CN101974594A

CN101974594A - Method for extracting tortoise-plastron peptide contained in tortoise-plastron glue through enzyme method

Info

Publication number: CN101974594A
Application number: CN 201010540588
Authority: CN
Inventors: 叶传发; 张小东; 胡承四; 叶思
Original assignee: HUBEI YUANCHENG PHARMACEUTICAL CO Ltd
Current assignee: HUBEI YUANCHENG PHARMACEUTICAL CO., LTD.
Priority date: 2010-11-11
Filing date: 2010-11-11
Publication date: 2011-02-16
Anticipated expiration: 2030-11-11
Also published as: CN101974594B

Abstract

The invention discloses a method for extracting tortoise-plastron peptide contained in tortoise-plastron glue through an enzyme method, comprising the following steps of: blending the tortoise-plastron glue as main raw materials with water, and warming and stewing; carrying out compound protease enzymolysis; inactivating enzymes; decoloring; separating a membrane; and finally drying obtained filter liquor to obtain a product, i.e. the tortoise-plastron peptide. The method has short production period and low cost without generating any toxic and harmful substance; and in addition the obtained product has high safety and can be widely used in the fields of heal care products, drugs, and the like.

Description

The method of tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis

Technical field

The inventive method relates to tortoiseshell deep process technology field, is specifically related to the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis, be a kind of be raw material with the Colla Plastri Testudinis, utilize biological enzyme enzymolysis Colla Plastri Testudinis, thereby extract the method for tortoiseshell peptide.

Background technology

Tortoiseshell is the plastron of Testudinidae animal tortoise, has the effect of nourishing and suppressing Yang, the strong bone of kidney-nourishing, the bushing of nourishing blood, and is one of traditional Chinese medical science rare medicinal herbs commonly used.Its main chemical ingredients is amino acid, protein, gelatin, trace element etc.Colla Plastri Testudinis is top grade in China's traditional Chinese medicine, has fabulous nourishing function, Colla Plastri Testudinis is carried out enzymolysis can make the tortoiseshell peptide, and the tortoiseshell peptide that obtains by enzyme incision technology not only has the fabulous characteristic of digesting and assimilating, also has more effective physiological function, the tortoiseshell peptide is dissolved in after drinking can improves the colours of wine quality, promote the character of wine.

Tortoiseshell peptide series product have the increased blood pressure of inhibition, protection stomach mucous membrane, antiulcer agent, the metabolism of promotion skin collagen, beautify effects such as skin, preventing osteoporosis.

From Colla Plastri Testudinis, extract the tortoiseshell peptide and adopt following method mostly: Colla Plastri Testudinis → alkaline purification → high temperature steaming → enzymolysis → filtration → concentrate → spraying drying.Use a large amount of alkaline purifications in the leaching process, not only environment was polluted but also there is potential safety hazard in human body, also destroyed proteinic composition; And concentration operation destroys the product phase of product easily, does not use membrane separation technique in the prior art, has influenced the quality of product.

Summary of the invention

At the deficiencies in the prior art, the object of the present invention is to provide the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis, need not to use alkaline purification, but utilize complex enzyme zymohydrolysis, and then by membrane separation technique desalination and concentrated, extract the tortoiseshell peptide, present method with short production cycle, easy and simple to handle, cost is low, safe without toxic side effect.

In order to realize above-mentioned purpose, the present invention adopts following technical measures:

The method of tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis, its step is as follows:

1, be raw material with Colla Plastri Testudinis (claiming Glue of Tortoise Plastron again), the 5-7 distilled water doubly that adds raw material weight stirs, and is warming up to 100 ℃ of boiling 5-7h;

2, be cooled to 45-60 ℃, adjust pH is 7-8, adds Sumizyme MP and trypsinase in the 1-3% of raw material Colla Plastri Testudinis weight and the ratio of 0.5-1.5% respectively then, keeps pH at 7-8 with sodium hydroxide solution in the reaction process, and constant temperature stirs enzymolysis 6-10h; The enzymolysis solution that obtains is warming up to 90-100 ℃, keeps the 10-20min enzyme that goes out;

3, with the centrifugal after-filtration of enzymolysis solution, remove proteolytic enzyme, add powdered active carbon in the filtrate and decolour, and then use the membrane sepn means to separate, concentrate, be drying to obtain tortoiseshell peptide dry product.

Sumizyme MP that uses in the inventive method and tryptic vigor are respectively 25U/mg and 35U/mg.

Compared with prior art, advantage of the inventive method and beneficial effect are as follows:

1, avoided effectively that chemical means is handled the environmental pollution that brought and to human injury's problem;

2, adopt the purity height of the tortoiseshell peptide product that the inventive method produces, molecular weight reaches more than 96% at the daltonian peptide content of 500-3000 in the gross protein of products obtained therefrom, and molecular weight is daltonian in 4% greater than 3000;

3, the tortoiseshell peptide that adopts the inventive method to produce, molecular weight is little, and human body absorbs easily, and specific absorption reaches more than 90%;

4, the present invention has adopted activated carbon decolorizing, residues in the product injury to human body when effectively having avoided using the hydrogen peroxide decolouring, and has guaranteed the natural colored degree of product;

5, the inventive method with short production cycle, easy and simple to handle, cost is low, safe without toxic side effect, can be widely used in fields such as multi-function health-care product, medicine.

Embodiment

Below by specific embodiment the inventive method is described in further detail.

