A ginseng peptide with anti-fatigue and constipation-relieving properties and its applications

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a ginseng peptide with anti-fatigue and constipation-relieving properties and its applications. Using ginseng as raw material, the ginseng peptide is prepared through a process involving pulverization, high-pressure homogenization, two-step enzymatic hydrolysis, membrane separation, and spray drying. The yield of the prepared ginseng peptide can reach over 18%, and the purity can reach over 94%. This invention significantly improves the yield and purity of the ginseng peptide by employing a two-step enzymatic hydrolysis process and eliminating the traditional alcohol precipitation purification step, making the entire preparation process more efficient. The ginseng peptide prepared by this invention has antioxidant, anti-fatigue, and constipation-relieving effects, and can be used to prepare products with these properties.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a ginseng peptide with anti-fatigue and constipation-relieving properties and its applications. Background Technology

[0002] In recent years, with the accelerating pace of society and increasing life pressure, the number of people who are chronically stressed, sleep-deprived, and have irregular eating habits has gradually increased, making them prone to sub-health issues such as fatigue and constipation. Fatigue not only manifests as a decline in physical strength and endurance but is also accompanied by energy metabolism disorders and weakened antioxidant capacity; constipation is often caused by slowed intestinal peristalsis, intestinal flora imbalance, and insufficient dietary fiber intake. Although these two types of problems are not considered serious illnesses, they significantly affect quality of life and can induce metabolic and digestive system diseases.

[0003] Ginseng ( Panax ginseng Ginseng (Callicarpa meyerii), known as the "King of Herbs," has a long history of medicinal use in my country. The *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) records that ginseng can "nourish the five internal organs, calm the mind, stop palpitations, dispel evil influences, brighten the eyes, improve intelligence, and prolong life with long-term use," highlighting its widely recognized medicinal value. As a traditional food and medicine plant, it contains various active substances such as ginsenosides, polysaccharides, peptides, and polyphenols, possessing multiple effects including tonifying qi and spirit, enhancing immunity, anti-oxidation, and anti-fatigue. Among these, ginseng peptides are low-molecular-weight polypeptides obtained from ginseng protein through enzymatic hydrolysis or microbial fermentation. They have stable molecular structures, good water solubility, high bioavailability, and are easily absorbed and utilized by the body. In recent years, they have shown good functional activity in anti-oxidation, neuroprotection, and metabolic regulation.

[0004] Chinese patent CN116240256A discloses a ginseng glycopeptide and its preparation process, the preparation process of which includes the following steps: (1) extracting ginseng glycopeptide, collecting the extract, performing alcohol precipitation to obtain ginseng glycoprotein, enzymatically hydrolyzing the ginseng glycoprotein, collecting the enzymatic hydrolysate, and then performing solid-liquid separation of the enzymatic hydrolysate to obtain ginseng glycopeptide; Chinese patent CN113151389B discloses a ginseng glycopeptide, its preparation method and medicinal use, using ginseng as raw material, and obtaining ginseng glycopeptide through steps such as decoction extraction, ethanol precipitation separation, double enzyme hydrolysis, dialysis, and freeze drying.

[0005] In existing technologies, ethanol precipitation is commonly used to prepare ginseng peptides to remove macromolecular impurities, polysaccharides, and some pigment components, thereby obtaining peptide components with higher purity. The ethanol precipitation process relies on controlled conditions such as ethanol concentration, temperature, and time. When the ethanol concentration is too high or the precipitation time is too long, some low-molecular-weight peptides are easily lost with the supernatant, leading to a decrease in the recovery rate of active ingredients. Furthermore, traditional ginseng peptide preparation processes often employ a single enzyme or a single enzymatic hydrolysis process. Although this method is simple to operate, due to the complex composition of ginseng raw materials, a single enzymatic hydrolysis often fails to fully expose the enzyme cleavage sites, resulting in incomplete protein degradation and consequently, a low yield of functional peptides.

[0006] Therefore, there is an urgent need to develop a method for preparing ginseng peptides that has high yield, high purity, and can effectively avoid the loss of activity caused by alcohol precipitation. Summary of the Invention

[0007] In view of the problems existing in the prior art, the present invention provides a ginseng peptide with anti-fatigue and constipation-improving properties and its applications.

