CN112970997A - Preparation method of selenoprotein powder and selenoprotein tablet - Google Patents
Preparation method of selenoprotein powder and selenoprotein tablet Download PDFInfo
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- CN112970997A CN112970997A CN202110362799.9A CN202110362799A CN112970997A CN 112970997 A CN112970997 A CN 112970997A CN 202110362799 A CN202110362799 A CN 202110362799A CN 112970997 A CN112970997 A CN 112970997A
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- selenoprotein
- powder
- selenium
- fish meat
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/385—Concentrates of non-alcoholic beverages
- A23L2/39—Dry compositions
- A23L2/395—Dry compositions in a particular shape or form
-
- 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
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1522—Inorganic additives, e.g. minerals, trace elements; Chlorination or fluoridation of milk; Organic salts or complexes of metals other than natrium or kalium; Calcium enrichment of milk
-
- 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
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- 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
- A23C9/18—Milk in dried and compressed or semi-solid form
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/20—Fish extracts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/16—Inorganic salts, minerals or trace elements
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/19—Dairy proteins
-
- 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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- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Inorganic Chemistry (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a preparation method and application of selenoprotein powder and selenoprotein tablets, belongs to the technical field of food processing, and specifically comprises the following steps: 1) pretreating raw materials; 2) fermenting; 3) extracting; 4) preparing a solution A; 5) preparing selenium protein powder; 6) and (4) preparing selenoprotein tablets. The product of the invention has better flavor and taste, can obviously improve the digestion and absorption utilization of human bodies, and is green, nontoxic and safe. Most importantly, the method can greatly reduce the use of inorganic selenium and other components in the prior art, reduce the use of additives and has higher added value of products. In addition, the invention can be drunk after being soaked in water, has better water solubility of the protein peptide, yellowish color and no peculiar smell, and is more suitable for middle-aged and elderly people.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a preparation method and application of selenoprotein powder and selenoprotein tablets.
Background
Selenium is a metal-like element and plays an important role in promoting the growth of organisms and maintaining the health of the organisms. Selenium is mainly absorbed in the small intestine, but inorganic selenium and organic selenium differ in absorption mechanism. After being ingested by animals, inorganic selenium is mainly absorbed in small intestine parts through a simple diffusion mode, and selenate and selenite are taken as two different inorganic selenium sources, and have great difference in body absorption due to different aspects such as chemical structures. The organoselenium-dependent neutral amino acid transport system is absorbed into the blood by the small intestine, where the duodenum and the anterior segment of the ileum are the major sites of selenium absorption. According to the metabolic form of selenium, it can be divided into selenoprotein and selenoprotein.
The intake range of organic selenium in human body is narrow, and too low or too high is not good for human health, while selenite is even a potential safety hazard due to large toxicity and dosage control. In the prior art, the selenium-rich products on the market mainly adopt food raw materials with higher selenium content, and the selenium content in the raw materials is different and difficult to enrich. In addition, the subsequent utilization rate of digestion and absorption of selenoprotein is low because the structural stability of selenoprotein is not researched by adopting fermented foods such as selenium yeast and the like.
Therefore, the problem to be solved by those skilled in the art is how to provide a selenoprotein food that is beneficial for the human body to absorb selenium.
Disclosure of Invention
In view of the above, the invention provides a preparation method of a selenium protein powder and a selenium protein tablet which are non-toxic, safe and easy to be absorbed by human body.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of selenium protein powder comprises the following steps:
1) pretreatment: cleaning fish meat, uniformly coating salt on the surface of the fish meat, cooling the fish meat to 4-6 ℃, refrigerating, washing the salt, draining water, chopping and beating to obtain fish meat paste;
2) fermentation: uniformly mixing the fish meat paste pulp with selenium-enriched yeast, carrying out aerobic fermentation, pumping the aerobic fermentation product to a certain vacuum degree, sealing, maintaining the internal air pressure at 0.02-0.04MPa, putting the aerobic fermentation product into a high-pressure container of 15-20MPa, carrying out sealed fermentation, and carrying out high-pressure sterilization treatment on the fermentation product to obtain the fermented fish meat paste pulp;
wherein the selenium-enriched yeast is commercial selenium-enriched yeast, the net content of the selenium-enriched yeast is 98%, and the concentration of the active selenium-enriched yeast is 7 IU/mg;
3) extraction: uniformly stirring the fermented fish meat paste pulp with water, centrifuging, and keeping supernatant;
4) adding salt into the whey protein solution, performing ultrasonic treatment, heating, and rapidly cooling to 4-6 deg.C to obtain solution A;
5) preparing selenium protein powder: and uniformly stirring the supernatant and the solution A to obtain a mixed solution B, slowly adding trehalose and corn oil into the mixed solution B at the speed of 10mL/min, uniformly stirring again, and spray-drying to obtain the selenoprotein powder.
