CN111944866A - Method for preparing yak hide small molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis - Google Patents
Method for preparing yak hide small molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis Download PDFInfo
- Publication number
- CN111944866A CN111944866A CN202010775142.0A CN202010775142A CN111944866A CN 111944866 A CN111944866 A CN 111944866A CN 202010775142 A CN202010775142 A CN 202010775142A CN 111944866 A CN111944866 A CN 111944866A
- Authority
- CN
- China
- Prior art keywords
- enzymolysis
- yak
- desolventizing
- collagen peptide
- rotary evaporation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
Abstract
The invention discloses a method for preparing yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis, which comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloring, deodorization and desalination, and (6) concentration and drying, and is characterized in that the enzymolysis in the step (3) comprises the following steps: carrying out first enzymolysis, ultramicro-refining treatment and carrying out second enzymolysis. According to the invention, through the combination of double enzymolysis and continuous rotary evaporation desolventizing, the enzymolysis time can be greatly shortened, the process efficiency is improved, and the prepared yak skin collagen peptide has high purity, is colorless and tasteless, has a small molecular weight, and is beneficial to direct absorption by a human body.
Description
Technical Field
The invention belongs to the technical field of collagen peptide extraction, and particularly relates to a method for preparing yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis.
Background
The Qinghai is the biggest yak production base in China, accounts for more than one third of the total number of yaks in China, and is popular among the yaks in the world. The unique plateau environment enables the yak to have extremely strong immunity, strong ultraviolet irradiation enables the yak to become a treasure house of sterile resources, and a natural stocking mode enables the yak to have unique nutritional value and reliable safety. Therefore, yaks called world ridges are excellent raw materials for extracting collagen peptides, the utilization value of yak skin is further increased, and the problem of slaughter wastes is reduced.
In the existing skin extraction raw materials, deep-sea fishes are mainly used, collagen peptides prepared from deep-sea fish skins are always the mainstream in the market, and particularly, the collagen peptides imported from deep-sea cod skins are popular among the public. Compared with fish skin collagen peptide, most of cow skin collagen peptide on the current market has yellow color and luster, has light fishy smell of cow leather, has certain influence on edible taste, and also influences the quality of a terminal product as an addition raw material. Because the yield of collagen peptide prepared by land vertebrates is lower than that of marine animals, and meanwhile, a large amount of unhydrolyzed debris is generated in the enzymolysis process to increase the filtration burden, the cost for producing small molecular peptide which is beneficial to human body absorption is quite high, so that many places for producing the bovine skin collagen peptide are not improved. Therefore, a more environment-friendly, low-cost and high-quality method for extracting bovine collagen peptide with high efficiency is needed to be developed, and the utilization value of the Qinghai yak skin and the market position of the bovine collagen peptide in China are enhanced.
Disclosure of Invention
In order to solve the defects in the content, the invention aims to provide a method for preparing yak skin small-molecule collagen peptide by continuous rotary evaporation desolventizing and double enzymolysis.
The technical scheme of the invention is as follows.
A method for preparing yak hide small molecule collagen peptide by continuous rotary steaming, desolventizing and double enzymolysis comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloration, deodorization and desalination, (6) concentration and drying, and is characterized in that the enzymolysis in the step (3) comprises the following steps:
carrying out first enzymolysis: adding pepsin into the yak skin slurry obtained by homogenizing in the step (2), and then performing rotary evaporation, desolventizing and enzymolysis for 4-6 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 15-60 r/min and the temperature of 35-40 ℃ to obtain a first enzymolysis liquid;
ultra-fine treatment: inactivating enzyme of the first enzymolysis liquid, sieving with 200 mesh sieve, collecting undersize liquid, and micronizing with nanometer collider to obtain nanosized enzymolysis liquid with average particle size of less than 500 nm;
and (3) carrying out second enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to a proper range, adding a composite enzyme solution of alkaline protease and serine protease, and then performing rotary evaporation desolventizing enzymolysis for 2-3 hours at a vacuum degree of 0.85-0.98 MPa, a rotating speed of 15-60 r/min and a temperature of 45-55 ℃; the serine protease is any one of trypsin, subtilisin and elastase;
in the steps, the substrate concentration in the enzymolysis process can be maintained at a higher level by performing rotary vacuum desolventizing during enzymolysis, so that the overall enzymolysis rate is favorably improved, and the enzymolysis time is greatly shortened; in addition, the enzymolysis liquid is processed by a nano collider, so that the average particle size of the enzymolysis liquid containing the collagen reaches the nano level, binding sites of enzyme and a substrate can be increased, and the enzymolysis speed is improved.