Embodiment 1:

1, takes by weighing Colla Plastri Testudinis 1kg, add distilled water 6kg then, after stirring, be warming up to 100 ℃ of boiling 5h;

2, be cooled to 55 ℃, food grade hydroxide flake sodium solution with 0.1M is transferred pH to 7-8, add 25g Sumizyme MP and 10g trypsinase then, stir enzymolysis 10h down, keep the pH value stabilization of reaction system in the enzymolysis process with above-mentioned 0.1M sodium hydroxide solution at 55 ℃; Be warming up to 95 ℃ then, the insulation 15min enzyme that goes out;

3, enzymolysis solution is filtered, get supernatant liquor; Add the 25g powdered active carbon in the supernatant liquor, 95 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains the Powdered tortoiseshell peptide of 412g white (adopt activated carbon decolorizing among the present invention, the color and luster of products obtained therefrom is a white, is better than the faint yellow of national Specification), and the examining report of products obtained therefrom sees Table 1.

The examining report of table 1 embodiment 1 products obtained therefrom

Embodiment 2:

1, takes by weighing Colla Plastri Testudinis 2kg, add distilled water 11kg then, after stirring, be warming up to 100 ℃ of boiling 6h;

2, be cooled to 50 ℃, solution with the food grade flaky sodium hydrate of 0.1M is transferred pH to 7-8, add 20g Sumizyme MP and 10g trypsinase then, stir enzymolysis 8h down, keep the pH value stabilization of reaction system in the enzymolysis process with the sodium hydroxide solution of above-mentioned 0.1M at 50 ℃; Be warming up to 100 ℃ then, the insulation 10min enzyme that goes out;

3, enzymolysis solution is filtered, get supernatant liquor; Add the 44g powdered active carbon in the supernatant liquor, 95 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 831g white powder tortoiseshell peptide, and the examining report of products obtained therefrom sees Table 2.

The examining report of table 2 embodiment 2 products obtained therefroms

Embodiment 3:

1, takes by weighing Colla Plastri Testudinis 10kg, add distilled water 50kg then, after stirring, be warming up to 100 ℃ of boiling 7h;

2, be cooled to 55 ℃, solution with the food grade flaky sodium hydrate of 0.1M is transferred pH to 7-8, add 200g Sumizyme MP and 100g trypsinase then, stir enzymolysis 7h down, keep the pH value stabilization of reaction system in the enzymolysis process with the sodium hydroxide solution of above-mentioned 0.1M at 55 ℃; Be warming up to 90 ℃ then, the insulation 20min enzyme that goes out;

3, enzymolysis solution is filtered, get supernatant liquor; Add the 250g powdered active carbon in the supernatant liquor, 95 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 4.18kg white powder tortoiseshell peptide, and the examining report of products obtained therefrom sees Table 3.

The examining report of table 3 embodiment 3 products obtained therefroms

Embodiment 4:

1, takes by weighing Colla Plastri Testudinis 100kg, add distilled water 700kg then, after stirring, be warming up to 100 ℃ of boiling 6h;

2, be cooled to 60 ℃, solution with the food grade flaky sodium hydrate of 0.1M is transferred pH to 7-8, add 2kg Sumizyme MP and 10kg trypsinase then, stir enzymolysis 6h, keep the pH value stabilization of reaction system in the enzymolysis process with the sodium hydroxide solution of above-mentioned 0.1M at 60 times ℃; Be warming up to 95 ℃ then, the insulation 15min enzyme that goes out;

3, enzymolysis solution is filtered, get supernatant liquor; Add the 2000g powdered active carbon in the supernatant liquor, 95 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 42.1kg white powder tortoiseshell peptide, and the examining report of products obtained therefrom sees Table 4.

Table 4 embodiment 4 products obtained therefrom examining reports

Claims

1. the method for tortoiseshell peptide in the Enzymatic Extraction Colla Plastri Testudinis, step is as follows:

A, be raw material, add after distilled water stirs, be warming up to 100 ℃ of boilings with the Colla Plastri Testudinis;

B, be cooled to 45-60 ℃, adjust pH is 7-8, add the Sumizyme MP of 25U/mg and the trypsinase of 35U/mg in the 1-3% of raw material Colla Plastri Testudinis weight and the ratio of 0.5-1.5% respectively then, constant temperature stirs enzymolysis 6-10h, keeps the pH value stabilization of reaction system in the enzymolysis process with sodium hydroxide solution; Then the enzymolysis solution that obtains is warming up to the 90-100 ℃ of enzyme that goes out;

C, with the centrifugal after-filtration of enzymolysis solution, filtrate and then is come desalination and is concentrated by membrane sepn after powdered active carbon decolouring, promptly gets the tortoiseshell peptide after the gained concentrated solution drying.

2. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 is characterized in that: the add-on of described distilled water is 5-7 a times of raw material weight.

3. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 or 2, it is characterized in that: the time of described boiling is 5-7h.

4. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 or 2, it is characterized in that: the time of the described enzyme that goes out is 10-20min.

5. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 or 2, it is characterized in that: the add-on of described powdered active carbon is the 2-2.5% of raw material Colla Plastri Testudinis weight.

6. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 or 2 is characterized in that: what described membrane sepn operation was adopted is that the aperture is the nanofiltration membrane of 1nm.

7. the method for tortoiseshell peptide in a kind of Enzymatic Extraction Colla Plastri Testudinis as claimed in claim 1 or 2 is characterized in that: described decolouring is 95 ℃ of following whip attachment 1h decolourings.