CN113142592A - Fermentation type small molecule peptide for assisting in improving memory and preparation method and application thereof - Google Patents
Fermentation type small molecule peptide for assisting in improving memory and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a fermented small molecular peptide for assisting in improving memory and a preparation method and application thereof, belonging to the technical field of health-care product preparation. The invention provides a fermented small molecule peptide for assisting in improving memory, which comprises the following raw materials for preparation in parts by mass: 1-3 parts of ginseng, 1-5 parts of Chinese yam, 1-3 parts of rhizoma polygonati, 1-3 parts of poria cocos, 2-4 parts of radix puerariae, 1-3 parts of fructus alpiniae oxyphyllae, 5-10 parts of walnut and 5-10 parts of needle mushroom; the fermentation type small molecular peptide for assisting in improving memory is obtained by fermenting, carrying out enzymolysis and carrying out ultrafiltration on the raw materials. The raw materials for preparing the fermented micromolecule peptide for assisting in improving the memory comprise various intelligence-promoting medicinal and edible homologous components, the source is wide, the cost is low, the components are safe, no toxic or side effect is caused, and the fermented micromolecule peptide can improve the memory and can regulate and improve systems of human digestion, endocrine and the like after being taken continuously.
Description
Technical Field
The invention belongs to the technical field of health-care product preparation, and particularly relates to a fermentation type small molecular peptide for assisting in improving memory, and a preparation method and application thereof.
Background
In recent years, the development of the traditional Chinese medicine industry is very rapid, various traditional Chinese medicines play an important role in the treatment of difficult and complicated diseases, but the potential of the traditional Chinese medicines is not completely explored in the aspect of memory improvement, active ingredients with homology of medicine and food are not developed very efficiently, and in some researches on memory improvement at present, the preparation of natural product extracts by using sea cucumber flowers, fish roes and the like is facilitated, but the materials of the natural products are not fixed, and the uncontrollable factors of high cost, long preparation process period, complex technology and the like are accompanied. For the existing health care products for improving memory on the market, capsules, medicaments and the like containing DHA and other active ingredients still lack a reasonable quality control method. Therefore, how to select raw materials with high efficiency, low cost, short preparation period, simple and clear technical route and easy control of market circulation quality to process food or medicine so as to achieve obvious effects on improving human organism and auxiliary memory becomes a main problem in the technical field of traditional Chinese medicine component extraction at present.
Disclosure of Invention
In order to solve the problems, the invention provides a fermentation type small molecule peptide for assisting in improving memory and a preparation method and application thereof. The fermentation type small molecular peptide for assisting in improving the memory has a good promoting effect on improving the memory, and meanwhile, the preparation raw materials are wide in source, low in cost, safe in components, free of toxic and side effects, simple in preparation process, capable of being repeatedly operated and capable of realizing quantitative production.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a fermentation type small molecule peptide for assisting in improving memory, which comprises the following raw materials for preparation in parts by mass: 1-3 parts of ginseng, 1-5 parts of Chinese yam, 1-3 parts of rhizoma polygonati, 1-3 parts of poria cocos, 2-4 parts of radix puerariae, 1-3 parts of fructus alpiniae oxyphyllae, 5-10 parts of walnut and 5-10 parts of needle mushroom; the fermentation type small molecular peptide for assisting in improving memory is obtained by fermenting, carrying out enzymolysis and carrying out ultrafiltration on the raw materials.
The invention provides a preparation method of a fermentation type small molecule peptide for assisting in improving memory in the technical scheme, which comprises the following steps: mixing the raw materials with deionized water, adding composite probiotics for fermentation to obtain fermentation liquor; carrying out enzymolysis on the obtained fermentation liquor to obtain a protease hydrolysate; and (3) performing ultrafiltration on the protease hydrolysate to obtain the fermentation type small molecular peptide for assisting in improving the memory.
Preferably, the volume ratio of the mass of the raw materials to the deionized water is 1g (2-6) mL.
Preferably, the complex probiotics comprise lactobacillus acidophilus, bifidobacterium lactis, lactobacillus rhamnosus and lactobacillus casei; the number of the live bacteria of the composite probiotics is 100 hundred million-2000 hundred million CFU/g; the added mass of the composite probiotics accounts for 0.2-0.5% of the total mass of the raw materials and the deionized water.
Preferably, the fermentation temperature is 35-36 ℃, and the fermentation time is 36-48 h.
