A compound plant fermentation broth, its preparation method and application

Abstract

The present invention relates to the technical field of plant fermentation, and specifically discloses a compound plant fermentation liquid, a preparation method thereof and an application. A preparation method of a compound plant fermentation liquid comprises the following steps: Step 1: Mix polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate with the peel removed and mulberry leaves to obtain a fermentation substrate; Step 2, ferment with bacillus subtilis; Step 3, ferment with monascus purpureus; Step 4, ferment with lactiplantibacillus plantarum; Step 5: After filtering and sterilization, a compound plant fermentation liquid is obtained.

Description

Technical Field

[0001] The present invention relates to the technical field of plant fermentation, and particularly relates to a composite plant fermentation liquid, a preparation method thereof, and an application thereof. Background Art

[0002] Existing products for relieving fatigue have some limitations. Although chemically synthesized anti-fatigue products have the effect of quickly relieving fatigue, long-term use may cause damage to liver and kidney functions or other health problems. And anti-fatigue products extracted chemically may lead to the loss of active ingredients due to improper extraction processes, thus affecting the effect. In addition, although single-fermented products for relieving fatigue provide a certain degree of relief, due to the lack of diverse microbial actions, the types of active ingredients produced are limited, making it difficult to achieve a good relief effect.

[0003] Facing the growing health needs and the preference for natural therapies, it is particularly important to improve the defects of existing products for relieving fatigue. In order to overcome the defects of existing products for relieving fatigue and achieve a safer and more effective effect of relieving fatigue, it is necessary to further improve the preparation methods of existing products for relieving fatigue. Summary of the Invention

[0004] In order to achieve a safer and more effective effect of relieving fatigue, the present application provides a composite plant fermentation liquid, a preparation method thereof, and an application thereof.

[0005] In a first aspect, a preparation method of a composite plant fermentation liquid provided by the present application adopts the following technical solution:

[0006] A preparation method of a composite plant fermentation liquid includes the following steps:

[0007] Step 1, preparing a fermentation substrate: Mixing polygonatum sibiricum, millettia speciosa champ, wolfberry, licorice, ginseng, pomegranate without peel, and mulberry leaves to make a pulp, adjusting the water content of the pulp to 90% - 100%, and then sterilizing to obtain a fermentation substrate;

[0008] Step 2, fermenting with bacillus subtilis: Inoculating bacillus subtilis into the fermentation substrate, and performing aerobic fermentation at 40 - 42 °C for 20 - 24 hours to obtain a fermentation product of bacillus subtilis;

[0009] Step 3, fermenting with monascus purpureus: After sterilizing the fermentation product of bacillus subtilis again, inoculating monascus purpureus, and performing aerobic fermentation at 28 - 32 °C for 5 - 6 days to obtain a fermentation product of monascus purpureus;

[0010] Step 4, fermenting with lactobacillus plantarum: After sterilizing the fermentation product of monascus purpureus again, inoculating lactobacillus plantarum, and performing anaerobic fermentation at 35 - 37 °C for 24 - 28 hours to obtain a composite fermentation product;

[0011] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation broth.

[0012] In the above technical solution, polygonatum sibiricum is rich in polysaccharides, amino acids and trace elements. Millettia speciosa can tonify the lung and strengthen the tendons, with antioxidant and anti-inflammatory effects, improving respiratory and tendon problems. Lycium barbarum has the functions of enhancing immunity, promoting hematopoiesis, reducing blood lipid, anti-tumor and anti-aging. Licorice can invigorate the spleen and replenish qi, resolve phlegm and relieve cough, relieve pain and inflammation, and clear heat and detoxify. Ginseng contains rare saponin Rg3, with the effects of replenishing qi and nourishing blood, calming the nerves and improving intelligence. Pomegranate and mulberry leaves without peel are rich in antioxidant substances and trace elements, which help to improve the nutritional value and biological activity of the overall fermentation broth.

