CN113068762A - Preparation method of whey protein - Google Patents
Preparation method of whey protein Download PDFInfo
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- CN113068762A CN113068762A CN202110242373.XA CN202110242373A CN113068762A CN 113068762 A CN113068762 A CN 113068762A CN 202110242373 A CN202110242373 A CN 202110242373A CN 113068762 A CN113068762 A CN 113068762A
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- whey protein
- seed liquid
- fermentation
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- bifidobacterium lactis
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
- A23J1/205—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey from whey, e.g. lactalbumine
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
- A23J3/08—Dairy proteins
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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
-
- 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
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Nutrition Science (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Dairy Products (AREA)
Abstract
The invention discloses a preparation method of whey protein, and relates to the technical field of microbial fermentation. The preparation method of the whey protein comprises the following steps: (1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the bifidobacterium lactis strain and the streptococcus acidophilus strain into a seed culture medium for amplification culture, performing shake culture to respectively obtain a bifidobacterium lactis seed liquid and a streptococcus acidophilus seed liquid, and mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain a mixed seed liquid; (2) fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a preparation method of whey protein.
Background
Whey protein is a general term for various protein components in whey, and generally refers to whey protein from cow milk, the content of which in cow milk is second only to casein, and accounts for 18% -20% of the protein content in cow milk. Whey proteins include beta-lactoglobulin, alpha-lactalbumin, serum albumin, immunoglobulins, lactoferrin, glycosyl peptides and some growth factors. Whey protein products of different protein compositions can be obtained by different processing procedures for extracting proteins, concentrated whey protein is a commercial product with a protein content of 25-28%, and isolated whey protein generally has a protein content of more than 90%. The whey protein belongs to complete protein, is easy to digest and absorb, has high nutritional value, and is widely applied to infant formula food.
However, the main components of the whey protein are beta-Lg and alpha-La, the beta-Lg and the alpha-La account for about 70-80% of the total amount of the whey protein and are main sensitization factors of cow's milk allergy of infants, so that the sensitization structure of the beta-Lg and the alpha-La can be effectively destroyed by properly modifying the whey protein, and the sensitization is reduced or eliminated. The method for modifying whey protein mainly utilizes physical factors (such as heat, rays, mechanical oscillation and the like), chemical factors (such as chemical reagents and the like) or enzyme preparations to destroy the spatial structure of the protein, so that certain changes occur in polypeptide chains and amino acid residues. The currently common whey protein modification methods mainly include heating treatment, glycosylation reaction, fermentation, enzymolysis and the like, and are introduced in the following ways:
heating treatment: mainly changes the mutual conformation of the proteins, expands the structure of the proteins and breaks disulfide bonds; and (3) glycosylation modification: i.e., a chemical reaction in which a carbohydrate is covalently bonded to an alpha or epsilon amino group on a protein molecule to form a glycosylated protein. Researches show that the beta-Lg modified by glycosylation has effectively reduced anaphylaxis and improved thermal stability and emulsification property; enzymolysis: protease-catalyzed hydrolysis of whey protein is an effective method for reducing allergic reactions. The protease can eliminate or reduce the antigenicity of the whey protein by hydrolyzing the epitope of the whey protein, and reduce the cow milk allergy. Different proteases have different specificities, so that the degradation of the enzymolysis products and antigenicity are different according to the enzymes used and the enzymolysis conditions.
However, the above 3 methods for modifying whey protein all have some disadvantages: heating treatment: because the protein and lactose form a complex structure in the heating process, the loss of lysine is increased, and the nutritional value of the protein is reduced. An enzymolysis method comprises the following steps: has high production cost and produces bitter peptides. The reason for the high cost is the large amount of enzyme used in the liquid enzymatic hydrolysis process and the high cost of the purchased enzyme. The bitter taste is caused because hydrophobic amino acids in the peptide chain are exposed due to the breakage of the peptide chain during the enzymolysis process, so that the hydrolysate is bitter. And (3) glycosylation modification: glycosylation modifications can affect the nutritional value and even the safety of proteins. Glycosylation can destroy essential amino acids, especially resulting in large losses of lysine.
