Preparation method of fish skin collagen

A technology of fish skin collagen and collagen, which is applied to the preparation method of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of low yield, high cost, hydrolysis of peptide bonds, etc., and achieve high yield , easy drying effect

Inactive Publication Date: 2011-01-19
SUZHOU UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0006] 1. The Chinese invention patent with the patent number 02111683.0 discloses a kind of collagen with good biocompatibility using deep-sea fish skin as raw material and its preparation method: the fish skin is treated with an alkaline solution, and then the above-mentioned collagen is treated with a restricted enzymatic hydrolysis method. For fish skin, degradable biocompatible fish skin collagen can be obtained by purifying the enzymatic supernatant; Improve, and, the yield of collagen protein is not high in this technical scheme, is 5%~8% by mass percentage, and yield is not high
[0007] 2. The Chinese Invention Patent No. 200710024384.0 discloses a preparation method of freshwater fish skin collagen, which belongs to the field of aquatic product processing technology: after the fish skin is washed, drained, and shredded; first, it contains 0.005%-2% (V / V) Hydrogen peroxide 0.001-0.2M lye soaked for 3-48H; after high-speed stirring, then cleaned and soaked with organic reagents; -5.0 acid solution extraction for 12-64H, the resulting system was filtered with a 30-100MM membrane, and then filtered with a 1-30MM membrane; finally, a 5M NaCl solution was added to the filtrate to 0.8-2.0M, and the precipitate obtained by centrifugation was washed with a 0.001- Dissolve in 0.1M acetic acid, dialyze in distilled water, and freeze-dry to obtain a white and odorless collagen solid; in this technical solution, the use of enzymes increases the cost
[0014] Moreover, the above four schemes did not point out that the obtained fish skin collagen has unexpected effects as a fish cell culture carrier

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0039]Embodiment one, fish skin collagen preparation method

[0040] Use air-dried tilapia as raw material, after crushing into 2mm×2mm×2mm pieces, do the following:

[0041] (1) According to the solid: liquid ratio of 1:20, add 0.05M NaOH, soak the fish skin fragments at 4°C for 4 hours, filter the fish skin residue with a 20-mesh sieve, and air-dry it; Double the volume of anhydrous ether, reflux in a water bath at 40°C for 3-4 hours, remove fat, and air-dry to obtain fat-removed fish skin residue;

[0042] (2) Add 0.5M acetic acid to the degreased fish skin residue according to the solid:liquid ratio of 1:60, extract at 4°C for 72 hours, and filter with a 40-mesh sieve to obtain the filtrate; repeat once;

[0043] (3) Add NaCl powder to the filtrate to a final concentration of 2.5M to salt out the collagen; centrifuge at 4°C and 6500rpm for 30min to collect the precipitate;

[0044] (4) Add 0.5M acetic acid to dissolve 3 times the volume of the precipitate to obtain a col...

Embodiment 2

[0046] Embodiment 2, referring to Embodiment 1, the difference is that in step (2), the solid-to-liquid ratio is 1:50, the concentration of acetic acid is 0.6M, and the leaching time is 60h.

Embodiment 3

[0047] Embodiment 3, referring to Embodiment 1, the difference is that in step (2), the solid-to-liquid ratio is 1:70, the concentration of acetic acid is 0.5M, and the leaching time is 80h.

[0048] The results are shown in Table 1: the yields of fish skin collagen obtained in Examples 1 to 3 are 40.18 ± 0.052%, 39.76 ± 0.012%, 39.76 ± 0.013% respectively; the hydroxyproline content reflecting the purity of fish skin collagen is 8.47 ± 0.06 %, 8.33±0.16%, 8.3±0.2%, the collagen content of fish skin is 93.98±0.64%, 92.50±1.28%, 92.13±2.22% respectively;

[0049] Table 1: Yield and purity of fish skin collagen extracted by degreasing with anhydrous ether and 0.5-0.6M acetic acid

[0050] batch

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Abstract

The invention belongs to the field of processing fish skin collagen and particularly relates to a preparation method of fish skin collagen which can be used as a fish cell cultural carrier. The preparation method comprises the steps of raw material processing, extracting, salting and purification dialysis drying, wherein in the raw material processing step, absolute ether is refluxed at the temperature of 35 to 40 DEG C for 3 to 4h to remove the fat of fish skin, the fish skin is dried in the air, and fish skin residues are obtained; in the extraction steps, 0.5 to 0.6M acetic acid is added into the fish skin residues according to a solid to liquid ratio of 1:50 to 70 for leaching for 60 to 80h, and a 40-mesh sieve is used for filtering to obtain filtrate containing the fish skin collagen; and the steps are repeated once. In the invention, yield is improved on the basis of keeping the biological nature and pureness of the fish skin collagen; when the fish skin collagen obtained is used as a fish cell cultural carrier, cellular affinity is good, adherence speed is fast, cell growth is excellent, and stand density is high; therefore, the fish skin collagen is more advanced than cow skin collagen or mouse tail collagen.

Description

technical field [0001] The invention belongs to the field of fish skin collagen processing, and in particular relates to a method for preparing fish skin collagen that can be used as a fish cell culture carrier. Background technique [0002] Collagen is an important functional protein. Collagen contains a unique amino acid-hydroxyproline (Hyp). Collagen is closely related to cell proliferation, differentiation, exercise, immunity, joint lubrication, and wound healing. Due to the special functions of collagen, it has been widely used in medicine, food, daily chemical industry, biosynthesis and other industrial fields, such as medical capsules, surgical materials, edible gelatin, photographic gelatin, cosmetics, etc. [0003] The raw materials of collagen currently produced are mainly the skin, bone and tendon of terrestrial mammals such as pigs and cattle. Due to the occurrence of mad cow disease, foot-and-mouth disease and other epidemics in recent years, collagen has been e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/14C12N5/071
Inventor 叶元土蔡春芳戴如燕唐崭姚仕彬宋亮
Owner SUZHOU UNIV
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