As a preferred technical scheme, when the first enzymolysis and the second enzymolysis in the step (3) are carried out by rotary evaporation, desolventizing and enzymolysis, ultrasonic waves are applied simultaneously.
As a specific technical scheme, the yak skin pretreatment in the step (1) specifically comprises the following steps: weighing fresh yak skin, unhairing, chopping, adding 3-5% sodium carbonate into the weighed fresh yak skin according to the mass ratio of the feed liquid to 1: 3-7 kg/kg at room temperature, soaking for 6-8 h, washing the yak skin to be neutral by clear water after the soaking is finished, adding the yak skin into 3-5% salt according to the mass ratio of the feed liquid to 1: 3-7 kg/kg, soaking for 6-8 h again, and washing the yak skin to be neutral by clear water after the soaking is finished.
As a specific technical scheme, the homogenate in the step (2) is specifically as follows: adding 0.05M acetic acid solution into the pretreated yak skin according to the mass ratio of 1: 4-8 kg/kg of the feed liquid, and fully homogenizing by using a colloid mill to obtain yak skin homogenate.
As a specific technical scheme, the centrifugal separation in the step (4) is specifically as follows: and (4) boiling the enzymatic hydrolysate obtained in the step (3) at high temperature to inactivate enzymes and sterilize, centrifuging at 8000-12000 rpm to remove residues, and performing microfiltration to obtain the collagen peptidase hydrolysate.
As a specific technical scheme, the decoloring, deodorization and desalting in the step (5) specifically comprises the following steps: and (3) adding 0.1-0.3% m/v of activated carbon into the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (4), stirring for 2-3 h at room temperature at 40-80 r/min, then performing suction filtration to remove the activated carbon, and further performing decoloration, desalination and deodorization through an ion exchange resin column.
As a specific technical scheme, the yak skin in the step (1) is Qinghai Chaaida wood yak skin in a basin.
According to the specific technical scheme, the compound enzyme solution is composed of elastase and alkaline protease, the elastase is introduced in the second enzymolysis to further degrade collagen fibers and collagen to swell, the synergistic effect is achieved in the enzymolysis process of the alkaline protease, the second enzymolysis is greatly promoted, and the yak skin is sufficiently hydrolyzed.
As a specific technical scheme, the pepsin used in the first enzymolysis in the step (3) is food-grade protease extracted from animal mucosa, and the enzyme activity is 10 ten thousand; the alkaline protease used in the second enzymolysis in the step (3) is food-grade protease extracted from bacillus licheniformis, the enzyme activity is 20 ten thousand, the elastase is food-grade protease extracted from pancreas, the enzyme activity is 10 ten thousand, and the optimal enzymolysis effect is achieved by the cooperation of the two preferable enzymes.
As a specific technical scheme, the active carbon is 200-mesh wood active carbon, and the ion exchange resin adopts strong-acid styrene cation exchange resin and macroporous acrylic weak-base anion exchange resin.
As a specific technical scheme, in the yak skin collagen peptide prepared by the method, the content of collagen peptide with the molecular weight of less than 3000Da is more than 95%, the content of oligopeptide with the molecular weight of less than 2000Da is more than 90%, and the content of oligopeptide with the molecular weight of less than 1000Da is more than 70%.
The invention has the beneficial effects that:
1. the invention adopts fresh yak hide in Qinghai Chailada basin as raw material, has natural material advantages, and greatly reduces the influence of fat and foreign protein on products and subsequent processes by adopting the pretreatment of sodium carbonate and salt.
2. According to the invention, a colloid mill is selected for carrying out homogenization treatment on yak skin, the cow skin is fully crushed, and the double enzymolysis technology is added, so that only acetic acid and NaOH which maintain the enzymolysis pH environment are contained in the hydrolysis process, a specific buffer system is not needed, after pepsin extracts collagen, elastase is introduced in the second enzymolysis for further degrading collagen fibers and expanding collagen, a synergistic effect is achieved in the enzymolysis process of alkaline protease, the enzymolysis of the alkaline protease is greatly promoted, the yak skin is fully hydrolyzed, only trace crushed cow hair is left in the residual residues, and the filtration burden is greatly reduced. In addition, the invention combines double enzymolysis and continuous rotary evaporation desolventizing, thereby greatly shortening the enzymolysis time and improving the process efficiency.
