A kind of feed protein raw material and preparation method thereof

Abstract

The present invention relates to the field of feed, and in particular to a feed protein raw material and a preparation method thereof, comprising the following raw materials: protein-seaweed polyphenol complex, beta-carotene, cysteine, whey protein hydrolyzate, vitamins, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, linseed oil, amylase, sodium chloride, corn fiber, lactic acid bacteria, superoxide dismutase and glutathione peroxidase-zinc complex. The present invention removes peroxides in cells by utilizing glutathione, and protein-seaweed polyphenols directly remove free radicals, protecting cell components and glutathione reductase. Cysteine ​​supplies glutathione to form a dynamic equilibrium circulation system.

Description

Technical Field

[0001] The invention relates to the field of feed, and in particular to a feed protein raw material and a preparation method thereof. Background Art

[0002] As an important part of modern animal husbandry, feed protein raw materials play a key role in animal nutrition. However, there are still many shortcomings.

[0003] First, the supply of traditional protein sources such as soybean meal and fish meal is unstable. Affected by climate change, fishing restrictions and international trade policies, prices fluctuate greatly, increasing the operating risks of the aquaculture industry. Secondly, the amino acid composition of some protein raw materials is not ideal. They may lack certain essential amino acids or have insufficient content, requiring additional synthetic amino acids to balance, increasing production costs and complexity. Furthermore, some plant protein raw materials contain anti-nutritional factors, such as trypsin inhibitors and phytic acid, which affect the digestion and absorption of protein and other nutrients by animals and reduce feed utilization efficiency.

[0004] In addition, the production process of traditional protein raw materials may have a negative impact on the environment. For example, large-scale soybean cultivation leads to deforestation, and overfishing threatens the ecological balance of the ocean. In terms of processing, some proteins are prone to denaturation during the processing process, which affects their nutritional value and functional properties, requiring special processes and measures, increasing the difficulty and cost of production. The palatability issue is also a challenge. Some protein raw materials may have poor palatability, which affects the animal's feed intake and thus affects growth performance. Biosafety risks should not be ignored either. Some animal-derived proteins, such as meat and bone meal, may have the risk of disease transmission and are banned in some countries and regions. Finally, the high price of high-quality protein raw materials such as fish meal limits its widespread application. Summary of the invention

[0005] (1) Technical issues to be resolved

[0006] The purpose of the present invention is to provide a feed protein raw material and a preparation method thereof, so as to reduce oxidative stress in animals, improve health level and disease resistance, and improve growth performance of animals.

[0007] (2) Technical solution

[0008] To achieve the above-mentioned object, on the one hand, the present invention provides a protein raw material for feeding, comprising the following raw materials in parts by weight: 43-72 parts of feed base, 35-45 parts of protein-seaweed polyphenol complex, 0.3-0.7 parts of glutathione peroxidase-zinc complex, and 0.5-1.5 parts of cysteine; the protein-seaweed polyphenol complex is the following raw materials in parts by weight: 33-43 parts of whey protein and 1-3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ion through coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0009] Furthermore, the feed protein raw material described herein is a feed base material composed of the following raw materials in parts by weight: 0.05-0.2 parts of beta-carotene, 15-25 parts of whey protein hydrolyzate, 0.5-1.5 parts of vitamin C, 1.5-2.5 parts of lysine, 0.8-1.2 parts of methionine, 0.3-0.7 parts of vitamin E, 0.03-0.07 parts of copper sulfate, 0.03-0.07 parts of manganese sulfate, 3-7 parts of linseed oil, 0.05-0.15 parts of amylase, 0.8-1.2 parts of sodium chloride, 20-30 parts of corn fiber, 0.05-0.15 parts of lactic acid bacteria, and 0.05-0.15 parts of superoxide dismutase.

[0010] Furthermore, the method for preparing the protein-seaweed polyphenol complex comprises:

[0011] S11. Weigh whey protein powder, dissolve it in phosphate buffer, stir it at room temperature for 25-30 min using a magnetic stirrer, filter it with a 0.22 μm filter membrane to remove insoluble matter, and obtain a whey protein solution;

[0012] S12. Weigh seaweed polyphenol powder, dissolve it in 50% to 60% ethanol aqueous solution, ultrasonically treat it for 10 to 15 minutes, and filter it with a 0.22 μm filter membrane to remove insoluble matter to obtain a seaweed polyphenol solution;

[0013] S13. In a beaker, slowly drop the seaweed polyphenol solution into the whey protein solution while maintaining gentle stirring, adjust the pH to 6.5-7.0 using dilute NaOH or HCl, and continue stirring at 25-30° C. for 2-3 hours to obtain a first mixed solution;

[0014] S14. The first mixed solution is transferred to an ultrafiltration device with a molecular weight cutoff of 10 kDa, and ultrafiltration is performed at 4 to 5° C. to concentrate the first mixed solution to 1 / 3 to 1 / 2 of the original volume to obtain a concentrated solution;

[0015] S15. The concentrated solution is divided into freeze-drying bottles, pre-frozen at -80~-90°C for 2~3 hours, and freeze-dried for 48~50 hours until a dry protein-seaweed polyphenol complex is obtained.