Preferably, the enzymolysis is carried out by sequentially using hemicellulase and trypsin; the adding amount of the hemicellulase accounts for 0.1-0.4% of the mass percent of the fermentation broth; the adding amount of the trypsin accounts for 0.1 to 0.4 percent of the mass percentage of the fermentation liquor; the enzymatic activity of the hemicellulase is 10 ten thousand u/g, and the enzymatic activity of the trypsin is 10 ten thousand u/g; the enzymolysis conditions of the hemicellulase are as follows: the temperature is 45-55 ℃, the pH value is 4.0-5.5, and the time is 1-2 h; the enzymolysis conditions of the trypsin are as follows: the temperature is 36-37 ℃, the pH value is 7.8-8.5, and the time is 1-2 h.
Preferably, the ultrafiltration comprises the steps of: and (3) performing ultrafiltration on the protein enzymolysis liquid through a filter membrane with the molecular weight of 2000Da, and collecting filtrate to obtain the fermentation type micromolecule peptide.
Preferably, after ultrafiltration, the filtrate is subjected to vacuum spray drying to obtain the fermentation type small molecular peptide powder.
The invention provides application of the fermented small molecule peptide for assisting in improving memory in the technical scheme or the fermented small molecule peptide for assisting in improving memory obtained by the preparation method in the technical scheme in preparation of an intelligence-improving health-care product.
Preferably, the types of the intelligence-developing health-care product comprise small-molecule peptide powder, small-molecule peptide rods, small-molecule peptide oral liquid, small-molecule peptide liquid beverage and small-molecule peptide biscuits.
Has the advantages that:
the invention provides a fermentation type small molecule peptide for assisting in improving memory, which comprises the following raw materials for preparation in parts by mass: 1-3 parts of ginseng, 1-5 parts of Chinese yam, 1-3 parts of rhizoma polygonati, 1-3 parts of poria cocos, 2-4 parts of radix puerariae, 1-3 parts of fructus alpiniae oxyphyllae, 5-10 parts of walnut and 5-10 parts of needle mushroom; the fermentation type small molecular peptide for assisting in improving memory is obtained by fermenting, carrying out enzymolysis and carrying out ultrafiltration on the raw materials. The basic preparation substance of the fermented micromolecule peptide for assisting in improving memory provided by the invention contains various intelligence-promoting medicinal and edible homologous components, has the advantages of wide sources, low cost, safe components and no toxic or side effect, and can improve memory, regulate and improve systems of human digestion, endocrine and the like after being taken continuously. The results of the examples show that the fermented small molecule peptide for assisting in improving the memory can obviously improve the memory ability and learning cognitive ability of mice.
Drawings
FIG. 1 is a schematic view of a water maze; a is a camera, B is a platform, and C is a water pool.
FIG. 2 is a graph showing the positioning times of different processing groups in example 1;
FIG. 3 is a graph showing the positioning times of different processing groups in example 2;
FIG. 4 is a graph showing the positioning times of different processing groups in example 3;
FIG. 5 is a graph of comparative lines for different concentrations of a fermentable small molecule peptide.
Detailed Description
The invention provides a fermentation type small molecule peptide for assisting in improving memory, which comprises the following raw materials for preparation in parts by mass: 1-3 parts of ginseng, 1-5 parts of Chinese yam, 1-3 parts of rhizoma polygonati, 1-3 parts of poria cocos, 2-4 parts of radix puerariae, 1-3 parts of fructus alpiniae oxyphyllae, 5-10 parts of walnut and 5-10 parts of needle mushroom; the fermentation type small molecular peptide for assisting in improving memory is obtained by fermenting, carrying out enzymolysis and carrying out ultrafiltration on the raw materials.
The raw material of the fermentation type small molecular peptide for assisting in improving memory comprises 1-3 parts of ginseng, preferably 2-3 parts, and more preferably 3 parts by mass. The invention has no special requirements on the source of the ginseng and can be obtained by adopting common commercial sources. The ginseng of the invention can regulate the central nervous system, treat neurasthenia, relieve or eliminate symptoms such as headache, palpitation, insomnia, amnesia, general weakness and the like, improve the memory of people, improve the working efficiency and enhance the body endurance and the accuracy of moving organs.
The raw material of the fermented small molecular peptide for assisting in improving memory comprises, by mass, 1-5 parts of Chinese yam, preferably 3-5 parts, and more preferably 5 parts. The invention has no special requirements on the source of the Chinese yam and can be obtained by adopting common commercial sources. The Chinese yam is mainly used for treating impairment of middle energizer, tonifying deficiency and winning, removing cold and heat evil qi, tonifying middle energizer and benefiting strength; the product can be taken for a long time, and has effects of improving hearing and eyesight, reducing weight, and prolonging life; tonify the five consumptive diseases and seven injuries, remove cold wind, calm the heart, tranquilize mind and tonify heart-qi deficiency.
The raw material of the fermentation type small molecular peptide for assisting in improving memory comprises 1-3 parts of rhizoma polygonati, preferably 2-3 parts of rhizoma polygonati, and more preferably 3 parts of rhizoma polygonati. The invention has no special requirements on the source of the rhizoma polygonati and only needs to adopt common commercial sources. Rhizoma Polygonati has effects of strengthening tendons and bones, replenishing essence, and treating weakness of spleen and stomach, and asthenia.