Has the advantages that: the selenium-enriched protein powder provided by the invention adopts a method of combining aerobic fermentation and low-pressure fermentation, so that a fermentation product and selenium form an organic combination, the structural stability of the selenium protein is enhanced, the bioavailability is improved, in the low-pressure fermentation process, the inside of a sealing bag is in a low-pressure environment, the outside of the sealing bag has 15-20MPa, the pressure difference environment can induce yeast to generate different metabolites, and the product is favorable for the organic combination of the selenium and the protein. In addition, the selenoprotein provided by the invention can enhance the structural stability in gastrointestinal digestion and improve the digestion and absorption rate.
Preferably, the fish meat in the step 1) is large yellow croaker meat, and the head, tail, bones, skin and internal organs of the fish meat are removed; the mass ratio of the fish meat to the salt is 1: 0.015-0.018; the refrigerating time is 80-100 min.
Has the advantages that: the pretreatment process of the raw materials can prepare for subsequent accurate nutrition and flavor.
Preferably, the adding amount of the selenium-enriched yeast in the step 2) is 0.3-0.4mg/Kg based on the weight of the fish meat; the aerobic fermentation temperature is 30-32 ℃, and the fermentation time is 3-3.5 hours; the low-pressure sealed fermentation pressure is 30-35MPa, and the fermentation time is 40-50 min; the high-pressure sterilization pressure is 280-350MPa, and the high-pressure sterilization time is 10-12 min.
Has the advantages that: the structural stability of selenoprotein is promoted by high pressure technology.
Preferably, the mass ratio of the fermented fish meat emulsion slurry to water in the step 3) is (6-8): 1.
Has the advantages that: the proper feed-liquid ratio is beneficial to the subsequent preparation of the protein.
Preferably, the stirring speed in the step 3) is 300-400r/min, and the stirring time is 10-15 min; the centrifugal temperature is 15-25 ℃, the centrifugal force is 5500-6500 Xg, and the centrifugal time is 15-20 min.
Has the advantages that: the setting of the process parameters can ensure high extraction rate of the protein.
Preferably, the mass ratio of the whey protein to the water in the whey protein solution in the step 4) is 1:10, and the pH value is 2.5-3.0; the weight ratio of the whey protein to the salt is 1 (0.002-0.003).
Preferably, the ultrasonic power in the step 4) is 400-600W, and the ultrasonic time is 30-40 min; the heating temperature is 85-90 ℃, and the heating time is 100-120 min; the cooling speed is 20 ℃/min.
Preferably, the volume ratio of the supernatant to the solution A in the step 5) is 1: 3; the adding ratio of the solution B to the trehalose and the corn oil is 100 mL: (0.5-0.7) g: (1.2-1.4) mL.
Preferably, the stirring speed in the step 5) is 300-400r/min, and the stirring time is 10-15 min; the stirring is carried out for 800-900r/min again and 20-25 min.
Has the advantages that: the setting of the process parameters can promote the structure of the selenoprotein to be more stable.