[0016] Furthermore, the preparation method of the glutathione peroxidase-zinc complex comprises:

[0017] S21. Dissolve the purified glutathione peroxidase in 50 mM pH 7.4 Tris-HCl buffer, adjust the concentration to 1 mg / mL, and filter using a 0.22 μm filter membrane to obtain a glutathione peroxidase solution;

[0018] S22. Prepare a 10 mM ZnCl2 solution, dissolve it in ultrapure water, and filter it with a 0.22 μm filter membrane to sterilize it to obtain a ZnCl2 solution;

[0019] S23. In a centrifuge tube, mix the glutathione peroxidase solution and the ZnCl2 solution, gently vortex to mix, and keep in contact at 4-5°C for 2-3 hours to obtain a second mixed solution;

[0020] S24. The second mixed solution is transferred to a pretreated dialysis bag, and dialyzed against 50 mM pH 7.4 Tris-HCl buffer at 4-5°C for 4 times, each time for 4 hours, and fresh buffer is replaced each time to obtain a dialyzed solution;

[0021] S25. The dialyzed solution was transferred to a centrifuge tube, and centrifuged at 10,000 g for 15 min at 4-5°C to remove the formed precipitate to obtain a centrifuged solution;

[0022] S26. Use an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa to concentrate the sample at 4~5℃ and centrifuge to the desired concentration to obtain glutathione peroxidase-zinc complex.

[0023] On the other hand, based on the same inventive concept, the present invention also provides a method for preparing a feed protein raw material, which is applied to the feed protein raw material, comprising the following steps:

[0024] S31. Preparation of protein-seaweed polyphenol complex;

[0025] S32. Preparation of glutathione peroxidase-zinc complex;

[0026] S33. β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, amylase, sodium chloride, corn fiber, lactic acid bacteria, whey protein hydrolysate, superoxide dismutase and protein-seaweed polyphenol complex were sieved through a 60-mesh sieve, and linseed oil and glutathione peroxidase-zinc complex were preheated to 25°C;

[0027] S34. In a mixer, add β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, sodium chloride and protein-seaweed polyphenol complex according to the formula ratio, mix for 15 minutes, slowly add preheated linseed oil, and continue mixing for 10 minutes to obtain a first mixture;

[0028] S35. Dissolving amylase, glutathione peroxidase-zinc complex and superoxide dismutase in distilled water to obtain an enzyme solution, spraying the enzyme solution evenly onto the first mixture, and continuing to mix for 5 min to obtain a second mixture;

[0029] S36. Premixing the lactic acid bacteria powder with a small amount of corn fiber and whey protein hydrolyzate to obtain a premix, and evenly sprinkling the premix into the second mixture, mixing for 2 minutes to obtain a third mixture;

[0030] S37. The third mixture is transferred to a granulator at a temperature of 40 to 45°C and a humidity of 15 to 18%, and granulated to obtain a granular feed protein raw material having a diameter of 2 to 3 mm;

[0031] S38. The granular protein raw material for feed is dried using a fluidized bed dryer, the drying temperature is set to 50-55°C, the time is 60-70min, and the final moisture content is controlled between 8-10% to obtain a dry protein raw material for feed;

[0032] S39. The dried feed protein raw material is transferred to a cooler, cooled to room temperature, and sieved using an 80-mesh sieve to remove fine powder and oversized particles to obtain a feed protein raw material.

[0033] The mechanism of action of the above raw material components is as follows:

[0034] Protein-seaweed polyphenol complex: Seaweed polyphenols are powerful antioxidants that can directly scavenge free radicals and prevent lipid peroxidation. Combining with protein may enhance its stability and bioavailability.

[0035] β-Carotene: As a fat-soluble antioxidant, it can scavenge singlet oxygen and peroxyl free radicals. It is a precursor of vitamin A and helps maintain immune system function.

[0036] Cysteine: It is the precursor of glutathione, which is an important antioxidant in the body. It contains sulfhydryl groups, which can directly react with free radicals to play an antioxidant role.

[0037] Whey protein hydrolysate: After whey protein is hydrolyzed, peptides with antioxidant activity are produced. These peptides can exert antioxidant effects through a variety of mechanisms, including scavenging free radicals, chelating metal ions, and inhibiting oxidase activity.

[0038] Vitamin C: A water-soluble antioxidant that can directly scavenge free radicals. It can regenerate other antioxidants, such as vitamin E.