[0013] In this application, polygonatum sibiricum, millettia speciosa, lycium barbarum, licorice, ginseng, pomegranate without peel and mulberry leaves are used as fermentation substrates, and three-stage sequential fermentation is carried out using bacillus subtilis, monascus purpureus and lactobacillus plantarum, realizing the synergistic effect among components. First of all, polygonatum sibiricum provides rich polysaccharides, ginseng contains various saponin compounds, peeled pomegranate is rich in antioxidants such as anthocyanins and ellagic acid, while mulberry leaves contribute rich flavonoids. This combination of fermentation substrates not only provides sufficient nutrient matrix for subsequent microbial fermentation, but also enhances the functionality of the overall fermentation product through the synergistic effect among components. Secondly, in the fermentation stage of this application, bacillus subtilis is first used to decompose complex macromolecules to provide a nutritional basis for subsequent fermentation, then monascus purpureus produces antioxidant metabolites during fermentation to further enhance the activity of the fermentation broth, and finally lactobacillus plantarum is used to increase the lactic acid content, promote intestinal health and inhibit the growth of harmful microorganisms. Through this multi-level microbial fermentation, this application not only optimizes the fermentation matrix, but also unexpectedly generates a fermentation broth rich in small molecule nutrients and various bioactive components, achieving the unexpected effect of effectively relieving physical fatigue.

[0014] Preferably, the mass ratio of polygonatum sibiricum, millettia speciosa, lycium barbarum, licorice, ginseng, pomegranate without peel and mulberry leaves is (4 - 6):(2 - 4):(0.5 - 1):(0.5 - 1):(0.5 - 1):(1 - 2):(1 - 2).

[0015] In the above technical solution, by controlling the dosage ratio of these raw materials, this application improves the bioavailability of these raw materials, ensures that each raw material can exert its best efficacy during fermentation, thereby promoting the synergistic effect among them, ensuring that there is sufficient nutrition for microbial metabolism during fermentation, and being able to generate small molecule active components that are more easily absorbed by the human body, thereby enhancing the effect of the composite plant fermentation broth in relieving physical fatigue.

[0016] Preferably, in step 2, the inoculation amount of bacillus subtilis is 0.3% - 0.5% of the weight of the fermentation substrate.

[0017] In the above technical solution, in the present application, by inoculating Bacillus subtilis in the fermentation substrate at a dosage ratio of 0.3% to 0.5%, it can ensure that there are a sufficient number of viable bacteria participating in the metabolic activities in the initial stage of fermentation, and it helps to optimize the fermentation environment, enabling Bacillus subtilis to effectively decompose the macromolecular substances in the substrate and convert them into small molecule nutrient components that are easy for subsequent microorganisms to utilize, thereby laying a good foundation for the next fermentation step.

[0018] Preferably, in step 3, the inoculation amount of Monascus purpureus is 0.5% to 1% of the weight of the fermentation product of Bacillus subtilis.

[0019] In the above technical solution, in the present application, by setting the inoculation amount of Monascus purpureus at 0.5% to 1%, it can ensure that there are sufficient Monascus purpureus spores participating in the fermentation process and quickly adapting to and utilizing the nutrient environment produced by the fermentation of Bacillus subtilis, promoting the growth and reproduction of Monascus purpureus and the accumulation of metabolites.

[0020] Preferably, in step 4, the inoculation amount of Lactiplantibacillus plantarum is 2% to 3% of the weight of the fermentation product of Monascus purpureus.

[0021] In the above technical solution, in the present application, by selecting an inoculation amount of Lactiplantibacillus plantarum of 2% to 3%, it can not only ensure that there are sufficient numbers of Lactiplantibacillus plantarum to achieve effective metabolic activities, but also better improve the quality characteristics of the composite plant fermentation broth, and further enhance the effect of the composite plant fermentation broth in relieving physical fatigue.

[0022] Preferably, step 1 is specifically as follows:

[0023] Step 1-1: Respectively chop Polygonatum sibiricum, Millettia speciosa, Lycium barbarum, Glycyrrhiza uralensis, Panax ginseng, pomegranate without peel, and Morus alba leaves, and then mix and beat them into a slurry.

[0024] Step 1-2: Add water to adjust the water content of the slurry to 90% to 100%, and sterilize it to obtain a fermentation substrate.