At present, the fermented whey protein is modified by lactobacillus fermentation, and the sensitization of cow milk protein can be greatly reduced by fermenting cow milk by lactobacillus. The main reasons are as follows: firstly, the sensitization protein conformation of the cow milk can be induced to change by sterilization treatment before the cow milk is fermented, so that the protein is inactivated, the epitope is masked, and the sensitization is reduced; secondly, in the fermentation process, lactic acid bacteria decompose lactose to generate lactic acid, so that the pH value is reduced, the charge is changed, the structure of milk protein is influenced, namely the conformation is changed, and the lactic acid bacteria are also a reason for reducing the allergenicity of the milk protein; thirdly, some protease can be produced by lactobacillus fermentation, and the sensitization of cow milk protein can be reduced by enzymolysis. However, the current lactic acid bacteria fermentation has the following disadvantages: in the current research at home and abroad, the antiallergic effect of lactic acid bacteria is greatly different.
Disclosure of Invention
Based on the above, the invention aims to overcome the defects of the prior art and provide a preparation method of whey protein, which has the advantages of mild fermentation conditions, low cost and less damage to essential amino acids and reduces the sensitization of the whey protein.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a preparation method of whey protein comprises the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the bifidobacterium lactis strain and the streptococcus acidophilus strain into a seed culture medium for amplification culture, performing shake culture to respectively obtain a bifidobacterium lactis seed liquid and a streptococcus acidophilus seed liquid, and mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain a mixed seed liquid;
(2) fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
The invention selects the bifidobacterium lactis and the streptococcus acidophilus as strains for preparing the mixed seed liquid, and the strains are stored in a glycerin tube at the temperature of between-75 and-85 ℃, wherein the bifidobacterium lactis and the streptococcus acidophilus are used as lactic acid bacteria which are symbiotic bacteria of intestinal micro-ecology, and can prevent and treat protein allergy in various aspects such as improving the proportion of host intestinal flora, enhancing the intestinal barrier function, reducing the release of inflammation media and the like. Meanwhile, the lactobacillus can also stimulate cells to generate immune response, reduce IgE production and relieve anaphylactic reaction. In addition, the bifidobacterium lactis and the streptococcus acidophilus have a synergistic effect, so that the fermentation time can be shortened, and the damage to essential amino acids can be reduced.
Preferably, in the step (1), the mass ratio of the bifidobacterium lactis to the streptococcus acidophilus in the mixed seed liquid is as follows: bifidobacterium lactis: acidophilic streptococcus 1: 1. Through a great deal of research, the inventor of the application finds that when the mass ratio of the bifidobacterium lactis to the streptococcus acidophilus is 1:1, the synergistic effect is optimal, and the damage to essential amino acid is minimum.
Preferably, in the step (2), the fermentation temperature is 29-31 ℃, the fermentation time is 2.5-3.5d, and the pH value is 7.0-8.0 during fermentation; the volume of the mixed seed liquid accounts for 3-5% of the total volume of the mixed seed liquid and the fermentation substrate. The inventor finds that the fermentation conditions are mild, the cost is low and the damage to essential amino acid is less when the temperature, time, pH and the like are adopted in the fermentation process through a large amount of creative experimental research.
Further preferably, in the step (2), the fermentation temperature is 29 ℃, the fermentation time is 2.5d, and the pH value during fermentation is 7.0; the volume of the mixed seed liquid accounts for 3% of the total volume of the mixed seed liquid and the fermentation substrate. The inventor finds that the fermentation process adopts the temperature, time, pH and the like, the fermentation conditions are mild, the cost is lowest, and the damage to essential amino acid is minimum through a great deal of creative experimental research.
Preferably, in the step (2),the concentration of Bifidobacterium lactis and Streptococcus acidophilus in the mixed seed solution is 10 × 107cfu/mL。
Preferably, in the step (1), the temperature for constant temperature culture is 25-30 ℃; the temperature of the amplification culture is 25-30 ℃, and the time of the amplification culture is 24-48 h; the temperature of the shaking culture is 25-30 ℃, the time of the shaking culture is 24-48h, and the rotation speed of the shaking culture is 120-180 r/min.