A selenoprotein tablet, characterized by comprising the selenoprotein powder of any one of claims 1-8, wherein the preparation method of the selenoprotein tablet comprises the following steps:
according to the weight portion, 15.2-16.4 portions of selenoprotein powder, 15-15.4 portions of milk powder, 0.2-0.4 portion of algae oil powder, 5.2-5.6 portions of maltodextrin, 0.02-0.04 portion of lactoferrin powder, 36-38 portions of disintegrating agent, 1.5-1.9 portions of aspartame, 2.6-2.8 portions of polyvinylpyrrolidone, 60004.4-4.8 portions of polyethylene glycol and 14.3-14.7 portions of mannitol are uniformly mixed and pressed into tablets, and then the selenoprotein tablets are obtained.
Has the advantages that: the invention improves the quality of the selenoprotein tablet in a micro-nano embedding mode, and improves the digestion and absorption utilization rate of the selenoprotein.
According to the technical scheme, compared with the prior art, the invention discloses a preparation method and application of selenium protein powder, and the selenium protein powder has the following beneficial effects:
1. compared with the currently common selenium supplement method and the currently mature technology in the leisure health food industry, the selenoprotein powder provided by the invention can obviously improve the digestion and absorption and utilization of human bodies, and the selenoprotein tablet has better flavor and mouthfeel, and is green, nontoxic and safe. Most importantly, the method can greatly reduce the use of inorganic selenium and other components in the prior art, reduce the use of additives and ensure higher added value of products.
2. The protein peptide tablet product can be drunk after being soaked in water, has better water solubility of protein peptide, light yellow color and no peculiar smell, and is more suitable for middle-aged and elderly people.
Detailed Description
The following will clearly and completely describe the technical solutions in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A preparation method of selenium protein powder comprises the following steps:
1. cleaning large yellow croaker, removing head and tail, bone, skin and viscera, adding salt 1.5 wt% of fish meat, uniformly coating the salt on the surface of fish body, and placing in a refrigerator at 4 deg.C for 100min to cool the central temperature of fish body to 5 deg.C. Washing the surface of the fish body with salt by using cold water at 4 ℃, and draining. Chopping fish, and beating in food beating machine (80 times/min) for 20min to obtain meat paste.
2. Adding selenium-enriched yeast into the fish meat paste pulp obtained in the step 1 according to 0.4mg/Kg, and slowly and uniformly stirring. Heating the fish meat paste pulp to 31 ℃, and keeping the temperature for 3.5 hours. Every 100 g of meat paste pulp is put into a sealed bag, vacuumized to 0.02-0.04MPa and sealed. Placing the sealed bag into a high static pressure device, and continuing fermenting under 32MPa for 50 min. Then pressurizing to 280MPa for sterilization, and maintaining for 12 min. And after pressure relief, taking out the sealing bag, and pouring out the minced fillet slurry.
3. Dissolving whey protein in 10 times of distilled water, adjusting pH to 2.7, adding salt 0.2% of the weight of whey protein, and stirring. Then subjecting the whey protein solution to 500W ultrasonic wave for 35 min. Placing into glass bottle, heating to 85 deg.C in water bath, and maintaining for 120 min. Taking out, adding ice water, and rapidly cooling to 4 deg.C.
4. Adding distilled water into the minced fillet slurry according to the weight ratio of 7:1, and stirring for 10min by a stirrer at 300 r/min. Weighing the minced fillet slurry, putting into a centrifugal bottle, setting the temperature at 20 ℃ to 6000 Xg, centrifuging for 17min, filtering and reserving supernate.
5. Adding the supernatant into the whey protein liquid obtained in the step 3 according to the volume ratio of 1:3, stirring for 10min at 300r/min, and gradually adding trehalose and corn oil during stirring, wherein 0.7 g of trehalose and 1.2mL of corn oil are added into 100mL of mixed liquid. The stirring speed is increased to 800r/min, and the stirring is carried out for 20 min. And (3) carrying out spray drying on the mixed solution, and collecting the sprayed dry powder to obtain the selenoprotein powder.
6. And (4) measuring the total nitrogen content of the selenoprotein powder in the step 5 by a Kjeldahl method. 15.2 percent of selenoprotein powder, 15.2 percent of milk powder, 0.3 percent of algae oil powder, 5.2 percent of maltodextrin, 0.04 percent of lactoferrin, 37 percent of disintegrant, 1.5 percent of aspartame, 2.7 percent of polyvinylpyrrolidone, 4.6 percent of polyethylene glycol 6000 and 14.7 percent of mannitol in the step 5.
