CN105063147A - Homogenization digestion method of collagen - Google Patents

Abstract

The invention belongs to the field of biotechnologies, and aims to provide a homogenization digestion method of collagen. According to the homogenization digestion method of the collagen, animal skin is minced and then subjected to detailed digestion treatment through a fining process so as to be used for extracting the collagen with a higher purity at the subsequent procedure. The method includes the steps that minced skin particle paste is homogenized three times according to the operation specifications of a tissue triturator till the minced skin particle paste is completely suspended, ultrapure water is added till the tissue content reaches 25 g/l, the ultrapure water and the minced skin particle paste are evenly mixed, glacial acetic acid is added, and a pepsin solution is transferred into the tissue homogenate till the pepsin solution content reaches 0.1 mg/ml. The process is simple and easy to implement; needless impurities in earlier-stage production of the collagen are removed, the minced animal skin is further triturated and homogenized perfectly, and therefore the prepared homogenate guarantees the high purity in later-stage production of the collagen.

Description

Collagen protein homogenate digestion method

Technical field

The invention belongs to biological technical field.

Background technology

The production initial stage of existing collagen protein, all need to carry out thinning processing to animal skin, bone etc., in order to the later stage produces content and the purity of collagen protein, the process in early stage is extremely crucial, current routine is fairly simple to the treatment process extracting collagen protein in animal skin, only that animal skin is rubbed, be made into pasty state, namely simple impurity elimination carries out the extraction of collagen protein, this kind of approach application is in the production of collagen protein, not only cause great waste, and impurities removal is not thorough, and much impurity finally may be had to be mixed in collagen protein.

Summary of the invention

The object of this invention is to provide by hand work, after animal skin being rubbed, carry out detailed digestion process, for the collagen protein homogenate digestion method of subsequent extracted high purity collagen more.

Step of the present invention is:

A, becoming the little iron spoon of the rubbing skin particle of pasty state to transfer in the glass cylinder of tissue mashing machine 3 spoonfuls, about 50 milliliters, adding the ultrapure water of 10 times of volumes, by the working specification homogenate 3 times of tissue mashing machine, to suspending completely, tissue homogenate being proceeded to plastic tank;

After the rubbing evenly of b, all tissue particles, plastic tank being added ultrapure water is 25 grams per liters to the content organized, mixing;

C, add Glacial acetic acid to suspensions of tissues plastic tank modulation strength of solution 0.5 mg/ml;

D, take stomach en-, stomach en-is dissolved in the pepsin solution making 10mg/ml in 0.5M acetic acid, is then transferred to tissue homogenate to 0.1mg/ml, namely add the pepsin solution of 1ml in every 100ml homogenate, mixing, room temperature digestion 7-10 days.

Present invention process is simple, processing ease, the present invention not only eliminates collagen protein and produces in earlier stage unnecessary impurity, and more perfectly smashs homogenate to pieces to rubbing animal skin further, and the production that the homogenate produced is later stage collagen protein provides high purity to ensure.

Embodiment,

Step of the present invention is:

A, becoming the little iron spoon of the rubbing skin particle of pasty state to transfer in the glass cylinder of tissue mashing machine 3 spoonfuls, about 50 milliliters, adding the ultrapure water of 10 times of volumes, by the working specification homogenate 3 times of tissue mashing machine, to suspending completely, tissue homogenate being proceeded to plastic tank;

Become the rubbing skin particle of pasty state to be that animal skin is simply rubbed technique and formed by existing common process, work of the present invention proceeds on this basis to smash impurity elimination work to pieces; Through repeatedly smashing to pieces of this step, the dilution of skin particle is also suspended completely with liquid;

After the rubbing evenly of b, all tissue particles, plastic tank being added ultrapure water is 25 grams per liters to the content organized, mixing; After smashing to pieces, continue dilution;

C, add Glacial acetic acid to suspensions of tissues plastic tank modulation strength of solution 0.5 mg/ml; By adding the acidity of Glacial acetic acid regulator solution, make its pH value to 2.0 ~ 3.0;

D, take stomach en-, stomach en-is dissolved in the pepsin solution making 10mg/ml in 0.5M acetic acid, is then transferred to tissue homogenate to 0.1mg/ml, namely add the pepsin solution of 1ml in every 100ml homogenate, mixing, room temperature digestion 7-10 days.Pepsin solution joins in the suspensions of tissues mixed up above, the collegen filament of natural triple-helix structure form can be digested to the collagen molecules of molecular weight about 300,000, excise the amino of collagen molecules and the bulbous region of C-terminal (terminal peptide) simultaneously, and maintain the natural spatially spiral section structure of collagen protein and biological activity, thus eliminate the potential immunogenicity of collagen protein.

Claims (1)

1. a collagen protein homogenate digestion method, is characterized in that: the steps include:

A, becoming the little iron spoon of the rubbing skin particle of pasty state to transfer in the glass cylinder of tissue mashing machine 3 spoonfuls, about 50 milliliters, adding the ultrapure water of 10 times of volumes, by the working specification homogenate 3 times of tissue mashing machine, to suspending completely, tissue homogenate being proceeded to plastic tank;

After the rubbing evenly of b, all tissue particles, plastic tank being added ultrapure water is 25 grams per liters to the content organized, mixing;

C, add Glacial acetic acid to suspensions of tissues plastic tank modulation strength of solution 0.5 mg/ml;

D, take stomach en-, stomach en-is dissolved in the pepsin solution making 10mg/ml in 0.5M acetic acid, is then transferred to tissue homogenate to 0.1mg/ml, namely add the pepsin solution of 1ml in every 100ml homogenate, mixing, room temperature digestion 7-10 days.