[0025] In the above technical solution, in the present application, by chopping and beating Polygonatum sibiricum, Millettia speciosa, peeled pomegranate, and Morus alba leaves, the uniform dispersion and enzymatic hydrolysis efficiency between the raw materials are increased, thereby improving the nutritional value and functionality; finally, the water content of the slurry is adjusted to 90% to 100% and sterilization treatment is carried out to ensure a good growth environment for microorganisms and the purity of the fermentation substrate during the fermentation process, laying a good foundation for subsequent fermentation.

[0026] In a second aspect, a composite plant fermentation broth provided by the present application adopts the following technical solution:

[0027] A composite plant fermentation broth is prepared by using the preparation method of the composite plant fermentation broth described in the first aspect.

[0028] In the above technical solution, through the above specific preparation process, the composite plant fermentation broth of the present application realizes the synergistic effect of various plant components and microorganisms, thereby generating a composite plant fermentation broth rich in small molecule nutrients and various bioactive components. Such a composite plant fermentation broth has a significant effect of relieving physical fatigue and has broad application prospects in the biomedical industry.

[0029] In the third aspect, the application of a composite plant fermentation broth provided by the present application adopts the following technical solution:

[0030] For the application of a composite plant fermentation broth, the composite plant fermentation broth is used in the biomedical industry.

[0031] In the above technical solution, the composite plant fermentation broth of the present application can not only effectively relieve physical fatigue, but also show broad application potential in the biomedical industry due to its natural ingredients and the characteristics of microbial fermentation.

[0032] In summary, the present application includes at least one of the following beneficial technical effects:

[0033] 1. Through the combination of specific plant raw materials such as polygonatum sibiricum, millettia speciosa, wolfberry fruit, liquorice root, ginseng, pomegranate without peel, and mulberry leaf and microbial fermentation, the present application realizes the synergistic effect among the raw materials, improves the bioavailability of these raw materials, optimizes the fermentation substrate, and ensures that each raw material can exert its best efficacy during the fermentation process, thereby enhancing the effect of the composite plant fermentation broth in relieving physical fatigue.

[0034] 2. By using three-stage sequential fermentation of bacillus subtilis, monascus purpureus, and lactobacillus plantarum, the present application realizes the synergistic effect among the components and the co-synergistic effect among the fermentation substrates, significantly enhancing the effect of the composite plant fermentation broth in relieving physical fatigue. Detailed Embodiments

[0035] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0036] Example 1: A method for preparing a composite plant fermentation broth, comprising the following steps:

[0037] Step 1, prepare the fermentation substrate:

[0038] Step 1-1: Chop polygonatum sibiricum, millettia speciosa, wolfberry fruit, liquorice root, ginseng, pomegranate without peel, and mulberry leaf respectively, then mix them. The mass ratio of polygonatum sibiricum, millettia speciosa, wolfberry fruit, liquorice root, ginseng, pomegranate without peel, and mulberry leaf is 4:2:0.5:0.5:0.5:1:1. Then beat them into a slurry to obtain a slurry.

[0039] Step 1-2: Add water to adjust the water content of the slurry to 90%, and sterilize it under high pressure at 121 °C for 15 minutes to obtain a fermentation substrate.

[0040] Step 2, Bacillus subtilis fermentation: Inoculate Bacillus subtilis into the fermentation substrate. The inoculation amount of Bacillus subtilis is 0.3% of the weight of the fermentation substrate. Then, carry out aerobic fermentation at 40 °C for 20 hours to obtain a Bacillus subtilis fermentation product.

[0041] Step 3, Monascus fermentation: After sterilizing the Bacillus subtilis fermentation product again, inoculate Monascus. The inoculation amount of Monascus is 0.5% of the weight of the Bacillus subtilis fermentation product. Then, carry out aerobic fermentation at 28 °C for 5 days to obtain a Monascus fermentation product.

[0042] Step 4, Lactiplantibacillus plantarum fermentation: After sterilizing the Monascus fermentation product again, inoculate Lactiplantibacillus plantarum. The inoculation amount of Lactiplantibacillus plantarum is 2% of the weight of the Monascus fermentation product. Then, carry out anaerobic fermentation at 35 °C for 24 hours to obtain a compound fermentation product.