7. Tabletting the mixed powder by a tabletting machine, sealing and packaging.
Example 2
A preparation method of selenium protein powder comprises the following steps:
1. cleaning large yellow croaker, removing head and tail, bone, skin and viscera, adding salt 1.65 wt% of fish meat, uniformly coating the salt on the surface of fish body, and placing in a refrigerator at 4 deg.C for 90min to cool the center temperature of fish body to 6 deg.C. Washing the surface of the fish body with salt by using cold water at 4 ℃, and draining. Chopping fish, and beating in food beating machine (80 times/min) for 22min to obtain meat paste.
2. Adding selenium-enriched yeast according to the weight of 0.35mg/Kg to the fish meat paste pulp in the step 1, and slowly stirring uniformly. Heating the fish meat paste pulp to 30 ℃, and keeping the temperature for 3 hours. Every 100 g of meat paste pulp is put into a sealed bag, vacuumized to 0.02-0.04MPa and sealed. Placing the sealed bag into a high static pressure device, and continuing fermenting under 35MPa for 40 min. Then pressurizing to 350MPa for sterilization, and maintaining for 10 min. And after pressure relief, taking out the sealing bag, and pouring out the minced fillet slurry.
3. Dissolving whey protein in 10 times of distilled water, adjusting pH to 2.5, adding salt 0.25% of the weight of whey protein, and stirring. Then subjecting the whey protein solution to ultrasonic treatment at 400W for 40 min. Placing into glass bottle, heating to 87 deg.C in water bath, and maintaining for 110 min. Taking out, adding ice water, and rapidly cooling to 4 deg.C.
4. Adding distilled water into the minced fillet slurry according to the weight ratio of 6:1, and stirring for 10min by a stirrer at 300 r/min. Weighing minced fillet slurry, placing into a centrifugal bottle, setting the temperature at 20 ℃ to 6000 Xg, centrifuging for 20min, filtering and reserving supernate.
5. Adding the supernatant into the whey protein liquid obtained in the step 3 according to the volume ratio of 1:3, stirring for 10min at 300r/min, and gradually adding trehalose and corn oil during stirring, wherein 0.6 g of trehalose and 1.3mL of corn oil are added into 100mL of mixed liquid. The stirring speed is increased to 800r/min, and the stirring is carried out for 20 min. And (3) carrying out spray drying on the mixed solution, and collecting the sprayed dry powder to obtain the selenoprotein powder.
6. And (4) measuring the total nitrogen content of the selenoprotein powder in the step 5 by a Kjeldahl method. 15.8 percent of selenoprotein powder, 15.4 percent of milk powder, 0.4 percent of algae oil powder, 5.6 percent of maltodextrin, 0.02 percent of lactoferrin, 36 percent of disintegrant, 1.9 percent of aspartame, 2.6 percent of polyvinylpyrrolidone, 4.4 percent of polyethylene glycol 6000 and 14.5 percent of mannitol in the step 5.
7. Tabletting the mixed powder by a tabletting machine, sealing and packaging.
Example 3
A preparation method of selenium protein powder comprises the following steps:
1. cleaning large yellow croaker, removing head and tail, bone, skin and viscera, adding salt 1.8 wt% of fish meat, uniformly coating the salt on the surface of fish body, and placing in a refrigerator at 4 deg.C for 80min to cool the central temperature of fish body to 4 deg.C. Washing the surface of the fish body with salt by using cold water at 4 ℃, and draining. Chopping fish, and beating in food beating machine (80 times/min) for 25min to obtain meat paste.
2. Adding selenium-enriched yeast according to the weight of 0.3mg/Kg to the fish meat paste pulp in the step 1, and slowly stirring uniformly. Heating the fish meat paste pulp to 32 ℃, and keeping the temperature for 3.25 hours. Every 100 g of meat paste pulp is put into a sealed bag, vacuumized to 0.02-0.04MPa and sealed. Placing the sealed bag into a high static pressure device, and continuing fermenting under 30MPa for 45 min. Then pressurizing to 310MPa for sterilization, and maintaining for 11 min. And after pressure relief, taking out the sealing bag, and pouring out the minced fillet slurry.
