A neurotrophic protein peptide composition and its application

Abstract

This invention discloses a neurotrophic brain protein peptide composition and its applications, belonging to the field of food technology. The components of the composition provided by this invention have synergistic effects. On the one hand, they reduce the damage of oxidative stress to the nervous system, improve sleep quality, and create a favorable environment for the repair and regeneration of the nervous system. On the other hand, they provide the raw materials needed for the maintenance, repair, and regeneration of nerve cells, as well as the synthesis of neurotransmitters, ensuring energy supply and the activity of the required enzyme systems, thereby achieving the effect of effectively nourishing / repairing nerves and / or promoting nerve regeneration. The provided composition does not use organic reagents, is environmentally friendly, and has high safety; it has low preparation cost and a good taste.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of food technology, in particular to a brain protein peptide composition with neurotrophic activity and application thereof. BACKGROUND

[0002] The nervous system regulates all unconscious life activities such as heartbeat, breathing, body temperature, and is also the basis for the body to seek benefits and avoid harm (allowing people to perceive danger such as pain and heat and make the body react quickly), so protecting the nervous system is the foundation of life; high-quality cognitive functions such as memory, thinking, language, and decision-making also depend on healthy neural networks, and protecting the nervous system can keep a clear head and efficient thinking in learning, work, and life, which is directly related to people's professional ability, social skills, and ability to live independently; delaying the natural aging of the nervous system can reduce the risk of diseases such as Alzheimer's disease and Parkinson's disease, which not only prolongs the life of the body, but also prolongs the healthy life, and is one of the fundamental factors for maintaining dignity and self-care in old age. In addition, protecting the nervous system also helps maintain normal levels of substances such as dopamine and serotonin, thereby better coping with stress and preventing emotional disorders such as depression and anxiety.

[0003] Although pig brain contains rich and high-quality protein and minerals, studies have shown that pig brain also contains a large amount of cholesterol (content up to 2571mg / 100g) and saturated fatty acids, according to the Dietary Guidelines for Chinese Residents, the daily cholesterol intake of adults should be less than 300mg, which means that as long as a small piece of pig brain is eaten, the cholesterol intake may exceed the standard. Excessive intake of cholesterol and saturated fat can increase blood viscosity, accelerate atherosclerosis, block blood vessels, and cause serious health problems. For people with a history of hypercholesterolemia, hyperlipidemia, and cardiovascular and cerebrovascular diseases (such as coronary heart disease and stroke), or high-risk groups, pig brain is definitely recommended to avoid. SUMMARY

[0004] The purpose of the present application is to provide a brain protein peptide composition with neurotrophic activity and application thereof, which provides a safe, low-cost, and effective composition with neurotrophic activity for the repair and daily maintenance of the nervous system.

[0005] To achieve the above-mentioned purpose, the present application provides a brain protein peptide composition with neurotrophic activity, which is composed of brain protein peptides and spina date seed extract; the brain protein peptides include two parts: extract I and extract II; under a protective gas atmosphere, the extract I, the extract II, and the spina date seed extract are mixed and passed through a 0.22μm filter membrane to obtain the brain protein peptide composition with neurotrophic activity; the composition can also be freeze-dried into tablets.

[0006] Preferably, the method for preparing the brain protein peptide is as follows: S1. Freeze fresh pig brains, cut them into pieces, and then grind them into powder to obtain pig brain powder; S2. Add the pig brain powder obtained in S1 to drinking water and mix well. Then treat at 50-60℃ for 6-8 hours. S3. After adjusting the pH of the mixture from S2, add the protease, stir and react for 10-20 minutes for enzymatic hydrolysis, then homogenize, then centrifuge and collect the supernatant into a sterile container for later use, which is recorded as extract I. S4. After drying the precipitate obtained in S3, grind it into powder, add fresh walnut oil, and extract under a protective atmosphere for 60-90 minutes. After the protective atmosphere has cooled to room temperature, centrifuge to collect the supernatant, which is recorded as extract II.

[0007] Preferably, the fresh pig brain in S1 is the brain tissue of a healthy 1-year-old, freshly slaughtered pig; the freezing and cutting process is as follows: the fresh pig brain tissue is frozen at -20°C for 40-60 minutes, and then cut into pieces with a maximum diameter of no more than 0.5 cm on ice; the grinding process is as follows: the cut pig brain tissue is ground in liquid nitrogen until there is no powder.