[0043] Step 5: Filter the compound fermentation product, take the filtrate and sterilize it to obtain a compound plant fermentation broth.

[0044] Among them, the Bacillus subtilis is Bacillus subtilis CICC 10023 purchased from the China Center for Industrial Culture Collection.

[0045] Among them, the Monascus is Monascus CICC 40712 purchased from the China Center for Industrial Culture Collection.

[0046] Among them, the Lactiplantibacillus plantarum is Lactiplantibacillus plantarum CICC 6009 purchased from the China Center for Industrial Culture Collection.

[0047] Example 2: A method for preparing a compound plant fermentation broth, comprising the following steps:

[0048] Step 1, prepare a fermentation substrate:

[0049] Step 1-1: Chop polygonatum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaves respectively, then mix them. The mass ratio of polygonatum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaves is 5:3:0.8:0.8:0.8:1.5:1.5. Then, make a slurry.

[0050] Step 1-2: Add water to adjust the water content of the slurry to 95%, and sterilize it under high pressure at 121 °C for 15 minutes to obtain a fermentation substrate.

[0051] Step 2, Bacillus subtilis fermentation: Inoculate Bacillus subtilis into the fermentation substrate. The inoculation amount of Bacillus subtilis is 0.4% of the weight of the fermentation substrate. Then, perform aerobic fermentation at 41 °C for 22 hours to obtain the Bacillus subtilis fermentation product.

[0052] Step 3, Monascus fermentation: After sterilizing the Bacillus subtilis fermentation product again, inoculate Monascus. The inoculation amount of Monascus is 0.7% of the weight of the Bacillus subtilis fermentation product. Then, perform aerobic fermentation at 30 °C for 5.5 days to obtain the Monascus fermentation product.

[0053] Step 4, Lactiplantibacillus plantarum fermentation: After sterilizing the Monascus fermentation product again, inoculate Lactiplantibacillus plantarum. The inoculation amount of Lactiplantibacillus plantarum is 2.5% of the weight of the Monascus fermentation product. Then, perform closed fermentation at 36 °C for 26 hours to obtain the composite fermentation product.

[0054] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation broth.

[0055] Example 3: A method for preparing a composite plant fermentation broth, comprising the following steps:

[0056] Step 1, Prepare the fermentation substrate:

[0057] Step 1-1: Chop polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel, and mulberry leaves respectively, then mix them. The mass ratio of polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel, and mulberry leaves is 6:4:1:1:1:2:2. Then, make a pulp to obtain the slurry.

[0058] Step 1-2: Add water to adjust the water content of the slurry to 100%, and perform high-pressure sterilization at 121 °C for 15 minutes to obtain the fermentation substrate.

[0059] Step 2, Bacillus subtilis fermentation: Inoculate Bacillus subtilis into the fermentation substrate. The inoculation amount of Bacillus subtilis is 0.5% of the weight of the fermentation substrate. Then, perform aerobic fermentation at 42 °C for 24 hours to obtain the Bacillus subtilis fermentation product.

[0060] Step 3, Monascus fermentation: After sterilizing the Bacillus subtilis fermentation product again, inoculate Monascus. The inoculation amount of Monascus is 1% of the weight of the Bacillus subtilis fermentation product. Then, perform aerobic fermentation at 32 °C for 6 days to obtain the Monascus fermentation product.

[0061] Step 4, Lactiplantibacillus plantarum fermentation: After sterilizing the Monascus fermentation product again, inoculate Lactiplantibacillus plantarum. The inoculation amount of Lactiplantibacillus plantarum is 3% of the weight of the Monascus fermentation product. Then, perform closed fermentation at 37 °C for 28 hours to obtain the composite fermentation product.

[0062] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0063] Comparative Example 1: A preparation method of a composite plant fermentation liquid, which is different from Example 1 in that the fermentation sequence of the strains in the fermentation stage is inconsistent, and includes the following steps:

[0064] Step 1, prepare the fermentation substrate:

[0065] Step 1-1: Chop polygonatum, millettia speciosa champ, wolfberry, licorice, ginseng, pomegranate without peel and mulberry leaves respectively, then mix them. The mass ratio of polygonatum, millettia speciosa champ, wolfberry, licorice, ginseng, pomegranate without peel and mulberry leaves is 4:2:0.5:0.5:0.5:1:1. Then beat them into a slurry.