The raw material of the fermentation type small molecular peptide for assisting in improving memory comprises 1-3 parts of poria cocos, preferably 2-3 parts of poria cocos, and more preferably 3 parts of poria cocos. The invention has no special requirements on the source of the tuckahoe, and the tuckahoe can be obtained by adopting common commercial sources. The tuckahoe can promote diuresis, eliminate dampness, invigorate spleen and calm heart, and has certain regulation effect on spleen deficiency, anorexia, uneasiness, palpitation and insomnia.
The raw material of the fermentation type small molecular peptide for assisting in improving memory comprises, by mass, 2-4 parts of kudzu vine root, more preferably 3-4 parts, and even more preferably 4 parts. The invention has no special requirements on the source of the kudzuvine root and can be obtained by adopting common commercial sources. Radix Puerariae has effects of relieving muscles and fever, promoting eruption, promoting fluid production to quench thirst, invigorating yang and relieving diarrhea.
The raw material of the fermented small molecular peptide for assisting in improving memory comprises, by mass, 1-3 parts of fructus alpiniae oxyphyllae, more preferably 2-3 parts, and still more preferably 3 parts. The invention has no special requirements on the source of the sharpleaf galangal fruit and can be obtained by adopting common commercial sources. Fructus Alpinae Oxyphyllae is mainly used for treating deficiency-cold of spleen and stomach and has effect on cardiovascular system.
The raw material of the fermentation type small molecular peptide for assisting in improving memory comprises, by mass, 5-10 parts of walnuts, preferably 7-10 parts, and more preferably 10 parts. The invention has no special requirements on the source of the walnut and can be obtained by adopting common commercial sources. Walnuts contain a variety of free essential amino acids; isoleucine, leucine, tryptophan, phenylalanine, valine, threonine, lysine, and the like. Sweet in taste and warm in nature, entering kidney, lung and large intestine meridians; has effects of invigorating kidney, consolidating semen, strengthening waist, warming lung, relieving asthma, loosening bowel, and relieving constipation, and has rich nutrition and medicinal value.
The raw materials of the fermentation type small molecular peptide for assisting in improving memory comprise, by mass, 5-10 parts of needle mushroom, preferably 7-10 parts of needle mushroom, preferably 10 parts of needle mushroom, wherein the needle mushroom is rich in protein and multiple vitamins, the amino acid content is very rich, and especially the essential amino acidArginineAndlysineContent (wt.)High in content, and contains various mineral substances such as calcium, phosphorus, iron and the like, and has an important effect on improving the intelligence of children.
The fermentation type small molecular peptide for assisting in improving memory preferably exists in the form of small molecular peptide powder, the small molecular peptide powder is stable in nutrient content, favorable for storage and high in purity, and can be conveniently prepared into other dosage forms, such as small molecular peptide rods, small molecular peptide oral liquid, small molecular peptide liquid beverage, small molecular peptide biscuits and the like.
The ginseng, the poria cocos, the Chinese yam and the walnut regulate the nervous system of the organism, improve symptoms such as amnesia and the like, and play a role in improving memory; the sealwort, the sharpleaf galangal fruit and the kudzuvine root regulate spleen and stomach and cardiovascular systems to enable an organism to reach a mental plump state, and are helpful for the growth and development of the brain.
In addition, the invention makes the prepared fermented small molecular peptide contain various effective bioactive components such as ginsenoside, flammulina velutipes polysaccharide and the like by fermenting and enzymolyzing the raw materials, has the functions of anti-inflammation, antioxidation and neuroprotection, and improves the accumulation of brain Abeta and cognitive behavior by inducing and clearing protein aggregates through autophagy. The fermented small molecular peptide for assisting in improving the memory provided by the invention has the advantages of wide raw material source, low raw material cost, safe components, wide market circulation degree and no toxic or side effect, has a good effect on improving the memory by being taken persistently, and has the function of improving and regulating systems such as human digestion and endocrine, so that the utilization rate of medicinal and edible raw materials is improved, the medicine cost for improving the memory and regulating the organism is reduced, and the bioavailability and the bioactivity of the memory of a human body are increased to a greater extent.
The invention provides a preparation method of a fermentation type small molecule peptide for assisting in improving memory in the technical scheme, which comprises the following steps: mixing the raw materials with deionized water, adding composite probiotics for fermentation to obtain fermentation liquor; carrying out enzymolysis on the obtained fermentation liquor to obtain a protease hydrolysate; and (3) performing ultrafiltration on the protease hydrolysate to obtain the fermentation type small molecular peptide for assisting in improving the memory.
