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HPLC (high performance liquid chromatography) and GPC (gel permeation chromatography) based separation and purification method for marine organism sourced polypeptide

A high-performance liquid chromatography and gel permeation chromatography technology, which is applied in the field of separation and purification of marine biologically active polypeptides, can solve the problems of insufficient wild resources and high fishing intensity, and achieve advanced purification technology, significant ACE inhibitory activity and high accuracy. Effect

Inactive Publication Date: 2015-12-30
MARINE FISHERIES RES INST OF ZHEJIANG
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Problems solved by technology

However, due to excessive fishing intensity in recent years, wild resources have become insufficient

Method used

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  • HPLC (high performance liquid chromatography) and GPC (gel permeation chromatography) based separation and purification method for marine organism sourced polypeptide
  • HPLC (high performance liquid chromatography) and GPC (gel permeation chromatography) based separation and purification method for marine organism sourced polypeptide
  • HPLC (high performance liquid chromatography) and GPC (gel permeation chromatography) based separation and purification method for marine organism sourced polypeptide

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Embodiment

[0025] Example: The biologically active peptide derived from the viscera of C. monocylicus, specifically a short peptide with the following sequence: Gln-Pro-Met-Thr-Phe.

[0026] The preparation steps are as follows: (1) The viscera are cut into pieces and then homogenized with a tissue blender. Stir and degrease overnight with acetone, discard the supernatant by centrifugation, dry the precipitate in a blast drying oven at 40°C overnight, and then pulverize it. Add distilled water to the viscera powder of P. monoringensis at a ratio of 1:20 (mass volume ratio, 50g: 1000mL), heat it in a constant temperature water bath at 80°C for 4 hours, centrifuge to get the supernatant, repeat the above steps three times, and finally obtain the supernatant The supernatant is the experimental sample, concentrated. Add distilled water to the sample at a ratio of 1:5 (mass volume ratio, 10g:50mL), then adjust the pH to 6.8, add 2% (mass fraction) papain, heat in a 60°C water bath for 4 hour...

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Abstract

The invention relates to an HPLC (high performance liquid chromatography) and GPC (gel permeation chromatography) based separation and purification method for marine organism sourced polypeptide. According to the method, urechis unicinctus viscera are selected and crushed, and urechis unicinctus visceral meal is prepared; the urechis unicinctus visceral meal is subjected to enzymatic hydrolysis after concentration; a product of the urechis unicinctus visceral meal subjected to enzymatic hydrolysis is concentrated, frozen and dried and then is subjected to Sephadex G-15 gel chromatography and HPLC separation and purification, and Gln-Pro-Met-Thr-Phe is obtained. The method has the advantages that an extraction and purification technology is more advanced, extraction and purification are fast and high in accuracy, and ACE (angiotensin I-converting enzyme) inhibition activity of the prepared polypeptide is remarkable.

Description

technical field [0001] The invention relates to a method for separating and purifying marine biologically active polypeptides, in particular to a method for separating and purifying marine biologically derived polypeptides by high performance liquid chromatography and gel permeation chromatography. Background technique [0002] Urechisunicinctus (Urechisunicinctus), commonly known as sea intestines and sea intestines, belongs to the phylum Urechisunicinctus, the order Urechisunicinctus, the order Acanthus, and the family Urechisidae. Large body, about 100-300mm long, 25-27mm wide, 200mm-250mm long, covered with granular protrusions of different sizes, conical snout; 1 pair of abdominal setae, thick; 9-13 around the anus Tail setae brown. Distributed in Russia, Japan, North Korea and the coasts of the Yellow and Bohai Seas in my country, it is a common species of benthic organisms in the intertidal subtidal zone and subtidal shallow water zone on the mud and sandy coast of n...

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 李佩佩张小军严忠雍喻亮龙举陈荫
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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