Chinese wolfberry fruit enzyme composition capable of tonifying qi and blood as well as preparation method and application of Chinese wolfberry fruit enzyme composition

Abstract

The invention relates to a Chinese wolfberry fruit enzyme composition capable of tonifying qi and blood as well as a preparation method and application thereof, and belongs to the technical field of biological fermentation. The Chinese wolfberry enzyme composition with the function of tonifying qi and blood is prepared from Chinese wolfberry fermentation liquor, maltitol, xylitol, xylooligosaccharide, corn oligopeptide powder and D-sodium erythorbate, can increase the number of peripheral red blood cells, the number of peripheral platelets, the content of peripheral hemoglobin, the number of bone marrow nucleated cells and the content of hepatic glycogen, has the effect of tonifying the blood, and is free of toxic and side effects. The flavor and the taste are excellent. By utilizing a composite efficient strain and peptide-based culture medium fermentation promotion technology, the utilization rate of the raw materials is increased, the nutrient loss is reduced, and the original flavor of the fruit and vegetable raw materials is better reserved.

Description

Technical Field

[0001] The present invention relates to a wolfberry enzyme composition with the functions of supplementing qi and blood, a preparation method thereof, and an application thereof, belonging to the technical field of biological fermentation. Background Art

[0003] Enzymes are a class of proteins that naturally exist in living organisms and have the function of catalyzing biochemical reactions, and are called "catalysts of life". In living organisms, enzymes participate in key physiological processes such as digestion, metabolism, and energy conversion (such as amylase decomposing carbohydrates and protease decomposing proteins). The common "enzyme products" on the market (such as fruit fermentation broth) are not pure enzyme preparations, but mixtures made by microbial fermentation, containing enzymes, organic acids, probiotics, and metabolites, etc., and belong to the category of food. Compared with ordinary fruit juices, prebiotics (such as fructooligosaccharides) and short-chain fatty acids (such as butyric acid) will be produced during the fermentation preparation process of enzyme drinks. Among them, prebiotics can promote the proliferation of intestinal probiotics and thus improve constipation, and short-chain fatty acids can nourish immune cells and then enhance immunity. At the same time, the organic acids produced by fermentation can stimulate gastric acid secretion to promote digestion, and the dietary fiber in enzyme drinks can delay sugar absorption and interfere with cholesterol synthesis, thereby achieving the effect of regulating blood sugar / blood lipid.

[0004] Chinese Patent Document CN118947890A discloses a preparation method and application of black wolfberry enzyme. This patent document uses black wolfberry as a raw material to produce enzyme products. During the production of the products, high-temperature heating is avoided, and gellan gum with high acyl content is added, effectively preventing the thermal decomposition and oxidation of anthocyanins, and greatly retaining the nutrients in the black wolfberry itself. Then, the active substances produced by the fermentation of yeast and lactic acid bacteria strains are used to improve the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase, and finally improve the therapeutic effect on alcoholic liver injury. Chinese Patent Document CN112535284A discloses a preparation method of wolfberry enzyme. The preparation method of this wolfberry enzyme includes the following steps: washing wolfberries and drying them to a constant weight; grinding semen ziziphi spinosae, flos albiziae, poria cocos, semen platycladi, and radix polygalae into powders and mixing them; adding wolfberries into a solvent, and then adding sugar, ascorbic acid, tea water, and the mixed powder to prepare a fermentation medium, and inoculating fermentation strains after sterilization and fermenting for 5 to 30 days; filtering the obtained fermentation broth to remove residues and collecting the supernatant to obtain wolfberry enzyme; the wolfberry enzyme prepared by this patent document has no chemical additives, improves the fermentation efficiency and the content of functional active ingredients, and has antioxidant, anti-aging, anti-insomnia and depression, and calming functions. Chinese Patent Document CN108850759A discloses a preparation method of a wolfberry enzyme beverage with the function of moistening the intestines and relieving constipation. The preparation method of the wolfberry enzyme beverage disclosed in this patent document includes the following steps: washing peach kernels and apricot kernels, putting them into a pulping machine to make pulp, and then putting them into a steam explosion tank for treatment to obtain peach kernel and apricot kernel slurry, and then adding yeast for fermentation to obtain peach kernel and apricot kernel fermentation broth; washing bananas and wolfberries, soaking them in water, grinding them through a colloid mill, then pressurizing for homogenization and crushing, and then enzymolyzing with cellulase to obtain banana and wolfberry slurry; sterilizing the banana and wolfberry slurry, and then performing three-stage temperature-holding fermentation to obtain banana and wolfberry fermentation broth. After mixing the peach kernel and apricot kernel fermentation broth with the banana and wolfberry fermentation broth, adding syrup and honey and mixing evenly to obtain a wolfberry enzyme beverage with the function of moistening the intestines and relieving constipation. This patent document uses raw materials such as peach kernels, red dates, apricot kernels, and wolfberries to ferment to produce metabolites and expand and culture probiotics respectively, so that the product has the health care effect of moistening the intestines and relieving constipation.