[0039] Lysine and methionine: These essential amino acids can produce antioxidant substances such as glutathione and S-adenosylmethionine during metabolism in the body, which play an important role in the antioxidant defense of cells.

[0040] Vitamin E: A fat-soluble antioxidant that mainly protects cell membranes from damage caused by lipid peroxidation. It can work synergistically with vitamin C to enhance the antioxidant effect.

[0041] Copper and manganese: These trace elements are cofactors of various antioxidant enzymes, such as superoxide dismutase and glutathione peroxidase, which play an important role in the cell's antioxidant defense system.

[0042] Flaxseed oil: Rich in omega-3 fatty acids, it has anti-inflammatory and indirect antioxidant properties.

[0043] Amylase: Amylase itself does not directly have an antioxidant effect, but it may indirectly affect energy metabolism and antioxidant status in the body by helping digestion and absorption of carbohydrates.

[0044] Sodium, corn fiber: Corn fiber, as a dietary fiber, can play an antioxidant role by promoting intestinal health and reducing oxidative stress. Sodium is an important mineral for maintaining electrolyte balance and cell function.

[0045] Lactic acid bacteria: As probiotics, lactic acid bacteria can indirectly exert antioxidant effects by regulating the balance of intestinal microecology, enhancing immunity and inhibiting the growth of pathogens.

[0046] Glutathione peroxidase-zinc complex: Glutathione peroxidase is an important antioxidant enzyme that can catalyze the decomposition of hydrogen peroxide. Zinc acts as a cofactor to enhance the activity and stability of the enzyme.

[0047] Superoxide dismutase: Superoxide dismutase is an important antioxidant enzyme that catalyzes the dismutation of superoxide anion radicals into oxygen and hydrogen peroxide, thereby protecting cells from damage by reactive oxygen species.

[0048] (3) Beneficial effects

[0049] Compared with the prior art, the present invention has the following beneficial effects:

[0050] 1. The present invention forms a multi-level, dynamically balanced antioxidant system through the synergistic effect of glutathione peroxidase-zinc complex, protein-seaweed polyphenol complex and cysteine. This system not only provides immediate protection, but also regulates long-term antioxidant capacity, greatly improving the overall antioxidant effect of the product.

[0051] 2. By synergistically regulating the Nrf2 pathway, the present invention enhances the long-term antioxidant capacity in animals, which is difficult to achieve with traditional antioxidants.

[0052] 3. By adding lysine, methionine and cysteine, the present invention optimizes the amino acid composition of the protein, reduces the need for additional addition of synthetic amino acids, and reduces production costs and complexity. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 This is a morphological diagram of the feed protein raw material according to Example 1 of the present invention. DETAILED DESCRIPTION

[0054] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0055] The test equipment and preparations of the examples described below are as follows:

[0056] Electronic balance (Sartorius, Germany), electric blast constant temperature dryer (Shanghai Formosa Experimental Equipment), stainless steel reactor (Shanghai Laibei), electric constant temperature water bath (Jiangsu Kedao), rotary evaporator (Shanghai Darlo Scientific Instruments), pulverizer (Shandong Tianfang Machinery), magnetic stirrer (Shanghai Meiyingpu), ball mill (Henan Zhengkuang), ultrasonic reactor (Shanghai Ouhe Machinery Equipment Co., Ltd.); glutathione peroxidase was purchased from Shanghai Kanglang Biotechnology Co., Ltd., seaweed polyphenols were purchased from Shaanxi Snot Biotechnology Co., Ltd., and chemicals and reagents were purchased from Sigma-Aldrich.

[0057] Example 1: This example discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.86 parts of feed base, 40 parts of protein-seaweed polyphenol complex, 0.6 parts of glutathione peroxidase-zinc complex, and 1.2 parts of cysteine;

[0058] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0059] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0060] The glutathione peroxidase-zinc complex catalyzes the removal of peroxides by glutathione. Zinc ions, as cofactors, enhance the activity of glutathione peroxidase and can also indirectly regulate the synthesis of glutathione by affecting the metabolism of cysteine ​​by affecting multiple enzymes related to cysteine ​​metabolism.

