Method for preparing I type collagen and V type collagen

A collagen, type I technology, applied in the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., can solve problems such as the difficulty of purification of collagen, achieve high extraction efficiency, extend the industrial chain, and reduce environmental pollution Effect

Active Publication Date: 2014-06-25
HANGZHOU NUTRITION BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, people have extracted crude collagen from a variety of aquatic product processing wastes (skin, scale, bone, fin). Purification of protein is quite difficult, and there are few reports in the literature

Method used

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  • Method for preparing I type collagen and V type collagen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Wash the fish skin from which scales and subcutaneous tissue have been removed, drain it, and cut it into pieces (about 1 cm 2 ), dried (55°C), pulverized (50 mesh), weighed 200 g and soaked in 15 L of 0.1 mol / L NaOH solution at room temperature, stirred continuously for 30 h, centrifuged (2,500 g × 10 min) to collect the precipitate, and used Wash with tap water to pH 7.0 and dry (55°C).

[0031] Take 100 g (on a dry basis) of pretreated fish skin and place it in 10 L of pre-cooled water, add 150 mL of glacial acetic acid and 10 g of pepsin, stir continuously for 42 h, centrifuge (100,000 g × 60 min) and take Slowly add 1.25 Kg NaCl powder to the supernatant to 2.0 mol / L, salt out overnight, centrifuge (10,000 g × 15 min), and the obtained precipitate is crude collagen.

[0032] Dissolve the crude collagen in 7.5 L of 1.0 mol / L NaCl (0.05 mol / L Tris-HCl, pH 7.5) solution (use 1.0 mol / L NaOH to adjust the pH to 7.5), centrifuge (100,000 g × 60 min) to take To the supe...

Embodiment 2

[0036] Wash the fish skin from which scales and subcutaneous tissue have been removed, drain it, and cut it into pieces (about 1 cm 2 ), dried (50°C), crushed (20 mesh), weighed 200 g at room temperature, soaked in 10 L 0.1 mol / L NaOH solution, stirred continuously for 36 h, centrifuged (2,500 g × 10 min) to collect the precipitate, and used Wash with tap water to pH 6.0 and dry (50°C).

[0037] Take 100 g (on a dry basis) of pretreated fish skin and place it in 7.5 L of pre-cooled water, add 45 mL of glacial acetic acid and 7.5 g of pepsin, stir continuously for 48 h, and centrifuge (100,000 g × 60 min) to take Slowly add 1.0 Kg NaCl powder to the supernatant to 2.0 mol / L, salt out overnight, centrifuge (10,000 g × 15 min), and the obtained precipitate is crude collagen.

[0038] Dissolve the crude collagen in 5 L of 1.0 mol / L NaCl (0.05 mol / L Tris-HCl, pH 7.5) solution (use 1.0 mol / L NaOH to adjust the pH to 7.5), centrifuge (100,000 g × 60 min) and take To the supernatant...

Embodiment 3

[0042] Wash the fish skin from which scales and subcutaneous tissue have been removed, drain it, and cut it into pieces (about 1 cm 2 ), dried (60°C), crushed (100 mesh), weighed 200 g at room temperature, soaked in 20 L 0.1 mol / L NaOH solution, stirred continuously for 24 h, centrifuged (2,500 g × 10 min) to collect the precipitate, and used Wash with tap water to pH 8.0 and dry (60°C). Take 100 g (on a dry basis) of pretreated fish skin and place it in 15 L of pre-cooled water, add 450 mL of glacial acetic acid and 15 g of pepsin, stir continuously for 36 h, centrifuge (100,000 g × 60 min) and take Slowly add 1.85 Kg NaCl powder to the supernatant to 2.0 mol / L, salt out overnight, centrifuge (10,000 g × 15 min), and the obtained precipitate is crude collagen.

[0043] Dissolve the crude collagen in 10 L of 1.0 mol / L NaCl (0.05 mol / L Tris-HCl, pH 7.5) solution (use 1.0 mol / L NaOH to adjust the pH to 7.5), centrifuge (100,000 g × 60 min) to take To the supernatant, slowly ad...

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Abstract

The invention belongs to the field of food biotechnology, and particularly relates to a method for preparing I type collagen and V type collagen. The method comprises main steps of cleaning, draining off, cutting up and drying fishskin with scale and subcutaneous tissue removed, smashing, then steeping in 0.1M NaOH solution, stirring, then centrifuging to obtain precipitate, washing to be almost neutral, then putting in precooling water, adding acetic acid and pepsase and stirring, centrifuging to obtain the supernate, adding NaCl, then salting out, centrifuging, dissolving the precipitate into Tris-HCl buffer solution containing NaCl, centrifuging, then obtaining supernate, adding NaCl to salt out, centrifuging, then collecting precipitate and supernate respectively, dissolving, dialyzing, freeze-drying the precipitate so as to obtain the I type collagen, adding NaCl into the supernate, then salting out, centrifuging, then taking the precipitate, dissolving, dialyzing and freeze-drying so as to obtain the V type collagen, wherein all the steps are carried out at 4DEG C. The I type collagen and V type collagen are prepared through fractional salting out; the method is simple, and has strong operability, low cost and high efficiency.

Description

technical field [0001] The invention belongs to the field of food biotechnology, and in particular relates to a preparation method of type I and type V collagen. technical background [0002] my country is a large fishery country, but its processing technology is low, and a large amount of processing waste has caused serious waste of resources and environmental pollution. Collagen can be widely used in food, cosmetics, medicine, materials and other fields, but in recent years, frequent outbreaks of infectious diseases such as mad cow disease, foot-and-mouth disease, Streptococcus suis, and avian influenza have made the safety of collagen from traditional livestock and poultry sources more and more unsatisfactory. At the same time, religious customs also make its application subject to certain restrictions. Therefore, using aquatic product processing waste to produce collagen that can replace livestock and poultry sources is of great significance for reducing environment...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/36C07K1/34C07K1/30
Inventor 王林梁秋芳徐军民王振斌马海乐
Owner HANGZHOU NUTRITION BIOTECH
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