[0005] The wolfberry enzyme compositions disclosed in the above patent documents have the following defects: (1) After the fruit and vegetable raw materials are pulped, the viscosity is too high, resulting in insufficient fermentation, and the final drink has a poor taste and flavor. (2) The traditional fermentation process takes a long time and has a high risk of fermentation pollution. (3) The utilization rate of fermented fruit and vegetable raw materials is low, and the nutritional loss is serious, affecting the efficacy of the enzyme beverage. In addition, the functions of the wolfberry enzyme compositions disclosed in the above patent documents are too single and do not have the function of supplementing qi and blood. Summary of the Invention

[0006] The purpose of the present invention is to provide a wolfberry enzyme composition with the function of supplementing qi and blood, its preparation method and application, so as to solve the problem that the current wolfberry enzyme composition lacks the function of supplementing qi and blood.

[0007] The present invention provides a preparation method of a wolfberry enzyme composition with the function of tonifying qi and blood, comprising the following steps:

[0008] (1) Mix wolfberry pulp and enzyme preparation, and then carry out enzymatic hydrolysis reaction to obtain an initial enzymatic hydrolysis product; mix the initial enzymatic hydrolysis product, white granulated sugar and brown sugar, and then carry out sterilization to obtain a sterilized liquid; mix the sterilized liquid and a fermenting agent, and then carry out fermentation to obtain a wolfberry fermentation liquid;

[0009] (2) Mix the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium to obtain a wolfberry enzyme composition with the function of tonifying qi and blood; the mass ratio of the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:20-25:12-15:2-3:0.4-0.8:0.04-0.07.

[0010] Preferably, water is further added in step (2), and the mass ratio of the wolfberry fermentation liquid, water, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:30-35:20-25:12-15:2-3:0.4-0.8:0.04-0.07.

[0011] Preferably, the wolfberry pulp is prepared by soaking dried wolfberries in water and then grinding; the mass ratio of the dried wolfberries to water is 1:4-5, and the soaking time is 6-8 h.

[0012] Preferably, the enzyme preparation is composed of pectinase and cellulase, the mass of the pectinase is 0.015-0.03% of the mass of the wolfberry pulp, and the mass of the cellulase is 0.05-0.08% of the mass of the wolfberry pulp.

[0013] Preferably, the temperature of the enzymatic hydrolysis reaction is 48±2°C, and the time is 3-5 h.

[0014] Preferably, the mass of the white granulated sugar is 5-7% of the mass of the initial enzymatic hydrolysis product, and the mass of the brown sugar is 5-7% of the mass of the initial enzymatic hydrolysis product.

[0015] Preferably, the fermenting agent is composed of equal masses of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum and Bifidobacterium; the mass of the fermenting agent is 10-15% of the mass of the sterilized liquid.

[0016] Preferably, the temperature of the fermentation is 25±2°C, and the time is 45 days; continuous stirring is carried out on the 1st to 7th day of fermentation; stirring is carried out once a day for 2 h on the 8th to 30th day of fermentation; stirring is carried out once every 5 days for 2 h on the 31st to 45th day of fermentation.

[0017] The present invention provides a wolfberry enzyme composition with the function of supplementing qi and blood, which is composed of wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium. The mass ratio of the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:20-25:12-15:2-3:0.4-0.8:0.04-0.07. The preparation method of the wolfberry fermentation liquid is as follows: mix wolfberry pulp and enzyme preparation and carry out enzymatic hydrolysis reaction to obtain an initial enzymatic hydrolysis product; mix the initial enzymatic hydrolysis product, white granulated sugar and brown sugar and carry out sterilization to obtain a sterilized liquid; mix the sterilized liquid and a fermenting agent and carry out fermentation to obtain wolfberry fermentation liquid.