3. The invention adopts the double enzymolysis of pepsin and compound enzyme liquid without ultrafiltration separation, and the prepared yak skin collagen peptide has the content of collagen peptide with the molecular weight of less than 3000Da of more than 95 percent, the content of oligopeptide with the molecular weight of less than 2000Da of more than 90 percent and the content of oligopeptide with the molecular weight of less than 1000Da of more than 70 percent, so that the product is completely beneficial to the direct absorption of a human body, the production cost is greatly reduced, and the production process is simplified.
4. The invention adopts 200-mesh woody activated carbon and anion-cation resin to combine to achieve the best effects of decoloring, desalting and deodorizing, can enhance the quality of yak skin collagen peptide and has better taste.
5. The yak skin collagen peptide obtained by the invention has high purity, is colorless and tasteless, has obvious advantages compared with most of yak skin collagen peptide products in the market, can be widely applied to foods, health products and cosmetics, and does not influence the quality of terminal products.
Drawings
Fig. 1 is a 12% SDS-PAGE gel electrophoresis analysis diagram of yak skin collagen peptide prepared by the method of example 3, lane 1 is a collagen sample extracted by pepsin in the first enzymolysis, lane 2 is a yak skin collagen peptide sample after composite enzyme liquid enzymolysis in the second enzymolysis;
FIG. 2 is a size exclusion chromatogram of yak skin collagen peptide prepared by the method of example 3 and molecular weight distribution of collagen peptide, wherein a single peak in the chromatogram is retention time of standard peptide;
FIG. 3 is a mass spectrum of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) of yak skin collagen peptide prepared by the method of example 3.
Detailed Description
The present invention will be further described with reference to the following detailed description, which should be construed as illustrative only, and not limiting the scope of the invention, which is to be given the full breadth of the appended claims, and all changes that can be made by those skilled in the art and which are, therefore, intended to be embraced therein.
Example 1
A method for preparing yak hide small molecule collagen peptide by continuous rotary steaming, desolventizing and double enzymolysis comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloration, deodorization and desalination, (6) concentration and drying, and the specific steps are as follows:
(1) pretreatment: weighing 50g of fresh yak skin, unhairing, cutting into blocks of 1 × 1cm, adding soda with the mass concentration of 5% into the blocks according to the feed liquid mass ratio of 1:5kg/kg at room temperature, soaking for 8 hours, washing the yak skin to be neutral by clear water after soaking is finished, then adding the yak skin into salt with the mass concentration of 5% according to the feed liquid mass ratio of 1:5kg/kg, soaking for 8 hours again, and washing the yak skin to be neutral by clear water after soaking is finished so as to achieve the purposes of degreasing and impurity removal;
(2) homogenizing: adding 0.05M acetic acid solution into the pretreated yak skin according to the mass ratio of 1:5kg/kg of the feed liquid, and fully homogenizing by using a colloid mill to obtain yak skin homogenate;
(3) enzymolysis:
carrying out first enzymolysis: adding 1.5g of pepsin with 10 ten thousand of enzyme activity into the yak skin slurry obtained by homogenizing in the step (2), transferring the mixture into rotary evaporation equipment, and performing rotary evaporation desolventizing enzymolysis for 6 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 20r/min and the temperature of 35-40 ℃ to obtain a first enzymolysis liquid;
ultra-fine treatment: inactivating enzyme of the first enzymolysis liquid, sieving with 200 mesh sieve, collecting undersize liquid, and micronizing with nanometer collider to obtain nanosized enzymolysis liquid with average particle size of less than 500 nm;
and (3) carrying out second enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to 9 by adopting NaOH, adding 0.3g of alkaline protease with 20 ten thousand of enzyme activity and 0.03g of elastase with 10 ten thousand of enzyme activity, transferring the obtained mixture into rotary evaporation equipment, and performing rotary evaporation and desolventizing enzymolysis for 3 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 60r/min and the temperature of 50 ℃;
(4) centrifugal separation: and (4) boiling the enzymolysis liquid obtained in the step (3) at high temperature for 30min for enzyme deactivation and sterilization, centrifuging at 10000rpm to remove residues, and performing microfiltration to obtain the collagen peptidase enzymolysis liquid.