Preferably, in the step (1), the seed culture medium is MRS culture medium; the MRS culture medium comprises the following components in parts by weight: 10 parts of peptone, 10 parts of beef extract, 5 parts of yeast extract, 20 parts of glucose, 5 parts of sodium acetate, 2 parts of diamine citrate, 1.0 part of tween, 0.4 part of dipotassium hydrogen phosphate, 0.5 part of magnesium sulfate, 0.29 part of manganese sulfate, 20 parts of calcium carbonate, 15 parts of agar and 1 x 10 parts of6Distilled water is divided; the pH value of the MRS culture medium is 6-6.5.
Preferably, in the step (2), the fermentation substrate needs to be subjected to moist heat sterilization at a temperature of 110-.
Preferably, in the step (2), the obtained whey protein has an allergen content of 80-92 μ g/mL, calculated as the allergen content (in terms of β -lactoglobulin content).
In addition, the invention provides hypoallergenic whey protein prepared by the fermentation method for reducing the allergenicity of whey protein.
Compared with the prior art, the invention has the beneficial effects that: the invention selects the bifidobacterium lactis and the streptococcus acidophilus as strains for preparing the mixed seed liquid, utilizes the synergistic effect of the two strains to reduce the anaphylaxis of the whey protein, and the whey protein obtained under the condition has low anaphylaxis and is still full-value protein, and in addition, the content of the essential amino acid is basically unchanged.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.
In the present application, examples 1 to 9 are provided, and in the components and parts by weight of the specific examples 1 to 9, four factors affecting fermentation are as follows: the fermentation temperature, pH, the ratio of the volume of the mixed seed liquid to the total volume of the mixed seed liquid and the fermentation substrate, the setting of the fermentation time and the detection result of the allergen content are shown in Table 1, the rest preparation processes are as follows, and the analysis of variance of the measurement result of the allergen content is shown in Table 2:
TABLE 1 fermentation temperature, pH, volume of mixed seed solution/total volume of mixed seed solution and fermentation substrate, setting of fermentation time, and measurement results of allergen content in examples 1-9
TABLE 2 analysis of variance of measurement results of allergen content
Examples
The embodiment of the application adopts the technical scheme that: a preparation method of whey protein comprises the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the strains in a seed culture medium for amplification culture, performing shake culture to respectively obtain bifidobacterium lactis seed liquid and streptococcus acidophilus seed liquid, mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain mixed seed liquid, wherein the mass ratio of bifidobacterium lactis to streptococcus acidophilus in the mixed seed liquid is as follows: bifidobacterium lactis: acidophilic streptococcus 1: 1;
the constant temperature culture temperature is 25 ℃; the temperature of the amplification culture is 25 ℃, and the time of the amplification culture is 24 h; the temperature of the shaking culture is 25 ℃, the time of the shaking culture is 24 hours, and the rotating speed of the shaking culture is 180 r/min.
(2) Fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
The allergen content (calculated by beta-lactoglobulin content) is used as an evaluation index, and the evaluation index can be obtained by an intuitive analysis method, and the factor sequence influencing the fermentation is pH > fermentation temperature > fermentation time > volume of the mixed seed liquid/total volume of the mixed seed liquid and the fermentation substrate. The method can be obtained by an analysis of variance method, only pH has significant influence on a fermentation process, so the optimal pH level is 7, because the fermentation temperature, the fermentation time and the volume of the mixed seed liquid/the total volume of the mixed seed liquid and a fermentation substrate have no significant influence on the fermentation process, the production cost, the production time and resources are considered, the optimal fermentation process conditions are that the fermentation temperature is 29 ℃, the fermentation time is 2.5d, the pH is 7.0 during fermentation, and the volume of the mixed seed liquid accounts for 3 percent of the total volume of the mixed seed liquid and the fermentation substrate.