[0018] The present invention provides an application of a wolfberry enzyme composition with the function of supplementing qi and blood in increasing the number of peripheral blood red blood cells, peripheral blood platelets, peripheral hemoglobin content, bone marrow nucleated cell count and liver glycogen content.

[0019] The present invention provides an application of a wolfberry enzyme composition with the function of supplementing qi and blood in alleviating the decrease of the number of peripheral blood red blood cells, peripheral blood platelets, peripheral hemoglobin content, bone marrow nucleated cell count and liver glycogen content caused by acetylphenylhydrazine.

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0021] (1) The present invention adopts a fermentation process of composite fermentation strains, which can greatly shorten the fermentation time and obtain a fermented drink with the effect of nourishing blood and excellent flavor and taste.

[0022] (2) The present invention uses cellulase and pectinase to solve the problem of too high viscosity after the raw materials are pulped.

[0023] (3) The present invention uses a composite high-efficiency strain and a peptide-based culture medium fermentation promotion technology to improve the utilization rate of raw materials, reduce nutrient loss and better retain the original flavor of fruit and vegetable raw materials. Description of the Drawings

[0024] Figure 1 It is a schematic flow chart of the preparation method of the wolfberry enzyme composition with the function of supplementing qi and blood according to the embodiment of the present invention;

[0025] Figure 2 It is a schematic diagram of the influence result of the wolfberry enzyme composition on the number of peripheral blood red blood cells in the qi and blood deficiency mouse model;

[0026] Figure 3 It is a schematic diagram of the influence result of the wolfberry enzyme composition on the number of peripheral blood platelets in the qi and blood deficiency mouse model;

[0027] Figure 4Schematic diagram of the effect of the wolfberry enzyme composition on the peripheral blood hemoglobin content in a mouse model with qi and blood deficiency;

[0028] Figure 5 Schematic diagram of the effect of the wolfberry enzyme composition on the number of nucleated bone marrow cells in a mouse model with qi and blood deficiency;

[0029] Figure 6 Schematic diagram of the effect of the wolfberry enzyme composition on the hepatic glycogen content in a mouse model with qi and blood deficiency. Detailed implementation manners

[0030] The following examples are intended to further illustrate the content of the present invention rather than limit the scope of protection of the present invention.

[0031] Specific examples of the wolfberry enzyme composition with qi and blood tonifying function and its preparation method of the present invention are as follows:

[0032] Example

[0033] The preparation method of the wolfberry enzyme composition with qi and blood tonifying function in this example is as Figure 1 shown, and includes the following steps:

[0034] (1) Soak dried wolfberries in water for 6 h, and then use a colloid mill to grind them into a wolfberry pulp with a fineness of 2 - 50 μm; the mass ratio of dried wolfberries to water is 1:4. The dried wolfberries in this example are Ningxia wolfberries, from a pollution-free raw material base or contracted planting farmers. The planting base should be guided by professional technical personnel in production, pesticides cannot be used, and the purchased raw materials must be tested for pesticide residues and used after passing the test.

[0035] (2) Stir the wolfberry pulp and the enzyme preparation evenly and carry out an enzymatic hydrolysis reaction at a temperature of 48 ± 2 °C for 3 h to obtain an initial enzymatic hydrolysis product; the enzyme preparation is composed of pectinase and cellulase. The mass of pectinase is 0.015% of the mass of the wolfberry pulp, and the mass of cellulase is 0.05% of the mass of the wolfberry pulp; both pectinase and cellulase are commercially available products;

[0036] Then heat the initial enzymatic hydrolysis product to 70 ± 2 °C through a shell-and-tube sterilizer and transport it to a sugar dissolving tank. Then add white granulated sugar and brown sugar to the sugar dissolving tank. After the sugar is fully dissolved, a sugar melting solution is obtained; the initial enzymatic hydrolysis product is heated from room temperature to 70 ± 2 °C in the shell-and-tube sterilizer in 10 - 15 min. The mass of white granulated sugar is 5% of the mass of the initial enzymatic hydrolysis product, and the mass of brown sugar is 5% of the mass of the initial enzymatic hydrolysis product;