[0066] Step 1-2: Adjust the water content of the slurry to 90% by adding water, and sterilize it at 121 °C for 15 minutes to obtain the fermentation substrate.

[0067] Step 2, ferment with Lactobacillus plantarum: Inoculate Lactobacillus plantarum into the fermentation substrate. The inoculation amount of Lactobacillus plantarum is 2% of the weight of the fermentation substrate. Then ferment it airtight at 35 °C for 24 hours to obtain the Lactobacillus plantarum fermentation product.

[0068] Step 3, ferment with Monascus purpureus Went: After sterilizing the Lactobacillus plantarum fermentation product again, inoculate Monascus purpureus Went. The inoculation amount of Monascus purpureus Went is 0.5% of the weight of the Lactobacillus plantarum fermentation product. Then ferment it aerobically at 28 °C for 5 days to obtain the Monascus purpureus Went fermentation product.

[0069] Step 4, ferment with Bacillus subtilis: After sterilizing the Monascus purpureus Went fermentation product again, inoculate Bacillus subtilis. The inoculation amount of Bacillus subtilis is 0.3% of the weight of the Monascus purpureus Went fermentation product. Then ferment it aerobically at 40 °C for 20 hours to obtain the composite fermentation product.

[0070] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0071] Comparative Example 2: A preparation method of a composite plant fermentation liquid, which is different from Example 1 in that the fermentation sequence of the strains in the fermentation stage is inconsistent, and includes the following steps:

[0072] Step 1, prepare the fermentation substrate:

[0073] Step 1-1: Chop polygonatum, millettia speciosa champ, wolfberry, licorice, ginseng, pomegranate without peel and mulberry leaves respectively, then mix them. The mass ratio of polygonatum, millettia speciosa champ, wolfberry, licorice, ginseng, pomegranate without peel and mulberry leaves is 4:2:0.5:0.5:0.5:1:1. Then beat them into a slurry.

[0074] Step 1-2: Adjust the water content of the slurry to 90% by adding water, and autoclave at 121 °C for 15 minutes to obtain a fermentation substrate.

[0075] Step 2, Monascus fermentation: Inoculate Monascus in the fermentation substrate, and the inoculation amount of Monascus is 0.5% of the weight of the fermentation substrate. Then, carry out aerobic fermentation at 28 °C for 5 days to obtain a Monascus fermentation product.

[0076] Step 3, Lactiplantibacillus plantarum fermentation: After sterilizing the Monascus fermentation product again, inoculate Lactiplantibacillus plantarum, and the inoculation amount of Lactiplantibacillus plantarum is 2% of the weight of the Monascus fermentation product. Then, carry out anaerobic fermentation at 35 °C for 24 hours to obtain a Lactiplantibacillus plantarum fermentation product.

[0077] Step 4, Bacillus subtilis fermentation: After sterilizing the Lactiplantibacillus plantarum fermentation product again, inoculate Bacillus subtilis, and the inoculation amount of Bacillus subtilis is 0.3% of the weight of the Lactiplantibacillus plantarum fermentation product. Then, carry out aerobic fermentation at 40 °C for 20 hours to obtain a compound fermentation product.

[0078] Step 5: Filter the compound fermentation product, take the filtrate and sterilize it to obtain a compound plant fermentation broth.

[0079] Comparative Example 3: A method for preparing a compound plant fermentation broth, which is different from Example 1 in that the fermentation sequence of the strains in the fermentation stage is inconsistent, and includes the following steps:

[0080] Step 1, Prepare a fermentation substrate:

[0081] Step 1-1: Chop Polygonatum sibiricum, Millettia speciosa Champ., Lycium barbarum, Glycyrrhiza uralensis Fisch., Panax ginseng, pomegranate without peel, and Morus alba L. respectively, then mix them. The mass ratio of Polygonatum sibiricum, Millettia speciosa Champ., Lycium barbarum, Glycyrrhiza uralensis Fisch., Panax ginseng, pomegranate without peel, and Morus alba L. is 4:2:0.5:0.5:0.5:1:1. Then, make a slurry to obtain a slurry.