[0061] Protein-seaweed polyphenols increase the activity of glutathione peroxidase in two ways: first, they form non-covalent interactions such as van der Waals forces and electrostatic interactions with glutathione peroxidase-zinc complexes to fine-tune the spatial conformation of the enzyme; second, they activate Nrf2 transcription factors and activate Nrf2 to upregulate the expression of glutathione peroxidase. The increase in glutathione peroxidase activity changes the redox state of cells, thereby affecting the activity of transcription factors such as Nrf2. Nrf2 regulates the expression of key enzymes in animals, such as cystathionine-β-synthetase and cystathionine-γ-lyase, and then regulates the content of cysteine. At the same time, the consumption of glutathione stimulates the synthesis and transport of cysteine ​​to maintain a stable level of glutathione. The polyphenol structure of protein-seaweed polyphenols not only directly scavenges free radicals, but also forms short-term hydrogen bonds or hydrophobic interactions with the sulfhydryl groups of cysteine. This transient molecular binding is called a "transient complex". When seaweed polyphenols form a complex with cysteine, they can redistribute the electron cloud. Polyphenol molecules are usually rich in electrons, especially their aromatic ring systems. These electrons are partially transferred to the sulfhydryl group (-SH) of cysteine, increasing the electron density around the sulfur atom. This increase in electron density reduces the tendency of cysteine ​​to lose electrons, changes the redox potential of cysteine, and further enhances the antioxidant capacity of cysteine. It can also change the redox potential of cysteine ​​through its polyphenol structure, thereby enhancing its antioxidant capacity.

[0062] In addition, cysteine ​​affects the activity of multiple redox-sensitive signaling proteins including Nrf2, NF-κB and MAPK by regulating the balance of thiol-disulfide bonds in cells, enabling cells to maintain redox balance to different degrees of oxidative stress and protect cells from oxidative damage.

[0063] In conclusion, the protein-algae polyphenol complex, glutathione peroxidase-zinc complex, and cysteine ​​form a highly integrated, interdependent antioxidant network that not only provides powerful immediate antioxidant protection but also regulates the long-term antioxidant capacity of cells through multi-level synergistic effects.

[0064] The method for preparing the protein-seaweed polyphenol complex comprises:

[0065] S11. Weigh whey protein powder, dissolve it in phosphate buffer, stir it at room temperature for 25-30 min using a magnetic stirrer, filter it with a 0.22 μm filter membrane to remove insoluble matter, and obtain a whey protein solution;

[0066] S12. Weigh seaweed polyphenol powder, dissolve it in 50% to 60% ethanol aqueous solution, ultrasonically treat it for 10 to 15 minutes, and filter it with a 0.22 μm filter membrane to remove insoluble matter to obtain a seaweed polyphenol solution;

[0067] S13. In a beaker, slowly drop the seaweed polyphenol solution into the whey protein solution while maintaining gentle stirring, adjust the pH to 6.5-7.0 using dilute NaOH or HCl, and continue stirring at 25-30° C. for 2-3 hours to obtain a first mixed solution;

[0068] S14. The first mixed solution is transferred to an ultrafiltration device with a molecular weight cutoff of 10 kDa, and ultrafiltration is performed at 4 to 5° C. to concentrate the first mixed solution to 1 / 3 to 1 / 2 of the original volume to obtain a concentrated solution;

[0069] S15. The concentrated solution is divided into freeze-drying bottles, pre-frozen at -80~-90°C for 2~3 hours, and freeze-dried for 48~50 hours until a dry protein-seaweed polyphenol complex is obtained.

[0070] The preparation method of the glutathione peroxidase-zinc complex comprises:

[0071] S21. Dissolve the purified glutathione peroxidase in 50 mM pH 7.4 Tris-HCl buffer, adjust the concentration to 1 mg / mL, and filter using a 0.22 μm filter membrane to obtain a glutathione peroxidase solution;

[0072] S22. Prepare a 10 mM ZnCl2 solution, dissolve it in ultrapure water, and filter it with a 0.22 μm filter membrane to sterilize it to obtain a ZnCl2 solution;

[0073] S23. In a centrifuge tube, mix the glutathione peroxidase solution and the ZnCl2 solution, gently vortex to mix, and keep in contact at 4-5°C for 2-3 hours to obtain a second mixed solution;

[0074] S24. The second mixed solution is transferred to a pretreated dialysis bag, and dialyzed against 50 mM pH 7.4 Tris-HCl buffer at 4-5°C for 4 times, each time for 4 hours, and fresh buffer is replaced each time to obtain a dialyzed solution;

[0075] S25. The dialyzed solution was transferred to a centrifuge tube, and centrifuged at 10,000 g for 15 min at 4-5°C to remove the formed precipitate to obtain a centrifuged solution;

[0076] S26. Use an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa to concentrate the sample at 4~5℃ and centrifuge to the desired concentration to obtain glutathione peroxidase-zinc complex.