[0037] Sterilize the sugar melting solution using a shell-and-tube UHT sterilizer to obtain a sterilized solution; the sterilization temperature is controlled at 125 ± 2 °C, the time is 15 s, and the outlet temperature of the shell-and-tube UHT sterilizer is controlled at 30 ± 2 °C;

[0038] Inject the bactericidal liquid into the specified fermenter (the fermenter should be cleaned in advance and sterilized with steam for 30 min), then heat the fermenter to 95 ± 2 °C and maintain it for 30 min for air sterilization; then use sterile air to press the prepared working starter in the starter tank into the fermenter, start stirring to make the working starter and the bactericidal liquid mix evenly, and at the same time control the fermentation temperature at 25 ± 2 °C and ferment for 45 days (the first 30 days are mainly the fermentation stage, and the last 15 days are mainly the flavor fusion stage) to obtain the wolfberry fermentation liquid; continuous stirring must be carried out on the 1st - 7th day of fermentation; stirring is required once a day for 2 h on the 8th - 30th day; stirring is carried out once every 5 days for 2 h on the 31st - 45th day; during fermentation and extended storage, it is necessary to ensure that the pressure inside the tank is greater than the external pressure to prevent air from entering the tank and causing bacterial contamination. Among them, the working starter is composed of equal masses of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, and Bifidobacterium. The mass of the starter is 10% of the mass of the bactericidal liquid.

[0039] (3) Filter the wolfberry fermentation liquid through a 40 - mesh vibrating screen, then send it into a blending tank, and then add water, maltitol, xylitol, xylo - oligosaccharide, corn oligopeptide powder, and D - isoascorbic acid sodium into the blending tank to obtain a blended material; finally, sterilize the blended material with ultraviolet light for 30 min to obtain a wolfberry enzyme composition with the effect of tonifying qi and blood; when filling the prepared wolfberry enzyme composition, clean the filling glass bottles and sterilize the bottle caps with ozone for 30 min. During the filling process, it is necessary to ensure that the wolfberry enzyme composition liquid is uniform. After capping, the glass bottles enter the tunnel and adopt a back - pressure spraying sterilization process. Among them, the first stage is the heating process, and the temperature is controlled at 65 ± 2 °C; the second stage is the sterilization process, and the temperature is controlled at 90 ± 2 °C; the third stage is the cooling process, and the temperature is controlled at 65 ± 2 °C; the fourth stage is the cleaning process, and the temperature is controlled at 30 ± 2 °C; adjust the main motor speed to ensure that the sterilization time is more than 30 min. Softened water is used for sterilization, and it is quickly cooled after sterilization and dried by a hair dryer. Among them, the mass ratio of the wolfberry fermentation liquid, water, maltitol, xylitol, xylo - oligosaccharide, corn oligopeptide powder, and D - isoascorbic acid sodium is 45:30:20:12:2:0.4:0.04. Corn oligopeptide powder

[0040] Experimental example

[0041] There were three groups in this experiment, with 5 female Balb / c mice and 5 male Balb / c mice in each group. The first group was the blank control group, which was given saline by gavage every day for 7 days; the second group was the model group, in which a model of qi and blood deficiency was established by intraperitoneal injection of 2% acetylphenylhydrazine on the 1st and 4th days, and saline was given by gavage every day for 7 days; the third group was the enzyme group, in which a model of qi and blood deficiency was established by intraperitoneal injection of 2% acetylphenylhydrazine on the 1st and 4th days, and the prepared wolfberry enzyme composition was used to give the mice gavage at a dosage of 200 μL / 10 g every day for 7 days. The mice were sacrificed on the 8th day, and the number of red blood cells, platelets, hemoglobin content, nucleated bone marrow cell count and liver glycogen content in the peripheral blood of the mice were detected.

[0042] (1) Effect of wolfberry enzyme composition on the number of red blood cells in the peripheral blood of mice with qi and blood deficiency model

[0043] Experimental method: On the 8th day of the experiment, the eyeballs of the mice were removed to collect the peripheral blood of the mice into an anticoagulant tube, gently flicked and mixed evenly, 10 μL of peripheral blood was aspirated into 2 mL of red blood cell diluent (Solarbio G3610), blown and mixed evenly, 20 μL was aspirated into the cell counting chamber, and a cell counter (Countstar IC1000) was used for detection. The experimental results are shown in Table 1 and Figure 2 as follows. RBC in Table 1 and Figure 2 represents the content of red blood cells in the peripheral blood.

