Composite plant fermentation liquor as well as preparation method and application thereof

Abstract

The invention relates to the technical field of plant fermentation, and particularly discloses composite plant fermentation liquor as well as a preparation method and application thereof. The preparation method of the composite plant fermentation liquor comprises the following steps: step 1, mixing rhizoma polygonati, beautiful millettia roots, Chinese wolfberry fruits, licorice roots, ginseng, pomegranate without peels and mulberry leaves to obtain a fermentation substrate; step 2, bacillus subtillis fermentation; step 3, monascus fermentation; step 4, fermenting the plant milk with plant bacilli; and step 5, filtering and sterilizing to obtain the composite plant fermentation liquor.

Description

Technical Field

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

[0002] Existing fatigue-relieving products have some limitations. Although chemically synthesized anti-fatigue products have the effect of quickly relieving fatigue, long-term use may cause liver and kidney damage or other health problems, while chemically extracted anti-fatigue products may cause the loss of effective ingredients due to improper extraction process, thus affecting the effect. In addition, although single-fermentation fatigue-relieving products provide certain relief effects, due to the lack of diversified microbial effects, the types of active ingredients they produce are limited, making it difficult to achieve a good relief effect.

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

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

[0005] In the first aspect, the present application provides a method for preparing a composite plant fermentation liquid using the following technical scheme: A method for preparing a composite plant fermentation liquid comprises the following steps: Step 1, preparing a fermentation substrate: mixing and slurrying polygonatum, smilax glabra, wolfberry, liquorice, ginseng, pomegranates with peels removed, and mulberry leaves, adjusting the water content of the slurry to 90% to 100%, and sterilizing to obtain a fermentation substrate; Step 2, Bacillus subtilis fermentation: inoculating Bacillus subtilis in a fermentation substrate, and aerobically fermenting at 40-42° C. for 20-24 hours to obtain a Bacillus subtilis fermentation product; Step 3, Monascus fermentation: sterilize the Bacillus subtilis fermentation product again, inoculate Monascus, and ferment aerobically at 28-32° C. for 5-6 days to obtain a Monascus fermentation product; Step 4, fermentation with Bacillus plantarum: sterilize the Monascus fermentation product again, inoculate Bacillus plantarum, and perform closed fermentation at 35-37° C. for 24-28 hours to obtain a composite fermentation product; Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0006] In the above technical scheme, Polygonatum sibiricum is rich in polysaccharides, amino acids and trace elements, Rhizoma Dioscoreae can nourish the lungs and strengthen the tendons, resist oxidation and inflammation, and improve breathing and tendon problems, Lycium barbarum has the effects of enhancing immunity, promoting hematopoietic function, lowering blood lipids, anti-tumor and anti-aging; Licorice can nourish the spleen and replenish qi, eliminate phlegm and relieve cough, relieve pain and inflammation, clear away heat and detoxify, ginseng contains rare saponin Rg3, which has the effects of replenishing qi and blood, calming the nerves and improving intelligence, and the pomegranate and mulberry leaves without the peel are rich in antioxidants and trace elements, which help to improve the nutritional value and biological activity of the overall fermentation liquid.

[0007] This application uses polygonatum, oxalis, wolfberry, licorice, ginseng, pomegranates with peels removed, and mulberry leaves as fermentation substrates, and uses Bacillus subtilis, Monascus, and plant lactobacillus to perform three-stage orderly fermentation to achieve synergistic effects among the ingredients. First, polygonatum provides rich polysaccharides, ginseng contains a variety of saponin compounds, peeled pomegranates are rich in antioxidants such as anthocyanins and ellagic acid, and 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 the ingredients. Secondly, during the fermentation stage, the present application first uses Bacillus subtilis to decompose complex macromolecules to provide a nutritional basis for subsequent fermentation, and then uses Monascus to produce antioxidant metabolites during the fermentation process to further enhance the activity of the fermentation liquid. Finally, Bacillus plantarum increases the lactic acid content to promote intestinal health and inhibit the growth of harmful microorganisms. Through this multi-level microbial fermentation, the present application not only optimizes the fermentation matrix, but also unexpectedly generates a fermentation liquid rich in small molecule nutrients and a variety of bioactive ingredients, achieving an unexpected and effective effect in relieving physical fatigue.

[0008] Preferably, the mass ratio of polygonatum, smilax china, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaf is (4-6): (2-4): (0.5-1): (0.5-1): (0.5-1): (1-2): (1-2).

[0009] In the above technical scheme, the present application improves the bioavailability of these raw materials by controlling the dosage ratio of these raw materials, ensuring that each raw material can exert its optimal efficiency during the fermentation process, thereby promoting a synergistic effect between them, ensuring that there are sufficient nutrients for microbial metabolism during the fermentation process, and being able to generate small molecule active ingredients that are easier for the human body to absorb, thereby enhancing the effect of the composite plant fermentation liquid in relieving physical fatigue.