At the same time, the present application is based on the following conditions of optimal fermentation: the fermentation temperature was 29 ℃, the fermentation time was 2.5d, the pH during fermentation was 7.0, the volume of the mixed seed solution accounted for 3% of the total volume of the mixed seed solution and the fermentation substrate, setting examples 10-11 as follows:
example 10
In an embodiment of the present application, a technical solution adopted in this embodiment is: a preparation method of whey protein comprises the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the strains in a seed culture medium for amplification culture, performing shake culture to respectively obtain bifidobacterium lactis seed liquid and streptococcus acidophilus seed liquid, mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain mixed seed liquid, wherein the mass ratio of bifidobacterium lactis to streptococcus acidophilus in the mixed seed liquid is as follows: bifidobacterium lactis: acidophilic streptococcus 1: 1;
the constant temperature culture temperature is 25 ℃; the temperature of the amplification culture is 25 ℃, and the time of the amplification culture is 24 h; the temperature of the shaking culture is 25 ℃, the time of the shaking culture is 24 hours, and the rotating speed of the shaking culture is 120 r/min.
(2) Fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting, wherein the fermentation temperature is 29 ℃, the fermentation time is 2.5 days, the pH value during fermentation is 7.0, and the volume of the mixed seed liquid accounts for 3% of the total volume of the mixed seed liquid and the fermentation substrate to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
Example 11
In an embodiment of the present application, a technical solution adopted in this embodiment is: a preparation method of whey protein comprises the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the strains in a seed culture medium for amplification culture, performing shake culture to respectively obtain bifidobacterium lactis seed liquid and streptococcus acidophilus seed liquid, mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain mixed seed liquid, wherein the mass ratio of bifidobacterium lactis to streptococcus acidophilus in the mixed seed liquid is as follows: bifidobacterium lactis: acidophilic streptococcus 1: 1;
the constant temperature culture temperature is 28 ℃; the temperature of the amplification culture is 28 ℃, and the time of the amplification culture is 36 h; the temperature of the shaking culture is 28 ℃, the time of the shaking culture is 36h, and the rotation speed of the shaking culture is 160 r/min.
(2) Fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting, wherein the fermentation temperature is 29 ℃, the fermentation time is 2.5 days, the pH value during fermentation is 7.0, and the volume of the mixed seed liquid accounts for 3% of the total volume of the mixed seed liquid and the fermentation substrate to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
Example 12
In an embodiment of the present application, a technical solution adopted in this embodiment is: a preparation method of whey protein comprises the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the strains in a seed culture medium for amplification culture, performing shake culture to respectively obtain bifidobacterium lactis seed liquid and streptococcus acidophilus seed liquid, mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain mixed seed liquid, wherein the mass ratio of bifidobacterium lactis to streptococcus acidophilus in the mixed seed liquid is as follows: bifidobacterium lactis: acidophilic streptococcus 1: 1;
the constant temperature culture temperature is 30 ℃; the temperature of the amplification culture is 30 ℃, and the time of the amplification culture is 48 h; the temperature of the shaking culture is 30 ℃, the time of the shaking culture is 48h, and the rotating speed of the shaking culture is 180 r/min.
(2) Fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting, wherein the fermentation temperature is 29 ℃, the fermentation time is 2.5 days, the pH value during fermentation is 7.0, and the volume of the mixed seed liquid accounts for 3% of the total volume of the mixed seed liquid and the fermentation substrate to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
Effect verification 1 allergen content detection
And (3) detection process: the residual antigenicity of the whey protein hydrolysate is measured by a competitive enzyme-linked immunosorbent assay, which indirectly reflects whether the allergy of the whey protein is reduced. The assays and formula calculations were performed with reference to kit instructions.
And (3) detection results: the allergen contents measured in examples 10 to 12 of the present application were 80.2. mu.g/mL, 81.7. mu.g/mL, and 80.3. mu.g/mL, respectively, the average allergen content was 80.7. mu.g/mL, the initial allergen content was 190.5. mu.g/mL, which was a 42.4% reduction.
Effect verification 2 detection of amino acid composition and content
The detection device comprises: the composition and content of the whey protein amino acid are measured by a Hitachi 835-50 type amino acid automatic analyzer.