[0044] Table 1 Effect of wolfberry enzyme composition on the number of red blood cells in the peripheral blood of mice with qi and blood deficiency model

[0045] Group <![CDATA[RBC(10 12 / L)]]> Blank Group 2.065±0.399 Model Group <![CDATA[1.431±0.109 ## > Wolfberry Enzyme Group <![CDATA[1.929±0.113 *** >

[0046] Note: ## P < 0.01 vs blank group; #### P < 0.0001 vs blank group; * P < 0.05 vs model group; *** P < 0.001 vs model group

[0047] As can be seen from Table 1 and Figure 2 , compared with the blank group, after intraperitoneal administration of 2% acetylphenylhydrazine to the mice, the number of red blood cells in the peripheral blood of the model group mice decreased significantly; compared with the model group, the number of red blood cells in the peripheral blood of the mice treated with wolfberry enzyme composition by gavage increased significantly, indicating that the prepared wolfberry enzyme composition of the present invention has the function of increasing the number of red blood cells in the peripheral blood and can relieve the decrease in the number of red blood cells in the peripheral blood caused by acetylphenylhydrazine.

[0048] (2) Effect of wolfberry enzyme composition on the number of platelets in the peripheral blood of mice with qi and blood deficiency model

[0049] Experimental method: On the 8th day of the experiment, the eyeballs of the mice were removed, and the peripheral blood of the mice was collected into anticoagulant tubes. The tubes were gently flicked to mix evenly. Then, 10 μL of the peripheral blood was aspirated into 0.38 mL of platelet diluent (Solarbio G3590), and the mixture was pipetted up and down to mix evenly. Next, 20 μL of the mixture was aspirated into a hemocytometer, and a cell counter (Countstar IC1000) was used for detection. The experimental results are shown in Table 2 and Figure 3 as follows. PLT in Table 2 and Figure 3 represents the platelet concentration in peripheral blood.

[0050] Table 2 Effects of wolfberry enzyme composition on the number of peripheral blood platelets in the qi and blood deficiency mouse model

[0051] Group <![CDATA[PLT(10 9 / L)]]> Blank Group 687.5±23.081 Model Group <![CDATA[359.3±24.994 #### > Wolfberry Enzyme Group <![CDATA[434.667±27.777 * >

[0052] Note: ## P < 0.01 vs blank group; #### P < 0.0001 vs blank group; * P < 0.05 vs model group; *** P < 0.001 vs model group

[0053] As can be seen from Table 2 and Figure 3 it can be known that compared with the blank group, after intraperitoneal administration of 2% acetylphenylhydrazine to the mice, the number of peripheral blood platelets in the model group of mice decreased significantly; compared with the model group, the number of platelets in the peripheral blood of the mice treated with wolfberry enzyme by gavage increased significantly, indicating that the wolfberry enzyme composition prepared by the present invention has the function of increasing the number of platelets in peripheral blood and can alleviate the decrease in the number of platelets in peripheral blood caused by acetylphenylhydrazine.

[0054] (3) Effects of wolfberry enzyme composition on the content of peripheral blood hemoglobin in the qi and blood deficiency mouse model

[0055] Experimental method: On the 8th day of the experiment, the eyeballs of the mice were removed, and the peripheral blood of the mice was collected into anticoagulant tubes. The tubes were gently flicked to mix evenly. After diluting the peripheral blood of the mice 80 times, 50 μL of the diluted blood was taken and added to 200 μL of working solution (Solarbio BC5585), and the reaction was carried out at room temperature for 5 minutes. Then, the absorbance value at 400 nm was measured. The experimental results are shown in Table 3 and Figure 4 as follows.