[0010] Preferably, in step 2, the inoculation amount of Bacillus subtilis is 0.3% to 0.5% of the weight of the fermentation substrate.

[0011] In the above technical scheme, the present application ensures that a sufficient number of live bacteria participate in metabolic activities in the early stage of fermentation by inoculating Bacillus subtilis in the fermentation substrate at a dosage ratio of 0.3% to 0.5%, and helps to optimize the fermentation environment, so that Bacillus subtilis can effectively decompose the macromolecules in the substrate and convert them into small molecule nutrients that are easy for subsequent microorganisms to utilize, thereby laying a good foundation for the subsequent fermentation steps.

[0012] Preferably, in step 3, the inoculation amount of Monascus is 0.5% to 1% of the weight of the Bacillus subtilis fermentation product.

[0013] In the above technical scheme, the present application sets the inoculation amount of Monascus at 0.5%~1%, which can ensure that there are sufficient Monascus spores participating in the fermentation process and quickly adapting to and utilizing the nutritional environment produced by the fermentation of Bacillus subtilis, thereby promoting the growth and reproduction of Monascus and the accumulation of metabolites.

[0014] Preferably, in step 4, the inoculation amount of Bacillus plantarum is 2% to 3% of the weight of the Monascus fermentation product.

[0015] In the above technical scheme, the present application selects a 2% to 3% inoculation amount of Bacillus plantarum, which can not only ensure that there is a sufficient number of Bacillus plantarum to achieve effective metabolic activities, but also better improve the quality characteristics of the composite plant fermentation liquid, and further enhance the effect of the composite plant fermentation liquid in relieving physical fatigue.

[0016] Preferably, step 1 is as follows: Step 1-1: chop polygonatum, smilax china, wolfberry, liquorice, ginseng, pomegranate and mulberry leaves from which the peel is removed, respectively, and then mix and beat to obtain a slurry; Step 1-2: Add water to adjust the water content of the slurry to 90%~100%, sterilize, and obtain the fermentation substrate.

[0017] In the above technical scheme, the present application improves the nutritional value and functionality by chopping and pulping Polygonatum sibiricum, Glechoma longituba, peeled pomegranate and mulberry leaves to increase the uniform dispersion and enzymatic hydrolysis efficiency among the raw materials; finally, the water content of the pulp is adjusted to 90%~100% and sterilized 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.

[0018] In the second aspect, the present application provides a composite plant fermentation liquid using the following technical solution: A composite plant fermentation liquid is prepared by the preparation method of the composite plant fermentation liquid as described in the first aspect.

[0019] In the above technical scheme, the composite plant fermentation liquid of the present application realizes the synergistic effect of multiple plant components and microorganisms through the above-mentioned specific preparation process, thereby generating a composite plant fermentation liquid rich in small molecule nutrients and multiple biologically active ingredients. Such a composite plant fermentation liquid has a significant effect of relieving physical fatigue and has broad application prospects in the food or biopharmaceutical industry.

[0020] In the third aspect, the application of a composite plant fermentation liquid provided in the present application adopts the following technical solution: An application of a composite plant fermentation liquid, wherein the composite plant fermentation liquid is used in the food or biopharmaceutical industry.

[0021] In the above technical scheme, the composite plant fermentation liquid of the present application can not only effectively relieve physical fatigue, but also show wide application potential in the food or biopharmaceutical industry due to its natural ingredients and the characteristics of microbial fermentation.

[0022] In summary, the present application includes at least one of the following beneficial technical effects: 1. This application achieves a synergistic effect among the raw materials by combining several specific plant raw materials, including polygonatum, smilax china, wolfberry, licorice, ginseng, pomegranate with peel removed, and mulberry leaves, with microbial fermentation, thereby improving the bioavailability of these raw materials, optimizing the fermentation substrate, and ensuring that each raw material can exert its optimal efficiency during the fermentation process, thereby enhancing the effect of the composite plant fermentation liquid in relieving physical fatigue.

[0023] 2. The present application realizes the synergistic effect between the components and the co-synergistic effect between the fermentation substrates by utilizing three-stage orderly fermentation of Bacillus subtilis, Monascus purpureus and Bacillus plantarum, thereby significantly enhancing the effect of the composite plant fermentation liquid in relieving physical fatigue. DETAILED DESCRIPTION

[0024] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0025] Embodiment 1: A method for preparing a composite plant fermentation liquid comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 4:2:0.5:0.5:0.5:1:1, and then beat them to obtain slurry.