[0082] Step 1-2: Adjust the water content of the slurry to 90% by adding water, and autoclave at 121 °C for 15 minutes to obtain a fermentation substrate.

[0083] Step 2, Monascus fermentation: Inoculate Monascus in the fermentation substrate, and the inoculation amount of Monascus is 0.5% of the weight of the fermentation substrate. Then, carry out aerobic fermentation at 28 °C for 5 days to obtain a Monascus fermentation product.

[0084] Step 3, Bacillus subtilis fermentation: After sterilizing the Monascus fermentation product again, inoculate Bacillus subtilis, and the inoculation amount of Bacillus subtilis is 0.3% of the weight of the Monascus fermentation product. Then, carry out aerobic fermentation at 40 °C for 20 hours to obtain a Bacillus subtilis fermentation product.

[0085] Step 4, fermentation by Lactiplantibacillus plantarum: After resterilizing the Bacillus subtilis fermentation product, inoculate Lactiplantibacillus plantarum with an inoculation amount of 2% of the weight of the Bacillus subtilis fermentation product, and then ferment in a sealed state at 35°C for 24 hours to obtain a composite fermentation product.

[0086] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain a composite plant fermentation broth.

[0087] Comparative Example 4: A method for preparing a composite plant fermentation broth, which is different from Example 1 in that the fermentation substrate does not contain pomegranates with the peel removed.

[0088] Comparative Example 5: A method for preparing a composite plant fermentation broth, which is different from Example 1 in that the fermentation substrate does not contain mulberry leaves.

[0089] Effect test for relieving physical fatigue:

[0090] Test samples: The composite plant fermentation broths of the above-mentioned examples and comparative examples (experimental groups) and distilled water (control group).

[0091] Detection method: Select 270 clean-grade male Kunming mice provided by the Henan Provincial Center for Experimental Animals, with a body weight of 18 - 22 g, and divide them into three major groups: a loaded swimming test group, a serum urea determination test group, and a liver glycogen determination test group, with 90 mice in each major group. Each major group is further evenly divided into 9 small groups, with 10 mice in each small group, and they are respectively administered the above-mentioned test samples. The dosage of the test sample is 0.12 mL / kg.bw. When used, it is diluted with water to 200 mL and administered by gavage once a day. After continuous use for 30 days, an effect test for relieving physical fatigue is carried out.

[0092] Loaded swimming test: 30 minutes after the last administration of the test substance to the mice, load a lead sheet with a weight of 5% of the body weight at the root of the mouse's tail, place it in a swimming tank to swim, with the water depth not less than 30 cm and the water temperature at 25°C ± 1.0°C. Record the time from the start of swimming to the death of the mouse as the mouse's loaded swimming time (seconds). If the swimming time of the experimental group is significantly longer than that of the control group and the difference is significant (P < 0.05), it can be determined that the experimental group has the effect of prolonging the loaded swimming time of the mice.

[0093] Serum urea determination test: 30 minutes after the last administration of the test substance to the mice, swim in water at 30°C without load for 90 min, rest, and immediately collect 0.5 mL of blood by eye socket puncture 60 minutes later. Let it stand at 4°C for 3 hours and then centrifuge at 2000 rpm for 15 minutes. Take the serum and measure the serum urea with an automatic biochemical analyzer. If the serum urea content of the experimental group is significantly lower than that of the control group and the difference is significant (P < 0.05), it can be determined that the test substance has the effect of reducing the urea production of fatigued mice.

[0094] Liver glycogen determination test: Anthrone method was used. 30 minutes after the last administration of the test substance to mice, the animals were killed, the liver was taken out, rinsed with physiological saline, and dried with filter paper. 100 mg of liver was weighed, and the remaining operations were carried out according to the instructions of the kit for liver glycogen determination. If the liver glycogen content of the test group was significantly higher than that of the control group, and the difference was significant (P<0.05), it can be determined that the test substance has the effect of promoting the liver glycogen reserve of mice.