[0077] A method for preparing a feed protein raw material, applied to the feed protein raw material, comprises the following steps:

[0078] S31. Preparation of protein-seaweed polyphenol complex;

[0079] S32. Preparation of glutathione peroxidase-zinc complex;

[0080] S33. β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, amylase, sodium chloride, corn fiber, lactic acid bacteria, whey protein hydrolysate, superoxide dismutase and protein-seaweed polyphenol complex were sieved through a 60-mesh sieve, and linseed oil and glutathione peroxidase-zinc complex were preheated to 25°C;

[0081] S34. In a mixer, add β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, sodium chloride and protein-seaweed polyphenol complex according to the formula ratio, mix for 15 minutes, slowly add preheated linseed oil, and continue mixing for 10 minutes to obtain a first mixture;

[0082] S35. Dissolving amylase, glutathione peroxidase-zinc complex and superoxide dismutase in distilled water to obtain an enzyme solution, spraying the enzyme solution evenly onto the first mixture, and continuing to mix for 5 min to obtain a second mixture;

[0083] S36. Premixing the lactic acid bacteria powder with a small amount of corn fiber and whey protein hydrolyzate to obtain a premix, and evenly sprinkling the premix into the second mixture, mixing for 2 minutes to obtain a third mixture;

[0084] S37. The third mixture is transferred to a granulator at a temperature of 40 to 45°C and a humidity of 15 to 18%, and granulated to obtain a granular feed protein raw material having a diameter of 2 to 3 mm;

[0085] S38. The granular protein raw material for feed is dried using a fluidized bed dryer, the drying temperature is set to 50-55°C, the time is 60-70min, and the final moisture content is controlled between 8-10% to obtain a dry protein raw material for feed;

[0086] S39. The dried feed protein raw material is transferred to a cooler, cooled to room temperature, and sieved using an 80-mesh sieve to remove fine powder and oversized particles to obtain a feed protein raw material.

[0087] Example 2: This example discloses a feed protein raw material, comprising the following raw materials in parts by weight: 72 parts of feed base, 45 parts of protein-seaweed polyphenol complex, and 0.7 parts of glutathione peroxidase-zinc complex;

[0088] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 43 parts of whey and 2 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by combining glutathione peroxidase and zinc ions through coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0089] The feed base material comprises the following raw materials in parts by weight: 0.2 parts of beta-carotene, 1.5 parts of cysteine, 25 parts of whey protein hydrolyzate, 1.5 parts of vitamin C, 2.5 parts of lysine, 1.2 parts of methionine, 0.7 parts of vitamin E, 0.07 parts of copper sulfate, 0.07 parts of manganese sulfate, 7 parts of linseed oil, 0.15 parts of amylase, 1.2 parts of sodium chloride, 30 parts of corn fiber, 0.15 parts of lactic acid bacteria, and 0.15 parts of superoxide dismutase.

[0090] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0091] Example 3: This example discloses a feed protein raw material, comprising the following raw materials in parts by weight: 43 parts of feed base, 35 parts of protein-seaweed polyphenol complex, 0.3 parts of glutathione peroxidase-zinc complex, and 0.5 parts of cysteine;

[0092] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 33 parts of whey protein and 2 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0093] The feed base material comprises the following raw materials in parts by weight: 0.05 parts of beta-carotene, 15 parts of whey protein hydrolyzate, 0.5 parts of vitamin C, 1.5 parts of lysine, 0.8 parts of methionine, 0.3 parts of vitamin E, 0.03 parts of copper sulfate, 0.03 parts of manganese sulfate, 3 parts of linseed oil, 0.05 parts of amylase, 0.8 parts of sodium chloride, 20 parts of corn fiber, 0.05 parts of lactic acid bacteria, and 0.05 parts of superoxide dismutase.

[0094] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0095] Example 4: This example discloses a feed protein raw material, comprising the following raw materials in parts by weight: 47.39 parts of feed base, 36 parts of protein-seaweed polyphenol complex, 0.4 parts of glutathione peroxidase-zinc complex, and 0.6 parts of cysteine;

[0096] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 35 parts of whey protein and 1 part of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by combining glutathione peroxidase and zinc ions through coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0097] The feed base material comprises the following raw materials in parts by weight: 0.06 parts of beta-carotene, 16 parts of whey protein hydrolyzate, 0.6 parts of vitamin C, 1.6 parts of lysine, 0.9 parts of methionine, 0.4 parts of vitamin E, 0.04 parts of copper sulfate, 0.04 parts of manganese sulfate, 4 parts of linseed oil, 0.08 parts of amylase, 0.9 parts of sodium chloride, 22 parts of corn fiber, 0.08 parts of lactic acid bacteria, and 0.09 parts of superoxide dismutase.

[0098] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0099] Example 5: This example discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.62 parts of feed base, 38 parts of protein-seaweed polyphenol complex, 0.6 parts of glutathione peroxidase-zinc complex, and 0.9 parts of cysteine;

[0100] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 35 parts of whey protein and 3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0101] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 21 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 1.8 parts of lysine, 1.2 parts of methionine, 0.6 parts of vitamin E, 0.06 parts of copper sulfate, 0.06 parts of manganese sulfate, 6 parts of linseed oil, 0.13 parts of amylase, 1.2 parts of sodium chloride, 29 parts of corn fiber, 0.14 parts of lactic acid bacteria, and 0.13 parts of superoxide dismutase.