(5) Decoloring, deodorizing and desalting: and (4) adding 1.5g of wood activated carbon of 200 meshes into the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (4), stirring for 2-3 h at room temperature at 60r/min, then performing suction filtration to remove the activated carbon, and further performing decoloration, desalination and deodorization through an ion exchange resin column.
(6) Concentrating and drying: and (3) concentrating the enzymolysis liquid treated in the last step by falling film, and then carrying out spray drying to prepare colorless and tasteless yak skin collagen peptide, wherein the final yield of the yak skin collagen peptide is 15.6%.
Example 2
A method for preparing yak hide small molecule collagen peptide by continuous rotary steaming, desolventizing and double enzymolysis comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloration, deodorization and desalination, (6) concentration and drying, and the specific steps are as follows:
(1) pretreatment: weighing 500g of fresh yak skin, unhairing, cutting into blocks of 1 × 1cm, adding soda with the mass concentration of 3% into the blocks according to the feed liquid mass ratio of 1:5kg/kg at room temperature, soaking for 8 hours, washing the yak skin to be neutral by clear water after soaking is finished, then adding the yak skin into salt with the mass concentration of 3% according to the feed liquid mass ratio of 1:5kg/kg, soaking for 8 hours again, and washing the yak skin to be neutral by clear water after soaking is finished so as to achieve the purposes of degreasing and impurity removal;
(2) homogenizing: adding 0.05M acetic acid solution into the pretreated yak skin according to the mass ratio of 1:5kg/kg of the feed liquid, and fully homogenizing by using a colloid mill to obtain yak skin homogenate;
(3) enzymolysis:
carrying out first enzymolysis: adding 15g of pepsin with 10 ten thousand of enzyme activity into the yak skin slurry obtained by homogenizing in the step (2), transferring the mixture into rotary evaporation equipment, and performing rotary evaporation desolventizing enzymolysis for 5 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 30r/min and the temperature of 35-40 ℃ to obtain a first enzymolysis liquid;
ultra-fine treatment: inactivating enzyme of the first enzymolysis liquid, sieving with 200 mesh sieve, collecting undersize liquid, and micronizing with nanometer collider to obtain nanosized enzymolysis liquid with average particle size of less than 500 nm;
and (3) carrying out second enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to 9 by adopting NaOH, adding 3g of alkaline protease with 20 ten thousand of enzyme activity and 0.3g of elastase with 10 ten thousand of enzyme activity, transferring the obtained mixture into rotary evaporation equipment, and carrying out rotary evaporation desolventizing enzymolysis for 2.5 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 40r/min and the temperature of 50 ℃;
(4) centrifugal separation: boiling the enzymolysis liquid obtained in the step (3) at high temperature for 30min for enzyme deactivation and sterilization, centrifuging at 10000rpm to remove residues, and performing microfiltration to obtain collagen peptidase enzymolysis liquid;
(5) decoloring, deodorizing and desalting: adding 1.5g of wood active carbon of 200 meshes into the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (4), stirring for 2-3 h at room temperature at 60r/min, then removing the active carbon by suction filtration, and further decoloring, desalting and deodorizing by a styrene cation exchange resin and a macroporous acrylic acid weak base anion exchange resin column which are filled in advance;
(6) concentrating and drying: and (3) concentrating the enzymolysis liquid treated in the last step by falling film, and then carrying out spray drying to prepare colorless and tasteless yak skin collagen peptide, wherein the final yield of the yak skin collagen peptide is 15.2%.