The raw materials are mixed with deionized water, and the compound probiotics are added for fermentation to obtain the fermentation liquor. In the present invention, the raw material is preferably used in a powder state; the particle size of the powder is preferably 100-200 meshes, more preferably 100 meshes, raw materials which are difficult to grind are removed, so that fermentation is more sufficient, active substances such as small molecular peptides are fully released, and the utilization rate of the raw materials is improved. In the invention, the ratio of the mass of the raw materials to the volume of the deionized water is preferably 1g (2-6) mL. The specific proportion of the invention is beneficial to the fermentation of the raw materials, on one hand, the raw materials can be fully and uniformly mixed in the aqueous solution, which is beneficial to the liquid fermentation, and on the other hand, the concentration of the fermentation liquor can not be diluted due to the addition of excessive water. The composite probiotics preferably comprise lactobacillus acidophilus, bifidobacterium lactis, lactobacillus rhamnosus and lactobacillus casei. In the invention, the number of the live bacteria of the composite probiotics is preferably 100 hundred million to 2000 hundred million CFU/g; more preferably 2000 hundred million CFU/mL. In the invention, the added mass of the composite probiotics preferably accounts for 0.2-0.5% of the total mass of the raw materials and the deionized water; further preferably 0.5%. The invention has no special requirement on the source of the composite probiotics. The composite probiotics used in the examples of the present invention were purchased from jacobia bioengineering ltd of jaboticaceae, Shandong; wherein the lactobacillus acidophilus strain number is: JYLA-16, the serial number of the lactobacillus rhamnosus strain is as follows: JYLR-005, the number of the bifidobacterium lactis strain is as follows: JYBR-190, the number of lactobacillus casei strain is: JYLC-132. In the invention, the fermentation temperature is preferably 35-36 ℃, and more preferably 36 ℃; the fermentation time is preferably 36-48 h, and more preferably 48 h. The invention can fully release the effective substances in the raw materials through fermentation to obtain the active small molecular peptides, thereby improving the absorption and conversion efficiency of the active ingredients and enabling the effective ingredients in the food homologous substances to be better absorbed by organisms.
And (3) after obtaining the fermentation liquor, carrying out enzymolysis on the obtained fermentation liquor to obtain protein enzymolysis liquid. In the invention, the enzymolysis is carried out by using hemicellulase and trypsin in sequence. The hemicellulase has good effect on breaking cell walls, and the trypsin is further subjected to enzymolysis to obtain the small molecular peptides. In the present invention, the enzymatic activity of the hemicellulase is preferably 10 wu/g; the enzymatic activity of the trypsin is preferably 10 ten thousand u/g. The invention has no special requirements on the sources of hemicellulase and trypsin, and can meet the enzyme activity requirements. Hemicellulase and trypsin used in the examples of the present invention were purchased from Henan Wako Biotech Ltd. In the invention, the adding amount of the hemicellulase accounts for 0.1-0.4% of the mass percentage of the fermentation liquor, and is more preferably 0.4%; the adding amount of the trypsin accounts for 0.1-0.4% of the mass percent of the fermentation liquor, and the preferable amount is 0.4%. The specific proportion of the invention can lead the enzymolysis efficiency to be higher. In the invention, the enzymolysis temperature of the hemicellulase is preferably 45-55 ℃, and is further preferably 55 ℃; the enzymolysis time of the hemicellulase is 1-2 hours, and the preferable time is 2 hours; the enzymolysis pH of the hemicellulase is 4.5-6.5, preferably 5.5-6.5, and more preferably 6.5. In the invention, the enzymolysis temperature of the trypsin is preferably 45-55 ℃, and further preferably 55 ℃; the enzymolysis time of the trypsin is 1-2 h, and the preferable time is 2 h; the enzymolysis pH of the trypsin is 4.5-6.5, preferably 5.5-6.5, and more preferably 6.5. The specific enzymolysis conditions of the invention ensure that the enzymolysis efficiency achieves the optimal effect.
After the protease hydrolysate is obtained, the invention ultrafilters the protease hydrolysate to obtain the fermentation type micromolecule peptide for assisting in improving the memory. In the present invention, the ultrafiltration preferably comprises the steps of: and (3) performing ultrafiltration on the protein enzymolysis liquid through a filter membrane with the molecular weight of 2000Da, and collecting filtrate to obtain the fermentation type micromolecule peptide. The invention can obtain the fermentation type micromolecule peptide with the molecular weight of 2000Da by ultrafiltration so as to improve the absorption and utilization rate of the organism and further improve the treatment effect.
After ultrafiltration, the filtrate is preferably dried to obtain the fermented small molecule peptide dry powder for assisting in improving the memory. In the present invention, the drying is preferably carried out by a spray drying method, and more preferably by vacuum spray drying. The invention prepares the fermentation type small molecular peptide into the fermentation type small molecular peptide dry powder, which is more beneficial to subsequent storage and processing.