[0008] The technical solution of the present invention to solve the above-mentioned technical problems is as follows:

[0009] A process for preparing ginseng peptides includes the following steps:

[0010] (1) The ginseng was ultrasonically cleaned twice, drained, and then crushed to obtain ginseng granules;

[0011] (2) Mix and stir the ginseng granules with deionized water, then homogenize under high pressure to obtain a liquid;

[0012] (3) Transfer the liquid to an enzymatic hydrolysis tank, add 1-2% by mass of dextranase or cellulase, and carry out enzymatic hydrolysis to obtain the first enzymatic hydrolysate;

[0013] (4) Centrifuge the first enzymatic hydrolysate to separate the supernatant and the residue. Collect the supernatant into the enzymatic hydrolysis tank. After high-pressure homogenization, add the residue into the enzymatic hydrolysis tank, raise the temperature and adjust the pH, add the complex protease for enzymatic hydrolysis, and then add 1-3% of the residue mass of trypsin to continue enzymatic hydrolysis to obtain the second enzymatic hydrolysate.

[0014] (5) The second enzymatic hydrolysate is separated by ultrafiltration membrane, the permeate is collected, and filtered through a three-stage filter cartridge sterilization system to obtain sterile peptide solution;

[0015] (6) The sterile peptide solution is spray-dried and then aseptically packaged in a clean area to obtain the ginseng peptide.

[0016] Furthermore, in step (2), ginseng granules and deionized water are mixed at a solid-liquid mass ratio of 1:20, and the high-pressure homogenization conditions are 1300-1500 bar pressure, and homogenization is performed 2-3 times.

[0017] Furthermore, in step (4), the centrifugation conditions are 4000-6000 rpm for 20-30 min.

[0018] Furthermore, in step (4), the temperature is raised to 60±0.2℃, and the pH is 7.5-8.5;

[0019] Furthermore, the complex protease consists of flavor protease and papain, with a mass ratio of flavor protease to papain of 1:1-2.

[0020] Furthermore, in step (5), the ultrafiltration membrane separation conditions are a molecular weight cutoff of 1000-5000 Da.

[0021] Furthermore, in step (6), the spray drying conditions are: inlet air temperature of 180±10℃, outlet air temperature of 90±5℃, feed rate of 300 mL / h, and feed speed of 3 mL / min.

[0022] This invention provides a method for preparing ginseng peptides.

[0023] This invention provides the application of the above-mentioned ginseng peptide in the preparation of antioxidant products.

[0024] This invention provides the application of the above-mentioned ginseng peptide in the preparation of anti-fatigue products.

[0025] This invention provides the application of the above-mentioned ginseng peptide in the preparation of products that improve constipation.

[0026] The products mentioned above include, but are not limited to, one or more of the following: pharmaceuticals, food, and health products.

[0027] The beneficial effects of this invention are as follows: Compared with traditional ginseng peptide preparation methods, this invention adopts a two-step enzymatic hydrolysis process and eliminates the traditional alcohol precipitation purification step. This improvement increases the yield and purity of ginseng peptides, making the entire preparation process more efficient and the process simpler. It also avoids the problem of low molecular weight peptides being lost with the supernatant under high ethanol concentration or long precipitation conditions, effectively reducing the loss of active ingredients, avoiding alcohol residues and environmental safety hazards. The process is more green and safe, and can be scaled up for industrial production.

[0028] The ginseng peptides prepared by this invention exhibit excellent performance in multiple bioactivities, including good antioxidant, anti-fatigue, and constipation-relieving effects. They can provide a feasible technical path and theoretical basis for their application in the research and development of functional foods, nutritional health products, and novel drugs, and have broad industrialization prospects. Attached Figure Description

[0029] Figure 1 This is a flow chart of the ginseng peptide preparation process described in this invention;

[0030] Figure 2 This is a diagram showing the results of the weighted swimming test in mice according to the present invention. Detailed Implementation

[0031] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, the reagents used in this invention are all conventional reagents in the art and can be obtained commercially. Unless otherwise specified, the experimental methods used in this invention are all conventional experimental methods in the art.

