CN112941137A - Production method of tortoise peptide - Google Patents
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The invention belongs to the technical field of biological preparation, and relates to a production method of tortoise peptide, which comprises the steps of taking adult live tortoise, slaughtering, cleaning, boning and crushing into tortoise meat paste; mixing the tortoise shell extract and the tortoise meat paste, homogenizing by ultrasonic waves, preparing into a substrate solution by using ultrapure water, and adding protease for enzymolysis; and (3) carrying out enzyme inactivation, sterilization, ultrafiltration separation and vacuum freeze drying on a reaction product obtained by enzymolysis to obtain soluble powdered turtle peptide. The invention takes a plurality of turtle species composite components as raw materials, extracts active peptide molecules in the turtle to the maximum extent, simultaneously preserves other bioactive components in the raw materials, and a plurality of bioactive components supplement each other, can effectively regulate immunity, resist oxidation, resist tumor, resist fatigue, regulate blood fat, improve sleep and delay aging, thereby achieving the effects of nourishing yin and strengthening yang, building up body, maintaining beauty and keeping young and prolonging life.
Description
Technical Field
The invention belongs to the technical field of biological preparation, relates to a production method of turtle peptide, and particularly relates to a production method of turtle peptide with multiple turtle sources and relatively uniformly distributed amino acid polymerization degrees.
Background
Modern scientific researches find that turtles with the meaning of 'longevity and luckiness' in Chinese traditional culture have longevity due to unique physiological structures and life habits, and because the turtles contain rich bioactive substances with the functions of regulating immunity, resisting oxidation, resisting tumors, resisting fatigue, regulating blood fat, improving sleep, delaying senescence and the like, such as trace elements of unsaturated fatty acid, vitamin E, selenium and the like, active protein, collagen, free fatty acid and the like, the turtles are survived from cancers, cardiovascular and cerebrovascular diseases, diabetes and the like which are the most common diseases of human beings.
The results of the study showed that peptides are important components constituting proteins. Meanwhile, as a basic material constituting all human cells, peptides are involved in a plurality of fields such as hormones, nerves, reproduction, and cell renewal, metabolism, growth, repair, etc. in the human body. Peptides also have the physiological function of regulating various systemic organs and cells in vivo. Nowadays, among peptides from various sources, the tortoise peptide is most likely to be crown in the peptide, and meanwhile, research results related to the tortoise peptide also provide powerful scientific evidence for long-term medicine and food traditions of Chinese turtle medicine and health care and health preservation, and also indicate directions for developing health food, special medical food and even biological medicines which are beneficial to human health and longevity.
At present, the method for preparing the turtle peptide by taking turtle organisms as raw materials adopts a single turtle species as a raw material, and prepares the turtle peptide by taking live turtle tissues as the raw material or taking tortoise-shell glue as the raw material through an enzymatic hydrolysis method, but the components of a turtle peptide product are limited, and the components of a whole turtle cannot be embodied.
Disclosure of Invention
The invention aims to provide a production method of turtle peptide, which is characterized in that a plurality of turtle species are matched, turtle tissues and turtle shells are respectively taken as raw material sources, the turtle peptide produced by an enzymolysis method is applied, active peptide molecules in the turtle are extracted to the maximum extent, other bioactive components in the raw materials are preserved, and a plurality of bioactive components supplement each other and can effectively regulate immunity, oxidation resistance, tumor resistance, fatigue resistance, blood fat regulation, sleep improvement and aging delay, so that the effects of nourishing yin and tonifying yang, building up the body, maintaining beauty and keeping young and prolonging life are achieved.
The technical scheme adopted by the invention is as follows:
a production method of turtle peptide comprises the following steps:
1. taking different kinds of adult live turtles, slaughtering, cleaning, boning, mixing the turtle meat according to the weight equal parts, mechanically crushing, and ultrasonically crushing to form turtle meat paste.
The different kinds of adult live tortoise are selected from three-line box turtle, stone water turtle, Chinese grass turtle, tortoise, crocodile turtle, yellow box turtle, Suka turtle, big head turtle, platysternon tortoise, box turtle, brocade turtle or mud turtle 3 or more.
The solid content in the turtle meat paste is not more than 10 meshes.
2. According to the weight ratio of 1: 1-10 of the method, mixing the tortoise shell extract (prepared by the method disclosed in patent CN 104946488A) and the tortoise meat paste, homogenizing, and preparing a substrate solution of 5-50% (W/V) by using ultrapure water.
3. Adding 0.1-1.0 mg/mL protease into the substrate solution, carrying out enzymolysis under a constant temperature condition, and adjusting the pH value to 5-8 (adjusting the pH value by using 0.01-0.5 mol/L NaOH or HCl).