[0102] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0103] Control group 1: The difference between this example and Example 1 is that the glutathione peroxidase-zinc complex replaces the protein-seaweed polyphenol complex.

[0104] This embodiment discloses a protein raw material for feeding, comprising the following raw materials in parts by weight: 63.86 parts of feed base, 40.6 parts of glutathione peroxidase-zinc complex, and 1.2 parts of cysteine;

[0105] The glutathione peroxidase-zinc complex is formed by the combination of glutathione peroxidase and zinc ion through coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0106] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0107] The preparation method of the glutathione peroxidase-zinc complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0108] Control group 2: This example differs from example 1 in that cysteine ​​replaces the protein-seaweed polyphenol complex.

[0109] This embodiment discloses a protein raw material for feeding, comprising the following raw materials in parts by weight: 63.86 parts of feed base, 0.6 parts of glutathione peroxidase-zinc complex, and 41.2 parts of cysteine;

[0110] The glutathione peroxidase-zinc complex is formed by the combination of glutathione peroxidase and zinc ion through coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0111] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0112] The preparation method of the glutathione peroxidase-zinc complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0113] Control group 3: This example is different from example 1 in that the protein-seaweed polyphenol complex replaces the glutathione peroxidase-zinc complex and cysteine.

[0114] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.86 parts of feed base material, 41.8 parts of protein-seaweed polyphenol complex;

[0115] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol.

[0116] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0117] The preparation method of the protein-seaweed polyphenol complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0118] Control group 4: This example differs from Example 1 in that the protein-seaweed polyphenol complex replaces the glutathione peroxidase-zinc complex.

[0119] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.86 parts of feed base, 40.6 parts of protein-seaweed polyphenol complex, and 1.2 parts of cysteine;

[0120] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol.

[0121] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0122] The preparation method of the protein-seaweed polyphenol complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0123] Control group 5: This example differs from Example 1 in that cysteine ​​replaces the glutathione peroxidase-zinc complex.

[0124] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.86 parts of feed base, 40 parts of protein-seaweed polyphenol complex, and 1.8 parts of cysteine;

[0125] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol.

[0126] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0127] The preparation method of the protein-seaweed polyphenol complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0128] Control group 6: This example differs from example 1 in that cysteine ​​replaces the glutathione peroxidase-zinc complex and the protein-seaweed polyphenol complex.

[0129] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 63.86 parts of feed base material, 41.8 parts of cysteine;

[0130] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0131] The method for preparing a feed protein raw material in this embodiment is consistent with that in Example 1.

[0132] Control group 7: This example differs from Example 1 in that the protein-seaweed polyphenol complex replaces cysteine.

[0133] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 62.66 parts of feed base, 41.2 parts of protein-seaweed polyphenol complex, and 0.6 parts of glutathione peroxidase-zinc complex;

[0134] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0135] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0136] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0137] Control group 8: This example differs from Example 1 in that glutathione peroxidase-zinc complex replaces cysteine.

[0138] This embodiment discloses a feed protein raw material, comprising the following raw materials in parts by weight: 62.66 parts of feed base, 40 parts of protein-seaweed polyphenol complex, and 1.8 parts of glutathione peroxidase-zinc complex;

[0139] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0140] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0141] The preparation methods of the protein-seaweed polyphenol complex and the glutathione peroxidase-zinc complex in this example are the same as those in Example 1. The preparation method of a feed protein raw material in this example is the same as that in Example 1.

[0142] Control group 9: This example differs from Example 1 in that glutathione peroxidase-zinc complex replaces cysteine ​​and protein-seaweed polyphenol complex.

[0143] This embodiment discloses a protein raw material for feed, comprising the following raw materials in parts by weight: 62.66 parts of feed base material, 41.8 parts of glutathione peroxidase-zinc complex;

[0144] The protein-seaweed polyphenol complex is composed of the following raw materials in parts by weight: 37 parts of whey protein and 3 parts of seaweed polyphenol; the glutathione peroxidase-zinc complex is formed by glutathione peroxidase and zinc ions being bound by coordination bonds, and the glutathione peroxidase-zinc complex has a tetramer structure.

[0145] The feed base material comprises the following raw materials in parts by weight: 0.1 part of beta-carotene, 22 parts of whey protein hydrolyzate, 1.3 parts of vitamin C, 2.1 parts of lysine, 1.1 parts of methionine, 0.5 parts of vitamin E, 0.05 parts of copper sulfate, 0.04 parts of manganese sulfate, 6 parts of linseed oil, 0.12 parts of amylase, 1.1 parts of sodium chloride, 28 parts of corn fiber, 0.13 parts of lactic acid bacteria, and 0.12 parts of superoxide dismutase.

