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Preparation method of pepsin-solubilized collagen from fish scale

A fish scale and collagen technology, applied in the field of preparation of fish scale enzymatic collagen, can solve the problems of complex process flow, low electrophoresis purity, long separation and purification cycle, etc., and achieves the guarantee of integrity, the preparation process flow is simple, reasonable and easy to operate, and the electrophoresis high purity effect

Inactive Publication Date: 2018-12-07
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of preparation process has problems such as complicated process flow and long separation and purification cycle, it is difficult to ensure the integrity of the collagen triple helix structure, and the electrophoretic purity is not high.

Method used

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  • Preparation method of pepsin-solubilized collagen from fish scale
  • Preparation method of pepsin-solubilized collagen from fish scale

Examples

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Embodiment 1

[0021] A preparation method of fish scale enzyme-soluble collagen, comprising the following steps:

[0022] (1) Fish scale pretreatment: remove the impurities in the fish scales, clean the fish scales, and dry them;

[0023] (2) Degreasing and decalcification of fish scales: put 0.5 kg of fish scales in a glass reactor, add 6L of 0.1M NaOH solution (pH=13), stir at 4°C for 24 hours, and change fresh NaOH solution every 12 hours during the stirring process. Then wash to neutral;

[0024] Add 6L 0.5M EDTA-2Na (pH=7.5) to the degreased fish scales for decalcification, stir at 4°C for 36 hours, wash until neutral, and add liquid nitrogen to the pulverizer after drying to crush the fish scales. The mesh size of the punching screen is 10 mesh (aperture is 2.5mm), obtain fish scale powder;

[0025] (3) Extraction of fish scale collagen: add 1L of 0.5M acetic acid solution to 100g of fish scale powder, then add 2g of pepsin, stir at 4°C for 24 hours for enzyme extraction, and then u...

Embodiment 2

[0029] A preparation method of fish scale enzyme-soluble collagen, comprising the following steps:

[0030] (1) Fish scale pretreatment: remove the impurities in the fish scales, clean the fish scales, and dry them;

[0031] (2) Degreasing and decalcification of fish scales: put 2.5kg of fish scales in a glass reactor, add 25L of 0.1M NaOH solution (pH=13), stir at 4°C for 12 hours, and change fresh NaOH solution every 6 hours during the stirring process. Then wash to neutral;

[0032] Add 25L 0.5M EDTA-2Na (pH=7.5) to the degreased fish scales for decalcification, stir at 4°C for 12 hours, wash until neutral, and add liquid nitrogen to the pulverizer after drying to crush the fish scales. The mesh size of the punching screen is 10 mesh (aperture is 2.5mm), obtain fish scale powder;

[0033] (3) Extraction of fish scale collagen: Add 2L of 0.5M acetic acid solution to 200g of fish scale powder, then add 3g of pepsin, stir at 4°C for 36h for enzyme extraction, and then use a ...

Embodiment 3

[0037] A preparation method of fish scale enzyme-soluble collagen, comprising the following steps:

[0038] (1) Fish scale pretreatment: remove the impurities in the fish scales, clean the fish scales, and dry them;

[0039] (2) Degreasing and decalcifying fish scales: put 5 kg of fish scales in a glass reactor, add 40L 0.1M NaOH solution (pH=13), stir at 4°C for 12 hours, change the fresh NaOH solution every 6 hours during the stirring process, and then Wash to neutral;

[0040] Add 40L 0.5M EDTA-2Na (pH=7.5) to the degreased fish scales for decalcification, stir at 4°C for 24 hours, wash until neutral, and add liquid nitrogen to the pulverizer after drying to crush the fish scales. The mesh size of the punching screen is 10 mesh (aperture is 2.5mm), obtain fish scale powder;

[0041] (3) Extraction of fish scale collagen: add 3L 0.5M acetic acid solution to 300g fish scale powder, then add 3g pepsin, stir at 4°C for 48h for enzyme extraction, and then use a large high-spee...

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Abstract

The invention discloses a preparation method of pepsin-solubilized collagen from fish scale. The method comprises the following steps: performing cleaning; performing degreasing and decalcification; and performing extraction, separation, purification and lyophilization to finally prepare the pepsin-solubilized collagen from the fish scale. The preparation method of the pepsin-solubilized collagenfrom the fish scale has the main advantages that liquid nitrogen is added in a pulverization process, and completeness of a triple helix structure of the collagen is ensured to the maximum extent; a whole preparation process has the advantages that the process is simple and reasonable, and is easy to operate; after the fish scale is preprocessed, low-temperature enzyme extraction, together with the techniques including separation, purification, lyophilization and the like is utilized, the pepsin-solubilized collagen with high triple helix degrees and high cataphoresis purity (more than or equal to 95 percent) is extracted, and moreover, the pepsin-solubilized collagen from the fish scale can be widely applied to the fields of bio-medicine materials, food, cosmetics, drugs and the like.

Description

technical field [0001] The invention relates to the technical field of collagen extraction, in particular to a method for preparing fish scale enzyme-soluble collagen. Background technique [0002] Collagen is the most abundant and widely distributed structural protein in animals. It is widely used in biomedicine due to its biological properties such as good biocompatibility, degradability, low immunogenicity, hemostasis and cell growth promotion. Materials, food, cosmetics and pharmaceuticals and other fields. In recent years, with the outbreak of mad cow disease, bird flu and other diseases, the safety of collagen from terrestrial animal sources has been questioned. Fish scales are rich in collagen, and the collagen extracted from fish scales has the characteristics of good safety and wide application prospects. At present, the preparation process of fish scale collagen is basically rough extraction, separation and purification steps after simple pretreatment of fish sca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/14C07K1/34
CPCC07K14/78
Inventor 肖美添王光宇叶静张娜
Owner HUAQIAO UNIVERSITY
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