The protease is one, two or more of trypsin, pepsin, papain, alkaline protease and neutral protease.
The constant temperature is 40-70 ℃.
The enzymolysis time is 1-24 h.
4. Inactivating enzyme of the enzymolysis product, sterilizing, sequentially filtering with ultrafiltration membranes with cut-off molecular weights of 9KD, 6KD and 3KD to obtain 4 kinds of turtle enzyme enzymolysis solutions with different molecular weights, wherein the molecular weights are respectively: greater than 9KD, 6-9 KD, 3-6 KD and less than 3 KD.
The enzyme deactivation process comprises the following steps: and (3) putting the reaction system into hot water of 90-100 ℃, and uniformly stirring for 5-30 min.
In the process of filtering the ultrafiltration membrane, carbon dioxide gas is adopted to protect a reaction system.
5. The obtained tortoise peptidase hydrolyzed liquid with each molecular weight segment is respectively frozen and dried in vacuum to obtain 4 kinds of peptide component crude products (white to light yellow soluble powder) with different molecular weights.
6. And uniformly mixing the prepared 4 kinds of peptide component crude products according to the weight ratio of 1:1:1:1 to obtain the turtle component turtle peptide.
The invention has the beneficial effects that:
(1) the active tortoise as the raw material is selected from 3 or more than 3, and the active peptide molecules in the adult tortoise are extracted to the maximum extent while the raw material is fed in a certain proportion, and other bioactive components in the raw material are preserved to ensure that the types and the contents of the functional components in the final product are balanced as much as possible, and a plurality of bioactive components supplement each other, so that the health-care turtle feed can effectively regulate immunity, oxidation resistance, tumor resistance, fatigue resistance, blood fat regulation, sleep improvement and aging delay, and further has the effects of nourishing yin and tonifying yang, building up the body, maintaining beauty and keeping young and prolonging life.
(2) The preparation of the tortoise peptide from the tortoise-shell glue has the advantage of high production efficiency. However, during the process of decocting tortoise-shell glue from tortoise shell, part of protein is inevitably denatured and decomposed at high temperature, and thus the bioactivity is lost. The invention adopts the live tortoise tissue and the tortoise-shell glue as raw materials to produce the tortoise peptide, thereby not only maximally utilizing protein resources in the live tortoise tissue, but also considering the production efficiency.
(3) The peptides with different molecular weights are intercepted from the enzymolysis products and prepared into final products according to the same weight ratio, and simultaneously, the medical requirements of balanced nutrition and the condition that small molecular peptides are easier to be absorbed by human bodies are met, so that the preparation method is a more advanced formula form.
Detailed Description
The following examples are given to further illustrate the embodiments of the present invention. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.
Example one
(1) Taking live tortoise of small water turtle, small stone water turtle and Chinese grass turtle, slaughtering, cleaning, boning, mixing turtle meat according to the weight ratio of 1:1:1, mechanically crushing and ultrasonically crushing the turtle meat respectively to form turtle meat paste, wherein the solid content is not more than 10 meshes.
(2) According to the method disclosed in patent CN104946488A, tortoise shell is taken to prepare tortoise shell extract.
(3) Mixing 1000g of minced tortoise meat slurry and 100g of tortoise shell extract, homogenizing, and preparing 10% (W/V) substrate solution with ultrapure water.
(4) Adding papain of 0.5mg/mL into the substrate solution, and carrying out enzymolysis at the constant temperature of 65 ℃ for 15h, wherein the pH is adjusted to 6.5 +/-0.1.
(5) And (3) inactivating the enzyme of the obtained enzymolysis product, sterilizing, and sequentially filtering with ultrafiltration membranes with cut-off molecular weights of 9KD, 6KD and 3KD to obtain 4 turtle enzyme enzymolysis solutions with different molecular weights (larger than 9KD, 6-9 KD, 3-6 KD and smaller than 3 KD).
(6) Respectively carrying out vacuum freeze drying on the obtained tortoise peptidase hydrolyzed solution with each molecular weight segment to obtain 4 kinds of white to light yellow soluble powder with different molecular weights, namely a crude product of the peptide component;
(7) considering the principle that peptides with smaller molecular weight are easier to be absorbed by human body, the prepared 4 crude peptide components are uniformly mixed according to the weight ratio of 1:1:1:1 to obtain 217g of tortoise peptide as a whole tortoise component.
Example two
(1) Taking live tortoise of small water turtle, crocodile turtle, yellow box turtle, platysternon tortoise and box turtle, slaughtering, cleaning, boning, mixing turtle meat according to the weight ratio of 1:1:1:1:1, mechanically crushing and ultrasonically crushing the mixed turtle meat into turtle meat paste, wherein the solid content is not more than 10 meshes.