[0032] See attached document Figure 1 -Appendix Figure 2 The present invention provides: Example 1

[0033] (1) Weigh the dried ginseng, add distilled water at 30 times the weight of the ginseng and place it in an ultrasonic cleaning device. Clean it twice at 30°C and 800W for 1 hour each time. After cleaning, drain the water from the ginseng and put it into a pulverizer. Pulverize it at 18000 rpm for 5 minutes to obtain uniform ginseng particles.

[0034] (2) Mix the ginseng granules from step (1) with deionized water at a solid-liquid mass ratio of 1:20, stir evenly, and then send the mixture into a high-pressure homogenizer. Homogenize twice under a pressure of 1300 bar to obtain the liquid.

[0035] (3) Transfer the liquid from step (2) into an enzymatic hydrolysis tank, add 1% by mass of dextranase at 50°C with an enzyme activity of 20 U / mL, and hydrolyze for 1.5 h to obtain the first enzymatic hydrolysate;

[0036] (4) The first enzymatic hydrolysate from step (3) is centrifuged at 4000 rpm for 30 min using an automatic unloading centrifuge to separate the supernatant and the residue. The supernatant is collected in an enzymatic hydrolysis tank. The residue is homogenized by a homogenizer and then enters the enzymatic hydrolysis tank. The temperature is raised to 59.8℃ and the pH is adjusted to 7.5. 3% of the residue mass of a complex protease is added. The complex protease is a mixture of flavor protease and papain in a mass ratio of 1:1. Enzymatic hydrolysis is performed for 2.5 h. Then, 1% of the substrate mass of trypsin is added and enzymatic hydrolysis is continued for 1 h to obtain the second enzymatic hydrolysate.

[0037] (5) The second enzymatic hydrolysate is sent into the ultrafiltration system and fractionated using an ultrafiltration membrane with a molecular weight cutoff of 1000 Da to remove large molecular weight proteins and insoluble components. The permeate is collected and then passed through a three-stage filter cartridge sterilization system (10 μm - 1 μm - 0.22 μm) to obtain sterile peptide solution.

[0038] (6) The sterile peptide solution in step (5) is spray-dried under the conditions of inlet air temperature of 170℃, outlet air temperature of 85℃, feed rate of 300 mL / h and feed speed of 3 mL / min, and then sterilely packaged in a clean area to obtain ginseng peptide. Example 2

[0039] (1) Weigh the dried ginseng, add distilled water at 30 times the weight of the ginseng and place it in an ultrasonic cleaning device. Clean it twice at 30°C and 800W for 1 hour each time. After cleaning, drain the water from the ginseng and put it into a pulverizer. Pulverize it at 20,000 rpm for 4 minutes to obtain uniform ginseng particles.

[0040] (2) Mix the ginseng granules from step (1) with deionized water at a solid-liquid mass ratio of 1:20, stir evenly, and then send the mixture into a high-pressure homogenizer. Homogenize twice under a pressure of 1400 bar to obtain the liquid.

[0041] (3) Transfer the liquid from step (2) into an enzymatic hydrolysis tank, add 2% by mass of dextranase at 50°C with an enzyme activity of 20 U / mL, and hydrolyze for 1.5 h to obtain the first enzymatic hydrolysate;

[0042] (4) The first enzymatic hydrolysate from step (3) is centrifuged at 5000 rpm for 25 min using an automatic unloading centrifuge to separate the supernatant and the residue. The supernatant is collected in an enzymatic hydrolysis tank. The residue is homogenized by a homogenizer and then enters the enzymatic hydrolysis tank. The temperature is raised to 60°C and the pH is adjusted to 8.0. 1% of the mass of the residue is added with a complex protease, which is a mixture of flavor protease and papain in a mass ratio of 1:2. The mixture is hydrolyzed for 2.5 h. Then, 2% of the substrate mass of trypsin is added and the mixture is hydrolyzed for another 1 h to obtain the second enzymatic hydrolysate.

[0043] (5) The second enzymatic hydrolysate is sent into the ultrafiltration system and fractionated using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da to remove large molecular weight proteins and insoluble components. The permeate is collected and then passed through a three-stage filter cartridge sterilization system (10 μm - 1 μm - 0.22 μm) to obtain sterile peptide solution.

