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Method for preparing stichopus japonicus selenka glycosaminoglycans

The technology of Apostichopus japonicus glycosaminoglycan and Apostichopus japonicus is applied in the field of preparation of Apostichopus japonicus glycosaminoglycan, which can solve the problems of large change in the number of residues, complex conformation, uneven microstructure and the like, and achieves enhanced release and improved The effect of purity

Inactive Publication Date: 2010-07-28
QINGDAO MUNICIPAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned preparation method cannot obtain high-purity glycosaminoglycans of sea cucumbers. The main reason is that the chemical structures of glycosaminoglycans have significant overlap, such as large molecular weight differences, inconsistent sulfation degrees, complex conformations, and inconsistent microstructures. Uniformity and large changes in the number of residues, etc., especially on a specific proteoglycan molecule, there are differences in the type and number of glycosaminoglycan chains contained in it, which makes it difficult to obtain purity by the existing preparation methods of sea cucumber glycosaminoglycans Apostichopus glycosaminoglycan

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A preparation method of sea cucumber glycosaminoglycans, which comprises the following contents:

[0035] (1) Fresh sea cucumber pretreatment: rinse the sea cucumber with distilled water quickly to remove the foreign matter adhered to the body surface of the sea cucumber, use a long knife to dissect the fresh sea cucumber from the abdomen near the anus forward, remove the internal organs immediately, and then quickly wash them off with double distilled water Dirt, after weighing, place the body wall of the sea cucumber in a clean beaker, and then cut the body wall of the sea cucumber into pieces. The above-mentioned process should be completed quickly and smoothly, and the obvious autolysis of the body wall of the sea cucumber should be avoided as much as possible.

[0036] (2) Alkaline hydrolysis of sea cucumber body wall: add 2 times the double distilled water of sea cucumber body wall weight, then add 2mol / L NaOH, stir while adding, avoid local overheating and alkali...

Embodiment 2

[0049] The preparation method of the sea cucumber glycosaminoglycan of the present embodiment differs from that of Example 1 in the following two parts, and the remaining contents are the same:

[0050] First, during a. trypsin enzymolysis in the double-enzyme hydrolysis process of the sea cucumber body wall in step (3), adjust the pH value to 9.0 with glacial acetic acid, then add the trypsin of 0.45% sea cucumber body wall quality, Water bath at 52°C for 5 hours, add 5mmol / L NaOH solution dropwise under full stirring, and keep the pH at 9.0;

[0051] Second, when ethanol-precipitating glycosaminoglycans of sea cucumber in step (5), 5 mmol / L NaOH was added dropwise to the supernatant to adjust the pH value to 7.2.

Embodiment 3

[0053] The preparation method of the sea cucumber glycosaminoglycan of the present embodiment differs from that of Example 1 in the following two parts, and the remaining contents are the same:

[0054] First, during a. trypsin enzymolysis in the double-enzyme hydrolysis process of the sea cucumber body wall in step (3), adjust the pH value to 8.5 with glacial acetic acid, then add the trypsin of 0.45% sea cucumber body wall quality, Water bath at 52°C for 5 hours, add 5mmol / L NaOH solution dropwise under full stirring, and keep the pH at 8.5;

[0055] Second, when ethanol-precipitating glycosaminoglycans of sea cucumber in step (5), 5 mmol / L NaOH was added dropwise to the supernatant to adjust the pH value to 7.0.

[0056] The preparation methods of sea cucumber glycosaminoglycans implemented in the above-mentioned embodiments can balance the relationship between the yield and purity of sea cucumber glycosaminoglycans, and improve the purity of sea cucumber glycosaminoglycans...

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Abstract

The invention discloses a method for preparing stichopus japonicus selenka glycosaminoglycans, which comprises the following steps: (1) carrying out pretreatment on fresh sea cucumber; (2) carrying out alkaline hydrolysis on body wall of the sea cucumber; (3) carrying out dual-enzyme enzymatic hydrolysis on the body wall of the sea cucumber; (4) using trichloroacetic acid for removing proteins in dual-enzyme enzymatic hydrolysis solution after completing the enzymatic hydrolysis; (5) using ethanol for precipitating the stichopus japonicus selenka glycosaminoglycans; (6) drying for obtaining crude stichopus japonicus selenka glycosaminoglycans; (7) using potassium acetate for precipitating the stichopus japonicus selenka glycosaminoglycans; (8) decoloring the stichopus japonicus selenka glycosaminoglycans; (9) using the potassium acetate for precipitating the stichopus japonicus selenka glycosaminoglycans; and (10) drying for obtaining fine stichopus japonicus selenka glycosaminoglycans. The method can balance the relationship between the yield and the purity of the stichopus japonicus selenka glycosaminoglycans and improve the purity of the stichopus japonicus selenka glycosaminoglycans.

Description

technical field [0001] The invention relates to a preparation method of japonicus glycosaminoglycan. It belongs to the field of biotechnology natural active substance extraction. Background technique [0002] Chronic hepatitis B is a serious health problem facing the world. China, Southeast Asia and Africa are areas with high incidence of HBV infection. At the same time, the incidence of HBV-related liver diseases such as primary liver cancer in these areas is also significantly higher than that in central and southern America. Low incidence area of ​​HBV infection. The treatment of chronic hepatitis B mainly includes anti-virus, immune regulation, anti-inflammatory and liver protection, anti-fibrosis and symptomatic treatment. Among them, effective anti-viral treatment and lasting suppression of HBV DNA below the routine detection range of PCR are the key to treatment. Sulfated polysaccharides have shown different degrees of antiviral, antitumor and immune system regulati...

Claims

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Application Information

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IPC IPC(8): C08B37/00C12S3/02
Inventor 辛永宁宣世英姜相君
Owner QINGDAO MUNICIPAL HOSPITAL
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