[0008] Preferably, the mass-to-volume ratio of pig brain powder to drinking water in S2 is 1g:1.5mL.

[0009] Preferably, in step S3, pH adjustment is performed by using NaOH and / or HCl to adjust the pH to 6.0; enzymatic hydrolysis is performed using flavored protease at 50-60°C under stirring conditions; and centrifugation is performed at 8000-10000 rpm for 10-15 minutes.

[0010] Preferably, in step S4, drying is performed at 50-60℃; grinding is performed until the particle size is no greater than 2mm; the amount of fresh walnut oil added is the same as the volume of the precipitated powder; and centrifugation is performed at 8000-10000 rpm for 10-15 minutes.

[0011] Preferably, the preparation method of the jujube seed extract is as follows: After drying the purchased jujube seeds, grind them into powder. Take the sieved powder and add it to drinking liquor. Extract the powder overnight at room temperature, in a sealed environment with stirring. Centrifuge to remove the supernatant and then remove the liquid by rotary evaporation.

[0012] Preferably, the drying process is carried out at 50°C for 30-60 minutes; the sieving process is carried out through a 60 or 80 mesh sieve; the alcohol content of the liquor is 52°; and the centrifugation process is carried out at 8000-10000 rpm for 10-15 minutes.

[0013] The use of the composition described above in the preparation of food, including special medical foods, special dietary foods, health foods, and ordinary foods.

[0014] Preferably, the health food uses the composition as a functional component.

[0015] The composition can also be used to prepare special dietary foods and as ordinary food.

[0016] Pig brain contains approximately 7-11% protein, with a complete range of amino acids. Studies have shown that 16 amino acids have been identified, and it is rich in essential amino acids, accounting for about 44%, making it a high-quality protein source. In addition, it contains abundant minerals (including phosphorus, potassium, sodium, calcium, magnesium, iron, zinc, selenium, and copper), vitamins (including niacin, riboflavin, and thiamine), unique fatty acids (including monounsaturated fatty acids with oleic acid as the main component and polyunsaturated fatty acids with DHA), and phospholipids. However, pig brain also contains extremely high levels of cholesterol (often referred to as the "king of cholesterol"), fat, and purines, making it a food with extremely high consumption risks and unsuitable for daily consumption.

[0017] Sour jujube kernels (the seeds of sour jujube) are also rich in nutrients, including basic substances that can provide energy for the body (including soluble protein and abundant vitamin C), as well as a variety of bioactive functional components (including flavonoids, saponins, nitrogen-containing compounds with various bioactivities, and phenolic compounds), and minerals, including potassium, sodium, calcium, zinc, iron, copper, and manganese.

[0018] The method for preparing brain protein peptides provided by this invention utilizes the characteristic that cholesterol is in a solid state at low temperatures. Physical centrifugation can effectively remove cholesterol and saturated fatty acids from pig brain, offering good safety and low cost. The entire preparation process uses low temperatures (maximum temperature 60℃), which significantly reduces the dissolution rate of purines in pig brain while effectively preserving vitamins and dissolved minerals. Simultaneously, the method provided by this invention can effectively obtain abundant amino acids, polypeptides, phospholipids, and unsaturated fatty acids from pig brain. Through multiple mechanical disruptions, even with the addition of a small amount of protease, effective protein degradation can still be achieved.

[0019] The method for preparing jujube seed extract provided by this invention can effectively obtain a variety of active ingredients in jujube seed, as well as a small amount of soluble protein, vitamins and minerals.

[0020] The composition provided by this invention, firstly, contains vitamin C, flavonoids, saponins, phenolic compounds, selenium, and nitrogen-containing compounds, which can effectively scavenge free radicals in the body, inhibit inflammation, resist oxidation, protect mitochondria, and reduce oxidative stress damage to the nervous system; the saponins and flavonoids in jujube seed have sedative and calming effects and improve sleep, which, together with the low level of oxidation, create a suitable environment for maintaining the health of the nervous system, repairing damaged nervous systems, and developing new nervous systems.