[0044] (6) The sterile peptide solution in step (5) is spray-dried under the conditions of inlet air temperature of 180℃, outlet air temperature of 90℃, feed rate of 300 mL / h, and feed speed of 3 mL / min. Then, it is aseptically packaged in a clean area to obtain ginseng peptide. Example 3

[0045] (1) Weigh the dried ginseng, add distilled water at 30 times the weight of the ginseng and place it in an ultrasonic cleaning device. Clean it ultrasonically at 30°C and 800W for 1 hour. Repeat the cleaning once. After cleaning, drain the water from the ginseng and put it into a pulverizer. Pulverize it at 22000 rpm for 3 minutes to obtain uniform ginseng particles.

[0046] (2) Mix the ginseng granules from step (1) with deionized water at a solid-liquid mass ratio of 1:20, stir evenly, and then send it into a high-pressure homogenizer. Homogenize three times under a pressure of 1500 bar to obtain the liquid.

[0047] (3) Transfer the liquid from step (2) into an enzymatic hydrolysis tank, add 2% cellulase at 50°C with an enzyme activity of 20 U / mL, and hydrolyze for 1.5 h to obtain the first enzymatic hydrolysate;

[0048] (4) The first enzymatic hydrolysate from step (3) is centrifuged at 6000 rpm for 20 min using an automatic unloading centrifuge to separate the supernatant and the residue. The supernatant is collected in an enzymatic hydrolysis tank. The residue is homogenized by a homogenizer and then enters the enzymatic hydrolysis tank. The temperature is raised to 60.2℃ and the pH is adjusted to 8.5. 3% of the residue mass of a complex protease is added. The complex protease is a mixture of flavor protease and papain in a mass ratio of 1:1. Enzymatic hydrolysis is performed for 3.5 h. Then, 3% of the residue mass of trypsin is added and enzymatic hydrolysis is continued for 1 h to obtain the second enzymatic hydrolysate.

[0049] (5) The second enzymatic hydrolysate is sent into the ultrafiltration system and fractionated using an ultrafiltration membrane with a molecular weight cutoff of 5000 Da to remove large molecular weight proteins and insoluble components. The permeate is collected and then passed through a three-stage filter cartridge sterilization system (10 μm - 1 μm - 0.22 μm) to obtain sterile peptide solution.

[0050] (6) The sterile peptide solution in step (5) is spray-dried under the conditions of inlet air temperature of 190℃, outlet air temperature of 95℃, feed rate of 300 mL / h and feed speed of 3 mL / min, and then sterilely packaged in a clean area to obtain ginseng peptide.

[0051] Comparative Example 1

[0052] The difference between Comparative Example 1 and Example 1 is that trypsin is not added in step (4), while the other steps are the same.

[0053] Comparative Example 2

[0054] The difference between Comparative Example 2 and Example 1 is that no complex protease is added in step (4), while the other steps are the same.

[0055] Comparative Example 3

[0056] The difference between Comparative Example 2 and Example 1 is that the complex protease in step (4) is a neutral protease and a papain, and the mass ratio of the neutral protease to the papain is 1:2. All other steps are the same.

[0057] Comparative Example 4

[0058] Ginseng polypeptides are prepared according to the preparation method described in patent CN114107418B.

[0059] The yield and purity of the ginseng peptides prepared in Examples 1-3 and Comparative Examples 1-4 were tested, and the results are shown in Table 1.

[0060] Table 1. Yield and purity results of ginseng peptides prepared in each example and comparative example.

[0061] Group Yield (%) purity(%) Example 1 19.81 96.2 Example 2 19.15 95.6 Example 3 18.53 94.0 Comparative Example 1 10.46 84.4 Comparative Example 2 7.92 78.5 Comparative Example 3 14.64 89.2 Comparative Example 4 11.33 87.1

[0062] The results are shown in Table 1. The yield of ginseng peptides prepared in Examples 1-3 of this invention was 18.53%-19.81%, and the purity was 94.0%-96.2%, both superior to the ginseng peptides prepared in the comparative examples. In contrast, the yields and purity of the products obtained in Comparative Examples 1 and 2 were lower, indicating that the ginseng raw material had a more complex composition and higher impurity content. A single enzymatic hydrolysis process was insufficient to achieve sufficient protein degradation, thus limiting the extraction rate and purity of ginseng peptides. The yield and purity of the ginseng peptides obtained in Comparative Example 4 were also lower than those in Examples 1-3. Although multiple enzymes were used for enzymatic hydrolysis, the ethanol elution capacity was limited, making it difficult to fully remove impurities, resulting in lower yields and purity of ginseng peptides.