(2) According to the method disclosed in patent CN104946488A, tortoise shell is taken to prepare tortoise shell extract.
(3) 2000g of minced turtle meat slurry and 400g of tortoise shell extract are uniformly mixed and homogenized, and then ultrapure water is used for preparing a 20% (W/V) substrate solution.
(4) Adding 0.8mg/mL compound protease into the substrate solution, carrying out enzymolysis for 20h at the constant temperature of 55 ℃, and adjusting the pH value to 5.5 +/-0.1.
(5) And (3) inactivating the enzyme of the obtained enzymolysis product, sterilizing, and sequentially filtering with ultrafiltration membranes with cut-off molecular weights of 9KD, 6KD and 3KD to obtain 4 turtle enzyme enzymolysis solutions with different molecular weights (larger than 9KD, 6-9 KD, 3-6 KD and smaller than 3 KD).
(6) Respectively carrying out vacuum freeze drying on the obtained tortoise peptidase hydrolyzed solution with each molecular weight segment to obtain 4 kinds of white to light yellow soluble powder with different molecular weights, namely a crude product of the peptide component;
(7) considering the principle that peptides with smaller molecular weight are easier to be absorbed by human body, the prepared 4 kinds of crude peptide components are uniformly mixed according to the weight ratio of 1:1:1:1 to obtain 584g of tortoise peptide as a whole tortoise component.
Example III,
(1) Taking live tortoise of stone small water turtle, Chinese grass turtle, crocodile turtle and yellow box turtle, slaughtering, cleaning, boning, mixing turtle meat according to the weight ratio of 1:1:1:1, mechanically crushing and ultrasonically crushing the turtle meat respectively to form turtle meat paste, wherein the solid content is not more than 10 meshes.
(2) According to the method disclosed in patent CN104946488A, tortoise shell is taken to prepare tortoise shell extract.
(3) 3000g of turtle meat paste and 1200g of turtle shell extract are uniformly mixed and homogenized, and then ultrapure water is used for preparing 35% (W/V) substrate solution.
(4) Adding 0.8mg/mL compound protease into the substrate solution, carrying out enzymolysis for 18h at a constant temperature of 50 ℃, and adjusting the pH value to 7.0 +/-0.1.
(5) And (3) inactivating the enzyme of the obtained enzymolysis product, sterilizing, and sequentially filtering with ultrafiltration membranes with cut-off molecular weights of 9KD, 6KD and 3KD to obtain 4 turtle enzyme enzymolysis solutions with different molecular weights (larger than 9KD, 6-9 KD, 3-6 KD and smaller than 3 KD).
(6) Respectively carrying out vacuum freeze drying on the obtained tortoise peptidase hydrolyzed solution with each molecular weight segment to obtain 4 kinds of white to light yellow soluble powder with different molecular weights, namely a crude product of the peptide component;
(7) considering the principle that peptides with smaller molecular weight are easier to be absorbed by human body, the crude 4 kinds of peptide components are mixed uniformly according to the weight ratio of 1:1:1:1 to obtain 916g of tortoise peptide.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. The production method of the turtle peptide is characterized by comprising the following steps:
1) taking different kinds of adult live turtles, slaughtering, cleaning and boning the adult live turtles, mixing the turtle meat according to the weight equal parts, mechanically crushing and ultrasonically crushing the turtle meat into turtle meat paste;
2) the weight ratio of 1: 1-10, uniformly mixing the tortoise shell extract and the tortoise meat paste, homogenizing, and preparing a substrate solution of 5-50% (W/V) by using ultrapure water;
3) adding 0.1-1.0 mg/mL protease into the substrate solution, carrying out enzymolysis under a constant temperature condition, and adjusting the pH value to 5-8;
4) and after enzyme deactivation and sterilization of the enzymolysis products, sequentially filtering the enzymolysis products by ultrafiltration membranes with cut-off molecular weights of 9KD, 6KD and 3KD to obtain 4 kinds of turtle enzyme enzymolysis liquid with different molecular weights, wherein the molecular weights are respectively as follows: greater than 9KD, 6-9 KD, 3-6 KD and less than 3 KD;
5) respectively carrying out vacuum freeze drying on the obtained tortoise peptidase hydrolyzed liquid with each molecular weight section to obtain 4 kinds of peptide component crude products with different molecular weights;
6) and uniformly mixing the prepared 4 kinds of peptide component crude products according to the weight ratio of 1:1:1:1 to obtain the turtle component turtle peptide.