Example 3
A method for preparing yak hide small molecule collagen peptide by continuous rotary steaming, desolventizing and double enzymolysis comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloration, deodorization and desalination, (6) concentration and drying, and the specific steps are as follows:
(1) pretreatment: weighing 20kg of fresh yak skin, unhairing, cutting into blocks of 1 × 1cm, adding soda with the mass concentration of 5% into the blocks according to the feed liquid mass ratio of 1:5kg/kg at room temperature, soaking for 8 hours, washing the yak skin to be neutral by clear water after soaking is finished, then adding the yak skin into salt with the mass concentration of 5% according to the feed liquid mass ratio of 1:5kg/kg, soaking for 8 hours again, and washing the yak skin to be neutral by clear water after soaking is finished so as to achieve the purposes of degreasing and impurity removal;
(2) homogenizing: adding 0.05M acetic acid solution into the pretreated yak skin according to the mass ratio of 1:5kg/kg of the feed liquid, and fully homogenizing by using a colloid mill to obtain yak skin homogenate;
(3) enzymolysis:
carrying out first enzymolysis: adding 0.6kg of pepsin with 10 ten thousand of enzyme activity into the yak skin slurry obtained by homogenizing in the step (2), transferring the mixture into rotary evaporation equipment, and performing rotary evaporation desolventizing enzymolysis for 4 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 20r/min and the temperature of 35-40 ℃ to obtain a first enzymolysis liquid;
ultra-fine treatment: inactivating enzyme of the first enzymolysis liquid, sieving with 200 mesh sieve, collecting undersize liquid, and micronizing with nanometer collider to obtain nanosized enzymolysis liquid with average particle size of less than 500 nm;
and (3) carrying out second enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to 9 by adopting NaOH, adding 0.12kg of alkaline protease with 20 ten thousand of enzyme activity and 0.012kg of elastase with 10 ten thousand of enzyme activity, transferring the obtained mixture into rotary evaporation equipment, and performing rotary evaporation desolventizing enzymolysis for 2 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 15-60 r/min and the temperature of 50 ℃;
(4) centrifugal separation: and (4) boiling the enzymolysis liquid obtained in the step (3) at high temperature for 30min for enzyme deactivation and sterilization, centrifuging at 10000rpm to remove residues, and performing microfiltration to obtain the collagen peptidase enzymolysis liquid.
(5) Decoloring, deodorizing and desalting: and (3) adding 0.1-0.3% m/v of wood activated carbon (200 meshes) into the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (4), stirring for 2-3 h at room temperature at 60r/min, then performing suction filtration to remove the activated carbon, and further performing decoloration, desalination and deodorization through an ion exchange resin column.
(6) Concentrating and drying: and (3) concentrating the enzymolysis liquid treated in the last step by falling film, and then carrying out spray drying to prepare colorless and tasteless yak skin collagen peptide, wherein the final yield of the yak skin collagen peptide is 13.2%.
Example 4
The characteristic analysis of the collagen peptide is carried out by taking the yak skin collagen peptide prepared by the method in the embodiment 3 of the invention as a sample.
1) As shown in fig. 1, the yak skin collagen extracted by the first enzymolysis, i.e., pepsin enzymolysis, in lane 1 is collagen with a complete triple-helical structure; in lane 2, collagen is further decomposed by the second enzymolysis, i.e., the enzymolysis of the complex enzyme solution, to obtain collagen peptide.
2) As can be seen from FIG. 2, according to the detection method of molecular weight distribution in the national food safety standard GB/T22729-. In addition, in the yak skin collagen peptide prepared by the preparation method, the content of the collagen peptide with the molecular weight of less than 3000Da is more than 95%, the content of the oligopeptide with the molecular weight of less than 2000Da is more than 90%, and the content of the oligopeptide with the molecular weight of less than 1000Da is more than 70%, so that the prepared product is beneficial to direct absorption of a human body.
3) As shown in FIG. 3, the molecular weight of the yak skin collagen peptide prepared in example 3 is mainly concentrated below 3000Da as shown by the identification result of MALDI-TOF-MS mass spectrum.
Claims (10)
1. A method for preparing yak hide small molecule collagen peptide by continuous rotary steaming, desolventizing and double enzymolysis comprises the steps of (1) yak hide pretreatment, (2) homogenate, (3) enzymolysis, (4) centrifugal separation, (5) decoloration, deodorization and desalination, (6) concentration and drying, and is characterized in that the enzymolysis in the step (3) comprises the following steps:
carrying out first enzymolysis: adding pepsin into the yak skin slurry obtained by homogenizing in the step (2), and then performing rotary evaporation, desolventizing and enzymolysis for 4-6 hours at the vacuum degree of 0.85-0.98 MPa, the rotating speed of 15-60 r/min and the temperature of 35-40 ℃ to obtain a first enzymolysis liquid;
ultra-fine treatment: inactivating enzyme of the first enzymolysis liquid, sieving with 200 mesh sieve, collecting undersize liquid, and micronizing with nanometer collider to obtain nanosized enzymolysis liquid with average particle size of less than 500 nm;
and (3) carrying out second enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to a proper range, adding a composite enzyme solution of alkaline protease and serine protease, and then performing rotary evaporation desolventizing enzymolysis for 2-3 hours at a vacuum degree of 0.85-0.98 MPa, a rotating speed of 15-60 r/min and a temperature of 45-55 ℃; the serine protease is any one of trypsin, subtilisin and elastase.