3. Dissolving whey protein in 10 times of distilled water, adjusting pH to 3.0, adding salt 0.3% of the weight of whey protein, and stirring. Then the whey protein solution is treated with ultrasonic wave at 600W for 30 min. Placing into glass bottle, heating to 90 deg.C in water bath, and maintaining for 100 min. Taking out, adding ice water, and rapidly cooling to 4 deg.C.
4. Adding distilled water into the minced fillet slurry according to the weight ratio of 8:1, and stirring for 10min by a stirrer at 300 r/min. Weighing minced fillet slurry, placing into a centrifugal bottle, setting the temperature at 20 ℃ to 6000 Xg, centrifuging for 15min, filtering and reserving supernate.
5. Adding the supernatant into the whey protein liquid obtained in the step 3 according to the volume ratio of 1:3, stirring for 10min at 300r/min, and gradually adding trehalose and corn oil during stirring, wherein 0.5 g of trehalose and 1.4mL of corn oil are added into 100mL of mixed liquid. The stirring speed is increased to 800r/min, and the stirring is carried out for 20 min. And (3) carrying out spray drying on the mixed solution, and collecting the sprayed dry powder to obtain the selenoprotein powder.
6. And (4) measuring the total nitrogen content of the selenoprotein powder in the step 5 by a Kjeldahl method. 16.4 percent of selenoprotein powder, 15 percent of milk powder, 0.2 percent of seaweed oil powder, 5.4 percent of maltodextrin, 0.03 percent of lactoferrin, 38 percent of disintegrant, 1.7 percent of aspartame, 2.8 percent of polyvinylpyrrolidone, 4.8 percent of polyethylene glycol 6000 and 14.3 percent of mannitol in the step 5.
7. Tabletting the mixed powder by a tabletting machine, sealing and packaging.
Comparative example 1
1. Taking pork leg meat as a raw material, cleaning, adding salt according to 1.8% of the weight of the pork, uniformly coating the salt on the surface of the pork, and then placing the pork in a refrigerator at 4 ℃ for 80min to reduce the central temperature of the pork to 4 ℃. Washing the pork surface with cold water of 4 deg.C, and draining. Chopping pork, and beating in food beating machine (80 times/min) for 25min to obtain meat paste.
2. Adding yeast according to the weight of 0.3mg/Kg to the fish meat paste pulp in the step 1, and slowly stirring uniformly. Heating the pork paste pulp to 32 ℃, and keeping the constant temperature for 3.25 hours. Every 100 g of meat paste pulp is filled into a sealed bag and is vacuumized and sealed. Placing the sealed bag into a high static pressure device, and continuing fermenting under 30MPa for 45 min. Then pressurizing to 310MPa for sterilization, and maintaining for 11 min. And after pressure relief, taking out the sealing bag, and pouring out the meat paste.
3. Dissolving whey protein in 10 times of distilled water, adjusting pH to 3.0, adding salt 0.3% of the weight of whey protein, and stirring. Then the whey protein solution is treated with ultrasonic wave at 600W for 30 min. Placing into glass bottle, heating to 90 deg.C in water bath, and maintaining for 100 min. Taking out, adding ice water, and rapidly cooling to 4 deg.C.
4. Adding distilled water into the meat paste pulp according to the weight ratio of 8:1, and stirring for 10min by a stirrer at 300 r/min. Weighing the meat paste pulp, putting into a centrifugal bottle, setting the temperature at 20 ℃ to 6000 Xg, centrifuging for 15min, filtering and reserving supernate.
5. Adding the supernatant into the whey protein liquid obtained in the step 3 according to the volume ratio of 1:3, stirring at 300r/min for 10min, and adding trehalose and corn oil at the speed of 10mL/min during stirring, wherein the addition amount of trehalose to 100mL of mixed liquid is 0.5 g, and the addition amount of corn oil is 1.4 mL. The stirring speed is increased to 800r/min, and the stirring is carried out for 20 min. And (3) carrying out spray drying on the mixed solution, and collecting the sprayed dry powder to obtain the protein powder.