[0021] Then, it provides sufficient raw materials for the repair of damage to the nervous system, daily maintenance, and the formation and development of new nerves. Various amino acids and phospholipids are key components for the formation of nerve fibers and also help maintain nerve signal transmission. B vitamins are important components required for the synthesis of neurotransmitters and energy metabolism. Magnesium prevents excitatory toxicity caused by over-excitation and protects neurons. Magnesium, zinc, and copper are also (co)factors for various enzymatic reactions, ensuring the normal progress of enzymatic reactions in the repair and regeneration of the nervous system.

[0022] Furthermore, the composition also ensures the transmission of nerve signals: saponins and flavonoids in jujube seed can increase the level of acetylcholine in the brain, and acetylcholine is a key neurotransmitter closely related to memory and learning; while DHA in brain protein peptides (partly from pig brain and partly from walnut oil) is a substance essential for maintaining the health of neuronal cell membranes; phosphorus is a key component of ATP, and copper is a cofactor of mitochondrial cytochrome c oxidase, providing energy for all neural activities (including maintaining resting potential, signal transduction, and axonal transport); sodium mediates the depolarization process of action potentials. Potassium is the direct driving force for the generation of nerve signals; potassium maintains the resting potential of nerve cells, mediates the repolarization process of action potentials, and ensures the normal generation and transmission of nerve signals; calcium triggers the release of neurotransmitters from nerve endings, enabling communication between neurons, and acts as an intracellular second messenger, regulating gene expression, synaptic plasticity, and learning and memory; iron is a cofactor required for the synthesis of myelin sheath and is crucial for maintaining axonal insulation; zinc is a cofactor for the synthesis of various neurotransmitter enzymes, which can affect the proliferation and differentiation of neural stem cells, promote axonal growth and synaptic connections, and thus regulate synaptic plasticity and learning and memory.

[0023] Therefore, the present invention provides a brain protein peptide composition with neurotrophic activity and its application, the specific technical effects of which are as follows: (1) The present invention provides a brain protein peptide composition with neurotrophic activity, which is composed of porcine brain protein peptide prepared by a specific method and jujube seed extract; the components in the composition have synergistic effects. On the one hand, they scavenge free radicals, reduce oxidation levels, reduce the damage of oxidative stress to the nervous system, improve the body's sleep quality, and create a good environment for the repair and regeneration of the nervous system; on the other hand, they provide the raw materials required for the maintenance, repair and regeneration of nerve cells and the synthesis of neurotransmitters, ensure energy supply and the required enzyme system activity, thereby achieving the effect of effectively nourishing / repairing nerves and / or promoting nerve regeneration. (2) The composition provided by the present invention does not use organic reagents, is environmentally friendly and has high safety; it effectively removes or reduces the content of dangerous components in pig brain by physical methods such as multiple mechanical crushing and the morphological differences of different components at specific temperatures, while reducing the amount of enzyme used, reducing the preparation cost and improving the edibility of pig brain; the added flavor protease can not only improve the taste of the composition, but the enzymatic hydrolysis conditions are also compatible with other technical solutions.

[0024] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0025] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 This is the statistical result of cell survival rate for each treatment in the efficacy testing section of this invention; wherein... Compared with the blank control group P <0.01. Detailed Implementation

[0027] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0028] To make the objectives, technical solutions, and advantages of this application clearer, more thorough, and more complete, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments. The following detailed descriptions are all illustrations of embodiments, intended to provide further detailed explanation of the present invention. Unless otherwise specified, all technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0029] The instruments, equipment, reagents and materials used in the embodiments were all obtained through commercial means; the methods and steps not described in detail in the embodiments are all conventional techniques in the art.

[0030] Example 1 The specific steps for preparing a brain protein peptide are as follows: S11. Take fresh pig brains from healthy, 1-year-old, freshly slaughtered pigs, freeze them at -20℃ for 60 minutes, cut them into pieces with a maximum diameter of no more than 0.5cm on the ice, and then freeze them at -20℃ to disperse them (during which time the pieces should be separated and not stick together as much as possible), and then grind them into powder in liquid nitrogen.