[0063] Efficacy verification

[0064] 1. Validation of antioxidant activity in vitro

[0065] DPPH free radical scavenging ability

[0066] The antioxidant activity of the ginseng peptide product prepared in Example 1 was evaluated using the DPPH method. A 0.2 mM DPPH working solution was prepared; ginseng peptide solutions were prepared at concentrations of 10, 15, 20, 25, and 30 μg / mL. 100 μL of ginseng peptide solution was pipetted into each well of a 96-well plate, and 100 μL of DPPH working solution was added to each well as a sample group. 100 μL of anhydrous ethanol was added to each well instead of the DPPH working solution as a control group. 100 μL of anhydrous ethanol was added to each well as a blank group. After mixing, the plates were incubated in the dark for 30 min. The absorbance of each well was measured at 517 nm using a microplate reader. The experiment was repeated three times. Ascorbic acid was used as a positive control, and the DPPH free radical scavenging rate was measured. The DPPH scavenging capacity W (%) was calculated as follows:

[0067] W (%) = [1-(A1-A2) / A0]×100%

[0068] The absorbance value of the blank group was A0, the absorbance value of the ginseng peptide solution was A1, and the absorbance value of the control group was A2.

[0069] Table 2. Effects of ginseng peptides on DPPH free radical scavenging ability

[0070] Ginseng peptide solution concentration (μg / mL) DPPH clearance rate (%) 10 26.78±0.74 15 34.12±0.88 20 53.15±1.58 25 62.40±1.57 30 71.23±2.14 ascorbic acid 97.34

[0071] The results are shown in Table 2. Within the experimental concentration range, the scavenging ability of the ginseng peptide prepared in this invention against DPPH free radicals increased with the increase of ginseng peptide concentration. At the highest concentration of 30 μg / mL, its scavenging rate was 71.23%.

[0072] 2. Anti-fatigue effect – Mouse weight-bearing swimming test

[0073] (1) Animal grouping and drug dosage:

[0074] Sixty male SPF mice were selected and housed in a standardized clean animal room at a temperature of (23±2)℃ and a relative humidity of 45%-65%. Their normal diet was not restricted during this period. After one week of acclimatization, the mice were randomly divided into five groups: a blank control group, a low-dose ginseng peptide group, a medium-dose ginseng peptide group, a high-dose ginseng peptide group, and a taurine positive control group (PC group), with 12 mice in each group. The low, medium, and high-dose groups were administered the ginseng peptide prepared in Example 1 by gavage at doses of 400, 800, and 1200 mg / kg, respectively. The positive control group was administered taurine at a dose of 400 mg / kg, and the blank control group was administered an equal volume of physiological saline. All administrations were once daily for 28 consecutive days.

[0075] (2) Weight-bearing swimming test for mice:

[0076] The swimming pool consisted of a plastic bucket with a diameter of 40 cm and a depth of 30 cm. The water temperature was controlled at (25±2)℃. A weight equivalent to 15% of the mouse's body weight was tied to the tail of the mouse, and the time from the surface to the bottom was recorded as the weighted swimming time. The mice were subjected to weighted swimming 60 minutes after the last drug administration. When the mice were observed to float on the surface, they were gently tapped with a glass rod to keep them swimming. When the mouse's head was submerged for more than 10 seconds and it was unable to float back up, it was considered to be in a state of exhaustion. At this point, the time from the start of swimming to exhaustion was recorded.

[0077] The results are attached. Figure 2 As shown, the results indicate that with the increase of ginseng peptide dosage, the weight-bearing swimming time of mice is prolonged, and the weight-bearing swimming time of mice in the medium and high dose ginseng peptide groups is higher than that of the taurine control group. The results show that the ginseng peptide prepared in this invention has an anti-fatigue effect.