6. And (3) measuring the total nitrogen content of the protein powder in the step 5 by a Kjeldahl method. 16.4 percent of protein powder, 15 percent of milk powder, 0.2 percent of seaweed oil powder, 5.4 percent of maltodextrin, 0.03 percent of lactoferrin, 38 percent of disintegrant, 1.7 percent of aspartame, 2.8 percent of polyvinylpyrrolidone, 4.8 percent of polyethylene glycol 6000 and 14.3 percent of mannitol in the step 5.
7. Tabletting the mixed powder by a tabletting machine, sealing and packaging.
Test example 1:
1) cytotoxicity assay (survival):
caco-2 cells were cultured at 1X 104The density of individual cells/well was plated in collagen-coated 96-well plates, which were incubated at 37 ℃ with CO2Incubate in incubator for 24 h. After removing the medium, 200. mu.L of samples of different concentrations were added to each well, and 8-10 parallel wells were prepared for each sample. After incubation at 37 ℃ for 20 hours in a CO2 incubator, 20. mu.L of MTT (detection reagent) (5mg/mL) was added to each well and after incubation for 4h, the precipitate was removed by centrifugation. Then, 150 μ L DMSO (dimethyl sulfoxide) was added to each well, and the cell culture plate was shaken at low speed for 10 min. Measuring with enzyme-linked immunosorbent assay at 490nmMeasure absorbance.
Cell survival rate (%) ═ B1-B0)*100/B2-B0
In the formula: b is0Absorbance without cells; b is1Is the absorbance of the added sample; b is2Absorbance without sample.
2) And constructing an intestinal cell model in vitro.
The cells were cultured at 1X 105The density of individual cells/well was seeded in 12-well plates. The medium was changed every other day in the first week and every day in the second week. After 21 days of culture, an in vitro intestinal cell model was obtained. Removing Caco-2 cell culture medium, washing HBSS (cell culture solution) without selenium buffer solution twice, preheating at 37 deg.C for 10min, removing HBSS buffer solution, adding samples with different concentrations, and culturing in incubator for different times. After the reaction, the reaction solution was washed twice with HBSS buffer at 4 ℃ to remove the residual extract, 500. mu.L of 0.1% (v/v) Triton X-100 cell lysate was added, repeated freezing and thawing were performed to lyse the cells, and then sonication was performed for 10min to obtain a cell suspension. Total protein mass concentration (g/L) was measured with BCA kit, another portion was removed from the cell suspension, centrifuged at 15000r/min for 10min, and the selenium ion content in the solution and alkaline protease (AKP) activity was determined by atomic absorption spectrophotometer using the AKP activity detection kit. In the experiment, the control group is cultured by adding only culture medium, the sodium selenite group is cultured by adding 0.02mg/mL solution, and the model group is cultured by adding selenium-free HBSS.
The cellular uptake rate (%) ═ C1/C2
In the formula: c1Is the intracellular selenium concentration; c2Is the total cell protein concentration.
TABLE 1 in vitro Small intestinal mucosal cancer cells (Caco-2)
As can be seen from Table 1, compared with sodium selenite, the total amount of selenium retained in mice is higher, and the organic selenium product provided by the invention can be absorbed in stomach and intestines, therefore, the product has wide absorption range in human body and is easier to be absorbed and utilized by human body. Compared with the comparative example 1, the selenium retention of the product provided by the invention is far higher than that of the comparative example using pork.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. The preparation method of the selenium protein powder is characterized by comprising the following steps:
1) pretreatment: cleaning fish meat, uniformly coating salt on the surface of the fish meat, cooling the fish meat to 4-6 ℃, refrigerating, washing the salt, draining water, chopping and beating to obtain fish meat paste;
2) fermentation: uniformly mixing the fish meat paste pulp with selenium-enriched yeast, carrying out aerobic fermentation, pumping the aerobic fermentation product to a certain vacuum degree, sealing, maintaining the internal air pressure at 0.02-0.04MPa, putting the aerobic fermentation product into a high-pressure container of 15-20MPa, carrying out sealed fermentation, and carrying out high-pressure sterilization treatment on the fermentation product to obtain the fermented fish meat paste pulp;
3) extraction: uniformly stirring the fermented fish meat paste pulp with water, centrifuging, and keeping supernatant;
4) adding salt into the whey protein solution, performing ultrasonic treatment, heating, and rapidly cooling to 4-6 deg.C to obtain solution A;
5) preparing selenium protein powder: and uniformly stirring the supernatant and the solution A to obtain a mixed solution B, slowly adding trehalose and corn oil into the mixed solution B at the speed of 10mL/min, uniformly stirring again, and spray-drying to obtain the selenoprotein powder.
