CN111575332A - Preparation method of cerebroprotein hydrolysate - Google Patents
Preparation method of cerebroprotein hydrolysate Download PDFInfo
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- CN111575332A CN111575332A CN202010587502.4A CN202010587502A CN111575332A CN 111575332 A CN111575332 A CN 111575332A CN 202010587502 A CN202010587502 A CN 202010587502A CN 111575332 A CN111575332 A CN 111575332A
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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
- C07K1/30—Extraction; Separation; Purification by precipitation
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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
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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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
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
Abstract
The invention relates to a preparation method of a cerebroprotein hydrolysate, which comprises the following steps: homogenizing brain tissue to obtain homogenate; carrying out enzymolysis treatment on the homogenate to obtain an enzymolysis liquid; adding a settling agent into the enzymatic hydrolysate, standing for layering, removing a grease layer on the surface, and collecting a settling supernatant; adding a flocculating agent into the settled supernatant, performing centrifugal separation, and collecting the centrifugal supernatant; adding activated carbon into the centrifugal supernatant, preserving the heat at 50-70 ℃ for 10-30 min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate. The preparation method of the invention does not use organic solvent, has simple and feasible process and low energy consumption, considers the treatment of waste residue, is very environment-friendly, accords with the concept of green production, and is convenient for workers to operate and popularize for large-scale industrial production.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a preparation method of a cerebroprotein hydrolysate.
Background
The cerebroprotein hydrolysate is a mixture mainly containing polypeptides, various amino acids and trace elements such as selenium, potassium, phosphorus, magnesium and the like, which is obtained by hydrolyzing animal brain tissues. It can act on human nervous tissue in various ways, and can provide amino acids for neuron repair, nourish nerve cells, regulate and improve neuron metabolism, promote synapse formation, induce neuron differentiation, and further protect nerve cells from damage caused by ischemia and neurotoxin. In clinic, the traditional Chinese medicine composition can be applied to the treatment of acute cerebral infarction, encephalatrophy, neonatal hypoxia-ischemic encephalopathy, diffuse axonal injury, senile dementia, epidemic encephalitis B, diabetic peripheral neuropathy, sudden deafness and the like.
The preparation of the cerebroprotein hydrolysate generally comprises the steps of pretreatment of animal brain tissues, cerebroprotein hydrolysis, separation and purification, raw material forming and the like, and each step has different processing methods. Therefore, a method for preparing a cerebroprotein hydrolysate by combining the steps of different treatment methods is often formed. The separation and purification step generally adopts traditional separation and purification methods such as filtration, centrifugation, ultrafiltration, column chromatography, electrodialysis and the like. The raw material forming means that the cerebroprotein hydrolysate is finally prepared into an available product form, such as cerebroprotein hydrolysate powder prepared by spray drying and heating drying, or the cerebroprotein hydrolysate powder is directly prepared into further oral liquid, freeze-dried preparations, injections and the like. The existing preparation method of the cerebroprotein hydrolysate has the defects of complicated steps, heavy pollution and high energy consumption.
Disclosure of Invention
Therefore, a method for preparing the cerebroprotein hydrolysate which is simple in process, free of pollution and low in energy consumption is needed.
A method for preparing brain protein hydrolysate comprises the following steps:
homogenizing brain tissue to obtain homogenate;
carrying out enzymolysis treatment on the homogenate to obtain an enzymolysis liquid;
adding a settling agent into the enzymatic hydrolysate, standing for layering, removing a grease layer on the surface, and collecting a settling supernatant;
adding a flocculating agent into the settled supernatant, performing centrifugal separation, and collecting the centrifugal supernatant;
adding activated carbon into the centrifugal supernatant, preserving the heat at 50-70 ℃ for 10-30 min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
In one embodiment, the enzymolysis liquid is heated to boiling before the sedimentation agent is added, and the enzymolysis liquid is continuously boiled for 10-20 min after the sedimentation agent is added and before standing for layering.
In one embodiment, the sedimentation agent is selected from one or more of talc, diatomaceous earth and calcium carbonate.