[0031] S12. Take 200g of the fresh pig brain powder obtained in step S11 and add it to 300mL of drinking water. Stir and mix well, then keep it at 50-60℃ for 8 hours. Then adjust the pH to 6.0 with 1M NaOH and / or 1M HCl. Then add 1g of flavor protease (enzyme activity 10000U), keep warm, stir at 200rpm for 15 minutes, then homogenize for 10 minutes, then centrifuge at 8000rpm for 15 minutes. Take the supernatant into a new sterile container for later use, and record it as extract I, totaling 365mL.

[0032] S13. After drying the precipitate obtained in step S12 at 60°C, grind it into powder until no large particles are visible (large particles refer to particles with a maximum diameter greater than 2 mm). Then add 1 volume of fresh walnut oil and extract for 1 hour under a nitrogen atmosphere at 50°C and 200 rpm. After the nitrogen atmosphere cools to room temperature, centrifuge at 8000 rpm for 15 minutes, collect the supernatant for later use, and designate it as extract II.

[0033] Example 2 The specific steps for preparing a jujube seed extract are as follows: The jujube seeds (free from insects and mold) purchased from a pharmacy were dried at 50℃ for 60 minutes, then pulverized and ground into powder. 30g of the powder, passing through an 80-mesh sieve, was added to 450mL of purchased 52° liquor and left to stand overnight (approximately 16 hours) at room temperature, sealed, and at 200rpm. The next day, the mixture was centrifuged at 8000rpm for 15 minutes, and the supernatant was collected and rotary evaporated to obtain the jujube seed extract, totaling 0.31g.

[0034] Example 3 The specific steps for preparing a brain protein peptide composition with neurotrophic activity are as follows: Under a nitrogen atmosphere, extracts I and II prepared in Example 1 were mixed, and then 0.2g of jujube seed extract prepared in Example 2 was added and mixed well. The mixture was then sealed and stored.

[0035] Effect test The effect of the neurotrophic brain protein peptide composition prepared in Example 3 on nerve cell proliferation was investigated. The specific steps are as follows: The purchased human neuroblastoma cells SH-SY5Y were cultured in DMEM / F12 (1:1) medium containing 10% fetal bovine serum at 37°C in a 5% CO2 incubator until the logarithmic growth phase, according to the instructions. Then, cells were cultured at 1.0 × 10⁻⁶ cells / year. 5 Inoculate 10 cells / well into a 96-well plate, add 1 mL of fresh culture medium, and then add hydrogen peroxide to a final concentration of 420 μmol / L. Incubate at 37°C in a 5% CO2 incubator for 24 hours to simulate oxidative stress damage to neuroblastoma.

[0036] After culturing, the culture medium in the cells was replaced with fresh culture medium, and the steps of adding fresh culture medium and centrifuging to discard the supernatant were repeated three times to ensure complete removal of hydrogen peroxide. Then, 1 mL of the composition prepared in Example 3 (referred to as Example 3 group), 1 mL of the brain protein peptide prepared by the method of Example 1 (with extract I and extract II vortexed and mixed before addition, referred to as Example 1 group), and 1 mL of fresh culture medium (referred to as blank control group) were added to the cells respectively. After mixing, the cells were incubated at 37°C in a 5% CO2 incubator. The culture medium for each treatment was changed every 24 hours for a total of 7 days. Five replicate wells were set up for each treatment and labeled.

[0037] After treatment, cell viability was determined using the MTT assay, and the results are as follows: Figure 1 As shown, treatment with 420 μmol / L hydrogen peroxide severely damaged human neuroblastoma cells SH-SY5Y, resulting in a cell viability of only 30.01 ± 5.33% even after 7 days of recovery culture. However, the composition prepared in Example 3 of this invention has the effect of repairing damaged neuroblastoma cells or promoting the proliferation of undamaged neuroblastoma cells. After 7 days of treatment, the survival rate of human neuroblastoma cells SH-SY5Y was 68.92 ± 3.20%, which is highly significant. P <0.01) higher than the blank control group. The brain protein peptide prepared in Example 1 also has a certain effect on repairing damaged neuroblastoma cells or promoting the proliferation of neuroblastoma cells, but the effect is much worse than that of the composition prepared in Example 3. The cell survival rate is 50.12±4.65%, which shows that it is consistent with the theory and that the components in the composition prepared in Example 3 have a synergistic effect.

