Extraction method and application of fish skin collagen

A fish skin collagen and extraction method technology, which is applied in the field of fish skin collagen extraction, can solve the problems of long extraction period, failure to ensure the integrity of collagen structure, and low extraction rate, and achieve short extraction time, simple and feasible technical methods, The effect of high extraction efficiency

Pending Publication Date: 2021-05-07
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional extraction method has problems such as long extraction cycle, low extraction rate, and inability to ensure the integrity of the collagen structure. How to improve the extraction rate and shorten the extraction time while ensuring the integrity of the collagen structure requires exploring new extraction methods.

Method used

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  • Extraction method and application of fish skin collagen
  • Extraction method and application of fish skin collagen
  • Extraction method and application of fish skin collagen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of fish skin collagen with ultrasonic treatment power of 150W, 250W, 350W, 450W, 550W for 20min

[0043] Silver carp skin raw materials such as figure 1 , the fish skin is obtained by an automatic skin peeling machine, and the remaining fish meat and black impurities are manually removed from the fish skin. Cut the fish skin into 5mm×5mm, wash it, drain it with coarse emery cloth, and weigh 163.93g.

[0044] Defat, protein removal:

[0045] (1) The volume of 0.01M NaOH is 3L, stir at 4°C for 24 hours, change the NaOH every 8 hours, rinse repeatedly with distilled water until neutral, and fully drain

[0046] (2) The volume of 10% isopropanol is 3L, stirred at 4°C for 8h, washed repeatedly with distilled water until neutral, fully drained, and weighed 365.69g.

[0047] Sonication:

[0048] Weigh 60.16g, 60.13g, 60.28, 60.35, 60.80g of degreased and protein-removed fish skin respectively, place them in 1L of 0.5M acetic acid solution and perform ...

Embodiment 2

[0056] Example 2 Preparation of fish skin collagen with ultrasonic treatment power of 150W, 250W, 350W, 450W, 550W for 30min

[0057] The fish skin is obtained by an automatic skin peeling machine, and the remaining fish meat and black impurities are manually removed from the fish skin. Cut the fish skin into 5mm×5mm, wash it, drain it with coarse emery cloth, and weigh 121.68g.

[0058] Defat, protein removal:

[0059] (1) The volume of 0.01M NaOH is 3L, stirred at 4°C for 24 hours, changing the NaOH every 8 hours, repeatedly washing with distilled water until neutral, and fully draining.

[0060] (2) The volume of 10% isopropanol is 3L, stirred at 4°C for 8h, washed repeatedly with distilled water until neutral, fully drained, and weighed 271.44g.

[0061] Sonication:

[0062] Weigh 60.04g, 60.35g, 60.07, 60.15, and 60.89g of fish skin after degreasing and removing impurities, and place them in 1L of 0.5M acetic acid solution for ultrasonic treatment; use an ultrasonic ce...

Embodiment 1、2

[0073] Collagen polyacrylamide gel electrophoresis (SDS-PAGE) analysis in embodiment 1,2

[0074] 7% separating gel and 4% stacking gel were selected, and the sample loading volume was 20 μL. Sample treatment: the sample is a 5mg / ml collagen solution, centrifuged at 10000r / min for 10min to remove insoluble matter, take the supernatant and mix it with sodium dodecyl sulfate (SDS) loading buffer at a volume ratio of 2:3, The collagen concentration was made 2 mg / ml. Electrophoresis conditions: initial voltage 80V, electrophoresis time 40min, then adjust the voltage to 110V. Electrophoresis time is about 90min, stop electrophoresis until the dye in the sample migrates to 2cm from the bottom. After electrophoresis, the gel was stained with Coomassie Brilliant Blue R-250 for 30 minutes, then decolorized with a mixed solution of ethanol: acetic acid: distilled water = 5:1:4 for 1 hour, and finally soaked in distilled water overnight.

[0075] The electrophoretic bands with differen...

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Abstract

The invention belongs to the technical field of collagen extraction, and particularly relates to an extraction method and application of fish skin collagen. The invention provides a method for rapidly extracting non-denatured fish skin collagen, which solves the problem of waste of freshwater fish processing byproducts and overcomes the defects of low extraction rate and long extraction time of the traditional collagen extraction method. The technical method disclosed by the invention is simple and feasible, short in extraction time and high in extraction efficiency, most importantly, the integrity of collagen macromolecular structures is ensured, and the extraction time is greatly shortened compared with other methods. According to the extraction method, the collagen is subjected to ultrasonic treatment before being extracted by a traditional acid-enzyme combination method, so that the natural activity of extracted fish skin collagen molecules is ensured, and the fish skin collagen obtained by measurement keeps the natural structure of type I collagen and is non-denatured type I collagen. The method is simple to operate, lower in energy consumption, high in extraction efficiency and easy to amplify and apply to production.

Description

technical field [0001] The invention belongs to the technical field of collagen extraction, and in particular relates to an extraction method and application of fish skin collagen. Background technique [0002] my country is rich in aquatic resources, and a large number of by-products such as fish skin and scales are produced during the processing of freshwater fish, accounting for about 40%-60% of the total amount of aquatic products. The collagen is mainly distributed in the by-products of fish skin and fish scales, among which the collagen content of fish skin is as high as 25%, and it is mainly high-value type I collagen. Due to sufficient sources, using fish skin as the main raw material for extracting type I collagen can not only avoid wasting a lot of precious resources, increase the added value of fish processing, and reduce environmental pollution; but also obtain good economic and social benefits; in addition, aquatic organisms The antigenicity of collagen is weak...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/14C07K1/36C07K1/34C07K1/30
CPCC12P21/06C07K14/78
Inventor 刘茹吴丹胡杨熊善柏
Owner HUAZHONG AGRI UNIV
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