In one embodiment, the mass ratio of the homogenate to the settling agent is 100 (0.5-4).
In one embodiment, the flocculant is selected from one or more of gelatin, chitosan, egg white powder, and 101 juice clarifiers.
In one embodiment, the mass ratio of the settled supernatant to the flocculant is 100 (0.06-0.09), and the mass ratio of the centrifugal supernatant to the activated carbon is 100 (1.5-5).
In one embodiment, the enzymatic treatment comprises the following steps: and mixing the homogenate with water and protease, and carrying out hydrolysis reaction for 8-24 hours at 40-45 ℃.
In one embodiment, the proteases include pepsin, papain, and trypsin.
In one embodiment, the pepsin is added in an amount of 3-5% of the brain tissue mass, and the trypsin is added in an amount of 4-6% of the brain tissue mass.
In one embodiment, the brain tissue is one or more of porcine brain tissue, ovine brain tissue, and bovine brain tissue.
The preparation method of the cerebroprotein hydrolysate has the following advantages:
the preparation method of the invention adds a settling agent after enzymolysis for treatment: 1) the settling time can be shortened; because animal brain residues are particularly sticky, the animal brain residues are easy to filter only after being thoroughly layered, so that the animal brain residues usually need to be settled for several hours after enzymolysis, the operations such as filtering and the like are easy to perform after being layered, and the animal brain residues can be thoroughly layered within half an hour after the settling agent is added; 2) layering can be more thorough; the layered interface is clear and flat, the supernatant is easy to separate and is not easy to get turbid, so that the filtration process can be completely replaced; if no settling agent is added, after long-time standing, because the pig brain residues are light and gradually gather and suspend in the liquid, although a solid-liquid state is formed at the moment, the time consumption is long, a solid-liquid interface is uneven, and the pig brain residues are shaken and shaken slightly and easily become turbid, so that the common preparation method only needs to select a filtering or centrifuging method; 3) the addition of the settling agent can change animal brain dregs from sticky state to dispersed state, is easy to pour out, is convenient to treat, and can be used for harmless treatment such as composting and the like; 4) through determination, the sedimentation agent can improve the yield of the cerebroprotein hydrolysate, probably because the sedimentation agent reduces the viscosity of animal brain residues, the effective components are easier to dissolve out, namely the sedimentation agent reduces the viscosity of the animal brain residues and is not easy to wrap the effective components, thereby realizing the effect of improving the content of the effective components; 5) after the sedimentation agent treatment, the obtained supernatant is clear and transparent, and can be directly subjected to subsequent treatment procedures, thereby avoiding complicated and energy-consuming operations such as filtration, centrifugation and the like.
The preparation method further adopts the flocculating agent to further treat the supernatant after the sedimentation agent is used for treatment, can remove macromolecular substances through flocculation and sedimentation, has high treatment speed, and does not need treatment such as ultrafiltration. The flocculation treatment is combined with the sedimentation treatment of the previous step, so that redundant macromolecular substances are further removed, the effect similar to that of the traditional ultrafiltration process is achieved, the required time is shorter, the operation is simpler and more convenient, and compared with the sedimentation-flocculation process, the total nitrogen content of the product is higher, namely the effective ingredients of the substances are better reserved.
The preparation method does not need to carry out an additional degreasing treatment step in the brain tissue treatment process, and the brain tissue is treated by the sedimentation agent, and then the grease layer (grease-like floater) on the surface is skimmed.
The preparation method of the invention does not use organic solvent, has simple and feasible process and low energy consumption, considers the treatment of waste residue, is very environment-friendly and accords with the concept of green production.
Detailed Description
In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The method for preparing the cerebroprotein hydrolysate according to the embodiment of the invention comprises the following steps of S1-S5:
s1, homogenizing the brain tissue to obtain homogenate.
And S2, carrying out enzymolysis treatment on the homogenate to obtain an enzymolysis liquid.
And S3, adding a settling agent into the enzymatic hydrolysate, standing for layering, removing a surface grease layer, and collecting a settling supernatant.
And S4, adding a flocculating agent into the settled supernatant, performing centrifugal separation, and collecting the centrifugal supernatant.