[0056] Table 3 Effects of wolfberry enzyme composition on the content of peripheral blood hemoglobin in the qi and blood deficiency mouse model

[0057]

[0058]

[0059] Note: ## P < 0.01 vs blank group; ####P < 0.0001 vs blank group; * P < 0.05 vs model group; *** P < 0.001 vs model group

[0060] As shown in Table 3 and Figure 4 it can be seen that compared with the blank group, after intraperitoneal administration of 2% acetylphenylhydrazine to mice, the hemoglobin content in the peripheral blood of the model group mice was significantly decreased; compared with the model group, the concentration of hemoglobin in the peripheral blood of the mice treated with wolfberry enzyme by gavage was significantly increased, indicating that the wolfberry enzyme composition prepared by the present invention has the function of increasing the hemoglobin concentration in the peripheral blood and can relieve the decrease in hemoglobin concentration in the peripheral blood caused by acetylphenylhydrazine.

[0061] (4) Effect of wolfberry enzyme composition on the number of nucleated bone marrow cells in the model mice with qi and blood deficiency

[0062] Experimental method: On the 8th day of the experiment, after the mice were sacrificed, their femurs were taken, the attached muscles and connective tissues were removed, and both ends of the femurs were excised. 2 mL of DPBS was aspirated with a 5 mL syringe, and the bone marrow cavity was rinsed several times, and the rinsing solution was collected in a test tube. The cells were ensured to be evenly dispersed by repeatedly pipetting with a pipette gun, and 20 μL was aspirated into a cell counting plate and detected using a cell counter (Countstar IC1000). The experimental results are shown in Table 4 and Figure 5 as follows. WBC in Table 4 and Figure 5 represents the number of nucleated bone marrow cells.

[0063] Table 4 Effect of wolfberry enzyme composition on the number of nucleated bone marrow cells in the model mice with qi and blood deficiency

[0064] Group <![CDATA[WBC(10 6 / Femur)]]> Blank Group 7.267±0.923 Model Group <![CDATA[3.43±0.300 #### > Wolfberry Enzyme Group <![CDATA[4.51±0.508 *** >

[0065] Note: ## P < 0.01 vs blank group; #### P < 0.0001 vs blank group; * P < 0.05 vs model group; *** P < 0.001 vs model group

[0066] As shown in Table 4 and Figure 5 it can be seen that compared with the blank group, after intraperitoneal administration of 2% acetylphenylhydrazine to mice, the number of nucleated bone marrow cells in the model group mice was significantly decreased; compared with the model group, the number of nucleated bone marrow cells in the mice treated with wolfberry enzyme by gavage was significantly increased, indicating that the wolfberry enzyme composition prepared by the present invention has the function of increasing the number of nucleated bone marrow cells and can relieve the decrease in the number of nucleated bone marrow cells caused by acetylphenylhydrazine.

[0067] (5) Effect of wolfberry enzyme composition on the liver glycogen content in the model mice with qi and blood deficiency

[0068] Experimental method: On the 8th day of the experiment, after the mice were sacrificed, their livers were taken. The operation was carried out according to the instruction manual of Solarbio BC0345. Specifically, 0.1 g of liver was cut into a 10 mL centrifuge tube, minced as much as possible with surgical scissors and homogenized. 0.75 mL of extraction solution was added, and the mixture was boiled in a boiling water bath for 20 min, shaken every 5 min to make it fully mixed. After all the tissues were dissolved, it was taken out and cooled, and the volume was fixed to 5 mL with distilled water. 60 μL of the supernatant was centrifuged and added to 50 μL of the working solution. Then 190 μL of concentrated sulfuric acid was added, mixed well, placed in a boiling water bath for 10 min, and then cooled. 200 μL of the reaction solution was transferred to a 96-well plate, and the absorbance was measured at a wavelength of 620 nm. The experimental results are shown in Table 5 and Figure 6 as follows. Table 5 and Figure 6 the Glycogen in

[0069] Table 5 Effects of Lycium barbarum enzyme composition on liver glycogen content in mice model with deficiency of both qi and blood

[0070] Group Glycogen(mg / g) Blank Group 34.952±3.698 Model Group <![CDATA[7.185±0.581 #### > Wolfberry Enzyme Group <![CDATA[11.489±2.091 * >

[0071] Note: ## P < 0.01 vs blank group; #### P < 0.0001 vs blank group; * P < 0.05 vs model group; *** P < 0.001 vs model group

[0072] It can be seen from Table 5 and Figure 6 that compared with the blank group, after intraperitoneal administration of 2% acetylphenylhydrazine to the mice, the liver glycogen content of the mice in the model group decreased significantly; compared with the model group, the liver glycogen content of the mice treated with intragastric administration of Lycium barbarum enzyme increased significantly, indicating that the Lycium barbarum enzyme composition prepared by the present invention has the function of increasing the liver glycogen content and can relieve the decrease of liver glycogen content caused by acetylphenylhydrazine.