2. The method for producing turtle peptide according to claim 1, wherein: the different kinds of adult live tortoise include three or more than 3 kinds of small water turtle, small stone water turtle, Chinese grass turtle, tortoise, crocodile tortoise, yellow box turtle, Suka tortoise, big head turtle, platysternon tortoise, box turtle, brocade tortoise or mud tortoise.
3. The method for producing turtle peptide according to claim 1, wherein: the solid content in the turtle meat paste is not more than 10 meshes.
4. The method for producing turtle peptide according to claim 1, wherein: the protease is one, two or more of trypsin, pepsin, papain, alkaline protease and neutral protease.
5. The method for producing turtle peptide according to claim 1, wherein: the constant temperature is 40-70 ℃.
6. The method for producing turtle peptide according to claim 1, wherein: the enzymolysis time is 1-24 h.
7. The method for producing turtle peptide according to claim 1, wherein: the enzyme deactivation process comprises the following steps: and (3) putting the reaction system into hot water of 90-100 ℃, and uniformly stirring for 5-30 min.
8. The method for producing turtle peptide according to claim 1, wherein: in the process of filtering the ultrafiltration membrane, carbon dioxide gas is adopted to protect a reaction system.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112931863A (en) * | 2021-03-25 | 2021-06-11 | 海南金龟岛生物科技有限公司 | Preparation method of tortoise-plastron glue |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974594A (en) * | 2010-11-11 | 2011-02-16 | 湖北远成药业有限公司 | Method for extracting tortoise-plastron peptide contained in tortoise-plastron glue through enzyme method |
| CN103397066A (en) * | 2013-07-29 | 2013-11-20 | 广西还珠海洋生物科技有限公司 | Method for extracting active peptides from Qinzhou yellow pond turtle |
| CN104531818A (en) * | 2015-01-21 | 2015-04-22 | 华南理工大学 | Preparation method of anti-tumor cuora trifasciata polypeptide |
| CN104673869A (en) * | 2015-03-31 | 2015-06-03 | 广西还珠海洋生物科技有限公司 | Method for extracting bioactive peptide from mauremys guangxiensis |
| CN104946488A (en) * | 2015-07-21 | 2015-09-30 | 海南金龟园实业发展有限公司 | Nutritious tortoise liquor and making method thereof |
| CN104962597A (en) * | 2015-07-09 | 2015-10-07 | 严宜昌 | Method for extracting collagen polypeptide from tortoise shells and application of collagen polypeptide in preparation of sleeping mask |
| CN105132503A (en) * | 2015-09-22 | 2015-12-09 | 广西北部湾海皇生物科技有限公司 | Refining process of pond turtle bioactive peptides and application of pond turtle bioactive peptides in preventing liver cirrhosis and resisting fatigue |
-
2021
- 2021-03-25 CN CN202110317049.XA patent/CN112941137A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974594A (en) * | 2010-11-11 | 2011-02-16 | 湖北远成药业有限公司 | Method for extracting tortoise-plastron peptide contained in tortoise-plastron glue through enzyme method |
| CN103397066A (en) * | 2013-07-29 | 2013-11-20 | 广西还珠海洋生物科技有限公司 | Method for extracting active peptides from Qinzhou yellow pond turtle |
| CN104531818A (en) * | 2015-01-21 | 2015-04-22 | 华南理工大学 | Preparation method of anti-tumor cuora trifasciata polypeptide |
| CN104673869A (en) * | 2015-03-31 | 2015-06-03 | 广西还珠海洋生物科技有限公司 | Method for extracting bioactive peptide from mauremys guangxiensis |
| CN104962597A (en) * | 2015-07-09 | 2015-10-07 | 严宜昌 | Method for extracting collagen polypeptide from tortoise shells and application of collagen polypeptide in preparation of sleeping mask |
| CN104946488A (en) * | 2015-07-21 | 2015-09-30 | 海南金龟园实业发展有限公司 | Nutritious tortoise liquor and making method thereof |
| CN105132503A (en) * | 2015-09-22 | 2015-12-09 | 广西北部湾海皇生物科技有限公司 | Refining process of pond turtle bioactive peptides and application of pond turtle bioactive peptides in preventing liver cirrhosis and resisting fatigue |
Non-Patent Citations (2)
| Title |
|---|
| 何胜洁等: "金钱龟酶解多肽的分离、纯化及抗肿瘤活性研究", 《现代食品科技》 * |
| 张文芳: "龟及其药用价值", 《中国药房》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112931863A (en) * | 2021-03-25 | 2021-06-11 | 海南金龟岛生物科技有限公司 | Preparation method of tortoise-plastron glue |
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