[0078] 3. Verification of its effect in improving constipation

[0079] (1) Experimental animals: Eight-week-old healthy male Kunming mice weighing 29-32g were used. The temperature was (25±2)℃ and the humidity was 50-60%. Throughout the experiment, the mice had free access to water and food. They were given an acclimatization period of 1 day before the experiment.

[0080] (2) Grouping and modeling: After the mice adapted to the environment for one day, they were randomly divided into a blank control group, a model group, a whey protein group (400 mg / kg) and low, medium and high dose ginseng peptide groups (200, 400 and 800 mg / kg). The ginseng peptide was the ginseng peptide prepared in Example 1. There were 10 mice in each group. The control group and the model group were given distilled water by gavage. The whey protein group and each dose of ginseng peptide group were given the corresponding dose of whey protein and ginseng peptide aqueous solution by gavage, 1 mL once a day for 7 consecutive days.

[0081] 3.1 Small Intestinal Motility Experiment:

[0082] Weigh 25 mg of compound diphenoxylate, grind it into powder using a mortar and pestle, add water to 100 mL, shake to mix well, and set aside. Take 2 mL of Indian ink and add 98 mL of distilled water to prepare a 2% (v / v) ink solution. The gavage dose is 0.1 mL / (10 g·bw). Prepare solutions by mixing ink with the aqueous solutions of each test sample, and maintain a total gavage volume of 1 mL.

[0083] Mice that were fasted for 7 days but allowed free access to water for 16 hours after intervention were randomly divided into two groups: the model group, the whey protein group, and the low, medium, and high dose ginseng peptide groups were administered compound diphenoxylate solution by gavage (5 mg / kg bw) to establish a mouse model of small intestinal peristalsis inhibition. The blank control group was administered an equal volume of distilled water by gavage. 30 minutes later, each dose group was administered ink containing the corresponding amount of the test sample aqueous solution by gavage, while the blank group and the model group were administered ink by gavage. 25 minutes after gavage, the animals were immediately euthanized by cervical dislocation, the abdominal cavity was opened, the mesentery was separated, and the intestinal segment from the upper pylorus to the lower ileocecal junction was cut. The small intestine was gently pulled into a straight line, and the length of the intestinal segment was measured as the "total length of the small intestine". The length from the pylorus to the leading edge of the ink was measured as the "ink propulsion length".

[0084] Ink propulsion rate P (%) = (Ink propulsion length (cm) / Total small intestine length (cm)) × 100%

[0085] 3.2 Effects of ginseng peptides on mouse defecation

[0086] Weigh 1 g of fuchsin dye and add it to 49 mL of distilled water to prepare a 2% (v / v) fuchsin solution. The gavage dose is 0.1 mL / 10 g·bw. Prepare a solution of fuchsin solution and test sample aqueous solution, and maintain a total gavage volume of 1 mL.

[0087] Mice that were fasted for 7 days but allowed free access to water for 16 hours after intervention were randomly divided into two groups: the model group, the whey protein group, and the low, medium, and high dose ginseng peptide groups. A mouse constipation model was established by gavage with compound diphenoxylate solution (10 mg / kg·bw), while the blank control group was gavage with distilled water. 30 minutes later, each dose group was gavage with fuchsin solution containing the corresponding test sample, while the blank control group and the model group were gavage with fuchsin solution. Subsequently, the time of the first red feces excreted by each animal, the number of red feces excreted within 6 hours, and the weight of the feces were recorded.

[0088] 3.3 Results of small intestinal motility test

[0089] Table 3 Results of ginseng peptide-induced small intestinal propulsion in mice

[0090] Group Ink propulsion rate (%) Blank control group 93.32±14.72 Model group 65.14±10.44 whey protein group 78.65±12.94 Ginseng peptide low-dose group 82.23±15.23 Ginseng peptide medium dose group 86.58±15.27 High-dose ginseng peptide group 90.27±13.74

[0091] The experimental results are shown in Table 3. The ink propulsion rate in the model group was significantly lower than that in the blank control group, indicating that the constipation model was successfully established. Compared with the ink propulsion rate in the model group, the ink propulsion rates in the whey protein group, the low-dose ginseng peptide group, the medium-dose ginseng peptide group, and the high-dose ginseng peptide group were increased. Compared with the ink propulsion rate in the whey protein group, the ink propulsion rate in the high-dose ginseng peptide group was significantly increased. The results indicate that ginseng peptide has a positive effect on the small intestinal motility of mice.