Claims

1. A preparation method of a wolfberry enzyme composition for supplementing qi and blood, characterized in that, It includes the following steps: (1) Mix wolfberry pulp and enzyme preparation and carry out enzymatic hydrolysis reaction to obtain an initial enzymatic hydrolysis product; Mix the initial enzymatic hydrolysis product, granulated sugar and brown sugar and carry out sterilization to obtain a sterilized liquid; Mix the sterilized liquid and a starter and carry out fermentation to obtain a wolfberry fermentation liquid; (2) Mix the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium to obtain a wolfberry enzyme composition with the function of supplementing qi and blood; the mass ratio of the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:20-25:12-15:2-3:0.4-0.8:0.04-0.

07.

2. The preparation method of the wolfberry enzyme composition with qi and blood nourishing function according to claim 1, characterized in that, Water is also added in step (2), and the mass ratio of the wolfberry fermentation liquid, water, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:30-35:20-25:12-15:2-3:0.4-0.8:0.04-0.

07.

3. The preparation method of the goji berry enzyme composition with the function of supplementing qi and blood according to claim 1 or 2, characterized in that, The wolfberry pulp is prepared by soaking dried wolfberries in water and then grinding; the mass ratio of dried wolfberries to water is 1:4-5, and the soaking time is 6-8 h.

4. The preparation method of the wolfberry enzyme composition with the function of supplementing qi and blood as claimed in claim 1, characterized in that, The enzyme preparation consists of pectinase and cellulase. The mass of pectinase is 0.015-0.03% of the mass of wolfberry pulp, and the mass of cellulase is 0.05-0.08% of the mass of wolfberry pulp.

5. The preparation method of the goji berry enzyme composition with qi and blood nourishing function as claimed in claim 1 or 4, characterized in that, The temperature of the enzymatic hydrolysis reaction is 48±2 °C, and the time is 3-5 h.

6. The preparation method of the wolfberry enzyme composition with the function of supplementing qi and blood according to claim 1, characterized in that, The mass of the granulated sugar is 5-7% of the mass of the initial enzymatic hydrolysis product, and the mass of the brown sugar is 5-7% of the mass of the initial enzymatic hydrolysis product.

7. The preparation method of the goji berry enzyme composition with the function of supplementing qi and blood as described in claim 1 or 6, characterized in that, The starter consists of equal masses of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum and Bifidobacterium; the mass of the starter is 10-15% of the mass of the sterilized liquid.

8. The preparation method of the wolfberry enzyme composition with the function of supplementing qi and blood as claimed in claim 1, characterized in that, The temperature of the fermentation is 25±2 °C, and the time is 45 days; continuous stirring is carried out on the 1st to 7th day of fermentation; stirring is carried out once a day for 2 h on the 8th to 30th day of fermentation; stirring is carried out once every 5 days for 2 h on the 31st to 45th day of fermentation.

9. A wolfberry enzyme composition for tonifying qi and nourishing blood, characterized in that, It consists of wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium. The mass ratio of the wolfberry fermentation liquid, maltitol, xylitol, xylo-oligosaccharide, corn oligopeptide powder and D-isoascorbic acid sodium is 45-50:20-25:12-15:2-3:0.4-0.8:0.04-0.07; the preparation method of the wolfberry fermentation liquid is as follows: Mix wolfberry pulp and enzyme preparation and carry out enzymatic hydrolysis reaction to obtain an initial enzymatic hydrolysis product; Mix the initial enzymatic hydrolysis product, granulated sugar and brown sugar and carry out sterilization to obtain a sterilized liquid; Mix the sterilized liquid and a starter and carry out fermentation to obtain a wolfberry fermentation liquid.

10. Application of a wolfberry enzyme composition with the function of supplementing qi and blood prepared by the preparation method of the wolfberry enzyme composition with the function of supplementing qi and blood according to any one of claims 1-8 in increasing the number of peripheral blood red blood cells, the number of peripheral blood platelets, the content of peripheral blood hemoglobin, the count of bone marrow nucleated cells, and the content of liver glycogen.

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