S5, adding activated carbon into the centrifugal supernatant, preserving the heat at 50-70 ℃ for 10-30 min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
The preparation method of the cerebroprotein hydrolysate has the following advantages:
the preparation method of the invention adds a settling agent after enzymolysis for treatment: 1) the settling time can be shortened; because animal brain residues are particularly sticky, the animal brain residues are easy to filter only after being thoroughly layered, so that the animal brain residues usually need to be settled for several hours after enzymolysis, the operations such as filtering and the like are easy to perform after being layered, and the animal brain residues can be thoroughly layered within half an hour after the settling agent is added; 2) layering can be more thorough; the layered interface is clear and flat, the supernatant is easy to separate and is not easy to get turbid, so that the filtration process can be completely replaced; if no settling agent is added, after long-time standing, because the pig brain residues are light and gradually gather and suspend in the liquid, although a solid-liquid state is formed at the moment, the time consumption is long, a solid-liquid interface is uneven, and the pig brain residues are shaken and shaken slightly and easily become turbid, so that the common preparation method only needs to select a filtering or centrifuging method; 3) the addition of the settling agent can change animal brain dregs from sticky state to dispersed state, is easy to pour out, is convenient to treat, and can be used for harmless treatment such as composting and the like; 4) through determination, the sedimentation agent can improve the yield of the cerebroprotein hydrolysate, probably because the sedimentation agent reduces the viscosity of animal brain residues, the effective components are easier to dissolve out, namely the sedimentation agent reduces the viscosity of the animal brain residues and is not easy to wrap the effective components, thereby realizing the effect of improving the content of the effective components; 5) after the sedimentation agent treatment, the obtained supernatant is clear and transparent, and can be directly subjected to subsequent treatment procedures, thereby avoiding complicated and energy-consuming operations such as filtration, centrifugation and the like.
The preparation method further adopts the flocculating agent to further treat the supernatant after the sedimentation agent is used for treatment, can remove macromolecular substances through flocculation and sedimentation, has high treatment speed, and does not need treatment such as ultrafiltration. The flocculation treatment is combined with the sedimentation treatment of the previous step, so that redundant macromolecular substances are further removed, the effect similar to that of the traditional ultrafiltration process is achieved, the required time is shorter, the operation is simpler and more convenient, and compared with the sedimentation-flocculation process, the total nitrogen content of the product is higher, namely the effective ingredients of the substances are better reserved.
The preparation method does not need to carry out an additional degreasing treatment step in the brain tissue treatment process, and the brain tissue is treated by the sedimentation agent, and then the grease layer (grease-like floater) on the surface is skimmed.
The preparation method of the invention does not use organic solvent, has simple and feasible process and low energy consumption, considers the treatment of waste residue, is very environment-friendly and accords with the concept of green production.
In a specific example, before adding the sedimentation agent, the enzymolysis liquid is heated to boiling, and after adding the sedimentation agent, the enzymolysis liquid is continuously boiled for 10-20 min before standing for layering. The sedimentation agent is added after heating and boiling and is continuously boiled for a period of time, so that not only can the enzyme added during enzymolysis be inactivated, but also the sedimentation agent can be better combined with the brain dregs, the viscosity of the animal brain dregs is reduced, the dissolution of effective components is improved, meanwhile, the sedimentation agent can be better and uniformly dispersed under the boiling state, the sedimentation agent is convenient to be better combined with the animal brain dregs, and the aim of rapid sedimentation is favorably fulfilled. If the supernatant is too late to immediately proceed to the next flocculation treatment, the supernatant can be heated to boil again for about 10min, and left overnight for the next day of treatment.
In a specific example, the settling agent is selected from one or more of talc, diatomaceous earth, and calcium carbonate. Preferably, the settling agent is diatomaceous earth.
In a specific example, the mass ratio of the homogenate to the sedimentation agent is 100 (0.5-4), so that a good sedimentation effect can be obtained. Optionally, the brain dregs are washed once with water when the supernatant is collected after the brain dregs are separated, so that the yield of the brain protein hydrolysate can be improved.
In one particular example, the flocculating agent is selected from one or more of gelatin, chitosan, egg white powder, and 101 juice clarifying agent. Preferably, the flocculant is chitosan. The best effect of chitosan is found by respectively comparing the effects of flocculants such as egg white (powder), gelatin, chitosan, 101 fruit juice clarifying agent and the like, and the chitosan effect is mainly reflected in two aspects, namely the clarity of the treated solution, whether the subsequent treatment is precipitated or not and whether the loss of the content of the effective components can be reduced as much as possible or not.
In a specific example, the mass ratio of the settled supernatant to the flocculating agent is 100 (0.06-0.09), a good flocculation effect can be obtained, and the mass ratio of the centrifuged supernatant to the activated carbon is 100 (1.5-5).
In one specific example, the enzymatic treatment comprises the following steps: and mixing the homogenate with water and protease, and carrying out hydrolysis reaction for 8-24 hours at 40-45 ℃. Therefore, the hydrolysis can be effectively carried out without adjusting the pH value during the enzymatic hydrolysis, if the hydrolysis is carried out in an acidic or alkaline environment, multiple steps of treatment such as centrifugation and filtration are required after the hydrolysis, acid-base neutralization during the hydrolysis can generate a large amount of salt, which can not only affect the taste of the product, but also is very unfavorable for human health due to high salt content, and the desalting and energy consumption (such as electrodialysis desalination and the like) are required.
In one particular example, the protease includes pepsin and trypsin. It is understood that the kind of the enzyme is not limited thereto, and for example, papain, hydroxylamine, serine protease, etc.
In a specific example, the adding amount of the pepsin is 3% -5% of the brain tissue mass, and the adding amount of the trypsin is 4% -6% of the brain tissue mass, so that a better enzymolysis effect can be obtained.
In a particular example, the brain tissue is one or more of porcine brain tissue, ovine brain tissue, and bovine brain tissue. It is understood that the kind of the brain tissue is not limited thereto, and the brain tissue of other various animals is acceptable.
In one embodiment, the method further comprises the following steps before the homogenization treatment: the brain tissue is washed clean to remove blood, impurities, etc.
In one specific example, the preparation method further comprises the following steps: spray drying the cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder or directly preparing into liquid preparation.
Embodiments of the present invention will be described in detail below with reference to specific examples.
Example 1
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding chitosan into the supernatant, wherein the mass ratio of the supernatant to the chitosan is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 2
Taking 50kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 50kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 2000g of pepsin and 2500g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 1000g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding chitosan into the supernatant, wherein the mass ratio of the supernatant to the chitosan is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 3
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to 80 deg.C, adding 100g of diatomaceous earth, and keeping the temperature at 80 deg.C for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding chitosan into the supernatant, wherein the mass ratio of the supernatant to the chitosan is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 4
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of pulvis Talci when slightly boiling, and boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding chitosan into the supernatant, wherein the mass ratio of the supernatant to the chitosan is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 5
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of calcium carbonate when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding chitosan into the supernatant, wherein the mass ratio of the supernatant to the chitosan is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 6
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding gelatin into the supernatant, wherein the mass ratio of the supernatant to the gelatin is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 7
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding egg white powder into the supernatant, wherein the mass ratio of the supernatant to the egg white powder is 100:0.075, stirring while adding, and heating. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Example 8
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding 101 fruit juice clarifier into the supernatant, wherein the mass ratio of the supernatant to the 101 fruit juice clarifier is 100:0.075, and stirring while adding. Centrifuging and collecting supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Comparative example 1
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Adding 100g of diatomite into the enzymolysis liquid for filtration assistance, stirring for 30min, then filtering, and collecting the filtrate.
And (3) performing ultrafiltration on the filtrate, collecting components with the molecular weight of less than 5000 daltons, and performing nanofiltration concentration by using a nanofiltration machine with the molecular weight cutoff of 90-150 daltons to obtain the cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Comparative example 2
Taking 5kg of pig brain, cleaning, and homogenizing to obtain homogenate.
And adding acetone with the volume of 3 times of the homogenate, stirring, standing, filtering to obtain filter residue, and drying to obtain the cerebroprotein powder.
Placing the cerebroprotein powder into a hydrolysis tank, adding 5kg of purified water, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Centrifuging the enzymolysis solution for 20min, and collecting supernatant to obtain cerebroprotein hydrolysate.
Spray drying the obtained cerebroprotein hydrolysate to obtain cerebroprotein hydrolysate powder.
Comparative example 3
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Heating the enzymolysis solution to boil, adding 100g of diatomite when slightly boiling, and continuously boiling for 20 min. After sedimentation and stratification, discharging pig brain residues at the bottom of the tank, and collecting supernatant for later use.
Adding activated carbon and no flocculant into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering and collecting filtrate, centrifuging, and spray-drying to prepare the cerebroprotein hydrolysate powder.
Comparative example 4
Taking 5kg of pig brain, cleaning, homogenizing, and placing in a hydrolysis tank.
Adding 5kg of purified water into a hydrolysis tank, starting heating and stirring functions, controlling the temperature to be 40-45 ℃, adding 200g of pepsin and 250g of trypsin, uniformly stirring, and maintaining hydrolysis for 8 hours.
Adding chitosan into the enzymolysis liquid, wherein the mass ratio of the enzymolysis liquid to the chitosan is 100:7.5, and stirring while adding. Standing for layering, discharging precipitate from the bottom, and collecting supernatant.
Adding 100g of diatomite into the supernatant, boiling for 20min, standing for layering, discharging the sediment at the bottom of the tank, and collecting the supernatant.
Adding active carbon into the supernatant according to the mass ratio of 100:2.5, stirring and heating at 60 ℃ for 20min, filtering, collecting filtrate, and spray drying to prepare the cerebroprotein hydrolysate powder.
The quality of the cerebroprotein hydrolysate powders prepared in the examples and comparative examples was tested as follows:
the technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A preparation method of brain protein hydrolysate is characterized by comprising the following steps:
homogenizing brain tissue to obtain homogenate;
carrying out enzymolysis treatment on the homogenate to obtain an enzymolysis liquid;
adding a settling agent into the enzymatic hydrolysate, standing for layering, removing a grease layer on the surface, and collecting a settling supernatant;
adding a flocculating agent into the settled supernatant, performing centrifugal separation, and collecting the centrifugal supernatant;
adding activated carbon into the centrifugal supernatant, preserving the heat at 50-70 ℃ for 10-30 min, filtering and collecting filtrate to obtain the cerebroprotein hydrolysate.
2. The preparation method according to claim 1, wherein the enzymolysis liquid is heated to boiling before adding the sedimentation agent, and boiling is continued for 10-20 min after adding the sedimentation agent before standing for layering.
3. The method of claim 1, wherein the sedimentation agent is selected from one or more of talc, diatomaceous earth, and calcium carbonate.
4. The preparation method of claim 3, wherein the mass ratio of the homogenate to the sedimentation agent is 100 (0.5-4).
5. The method of claim 1, wherein the flocculating agent is selected from one or more of gelatin, chitosan, egg white powder, and 101 juice clarifying agent.
6. The preparation method according to claim 5, wherein the mass ratio of the settled supernatant to the flocculant is 100 (0.06-0.09), and the mass ratio of the centrifuged supernatant to the activated carbon is 100 (1.5-5).
7. The method according to any one of claims 1 to 6, wherein the enzymatic treatment comprises the steps of: and mixing the homogenate with water and protease, and carrying out hydrolysis reaction for 8-24 hours at 40-45 ℃.
8. The method of claim 7, wherein the protease includes pepsin, papain, and trypsin.
9. The method according to claim 8, wherein the pepsin is added in an amount of 3 to 5% and the trypsin is added in an amount of 4 to 6% based on the mass of the brain tissue.
10. The method according to any one of claims 1 to 6, wherein the brain tissue is one or more of a porcine brain tissue, a ovine brain tissue, and a bovine brain tissue.
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Application publication date: 20200825 |