[0146] The preparation method of the glutathione peroxidase-zinc complex of this embodiment is consistent with that of embodiment 1. The preparation method of a feed protein raw material of this embodiment is consistent with that of embodiment 1.

[0147] Test verification:

[0148] 1. Determination of DPPH radical scavenging ability: The samples of each embodiment group and the control group were prepared into 5 concentration gradients: 0.025, 0.05, 0.1, 0.2, and 0.4 mg / mL, 0.1 mM DPPH solution was mixed with the sample solutions of each embodiment group and the control group, and reacted at room temperature in the dark for 30 minutes. The absorbance was measured at a wavelength of 517 nm using a spectrophotometer, and the DPPH radical scavenging rate was calculated.

[0149] 2. β-carotene bleaching method: The samples of each embodiment group and the control group were prepared into 5 concentration gradients: 0.025, 0.05, 0.1, 0.2, and 0.4 mg / mL to prepare β-carotene-linoleic acid emulsion, and the emulsion was mixed with the sample solution of each embodiment group and the control group, and reacted in a 50°C water bath for 2 hours. The absorbance was measured at a wavelength of 470 nm using a spectrophotometer every 15 minutes, and the antioxidant activity was calculated.

[0150] 3. Animal experiment evaluation: Cattle were selected as experimental animals and randomly divided into an embodiment group and a control group. The feeding period was 8 weeks, and the feed intake and body weight changes were recorded every day.

[0151]

[0152]

[0153]

[0154] The antioxidant properties of the feed protein raw materials of the embodiment groups and the control groups are shown in Tables 1 to 3. The embodiment groups 1 to 5 and the control groups 1 to 9 showed significant differences in DPPH free radical scavenging ability, β-carotene bleaching antioxidant activity and animal experimental results.

[0155] In terms of DPPH free radical scavenging ability, the scavenging rates of the example groups at each concentration gradient were generally higher than those of the control group, and the scavenging rates showed a more obvious upward trend as the concentration increased. Example 1 achieved a maximum scavenging rate of 95% at a concentration of 0.4 mg / mL, while the maximum scavenging rate in the control group was only 83%. 50 The values ​​also reflect this trend. 50 The value was significantly lower than that of the control group, indicating that the example group had stronger free radical scavenging ability.

[0156] The antioxidant activity results of β-carotene bleaching method were consistent with the results of DPPH test. The antioxidant activity of the example group at each concentration was higher than that of the control group. 50 The value is also significantly lower than that of the control group. This further proves that the example group has stronger antioxidant capacity.

[0157] It is worth noting that the results of control groups 1 to 9 showed that glutathione peroxidase-zinc complex, protein-seaweed polyphenol complex and cysteine ​​also had certain antioxidant capacity when used alone. However, the antioxidant activity of the example group was significantly higher than that of any single component or the combination of two components.

[0158] In order to quantitatively evaluate the synergistic effect between the three components, we can focus on the "Synergy Index" column in the table. The synergy index of the embodiment group is less than 1, while the synergy index of the control group is mostly close to or equal to 1. A synergy index less than 1 indicates a positive synergy, and the smaller the value, the stronger the synergy. This data directly proves that there is a significant synergistic antioxidant effect between the three components.

[0159] The results of animal experiments more intuitively reflect the effects of different formulas on animal growth. The average feed intake and weight gain of cattle in the Example group were significantly higher than those in the Control group. This shows that the formula of the Example group not only improves the palatability of the feed, increases the feed intake of animals, but also promotes the growth and development of animals. This significant improvement further supports the conclusion that there is a synergistic effect between the three ingredients.