2. The method for preparing yak hide small molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis as claimed in claim 1, wherein the yak hide pretreatment in the step (1) specifically comprises: weighing fresh yak skin, unhairing, chopping, adding 3-5% sodium carbonate into the weighed fresh yak skin according to the mass ratio of the feed liquid to 1: 3-7 kg/kg at room temperature, soaking for 6-8 h, washing the yak skin to be neutral by clear water after the soaking is finished, adding the yak skin into 3-5% salt according to the mass ratio of the feed liquid to 1: 3-7 kg/kg, soaking for 6-8 h again, and washing the yak skin to be neutral by clear water after the soaking is finished.
3. The method for preparing yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to claim 1, wherein the homogenate in the step (2) is specifically as follows: adding 0.05M acetic acid solution into the pretreated yak skin according to the mass ratio of 1: 4-8 kg/kg of the feed liquid, and fully homogenizing by using a colloid mill to obtain yak skin homogenate.
4. The method for preparing yak hide small molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis as claimed in claim 1, wherein the centrifugal separation in the step (4) is specifically: and (4) boiling the enzymatic hydrolysate obtained in the step (3) at high temperature to inactivate enzymes and sterilize, centrifuging at 8000-12000 rpm to remove residues, and performing microfiltration to obtain the collagen peptidase hydrolysate.
5. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis as claimed in claim 1, wherein the decoloring, deodorization and desalting in the step (5) specifically comprises the following steps: and (3) adding 0.1-0.3% m/v of activated carbon into the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (4), stirring for 2-3 h at room temperature at 40-80 r/min, then performing suction filtration to remove the activated carbon, and further performing decoloration, desalination and deodorization through an ion exchange resin column.
6. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to claim 1, which is characterized by comprising the following steps: the yak skin in the step (1) is Qinghai Chaaida wood yak skin in a basin.
7. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to claim 1, which is characterized by comprising the following steps: in the step (3), the compound enzyme solution is elastase and alkaline protease.
8. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to claim 7, which is characterized in that: the pepsin used in the first enzymolysis in the step (3) is food-grade protease extracted from animal mucosa, and the enzyme activity is 10 ten thousand; the alkaline protease used in the second enzymolysis in the step (3) is food-grade protease extracted from bacillus licheniformis, the enzyme activity is 20 ten thousand, the elastase is food-grade protease extracted from pancreas, and the enzyme activity is 10 ten thousand.
9. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to claim 5, which is characterized by comprising the following steps: the active carbon is 200-mesh wood active carbon, and the ion exchange resin adopts strong-acid styrene cation exchange resin and macroporous acrylic weak base anion exchange resin.
10. The method for preparing the yak hide small-molecule collagen peptide by continuous rotary evaporation, desolventizing and double enzymolysis according to any one of claims 1 to 9, which is characterized by comprising the following steps: in the yak skin collagen peptide prepared by the method, the content of collagen peptide with the molecular weight of less than 3000Da is more than 95%, the content of oligopeptide with the molecular weight of less than 2000Da is more than 90%, and the content of oligopeptide with the molecular weight of less than 1000Da is more than 70%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010775142.0A CN111944866B (en) | 2020-07-31 | 2020-07-31 | Method for preparing small molecular collagen peptide of yak leather by continuous rotary steaming desolventizing double enzymolysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010775142.0A CN111944866B (en) | 2020-07-31 | 2020-07-31 | Method for preparing small molecular collagen peptide of yak leather by continuous rotary steaming desolventizing double enzymolysis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111944866A true CN111944866A (en) | 2020-11-17 |