[0095] The above test results are averaged and are shown in Table 1.

[0096] Table 1:

[0097] Specifically combining the above-mentioned embodiments and comparative examples and the data analysis in Table 1, it can be seen that the composite plant fermentation liquid of Embodiment 1, Embodiment 2 and Embodiment 3 showed significant effects in prolonging the weight-bearing swimming time of mice, reducing the urea production of fatigued mice and promoting the liver glycogen reserve of mice. Compared with the control group, the swimming time of the three embodiments was extended by 74.1 seconds, 78.6 seconds and 76.2 seconds, respectively, the urea content was reduced by 0.72 mmol / L, 0.85 mmol / L and 0.81 mmol / L, respectively, and the liver glycogen content was increased by 664.1 mg / 100 g, 755.8 mg / 100 g and 731.6 mg / 100 g, respectively, which shows that the composite plant fermentation liquid provided in the present application has a significant effect of relieving physical fatigue.

[0098] Combined with the analysis of Example 1 and Comparative Example 1, Comparative Example 2 and Comparative Example 3, due to the change in the order of bacterial fermentation, Example 1 has more advantages in relieving physical fatigue. This further illustrates the importance of the bacterial fermentation order in the present application. Only by adopting the bacterial fermentation order of the present application can the effect of the composite plant fermentation liquid in relieving physical fatigue be significantly improved.

[0099] In addition, further analysis based on Example 1 and Comparative Examples 4 and 5 shows that, since the pomegranate and mulberry leaves with peels removed were removed from the fermentation substrate, Example 1 has more advantages in relieving physical fatigue. This also proves the importance of the formula of the fermentation substrate in the present application for improving the effect of the composite plant fermentation liquid.

[0100] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A preparation method of a composite plant fermentation broth, characterized in that, It includes the following steps: Step 1, prepare the fermentation substrate: Mix polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaves and make them into pulp. After adjusting the water content of the pulp to 90% - 100%, sterilize it to obtain the fermentation substrate; Step 2, Bacillus subtilis fermentation: Inoculate Bacillus subtilis into the fermentation substrate and perform aerobic fermentation at 40 - 42 °C for 20 - 24 hours to obtain the Bacillus subtilis fermentation product; Step 3, Monascus fermentation: After sterilizing the Bacillus subtilis fermentation product again, inoculate Monascus and perform aerobic fermentation at 28 - 32 °C for 5 - 6 days to obtain the Monascus fermentation product; Step 4, Lactiplantibacillus plantarum fermentation: After sterilizing the Monascus fermentation product again, inoculate Lactiplantibacillus plantarum and perform closed fermentation at 35 - 37 °C for 24 - 28 hours to obtain the compound fermentation product; Step 5: Filter the compound fermentation product, take the filtrate and sterilize it to obtain the compound plant fermentation liquid; The mass ratio of polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaves is (4 - 6):(2 - 4):(0.5 - 1):(0.5 - 1):(0.5 - 1):(1 - 2):(1 - 2); In Step 2, the inoculation amount of Bacillus subtilis is 0.3% - 0.5% of the weight of the fermentation substrate; In Step 3, the inoculation amount of Monascus is 0.5% - 1% of the weight of the Bacillus subtilis fermentation product; In Step 4, the inoculation amount of Lactiplantibacillus plantarum is 2% - 3% of the weight of the Monascus fermentation product.

2. The preparation method of a composite plant fermentation broth according to claim 1, characterized in that, Step 1 is specifically as follows: Step 1 - 1: Chop polygonatum sibiricum, millettia speciosa champ, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaves respectively, then mix and make them into pulp to obtain the pulp; Step 1 - 2: Add water to adjust the water content of the pulp to 90% - 100% and sterilize it to obtain the fermentation substrate.

3. A composite plant fermentation broth, characterized in that, It is prepared by using the preparation method of the compound plant fermentation liquid described in Claim 1 or 2.

4. Use of the compound plant fermentation liquid described in Claim 3 in the preparation of anti - fatigue drugs.

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