[0092] 3.4 Effects of ginseng peptides on mouse defecation

[0093] Table 4. Effects of ginseng peptides on mouse defecation

[0094] Group Time to first red stool passage (min) Number of fecal particles within 6 hours Fecal weight (g) within 6 hours Blank control group 110.50±8.84 7.80±1.74 0.13±0.02 Model group 256.70±21.69 5.80±1.14 0.09±0.03 whey protein group 271.90±23.26 5.40±1.52 0.08±0.02 Ginseng peptide low-dose group 294.40±33.40 6.20±1.11 0.10±0.02 Ginseng peptide medium dose group 262.60±28.79 6.60±1.21 0.11±0.03 High-dose ginseng peptide group 229.30±28.36 7.10±1.82 0.12±0.03

[0095] The results are shown in Table 4. Compared with the blank control group, the time to first stool excretion in the model group mice was significantly prolonged, and the number of fecal particles and the weight of fecal matter within 6 hours were significantly reduced, indicating successful model establishment. Compared with the blank control group, the whey protein group showed a significant reduction in the number of fecal particles and the weight of fecal matter within 6 hours, excluding the influence of additional protein intake. After administration of ginseng peptides, their defecation-promoting effect was dose-dependent. Compared with the model group, the high-dose ginseng peptide group showed a significantly shortened time to first stool excretion, and a significant increase in the number of fecal particles and the weight of fecal matter within 6 hours. These results indicate that the ginseng peptides prepared in this invention have constipation-relieving activity at high doses, effectively promoting intestinal peristalsis and improving defecation function.

[0096] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A preparation process of ginseng peptide, characterized in that, Includes the following steps: (1) The ginseng was ultrasonically cleaned twice, drained, and then crushed to obtain ginseng granules; (2) Mix and stir the ginseng granules with deionized water, then homogenize under high pressure to obtain a liquid; (3) Transfer the liquid to an enzymatic hydrolysis tank, add 1-2% by mass of dextranase or cellulase, and carry out enzymatic hydrolysis to obtain the first enzymatic hydrolysate; (4) Centrifuge the first enzymatic hydrolysate to separate the supernatant and the residue. Collect the supernatant into the enzymatic hydrolysis tank. After high-pressure homogenization, add the residue into the enzymatic hydrolysis tank, raise the temperature and adjust the pH, add the complex protease for enzymatic hydrolysis, and then add 1-3% of the residue mass of trypsin to continue enzymatic hydrolysis to obtain the second enzymatic hydrolysate. (5) The second enzymatic hydrolysate is separated by ultrafiltration membrane, the permeate is collected, and filtered through a three-stage filter cartridge sterilization system to obtain sterile peptide solution; (6) The sterile peptide solution is spray-dried and then aseptically packaged in a clean area to obtain the ginseng peptide; In step (4), the temperature is raised to 60±0.2°C and the pH is 7.5-8.5; the complex protease is flavor protease and papain, and the mass ratio of flavor protease to papain is 1:1-2; In step (5), the ultrafiltration membrane separation conditions are a molecular weight cutoff of 1000-5000 Da.

2. The preparation process of ginseng peptide according to claim 1, characterized in that, In step (2), ginseng granules and deionized water are mixed at a solid-liquid mass ratio of 1:

20. The high-pressure homogenization conditions are 1300-1500 bar pressure and homogenization is performed 2-3 times.

3. The preparation process of ginseng peptide according to claim 1, characterized in that, In step (4), the centrifugation conditions are 4000-6000 rpm for 20-30 min.

4. The preparation process of ginseng peptide according to claim 1, characterized in that, In step (6), the spray drying conditions are: inlet air temperature of 180±10°C, outlet air temperature of 90±5°C, feed rate of 300 mL / h, and feed speed of 3 mL / min.

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