And (3) detection results: the results of the best process verification index measurement described in example 10 are shown in table 3, and the results of examples 10-12 are similar to those of example 10, which are not repeated here:
TABLE 3 measurement results of amino acid composition and content
| Essential amino acids | Content before fermentation (mg/g) | Content after fermentation (mg/g) |
| Isoleucine | 70.6 | 71.4 |
| Leucine | 116.9 | 118 |
| Lysine | 99.3 | 97.7 |
| Methionine + cystine | 55.2 | 56.9 |
| Phenylalanine + tyrosine | 58.9 | 61.8 |
| Threonine | 76 | 75.1 |
| Tryptophan | 19.3 | 18.9 |
| Valine | 60.4 | 59.9 |
The composition and content of amino acid of the obtained whey protein liquid are detected, and particularly as shown in table 3, the content of the amino acid is basically kept unchanged, so that the process has good feasibility and repeatability, and the damage to essential amino acid is small.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (9)
1. A preparation method of whey protein is characterized by comprising the following steps:
(1) preparing a mixed seed solution; respectively taking a bifidobacterium lactis strain and a streptococcus acidophilus strain, performing constant-temperature culture, then inoculating the bifidobacterium lactis strain and the streptococcus acidophilus strain into a seed culture medium for amplification culture, performing shake culture to respectively obtain a bifidobacterium lactis seed liquid and a streptococcus acidophilus seed liquid, and mixing the bifidobacterium lactis seed liquid and the streptococcus acidophilus seed liquid to obtain a mixed seed liquid;
(2) fermenting; mixing the mixed seed liquid obtained in the step (1) with a fermentation substrate, and fermenting to obtain the whey protein; wherein the fermentation substrate is whey protein solution.
2. The method for preparing whey protein according to claim 1, wherein in step (1), the mass ratio of bifidobacterium lactis to streptococcus acidophilus in the mixed seed liquid is: bifidobacterium lactis: acidophilic streptococcus 1: 1.
3. The method for preparing whey protein according to claim 1, wherein in step (2), the fermentation temperature is 29-31 ℃, the fermentation time is 2.5-3.5 days, and the fermentation pH is 7.0-8.0; the volume of the mixed seed liquid accounts for 3-5% of the total volume of the mixed seed liquid and the fermentation substrate.
4. The method for producing whey protein according to claim 1, wherein in the step (2), the concentration of Bifidobacterium lactis and Streptococcus acidophilus in the mixed seed solution is 10 x 107cfu/mL。
5. The method for preparing whey protein according to claim 3, wherein in step (2), the fermentation temperature is 29 ℃, the fermentation time is 2.5 days, and the fermentation pH is 7.0; the volume of the mixed seed liquid accounts for 3% of the total volume of the mixed seed liquid and the fermentation substrate.
6. The method for producing whey protein according to claim 1, wherein in the step (1), the incubation temperature is 25 to 30 ℃; the temperature of the amplification culture is 25-30 ℃, and the time of the amplification culture is 24-48 h; the temperature of the shaking culture is 25-30 ℃, the time of the shaking culture is 24-48h, and the rotation speed of the shaking culture is 120-180 r/min.
7. The method for producing whey protein according to claim 1, wherein in step (1), the seed medium is MRS medium; the MRS culture medium comprises the following components in parts by weight: 10 parts of peptone, 10 parts of beef extract, 5 parts of yeast extract, 20 parts of glucose, 5 parts of sodium acetate, 2 parts of diamine citrate, 1.0 part of tween, 0.4 part of dipotassium hydrogen phosphate, 0.5 part of magnesium sulfate, 0.29 part of manganese sulfate, 20 parts of calcium carbonate, 15 parts of agar and 1 x 10 parts of6Distilled water is divided; the pH value of the MRS culture medium is 6-6.5.
8. The method for preparing whey protein according to claim 1, wherein in step (2), the allergen content of the obtained whey protein is 80-92 μ g/mL.
9. A whey protein produced by the method for producing a whey protein according to any one of claims 1 to 8.
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