| CN111944866B CN111944866B (en) | 2023-06-20 |
Family
ID=73339494
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010775142.0A Active CN111944866B (en) | 2020-07-31 | 2020-07-31 | Method for preparing small molecular collagen peptide of yak leather by continuous rotary steaming desolventizing double enzymolysis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111944866B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574294A (en) * | 2020-12-30 | 2021-03-30 | 青海瑞肽生物科技有限公司 | Collagen peptide and preparation method and application thereof |
| CN112569347A (en) * | 2020-12-30 | 2021-03-30 | 青海瑞肽生物科技有限公司 | Application of yak collagen peptide in hypoglycemic drugs or hypoglycemic health foods |
| CN113105823A (en) * | 2021-03-03 | 2021-07-13 | 清远市浩宇化工科技有限公司 | Polyurethane finish paint and preparation method and application thereof |
| CN113862321A (en) * | 2021-10-11 | 2021-12-31 | 齐鲁工业大学 | Method for extracting nutritional ingredients from cattle hair or cattle hide leftover |
| CN117736307A (en) * | 2024-02-19 | 2024-03-22 | 北京盛美诺生物技术有限公司 | III type small molecule collagen peptide capable of promoting III type and IV type collagen secretion and preparation method thereof |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008074846A (en) * | 2006-08-25 | 2008-04-03 | Nippon Meat Packers Inc | Elastin decomposition peptide and method for preparing elastin and its zymolytic peptide |
| CN102212107A (en) * | 2011-04-12 | 2011-10-12 | 江南大学 | Rice protein polypeptide and preparation method thereof |
| CN105695548A (en) * | 2016-03-30 | 2016-06-22 | 蔡庭守 | Preparation method of donkey-hide gelatin small molecular peptide |
| CN106011208A (en) * | 2016-06-29 | 2016-10-12 | 肖建喜 | Method for preparing small-molecular weight collagen active peptide through enzymolysis of yak bone and skin |
| WO2018014841A1 (en) * | 2016-07-19 | 2018-01-25 | 华南生物医药研究院 | Self-assembled collagen and preparation method therefor |
| CN107653291A (en) * | 2017-11-15 | 2018-02-02 | 西藏央金生态农牧科技有限公司 | The standby method for hiding Yak-skin Gelatin original albumen and collagen polypeptide of multi-step enzyme method coordinate system |
| CN109295145A (en) * | 2018-10-31 | 2019-02-01 | 西藏央金生态农牧科技有限公司 | A kind of preparation method of Ultra-low molecular weight yak collagen peptide |
| CN110229862A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of Yak-skin Gelatin preparation method promoting platelet activation |
| CN110592053A (en) * | 2019-09-29 | 2019-12-20 | 河南新仰韶生物科技有限公司 | Collagen hydrolysis complex enzyme, protein oligopeptide and preparation method thereof |
| CN110628857A (en) * | 2019-10-29 | 2019-12-31 | 南宁学院 | Method for extracting collagen from cow leather |
| CN111363772A (en) * | 2020-04-08 | 2020-07-03 | 平凉市华科生物技术有限公司 | Method for preparing collagen peptide by hydrolyzing bovine bone and collagen peptide thereof |
-
2020
- 2020-07-31 CN CN202010775142.0A patent/CN111944866B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008074846A (en) * | 2006-08-25 | 2008-04-03 | Nippon Meat Packers Inc | Elastin decomposition peptide and method for preparing elastin and its zymolytic peptide |
| CN102212107A (en) * | 2011-04-12 | 2011-10-12 | 江南大学 | Rice protein polypeptide and preparation method thereof |
| CN105695548A (en) * | 2016-03-30 | 2016-06-22 | 蔡庭守 | Preparation method of donkey-hide gelatin small molecular peptide |
| CN106011208A (en) * | 2016-06-29 | 2016-10-12 | 肖建喜 | Method for preparing small-molecular weight collagen active peptide through enzymolysis of yak bone and skin |
| WO2018014841A1 (en) * | 2016-07-19 | 2018-01-25 | 华南生物医药研究院 | Self-assembled collagen and preparation method therefor |
| CN107653291A (en) * | 2017-11-15 | 2018-02-02 | 西藏央金生态农牧科技有限公司 | The standby method for hiding Yak-skin Gelatin original albumen and collagen polypeptide of multi-step enzyme method coordinate system |
| CN109295145A (en) * | 2018-10-31 | 2019-02-01 | 西藏央金生态农牧科技有限公司 | A kind of preparation method of Ultra-low molecular weight yak collagen peptide |
| CN110229862A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of Yak-skin Gelatin preparation method promoting platelet activation |
| CN110592053A (en) * | 2019-09-29 | 2019-12-20 | 河南新仰韶生物科技有限公司 | Collagen hydrolysis complex enzyme, protein oligopeptide and preparation method thereof |