According to the preparation method of the fermentation type micromolecule peptide for assisting in improving the memory, disclosed by the invention, the composite probiotics are utilized to ferment various medicine-food homologous substances and then carry out enzymolysis, so that the effective substances in the raw materials are fully released, the absorption by a human body is facilitated, and the utilization rate of the raw materials is improved; meanwhile, the invention has the advantages of simple material taking, obvious effect, more reserved effective components, easy preparation, easy success of fermentation, repeatable operation and suitability for quantitative production.
The invention provides application of the fermented small molecule peptide for assisting in improving memory in the technical scheme or the fermented small molecule peptide for assisting in improving memory obtained by the preparation method in the technical scheme in preparation of an intelligence-improving health-care product. In the invention, the types of the intelligence-developing health care product preferably comprise small molecule peptide powder, small molecule peptide rods, small molecule peptide oral liquid, small molecule peptide liquid beverage and small molecule peptide biscuits; further preferably small molecular peptide powder, small molecular peptide oral liquid and small molecular peptide liquid beverage; still more preferably a small molecule peptide liquid beverage. The fermented small molecular peptide for assisting in improving memory provided by the invention can obviously improve memory and learning cognitive ability.
To further illustrate the present invention, the following examples are provided to describe a black food fermentation type complex polypeptide and its preparation method and application in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
A fermented small molecule peptide for assisting in improving memory is composed of the following raw materials in parts by mass: 1g of ginseng, 1g of Chinese yam, 1g of rhizoma polygonati, 1g of poria cocos, 2g of radix puerariae, 1g of fructus alpiniae oxyphyllae, 5g of walnuts and 5g of needle mushroom.
The preparation method comprises the following steps:
1) grinding ginseng, Chinese yam, rhizoma polygonati, poria cocos, radix puerariae, fructus alpiniae oxyphyllae, walnuts and the like into powder, sieving with a 100-mesh sieve, adding 100mL of deionized water for dissolving, adding 0.4g of composite probiotics, fermenting at 36 ℃ for 48 hours, and sterilizing to obtain fermentation liquor. Wherein the composite probiotics are purchased from Jiayi bioengineering GmbH of Shandong Zhongke; wherein the lactobacillus acidophilus strain number is: JYLA-16, the serial number of the lactobacillus rhamnosus strain is as follows: JYLR-005, the number of the bifidobacterium lactis strain is as follows: JYBR-190, the number of lactobacillus casei strain is: JYLC-132.
2, sequentially carrying out enzymolysis on the fermentation liquor by using 0.2g of hemicellulase and 0.2g of trypsin to obtain protein enzymolysis liquor. Hemicellulase and trypsin were purchased from wakawa biotechnology limited, and the enzymatic conditions for both enzymes were the same: the enzymolysis temperature is 55 ℃, the enzymolysis time is 2 hours, and the enzymolysis PH is 6.5; the enzymatic activity of the hemicellulase is 10 ten thousand u/g; the enzymatic activity of trypsin was 10 wu/g.
3) And (3) performing ultrafiltration on the fermentation liquor by using a filter membrane with the molecular weight cutoff of 2000Da through an ultrafiltration membrane, and collecting filtrate to obtain the fermentation type small molecular peptide for assisting in improving the memory. And carrying out vacuum concentration and spray drying on the obtained trapped fluid to obtain the fermented micromolecule peptide powder.
Example 2
A fermented small molecule peptide for assisting in improving memory is composed of the following raw materials in parts by mass: 2g of ginseng, 3g of Chinese yam, 2g of rhizoma polygonati, 2g of poria cocos, 3g of radix puerariae, 2g of fructus alpiniae oxyphyllae, 7g of walnuts and 7g of needle mushroom.
The preparation method comprises the following steps:
1) grinding ginseng, Chinese yam, rhizoma polygonati, poria cocos, radix puerariae, fructus alpiniae oxyphyllae, walnuts and the like into powder, sieving with a 100-mesh sieve, adding 100mL of deionized water for dissolving, adding 0.4g of composite probiotics, fermenting at 36 ℃ for 48 hours, and sterilizing to obtain fermentation liquor. The source of the complex probiotics is the same as that in example 1.
2) And sequentially carrying out enzymolysis on the fermentation liquor by using 0.2g of hemicellulase and 0.2g of trypsin to obtain protein enzymolysis liquor. The sources of hemicellulase and trypsin and the enzymatic conditions were the same as in example 1.
3) And (3) performing ultrafiltration on the fermentation liquor by using a filter membrane with the molecular weight cutoff of 2000Da through an ultrafiltration membrane, and collecting filtrate to obtain the fermentation type small molecular peptide for assisting in improving the memory. And carrying out vacuum concentration and spray drying on the obtained trapped fluid to obtain the fermented micromolecule peptide powder.