[0038] Therefore, the components in the composition provided by this invention have a synergistic effect. On the one hand, they reduce the damage of oxidative stress to the nervous system, improve the body's sleep quality, and create a favorable environment for the repair and regeneration of the nervous system. On the other hand, they provide the raw materials required for the maintenance, repair and regeneration of nerve cells, as well as the synthesis of neurotransmitters, ensuring energy supply and the required enzyme system activity, thereby achieving the effect of effectively nourishing / repairing nerves and / or promoting nerve regeneration. The composition provided does not use organic reagents, is environmentally friendly, and has high safety. It also has low preparation cost and good taste.

[0039] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A brain protein peptide composition with neurotrophic activity, characterized in that, The composition consists of brain protein peptides and jujube seed extract; the brain protein peptides include two parts: extract I and extract II; under a protective atmosphere, extract I, extract II and jujube seed extract are mixed and filtered through a 0.22 μm filter membrane to obtain a brain protein peptide composition with neurotrophic activity.

2. The neurotrophic brain protein peptide composition according to claim 1, characterized in that, The preparation method of the brain protein peptide is as follows: S1. Freeze fresh pig brains, cut them into pieces, and then grind them into powder to obtain pig brain powder; S2. Add the pig brain powder obtained in S1 to drinking water and mix well. Then treat at 50-60℃ for 6-8 hours. S3. After adjusting the pH of the mixture from S2, add the protease, stir and react for 10-20 minutes for enzymatic hydrolysis, then homogenize, then centrifuge and collect the supernatant into a sterile container for later use, which is recorded as extract I. S4. After drying the precipitate obtained in S3, grind it into powder, add fresh walnut oil, and extract under a protective atmosphere for 60-90 minutes. After the protective atmosphere has cooled to room temperature, centrifuge to collect the supernatant, which is recorded as extract II.

3. The neurotrophic brain protein peptide composition according to claim 2, characterized in that, The fresh pig brain in S1 is the brain tissue of a healthy 1-year-old, freshly slaughtered pig; the freezing and cutting process is as follows: the fresh pig brain tissue is frozen at -20℃ for 40-60 minutes, and then cut into pieces with a maximum diameter of no more than 0.5cm on the ice; the grinding process is as follows: the cut pig brain tissue is ground in liquid nitrogen until there is no powder.

4. The neurotrophic peptide composition according to claim 2, characterized in that: The mass-to-volume ratio of pig brain powder to drinking water in S2 is 1g:1.5mL.

5. The neurotrophic peptide composition according to claim 2, characterized in that: In S3, pH is adjusted to 6.0 using NaOH and / or HCl; enzymatic hydrolysis is performed using flavored protease at 50-60℃ with stirring; and centrifugation is performed at 8000-10000 rpm for 10-15 minutes.

6. The neurotrophic peptide composition according to claim 2, characterized in that: In step S4, drying is performed at 50-60℃; grinding is performed until the particle size is no greater than 2mm; the amount of fresh walnut oil added is the same as the volume of the precipitated powder; centrifugation is performed at 8000-10000 rpm for 10-15 minutes.

7. The neurotrophic brain protein peptide composition according to claim 1, characterized in that, The preparation method of the jujube seed extract is as follows: After drying the purchased jujube seeds, grind them into powder. Take the sieved powder and add it to drinking liquor. Extract the powder overnight at room temperature, in a sealed environment with stirring. Centrifuge to remove the supernatant and then remove the liquid by rotary evaporation.

8. The neurotrophic peptide composition according to claim 7, characterized in that: The drying process involves drying at 50℃ for 30-60 minutes; sieving is done through a 60 or 80 mesh sieve; the alcohol content of the liquor is 52°; and centrifugation is performed at 8000-10000 rpm for 10-15 minutes.

9. The use of the composition according to any one of claims 1-8 in the preparation of food, characterized in that: The food products mentioned include special medical foods, special dietary foods, health foods, and ordinary foods.

10. The use of the composition according to claim 9 in the preparation of food, characterized in that: The health food uses the composition as its functional component.

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