| CN110628857A (en) * | 2019-10-29 | 2019-12-31 | 南宁学院 | Method for extracting collagen from cow leather |
| CN111363772A (en) * | 2020-04-08 | 2020-07-03 | 平凉市华科生物技术有限公司 | Method for preparing collagen peptide by hydrolyzing bovine bone and collagen peptide thereof |
Non-Patent Citations (3)
| Title |
|---|
| HONGDONG SONG等: "Effect of Orally Administered Collagen Peptides from Bovine Bone on Skin Aging in Chronologically Aged Mice", 《NUTRIENTS》 * |
| 杨远帆等: "弹性蛋白酶提取牛蛙皮胶原蛋白的工艺优化", 《中国食品学报》 * |
| 陈渭: "碱性蛋白酶提取牦牛皮胶原蛋白的工艺研究", 《青海大学学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574294A (en) * | 2020-12-30 | 2021-03-30 | 青海瑞肽生物科技有限公司 | Collagen peptide and preparation method and application thereof |
| CN112569347A (en) * | 2020-12-30 | 2021-03-30 | 青海瑞肽生物科技有限公司 | Application of yak collagen peptide in hypoglycemic drugs or hypoglycemic health foods |
| CN112569347B (en) * | 2020-12-30 | 2023-09-22 | 青海瑞肽生物科技有限公司 | Application of yak collagen peptide in hypoglycemic drugs or hypoglycemic health-care foods |
| CN113105823A (en) * | 2021-03-03 | 2021-07-13 | 清远市浩宇化工科技有限公司 | Polyurethane finish paint and preparation method and application thereof |
| CN113105823B (en) * | 2021-03-03 | 2022-06-21 | 清远市浩宇化工科技有限公司 | Polyurethane finish paint and preparation method and application thereof |
| CN113862321A (en) * | 2021-10-11 | 2021-12-31 | 齐鲁工业大学 | Method for extracting nutritional ingredients from cattle hair or cattle hide leftover |
| CN117736307A (en) * | 2024-02-19 | 2024-03-22 | 北京盛美诺生物技术有限公司 | III type small molecule collagen peptide capable of promoting III type and IV type collagen secretion and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111944866B (en) | 2023-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111944866B (en) | Method for preparing small molecular collagen peptide of yak leather by continuous rotary steaming desolventizing double enzymolysis | |
| US9974321B2 (en) | Fish protein oligopeptide with low allergenicity and slight fishiness and industrial preparation method and application thereof | |
| CN103993061B (en) | A kind of production method of Isin glue collagen | |
| CN112062834B (en) | Deep sea fish skin collagen peptide and extraction and preparation method thereof | |
| CN106967169B (en) | Extraction method of fish collagen | |
| CN102174627A (en) | Method for preparing rapeseed bioactive peptide | |
| CN112608968B (en) | Method for producing fish collagen peptide by using tilapia mossambica scale as raw material | |
| CN1478421A (en) | Seacucumbus whole powder food and its preparation method | |
| CN104480168A (en) | Method for preparing chondroitin sulfate and co-producing collagen peptide and biological calcium by using fishbone | |
| CN101869169B (en) | Method for preparing fish oligopeptide from gurry by combining fermentation and membrane technology | |
| CN110564802A (en) | Extraction method of yak achilles tendon bone collagen | |
| CN110699411A (en) | Preparation method of eggshell membrane polypeptide | |
| CN106906199A (en) | A kind of fast purifying squid viscera albumen enzyme method and application | |
| CN108728507A (en) | The preparation method of fishskin collagen polypeptide | |
| CN111493205A (en) | Fish protein hydrolysate and preparation method thereof | |
| CN110577975B (en) | Method for extracting and preparing swimming bladder collagen oligopeptide | |
| CN104450845A (en) | Method for preparing sea cucumber polypeptide from sea cucumber endogenous protease | |
| CN105039481A (en) | Rice bran polypeptide preparation method for improving polypeptide quality | |
| CN113563459A (en) | Protein extraction process in production process of collagen peptide | |
| CN105567772B (en) | A kind of high antioxidant protein peptides and the preparation method and application thereof | |
| CN109439716B (en) | Preparation method of silver carp protein peptide | |
| CN107089869A (en) | A kind of preparation method of the molten slurry Protein fertilizer of fish | |
| JP2870871B2 (en) | A method for treating crustacean shells using enzymes | |
| CN113136409B (en) | Preparation method of food-grade low-salinity ocean fish oligopeptide | |
| CN108277247A (en) | The method that antioxidant activity polypeptide is extracted from squid spawn tangled gland |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |