CN112106830A - Method for improving bioactivity of bovine colostrum freeze-dried powder immunoglobulin - Google Patents
Method for improving bioactivity of bovine colostrum freeze-dried powder immunoglobulin Download PDFInfo
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- CN112106830A CN112106830A CN202011050839.8A CN202011050839A CN112106830A CN 112106830 A CN112106830 A CN 112106830A CN 202011050839 A CN202011050839 A CN 202011050839A CN 112106830 A CN112106830 A CN 112106830A
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- freeze
- bovine colostrum
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- immunoglobulin
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- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C7/00—Other dairy technology
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
Abstract
A method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin belongs to the technical field of freeze-dried powder quality research. The invention aims to further improve the biological activity of immunoglobulin in bovine colostrum freeze-dried powder, and the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum; pre-freezing the bovine colostrum, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology; freeze-drying the pre-frozen bovine colostrum in a vacuum freeze dryer, and freeze-drying to obtain powder. By applying ultrasonic waves, the bovine colostrum can not form large ice crystals but only form fine ice crystals in the freezing process, the covalent bond of the immunoglobulin active site can not be damaged, and the immunoglobulin activity of the bovine colostrum powder freeze-dried by the method can be improved by 10-30%.
Description
Technical Field
The invention belongs to the technical field of freeze-dried powder research, and particularly relates to a method for improving the biological activity of bovine colostrum freeze-dried powder immunoglobulin.
Background
The milk secretion of cows is started 12 hours after calving, the cows can continuously express milk within 307-312 days later, 3 days before the modern pasture is generally specified are called as the colostrum period, and the milk of cows in the colostrum period is obviously different from that of ordinary cows and is called as bovine colostrum. Bovine colostrum has a high protein content and low fat and sugar content. The research finds that the bovine colostrum contains rich nutrient components, the average total dry matter is 14.4%, the protein is 5.0%, the fat is 4.3%, the ash content is 0.9%, and the bovine colostrum also contains rich vitamins and minerals. The contents of calcium, iron and zinc are respectively 2 times, 3-5 times and 2-4 times of the normal milk; the vitamin C and the vitamin E are 2-5 times of normal milk; the taurine is 5-6 times higher; but also contains a large amount of immune factors and growth factors, such as immunoglobulin, lactoferrin, lysozyme, insulin-like growth factor, epidermal growth factor, etc. Scientific experiments prove that the bovine colostrum has the physiological activity functions of regulating immunity, improving gastrointestinal tract, promoting growth and development, improving aging symptoms, inhibiting various germs and the like, and is known as health food in the 21 st century.
Because the active protein content in the bovine colostrum is very high, the heat stability is poor, the bovine colostrum is easy to coagulate and denature when being heated, and if a heating process is adopted, the biological activity of the product can be lost. Therefore, the large-scale development of bovine colostrum products requires that the heat-sensitive functional components, especially the biological activity of immunoglobulin such as IgG, in bovine colostrum be ensured in the preparation process. The common bovine colostrum on the market is produced by a spray drying process, the temperature of the production process reaches more than 60 ℃ within 3 minutes, and the process kills part of active immunoglobulin when the temperature is higher than 40 ℃, so that the content of the immunoglobulin is low and the immunoglobulin has no dynamic activity.
The whole process of freeze drying of the bovine colostrum comprises prefreezing, first-stage sublimation drying and second-stage re-drying. The volume is increased when the water is frozen in the pre-freezing process, so that some bonds connected by weak molecular force bonds in the active sites of the immunoglobulin are damaged, and the activity is lost; in addition, freezing of water causes an increase in the concentration of solutes and changes in pH due to inconsistent solubility of various solutes at different temperatures, resulting in inactivation or denaturation of the immunoglobulin due to changes in the environment in which it is placed.
Disclosure of Invention
The invention provides a method for improving the biological activity of immunoglobulin in bovine colostrum freeze-dried powder, which aims to further improve the biological activity of the immunoglobulin in the bovine colostrum freeze-dried powder.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin is as follows:
the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum;
step two: pre-freezing the bovine colostrum obtained in the step one, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology;
step three: and D, freeze-drying the pre-frozen bovine colostrum in the step two in a vacuum freeze-drying machine, and freeze-drying to obtain powder.
Compared with the prior art, the invention has the beneficial effects that: by applying ultrasonic waves, large ice crystals and fine ice crystals can not be formed in the process of freezing the bovine colostrum, covalent bonds of immunoglobulin active sites can not be damaged, and the immunoglobulin activity of the lyophilized bovine colostrum powder can be improved by 10-30%.
Detailed Description
The technical solutions of the present invention are further described below with reference to the following examples, but the present invention is not limited thereto, and any modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
The invention adopts a short-time centrifugal degreasing technology with higher temperature and lower rotating speed to achieve the degreasing effect of reducing protein loss, and retains protein components such as protective glycoprotein and immunocompetent protein, and can also retain fat-soluble nutrient substances. The radiation sterilization is carried out by adopting a special treatment mode and an advanced freeze drying process, so that abundant nutrient substances and bioactive substances except fat in the bovine colostrum are retained to the greatest extent, the damage of partial nutrient substances, protein, enzyme denaturation and inactivation of bioactive substances caused by high-temperature treatment are avoided, and meanwhile, special glycoprotein components such as a bovine colostrum trypsin inhibitor are retained, and the immunity and growth factors of the bovine colostrum can be protected from being damaged by trypsin in intestinal tracts. The bovine colostrum product contains seven kinds of vitamins (vitamin E, vitamin C, nicotinic acid, vitamin B1, vitamin B2, vitamin B6, vitamin B12), nineteen kinds of amino acids (taurine, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, tryptophan, arginine), and twelve kinds of minerals (barium, calcium, copper, chromium, iron, potassium, magnesium, manganese, sodium, phosphorus, strontium, and zinc) in addition to protein components such as protective glycoprotein and immunoactive protein. In the preparation method, no auxiliary materials, additives and other components are added in the processing process, and various nutritional ingredients, bioactive substances and the like are not damaged.
In the process of freeze drying, substances containing a large amount of moisture are cooled and frozen into solids, then water vapor is directly sublimated under the vacuum condition, and the substances are left in an ice rack during freezing, so that the volume of the dried substances is not changed, and loose and porous substances absorb heat during sublimation. The sublimation speed is reduced by causing the temperature of the product itself to decrease, and in order to increase the sublimation speed and shorten the drying time, the product must be properly heated. The entire drying is carried out at a relatively low temperature. In the preparation process, the vacuum freeze dryer is used, so that the moisture in the food can be directly sublimated from a solid state to be dried under the slurry and vacuum conditions without liquid evaporation. The vacuum freeze-drying machine can maximally preserve the original color, fragrance, taste and shape of food, and the original nutrient components and active substances are rarely damaged, and the vacuum freeze-drying machine can be quickly rehydrated.
The first embodiment is as follows: the embodiment describes a method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin, which comprises the following steps:
the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum; excessive fat can cause rancidity, which directly affects the quality and shelf life of the product.
Step two: pre-freezing the bovine colostrum obtained in the step one, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology;
step three: and D, freeze-drying the pre-frozen bovine colostrum in the step two in a vacuum freeze-drying machine, and freeze-drying to obtain powder. The step can make the product enter the dormant state quickly, prevent the change of the biological physicochemical and biological characteristics of the product, does not damage biological tissues, cell structures and cell characteristics, and effectively protects the stability of the effective components of the heat-sensitive biological product. The bovine colostrum produced by the freeze-drying process completely retains the immunoglobulin activity and other nutrient substances in the bovine colostrum.
The second embodiment is as follows: in the first step, the centrifugation is carried out at 5 ℃ and 6000r/min for 20 min; too high a centrifugation speed and too long a centrifugation time will result in increased protein precipitation, loss of immunoglobulins and protective glycoproteins, and fat-soluble nutrients such as: loss of vitamin E.
The third concrete implementation mode: in the second step of the method for improving the bioactivity of the immunoglobulin of the bovine colostrum freeze-dried powder, the frequency of the ultrasound is 20-60kHz, and the power density is 5-30W/cm2。
The fourth concrete implementation mode: in the second step, the pre-freezing temperature is-5 ℃ to-25 ℃ and the time is 8 to 12 minutes.
The fifth concrete implementation mode: the method for improving the bioactivity of the bovine colostrum freeze-dried powder immunoglobulin comprises the third step of controlling the vacuum degree to be 5-20 Pa.
The sixth specific implementation mode: in the third step, the freeze drying specifically comprises: slowly raising the temperature of the materials to-5 ℃ under vacuum and maintaining for 2-3 hours; the temperature of the materials is raised to 15-25 ℃, and the materials are maintained for 2-4 hours under vacuum to be completely freeze-dried. The structure of the active substance is unstable due to the rapid freeze-drying and the temperature shock in the rapid temperature rising process, the freeze-drying method adopts layered gradual freeze-drying, the activity loss of the active substance caused by the temperature shock is avoided, and the drying and crystallization can be more thorough.
The seventh embodiment: in a specific embodiment, the method for improving the biological activity of bovine colostrum lyophilized powder immunoglobulin comprises the third step of adding no additive into the obtained lyophilized powder, mixing, sieving, repeatedly stirring, filling capsules or powder, filling powder or directly tabletting powder into tablets, and finally carrying out finished product inspection.
Example 1:
a method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin is as follows:
the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum; centrifuging at 5 deg.C and 6000r/min for 20 min; too high a centrifugation speed and too long a centrifugation time will result in increased protein precipitation, loss of immunoglobulins and protective glycoproteins, and fat-soluble nutrients such as: loss of vitamin E.
Step two: pre-freezing the bovine colostrum obtained in the step one, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology; the frequency of the ultrasonic wave is 20kHz, and the power density is 25W/cm2. The pre-freezing temperature is-20 ℃ and the time is 10 minutes.
Step three: and D, freeze-drying the pre-frozen bovine colostrum in the step two in a vacuum freeze-drying machine, and freeze-drying to obtain powder. Slowly raising the temperature of the material to 0 ℃ under the vacuum of 8Pa, and maintaining for 2-3 hours; and (3) raising the temperature of the material to 15 ℃, maintaining the temperature for 2-4 hours under vacuum, completely freeze-drying, and encapsulating and filling the obtained freeze-dried powder. The step can make the product enter the dormant state quickly, prevent the change of the biological physicochemical and biological characteristics of the product, does not damage biological tissues, cell structures and cell characteristics, and effectively protects the stability of the effective components of the heat-sensitive biological product. The bovine colostrum produced by the freeze-drying process completely retains the activity of active immunoglobulin and other nutrient substances in the bovine colostrum.
The invention provides a preparation method of a bovine colostrum product, which adopts a low-temperature freeze-dried powder technology, and mainly freezes the moisture contained in the product to be dried at a low temperature, then the product is dried in a vacuum environment, and the moisture is directly sublimated from a solid state into water vapor and is removed from the product, so that the product is dried. In order to better enable the product to rapidly enter a dormant state and prevent the change of the biological physicochemical and biological characteristics of the product. In order to effectively retain bioactive substances in the bovine colostrum, the temperature in the whole production and processing process is not higher than 40 ℃. Meanwhile, the operation steps are carried out under the condition of purification or sterility meeting the requirements, then irradiation sterilization is carried out according to the requirements, ultraviolet lamps are installed in workshops of production, processing, semi-finished raw materials and the like, and sterilization is carried out at regular intervals.
Example 2:
a method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin is as follows:
the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum; excessive fat can cause rancidity, which directly affects the quality and shelf life of the product. Centrifuging at 5 deg.C and 6000r/min for 20 min;
step two: pre-freezing the bovine colostrum obtained in the step one, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology; the ultrasonic frequency is 60kHz, and the power density is 10W/cm2. Said prefreezingThe temperature was-25 ℃ and the time was 8 minutes.
Step three: and D, freeze-drying the pre-frozen bovine colostrum in the step two in a vacuum freeze-drying machine, and freeze-drying to obtain powder. Slowly raising the temperature of the materials to-5 ℃ under the vacuum of 20Pa, and maintaining for 2 hours; the batch temperature was raised to 15 ℃ and maintained under vacuum for 4 hours for complete lyophilization. The step can make the product enter the dormant state quickly, prevent the change of the biological physicochemical and biological characteristics of the product, does not damage biological tissues, cell structures and cell characteristics, and effectively protects the stability of the effective components of the heat-sensitive biological product. The bovine colostrum produced by the freeze-drying process completely retains the activity of active immunoglobulin and other nutrient substances in the bovine colostrum.
Claims (7)
1. A method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin is characterized in that: the method comprises the following steps:
the method comprises the following steps: centrifuging the bovine colostrum by a centrifugal machine, and removing fat to obtain defatted bovine colostrum;
step two: pre-freezing the bovine colostrum obtained in the step one, and applying ultrasonic waves to the frozen plate layer in the pre-freezing process of the bovine colostrum by utilizing an ultrasonic technology;
step three: and D, freeze-drying the pre-frozen bovine colostrum in the step two in a vacuum freeze-drying machine, and freeze-drying to obtain powder.
2. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to claim 1, wherein the method comprises the following steps: in the first step, the centrifugation is carried out at 4 ℃ at 7000r/min for 15 min.
3. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to claim 1, wherein the method comprises the following steps: in the second step, the ultrasonic frequency is 20-60kHz, and the power density is 5-30W/cm2。
4. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to claim 1, wherein the method comprises the following steps: in the second step, the pre-freezing temperature is-5 ℃ to-25 ℃, and the time is 8-12 minutes.
5. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to claim 1, wherein the method comprises the following steps: in the third step, the vacuum degree is 5-20 Pa.
6. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to claim 1, wherein the method comprises the following steps: in the third step, the freeze drying specifically comprises: slowly raising the temperature of the materials to-5 ℃ under vacuum and maintaining for 2-3 hours; the temperature of the materials is raised to 15-25 ℃, and the materials are maintained for 2-4 hours under vacuum to be completely freeze-dried.
7. The method for improving the bioactivity of bovine colostrum freeze-dried powder immunoglobulin according to any one of claims 1 to 6, wherein the method comprises the following steps: and in the third step, the obtained freeze-dried powder is mixed, sieved, repeatedly stirred, filled into capsules or powder, filled into powder or directly tableted into tablets, and finally finished product inspection is carried out.
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