[0160] Finally, it should be noted that: Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A feed protein raw material, characterized in that: The method comprises the following raw materials in parts by weight: 43-72 parts of feed base material, 35-45 parts of protein-seaweed polyphenol complex, 0.3-0.7 parts of glutathione peroxidase-zinc complex, and 0.5-1.5 parts of cysteine; The protein-seaweed polyphenol complex is prepared from the following raw materials in parts by weight: 33-43 parts of whey protein and 1-3 parts of seaweed polyphenol; The method for preparing the protein-seaweed polyphenol complex comprises: S11. Weigh whey protein powder, dissolve it in phosphate buffer, stir it at room temperature for 25 to 30 minutes using a magnetic stirrer, filter it with a 0.22 μm filter membrane to remove insoluble matter, and obtain a whey protein solution; S12. Weigh seaweed polyphenol powder, dissolve it in 50% to 60% ethanol aqueous solution, ultrasonically treat it for 10 to 15 minutes, and filter it with a 0.22 μm filter membrane to remove insoluble matter to obtain a seaweed polyphenol solution; S13. In a beaker, slowly drop the seaweed polyphenol solution into the whey protein solution while maintaining gentle stirring, adjust the pH to 6.5-7.0 using dilute NaOH or HCl, and continue stirring at 25-30° C. for 2-3 hours to obtain a first mixed solution; S14. The first mixed solution is transferred to an ultrafiltration device with a molecular weight cutoff of 10 kDa, and ultrafiltration is performed at 4 to 5° C. to concentrate the first mixed solution to 1 / 3 to 1 / 2 of the original volume to obtain a concentrated solution; S15. The concentrated solution is divided into freeze-drying bottles, pre-frozen at -80~-90°C for 2~3h, and freeze-dried for 48~50h until a dry protein-seaweed polyphenol complex is obtained; The glutathione peroxidase-zinc complex is formed by the combination of glutathione peroxidase and zinc ion through coordination bonds, and the glutathione peroxidase-zinc complex has a tetrameric structure; The preparation method of the glutathione peroxidase-zinc complex comprises: S21. Dissolve the purified glutathione peroxidase in Tris-HCl buffer, adjust the concentration to 1 mg / mL, and filter using a 0.22 μm filter membrane to obtain a glutathione peroxidase solution; S22. Prepare a 10 mM ZnCl2 solution, dissolve it in ultrapure water, and sterilize it by filtering it with a 0.22 μm filter membrane to obtain a ZnCl2 solution; S23. In a centrifuge tube, mix the glutathione peroxidase solution and the ZnCl2 solution, gently vortex to mix, and keep in contact at 4-5°C for 2-3 hours to obtain a second mixed solution; S24. The second mixed solution is transferred to a pretreated dialysis bag, and dialyzed with Tris-HCl buffer at 4-5°C for 4 times, each time for 4 hours, and fresh buffer is replaced each time to obtain a dialyzed solution; S25. The dialyzed solution was transferred to a centrifuge tube, and centrifuged at 10,000 g for 15 min at 4-5°C to remove the formed precipitate to obtain a centrifuged solution; S26. Use an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa to concentrate the sample at 4~5℃ and centrifuge to the desired concentration to obtain glutathione peroxidase-zinc complex.

2. A feed protein raw material according to claim 1, characterized in that: The feed base material comprises the following raw materials in parts by weight: 0.05-0.2 parts of beta-carotene, 15-25 parts of whey protein hydrolyzate, 0.5-1.5 parts of vitamin C, 1.5-2.5 parts of lysine, 0.8-1.2 parts of methionine, 0.3-0.7 parts of vitamin E, 0.03-0.07 parts of copper sulfate, 0.03-0.07 parts of manganese sulfate, 3-7 parts of linseed oil, 0.05-0.15 parts of amylase, 0.8-1.2 parts of sodium chloride, 20-30 parts of corn fiber, 0.05-0.15 parts of lactic acid bacteria, and 0.05-0.15 parts of superoxide dismutase.

3. A method for preparing a feed protein raw material, which is used to prepare the feed protein raw material as claimed in any one of claims 1 to 2, characterized in that: The method comprises the following steps: S31. Preparation of protein-seaweed polyphenol complex; S32. Preparation of glutathione peroxidase-zinc complex; S33. β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, amylase, sodium chloride, corn fiber, lactic acid bacteria, whey protein hydrolysate, superoxide dismutase and protein-seaweed polyphenol complex were sieved through a 60-mesh sieve, and linseed oil and glutathione peroxidase-zinc complex were preheated to 25°C; S34. In a mixer, add β-carotene, cysteine, vitamin C, lysine, methionine, vitamin E, copper sulfate, manganese sulfate, sodium chloride and protein-seaweed polyphenol complex according to the formula ratio, mix for 15 minutes, slowly add preheated linseed oil, and continue mixing for 10 minutes to obtain a first mixture; S35. Dissolving amylase, glutathione peroxidase-zinc complex and superoxide dismutase in distilled water to obtain an enzyme solution, spraying the enzyme solution evenly onto the first mixture, and continuing to mix for 5 min to obtain a second mixture; S36. Premixing lactic acid bacteria with a small amount of corn fiber and whey protein hydrolyzate to obtain a premix, and evenly spreading the premix into the second mixture, mixing for 2 minutes, to obtain a third mixture; S37. The third mixture is transferred to a granulator at a temperature of 40 to 45°C and a humidity of 15 to 18%, and granulated to obtain a granular feed protein raw material having a diameter of 2 to 3 mm; S38. The granular protein raw material for feed is dried using a fluidized bed dryer, the drying temperature is set to 50-55°C, the time is 60-70min, and the final moisture content is controlled between 8-10% to obtain a dry protein raw material for feed; S39. The dried feed protein raw material is transferred to a cooler, cooled to room temperature, and sieved using an 80-mesh sieve to remove fine powder and oversized particles to obtain a feed protein raw material.

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