Example 3
A fermented small molecule peptide for assisting in improving memory is composed of the following raw materials in parts by mass: 3g of ginseng, 5g of Chinese yam, 3g of rhizoma polygonati, 3g of poria cocos, 4g of radix puerariae, 3g of fructus alpiniae oxyphyllae, 10g of walnut and 10g of needle mushroom.
The preparation method comprises the following steps:
1) grinding ginseng, Chinese yam, rhizoma polygonati, poria cocos, radix puerariae, fructus alpiniae oxyphyllae, walnuts and the like into powder, sieving with a 100-mesh sieve, adding 100mL of deionized water for dissolving, adding 0.4g of composite probiotics, fermenting at 36 ℃ for 48 hours, and sterilizing to obtain fermentation liquor.
2) And sequentially carrying out enzymolysis on the fermentation liquor by using 0.2g of hemicellulase and 0.2g of trypsin to obtain protein enzymolysis liquor.
3) And (3) performing ultrafiltration on the fermentation liquor by using a filter membrane with the molecular weight cutoff of 2000Da through an ultrafiltration membrane, and collecting filtrate to obtain the fermentation type small molecular peptide for assisting in improving the memory. And carrying out vacuum concentration and spray drying on the obtained trapped fluid to obtain the fermented micromolecule peptide powder.
Comparative example 1
A ginseng fructus alpiniae oxyphyllae fermented small molecular peptide is composed of the following raw materials in parts by mass: 1g of ginseng and 1g of sharpleaf galangal fruit.
The preparation method is the same as that of example 1, except that the addition amount of deionized water is 12 mL; the amount of the composite probiotics is 0.05 g; the amount of hemicellulase was 0.05g and the amount of trypsin was 0.05 g.
Comparative example 2
A ginseng fructus alpiniae oxyphyllae fermented small molecular peptide is composed of the following raw materials in parts by mass: 2g of ginseng and 2g of sharpleaf galangal fruit.
The preparation method is the same as that of example 1, except that the addition amount of deionized water is 16 mL; the amount of the composite probiotics is 0.07 g; the amount of hemicellulase was 0.07g and the amount of trypsin was 0.07 g.
Comparative example 3
A ginseng fructus alpiniae oxyphyllae fermented small molecular peptide is composed of the following raw materials in parts by mass: 3g of ginseng and 3g of sharpleaf galangal fruit.
The preparation method is the same as that of example 1, except that the addition amount of deionized water is 14 mL; the amount of the composite probiotics is 0.08 g; the amount of hemicellulase was 0.08g and the amount of trypsin was 0.08 g.
Application example 1
The effect of the fermented small molecule peptides of example 1 and comparative example 1 on improving memory is examined, and the specific scheme is as follows:
(1) animals: 24 wistar mice were purchased from the animal laboratory center of white chen medical college, university of ji forest, housed in SPF-grade animal rooms for one week, during which time they had free access to water.
(2) The main reagents are as follows: the fermented small molecule peptide group hydrolysate in example 1, the ginseng fructus alpiniae oxyphyllae group hydrolysate in comparative example 1, normal saline and common feed; wherein the ratio of the hydrolysate is as follows: the volume of the deionized water is 1g and 0.5 mL.
(3) The test process comprises the following steps: mice were randomized into 3 groups: blank control group (A1), fermentation type small molecule peptide group (A2), and Ginseng radix fructus Alpinae Oxyphyllae group (A3). Each group of 8 mice (initial body weight 20 + -2.0 g, SPF grade) except the blank control group were subjected to intragastric administration of 1 mL/(one time) of the corresponding hydrolysate 2 times a day, i.e. once in the morning and evening. At the same time, the blank group was given an equal amount of physiological saline for 14 days. After 14 days, the Morris water maze test was performed, and FIG. 1 is a water maze pattern diagram; wherein A is a camera, B is a platform, and C is a pool.
Morris water maze test:
a water pool: circular, diameter of about 120cm, overall height of 30cm, water level height of 25 cm. A platform with the diameter of 20cm is arranged in the device, and the height of the platform is about 24 cm. Constant temperature treatment is carried out in the tank, the water temperature is controlled at 24 ℃, and the error range is +/-1 ℃. The column is a platform for the mouse to stay on and is slightly higher than the water surface; the camera records the test process.
The test is divided into a positioning navigation part and a space exploration part, wherein the positioning navigation part records the time when the mouse is placed into water from different points to find the platform, namely the positioning time. And the space exploration is to remove the platform after the positioning navigation is finished, record the space exploration times of the mouse from the original release point to the original position of the platform, and detect the learning and memory acquisition capacity of the mouse and the memory retention capacity of the platform on the space position. The test results are shown in table 1 and fig. 2.
