alcohol, filtered, concentrated, extracted, organic reagents are used to enrich polypeptides in Astragalus membranaceus and passed through alkyl Chemical treatment, mass spectrometry and De Novo analysis identified the disulfide bond polypeptides. The present invention successfully combines the traditional phytochemical analysis method, biochemical method and mass spectrometry method for polypeptide enrichment and detection without desalination, ultrafiltration separation, special chromatographic column packing and other operations, with few steps and easy operation. Peptide research provides technical reference and guidance.

Description

technical field [0001] The invention belongs to the field of separation analysis and biotechnology, in particular to a method for enriching and identifying disulfide bond polypeptides by combining a traditional plant extraction method with biotechnology. Background technique [0002] Astragalus is the dry root of Astragalus membranaceus (Fish.) Bge.var.mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fish.) Bge. First recorded in "Shen Nong's Materia Medica", it has the functions of invigorating qi and promoting yang, strengthening the surface and stopping sweating, diuresis and swelling, promoting body fluid and nourishing blood, promoting stagnation and numbness, supporting toxins and expulsion of pus, astringing sores and promoting muscle growth. Modern medical studies have shown that it has anti-tumor, anti-atherosclerosis, anti-virus, anti-injury, hypoglycemic, and therapeutic metabolic disorders. The most studied main active ingredients are flavonoids, saponins, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/30C07K1/14
CPCC07K1/145C07K1/30
Inventor 张晓哲何枚羲
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI