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Method for preparing urechis unicinctus glycosaminoglycan and antibacterial peptide simultaneously

A technology of monocyclic spiny glycosamine and glycosaminoglycan, which is applied in the field of bioengineering to achieve obvious antibacterial activity, high purity and mild reaction conditions

Inactive Publication Date: 2012-08-01
天津高蜚生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the researches on C. monoringensis focus on the morphological structure, embryonic development, life history, artificial breeding technology, nutritional composition analysis and thrombus-dissolving enzymes, etc. There is no report on the development and application of glycosaminoglycans and antimicrobial peptides of C.

Method used

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  • Method for preparing urechis unicinctus glycosaminoglycan and antibacterial peptide simultaneously
  • Method for preparing urechis unicinctus glycosaminoglycan and antibacterial peptide simultaneously
  • Method for preparing urechis unicinctus glycosaminoglycan and antibacterial peptide simultaneously

Examples

Experimental program
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Effect test

Embodiment 1

[0029] A method for simultaneously preparing monocyclic spiny glycosaminoglycans and antibacterial peptides, comprising the following steps:

[0030] (1) Raw material processing: After washing the spiny clam, homogenate to make a homogenate;

[0031] (2) Enzymolysis: Add Bacillus subtilis neutral protease to the homogenate, the amount of Bacillus subtilis neutral protease added is 1% of the homogenate mass, enzymolyze it in a water bath at 45°C for 5 hours, boil the enzymatic solution to kill the enzyme , centrifuged at 6000r / min for 10min to obtain supernatant;

[0032] (3) Ultrafiltration: the supernatant is separated by ultrafiltration through a hollow fiber membrane with a relative molecular mass of 10,000 to obtain liquids of two components with a relative molecular mass of less than 10,000 and a relative molecular mass of more than 10,000;

[0033] (4) Preparation of Glycosaminoglycans

[0034] ①Removal of protein: Concentrate the liquid of components with a relative m...

Embodiment 2

[0041] A method for simultaneously preparing monocyclic spiny glycosaminoglycans and antibacterial peptides, comprising the steps of:

[0042] (1) Raw material processing: After washing the raw material Echinococcus monoringensis, homogenate to make a homogenate;

[0043] (2) Enzymolysis: add Bacillus subtilis neutral protease to the homogenate, the added amount of Bacillus subtilis neutral protease is 0.8% of the homogenate quality, enzymolysis 6h under the condition of 43 ℃ water bath, enzymolysis solution is boiled out After the enzyme, centrifuge at 8000r / min for 10min to obtain the supernatant;

[0044] (3) ultrafiltration: the supernatant is separated by ultrafiltration through a hollow fiber membrane with a relative molecular mass of 10,000 to obtain liquids of two components with a relative molecular mass of less than 10,000 and a relative molecular mass of more than 10,000;

[0045] (4) Preparation of Glycosaminoglycans

[0046]①Removal of protein: Concentrate the l...

Embodiment 3

[0053] A method for simultaneously preparing monocyclic spiny glycosaminoglycans and antibacterial peptides, comprising the steps of:

[0054] (1) Raw material processing: After washing the raw material Echinococcus monoringensis, homogenate to make a homogenate;

[0055] (2) Enzymolysis: Add Bacillus subtilis neutral protease to the homogenate, the amount of Bacillus subtilis neutral protease added is 0.9% of the homogenate mass, enzymolysis is carried out at 45°C for 5 hours in a water bath, and the enzymolysis solution is boiled out After the enzyme, centrifuge at 5000r / min for 15min to obtain the supernatant;

[0056] (3) ultrafiltration: the supernatant is separated by ultrafiltration through a hollow fiber membrane with a relative molecular mass of 10,000 to obtain liquids of two components with a relative molecular mass of less than 10,000 and a relative molecular mass of more than 10,000;

[0057] (4) Preparation of Glycosaminoglycans

[0058] ①Removal of protein: Co...

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Abstract

The invention discloses a method for preparing urechis unicinctus glycosaminoglycan and antibacterial peptide simultaneously. The method includes: raw material process, producing raw material of urechis unicinctus into a homogenate; enzymolysis; ultrafitration; preparation of glycosaminoglycan, eliminating protein, ethanol precipitation and washing, anion exchange column chromatography, gel filtration chromatography to obtain glycosaminoglycan with high purity; and preparation of the antibacterial peptide. According to the method, the reaction condition is mild, the preparation process is environment-friendly and energy-saving, and the added value of products is high; the purity of the prepared glycosaminoglycan and the antibacterial peptide is high; a large amount of protein can be recycled, the comprehensive utilization of the urechis unicinctus is facilitated, the prepared glycosaminoglycan has obvious effects of anticoagulant and free radical scavenging, and the prepared antibacterial peptide has obvious bacteriostatic activity to staphylococcus aureus, Escherichia coli, Aeromonas hydrophila and the like; and the obtained glycosaminoglycan and the urechis unicinctus can provide fine immunopotentiators and novel antibacterial agents for marine rare life aquaculture.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to the preparation of glycosaminoglycans and antibacterial peptides, in particular to a method for simultaneously preparing monocyclic spiny glycosaminoglycans and antibacterial peptides. Background technique [0002] The common names of the single-ringed spinyweed are sea intestines and sea intestines, and they belong to the phylum Ceratozoa, the class Cicada, the order Acanthus, and the family Cicadae. Body thick, about 100-300mm long, about 25-27mm wide, the body surface is covered with granular protrusions of different sizes, snout conical; 1 pair of ventral setae, thick; there are 9-13 brown tail setae around the anus, which is A common species of benthic organisms in the intertidal subtidal zone and subtidal shallow water zone of the northern coastal muddy shore. At present, the researches on C. monocylicus focus on the morphological structure, embryonic development, life history, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/16C08B37/00
Inventor 袁春营崔青曼韩旭刘萍
Owner 天津高蜚生物科技发展有限公司
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