Table 1 example 1 fermentation type small molecule peptide mouse Morris water maze experiment
Note: the average of the water maze simulation data for 8 mice per group is shown in the table.
From the results in table 1, it can be seen that in the localization navigation test, the escape latency of the mice in the test group of the gavage fermentation type small molecular peptide hydrolysate is shorter and the time to reach the platform is shorter than those in the blank control group and the ginseng fructus alpiniae oxyphyllae group; and in the space exploration part, the track of the mice with the gastric perfusion fermentation type micromolecule peptide hydrolysate reaching the platform position shows certain memory characteristics, and the times are obviously more than those of mice in a blank control group and a ginseng fructus alpiniae oxyphyllae group. Therefore, the micromolecule peptide hydrolysate can be effectively proved to improve the memory capacity of the mouse and enhance the learning and cognition capacity.
Application example 2
The effect of the hydrolysate of example 2 and comparative example 2 on improving memory is examined, and the specific scheme is as follows:
(1) animals: 24 wistar mice were purchased from the animal laboratory center of white chen medical college, university of ji forest, housed in SPF-grade animal rooms for one week, during which time they had free access to water.
(2) The main reagents are as follows: the fermented small molecule peptide group hydrolysate in example 2, the ginseng fructus alpiniae oxyphyllae group hydrolysate in comparative example 2, normal saline and common feed. The ratio of hydrolysate was the same as in application example 1.
(3) The test process comprises the following steps: mice were randomized into 3 groups: a blank control group (B1), a fermentation type small molecular peptide group (B2) and a ginseng fructus alpiniae oxyphyllae group (B3). Each group of 8 mice (initial body weight 20 + -2.0 g, SPF grade) except the blank control group were subjected to intragastric administration of 1 mL/(one time) of the corresponding hydrolysate 2 times a day, i.e. once in the morning and evening. At the same time, the blank group was given an equal amount of physiological saline for 14 days. After 14 days the Morris water maze test was performed. The test results are shown in table 2 and fig. 3.
Table 2 example 2 fermentation type small molecule peptide mouse Morris water maze experiment
Note: the average of the water maze simulation data for 8 mice per group is shown in the table.
From the results in table 2, it can be seen that in the localization navigation test, the escape latency of the mice in the medium test group of the gavage fermentation type small molecular peptide hydrolysate is shorter and the time to reach the platform is shorter than those in the blank control group and the ginseng fructus alpiniae oxyphyllae group; and in the space exploration part, the track of the mice with the gastric perfusion fermentation type micromolecule peptide hydrolysate reaching the platform position shows certain memory characteristics, and the times are obviously more than those of mice in a blank control group and a ginseng fructus alpiniae oxyphyllae group. Therefore, the fermented micromolecule peptide hydrolysate can be effectively proved to improve the memory capacity of mice and enhance the learning and cognition capacity.
Application example 3
The effect of the hydrolysate of example 3 and comparative example 3 on improving memory is examined, and the specific scheme is as follows:
(1) animals: 24 wistar mice were purchased from the animal laboratory center of white chen medical college, university of ji forest, housed in SPF-grade animal rooms for one week, during which time they had free access to water.
(2) The main reagents are as follows: the fermented small molecule peptide group hydrolysate in example 3, the ginseng fructus alpiniae oxyphyllae group hydrolysate in comparative example 3, normal saline and common feed. The ratio of the hydrolysate was the same as in application example 1.
(3) The test process comprises the following steps: mice were randomized into 3 groups: a blank control group (C1), a fermentation type small molecule peptide group (C2) and a ginseng fructus alpiniae oxyphyllae group (C3). Each group of 8 mice (initial body weight 20 + -2.0 g, SPF grade) except the blank control group were subjected to intragastric administration of 1 mL/(one time) of the corresponding hydrolysate 2 times a day, i.e. once in the morning and evening. At the same time, the blank group was given an equal amount of physiological saline for 14 days. After 14 days the Morris water maze test was performed. The test results are shown in table 3 and fig. 4.
Table 3 example 3 fermentation type small molecule peptide mouse Morris water maze experiment
Note: the average of the water maze simulation data for 8 mice per group is shown in the table.