2. The method for preparing selenoprotein powder according to claim 1, wherein the fish meat in step 1) is large yellow croaker meat; the mass ratio of the fish meat to the salt is 1: 0.015-0.018; the refrigerating time is 80-100 min.
3. The method for preparing the selenium protein powder according to claim 1, wherein the selenium-enriched yeast is added in the step 2) in an amount of 0.3-0.4mg/Kg based on the weight of the fish meat; the aerobic fermentation temperature is 30-32 ℃, and the fermentation time is 3-3.5 hours; the low-pressure sealed fermentation pressure is 30-35MPa, and the fermentation time is 40-50 min; the high-pressure sterilization pressure is 280-350MPa, and the high-pressure sterilization time is 10-12 min.
4. The preparation method of the selenoprotein powder as claimed in claim 1, wherein the mass ratio of the fermented fish meat emulsion slurry to water in step 3) is (6-8): 1; the stirring speed is 300-400r/min, and the stirring time is 10-15 min; the centrifugal temperature is 15-25 ℃, the centrifugal force is 5500-6500 Xg, and the centrifugal time is 15-20 min.
5. The method for preparing the selenium protein powder as claimed in claim 1, wherein the mass ratio of the whey protein to the water in the whey protein solution in the step 4) is 1:10, and the pH value is 2.5-3.0; the weight ratio of the whey protein to the salt is 1 (0.002-0.003).
6. The method for preparing the selenium protein powder as claimed in claim 5, wherein the ultrasonic power in step 4) is 400-600W, and the ultrasonic time is 30-40 min; the heating temperature is 85-90 ℃, and the heating time is 100-120 min; the cooling speed is 20 ℃/min.
7. The method for preparing the selenium protein powder as claimed in claim 1, wherein the volume ratio of the supernatant to the solution A in the step 5) is 1: 3; the adding ratio of the solution B to the trehalose and the corn oil is 100 mL: (0.5-0.7) g: (1.2-1.4) mL.
8. The method for preparing selenium protein powder as claimed in claim 7, wherein the stirring speed in step 5) is 300-400r/min, and the stirring time is 10-15 min; the stirring is carried out for 800-900r/min again and 20-25 min.
9. A selenoprotein tablet, characterized by comprising the selenoprotein powder of any one of claims 1-8, wherein the preparation method of the selenoprotein tablet comprises the following steps:
according to the weight portion, 15.2-16.4 portions of selenoprotein powder, 15-15.4 portions of milk powder, 0.2-0.4 portion of algae oil powder, 5.2-5.6 portions of maltodextrin, 0.02-0.04 portion of lactoferrin powder, 36-38 portions of disintegrating agent, 1.5-1.9 portions of aspartame, 2.6-2.8 portions of polyvinylpyrrolidone, 60004.4-4.8 portions of polyethylene glycol and 14.3-14.7 portions of mannitol are uniformly mixed and pressed into tablets, and then the selenoprotein tablets are obtained.
10. The method of using a selenoprotein tablet as claimed in claim 9, comprising the steps of: the selenoprotein tablet prepared by the invention can be drunk after being put into warm boiled water with the temperature of 50-60 ℃ for 25-30 s; the weight ratio of the selenoprotein tablets to the warm boiled water is 1 (80-150).
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