From the results in table 3, it can be seen that in the localization navigation test, compared with the blank control group and the ginseng fructus alpiniae oxyphyllae group, the mice in the multiple test group of the gavage fermentation type small molecular peptide hydrolysate have shorter escape latency and shorter time to reach the platform; the results of the comparative examples 1 to 3 also show that the time for reaching the platform is shorter as the dosage concentration of the fermentation type micromolecule peptide hydrolysate is higher; and in the space exploration part, the track of the mice with the gastric perfusion fermentation type micromolecule peptide hydrolysate reaching the platform position shows certain memory characteristics, and the times are obviously more than those of mice in a blank control group and a ginseng fructus alpiniae oxyphyllae group. Fig. 5 is a comparison line graph of the fermented small molecule peptides of examples 1-3 with different concentrations, and it can be found that the higher the dosage concentration of the fermented small molecule peptide hydrolysate is, the more times the mouse reaches the platform position. Therefore, the fermented micromolecule peptide hydrolysate can be effectively proved to improve the memory capacity of mice and enhance the learning and cognition capacity.
The results of the above examples show that the fermented small molecule peptide for assisting in improving memory provided by the invention has a remarkable effect on improving memory, and has the advantages of wide raw material source, low cost, safe components and no toxic or side effect.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. The fermented small molecule peptide for assisting in improving memory is characterized by comprising the following raw materials for preparation in parts by mass: 1-3 parts of ginseng, 1-5 parts of Chinese yam, 1-3 parts of rhizoma polygonati, 1-3 parts of poria cocos, 2-4 parts of radix puerariae, 1-3 parts of fructus alpiniae oxyphyllae, 5-10 parts of walnut and 5-10 parts of needle mushroom; the fermentation type small molecular peptide for assisting in improving memory is obtained by fermenting, carrying out enzymolysis and carrying out ultrafiltration on the raw materials.
2. The method for preparing the fermented small molecule peptide for assisting in improving memory according to claim 1, comprising the following steps: mixing the raw materials with deionized water, adding composite probiotics for fermentation to obtain fermentation liquor; carrying out enzymolysis on the obtained fermentation liquor to obtain a protease hydrolysate; and (3) performing ultrafiltration on the protease hydrolysate to obtain the fermentation type small molecular peptide for assisting in improving the memory.
3. The preparation method of claim 2, wherein the volume ratio of the mass of the raw material to the deionized water is 1g (2-6) mL.
4. The preparation method according to claim 2, wherein the complex probiotics comprise lactobacillus acidophilus, bifidobacterium lactis, lactobacillus rhamnosus and lactobacillus casei; the number of the live bacteria of the composite probiotics is 100 hundred million-2000 hundred million CFU/g; the added mass of the composite probiotics accounts for 0.2-0.5% of the total mass of the raw materials and the deionized water.
5. The preparation method according to claim 2, wherein the fermentation temperature is 35-36 ℃, and the fermentation time is 36-48 h.
6. The preparation method according to claim 2, wherein the enzymolysis is carried out by using hemicellulase and trypsin in sequence; the adding amount of the hemicellulase accounts for 0.1-0.4% of the mass percent of the fermentation broth; the adding amount of the trypsin accounts for 0.1 to 0.4 percent of the mass percentage of the fermentation liquor; the enzymatic activity of the hemicellulase is 10 ten thousand u/g, and the enzymatic activity of the trypsin is 10 ten thousand u/g; the enzymolysis conditions of the hemicellulase are as follows: the temperature is 45-55 ℃, the pH value is 4.0-5.5, and the time is 1-2 h; the enzymolysis conditions of the trypsin are as follows: the temperature is 36-37 ℃, the pH value is 7.8-8.5, and the time is 1-2 h.
7. The method of claim 2, wherein the ultrafiltration comprises the steps of: and (3) performing ultrafiltration on the protein enzymolysis liquid through a filter membrane with the molecular weight of 2000Da, and collecting filtrate to obtain the fermentation type micromolecule peptide.
8. The preparation method of claim 7, further comprising the step of carrying out vacuum spray drying on the filtrate after ultrafiltration to obtain the fermented small molecule peptide powder.
9. Use of the fermented small molecule peptide for assisting in improving memory according to claim 1 or the fermented small molecule peptide for assisting in improving memory obtained by the preparation method according to any one of claims 2 to 8 in preparation of a health product for improving intelligence.
10. The use according to claim 9, wherein the types of said nootropic health products comprise small molecule peptide powder, small molecule peptide bar, small molecule peptide oral liquid, small molecule peptide liquid beverage and small molecule peptide biscuit.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117100774A (en) * | 2023-10-25 | 2023-11-24 | 山东中科嘉亿生物工程有限公司 | Application of lactobacillus acidophilus JYLA-16 in preparation of products for treating gall-stone |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117100774A (en) * | 2023-10-25 | 2023-11-24 | 山东中科嘉亿生物工程有限公司 | Application of lactobacillus acidophilus JYLA-16 in preparation of products for treating gall-stone |
| CN117100774B (en) * | 2023-10-25 | 2024-02-13 | 山东中科嘉亿生物工程有限公司 | Application of lactobacillus acidophilus JYLA-16 in preparation of products for treating gall-stone |
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