[0026] Step 1-2: Add water to adjust the water content of the slurry to 90%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0027] Step 2, Bacillus subtilis fermentation: Bacillus subtilis is inoculated into the fermentation substrate, the inoculation amount of Bacillus subtilis is 0.3% of the weight of the fermentation substrate, followed by aerobic fermentation at 40° C. for 20 hours to obtain a Bacillus subtilis fermentation product.

[0028] Step 3, Monascus fermentation: After the Bacillus subtilis fermentation product is sterilized again, Monascus is inoculated, and the inoculation amount of Monascus is 0.5% of the weight of the Bacillus subtilis fermentation product, followed by aerobic fermentation at 28° C. for 5 days to obtain the Monascus fermentation product.

[0029] Step 4, plant lactic acid Bacillus fermentation: after the Monascus fermentation product is sterilized again, plant lactic acid Bacillus is inoculated, and the inoculation amount of plant lactic acid Bacillus is 2% of the weight of the Monascus fermentation product, followed by closed fermentation at 35°C for 24 hours to obtain a composite fermentation product.

[0030] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0031] Among them, Bacillus subtilis was purchased from the China Industrial Microbiological Culture Collection Center (CICC 10023 Bacillus subtilis).

[0032] Among them, Monascus purpureus was purchased from the China Industrial Microbiological Culture Collection Center and is CICC 40712 Monascus purpureus.

[0033] Among them, Bacillus plantarum was purchased from the China Industrial Microbiology Culture Collection Center as CICC 6009 Bacillus plantarum.

[0034] Embodiment 2: A method for preparing a composite plant fermentation liquid comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 5:3:0.8:0.8:0.8:1.5:1.5, and then beat them to obtain slurry.

[0035] Step 1-2: Add water to adjust the water content of the slurry to 95%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0036] Step 2, Bacillus subtilis fermentation: Bacillus subtilis is inoculated into the fermentation substrate, the inoculation amount of Bacillus subtilis is 0.4% of the weight of the fermentation substrate, followed by aerobic fermentation at 41° C. for 22 hours to obtain a Bacillus subtilis fermentation product.

[0037] Step 3, Monascus fermentation: After the Bacillus subtilis fermentation product is sterilized again, Monascus is inoculated, and the inoculation amount of Monascus is 0.7% of the weight of the Bacillus subtilis fermentation product, followed by aerobic fermentation at 30° C. for 5.5 days to obtain the Monascus fermentation product.

[0038] Step 4, plant lactic acid Bacillus fermentation: after the Monascus fermentation product is sterilized again, plant lactic acid Bacillus is inoculated, and the inoculation amount of plant lactic acid Bacillus is 2.5% of the weight of the Monascus fermentation product, followed by closed fermentation at 36°C for 26 hours to obtain a composite fermentation product.

[0039] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0040] Embodiment 3: A method for preparing a composite plant fermentation liquid comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 6:4:1:1:1:2:2, and then beat them to obtain slurry.

[0041] Step 1-2: Add water to adjust the water content of the slurry to 100%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0042] Step 2, Bacillus subtilis fermentation: inoculate Bacillus subtilis in the fermentation substrate, the inoculation amount of Bacillus subtilis is 0.5% of the weight of the fermentation substrate, and then aerobically ferment at 42° C. for 24 hours to obtain a Bacillus subtilis fermentation product.

[0043] Step 3, Monascus fermentation: After the Bacillus subtilis fermentation product is sterilized again, Monascus is inoculated, and the inoculation amount of Monascus is 1% of the weight of the Bacillus subtilis fermentation product, followed by aerobic fermentation at 32° C. for 6 days to obtain the Monascus fermentation product.

[0044] Step 4, plant lactic acid Bacillus fermentation: after the Monascus fermentation product is sterilized again, plant lactic acid Bacillus is inoculated, and the inoculation amount of plant lactic acid Bacillus is 3% of the weight of the Monascus fermentation product, followed by closed fermentation at 37°C for 28 hours to obtain a composite fermentation product.

[0045] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0046] Comparative Example 1: A method for preparing a composite plant fermentation liquid, which is different from Example 1 in that the fermentation order of the strains in the fermentation stage is inconsistent, and comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 4:2:0.5:0.5:0.5:1:1, and then beat them to obtain slurry.

[0047] Step 1-2: Add water to adjust the water content of the slurry to 90%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0048] Step 2, fermentation with Bacillus plantarum: inoculating Bacillus plantarum in a fermentation substrate, wherein the inoculation amount of Bacillus plantarum is 2% of the weight of the fermentation substrate, followed by sealed fermentation at 35° C. for 24 hours to obtain a fermentation product of Bacillus plantarum.

[0049] Step 3, Monascus fermentation: sterilize the plant lactobacillus fermentation product again, and then inoculate Monascus, the inoculation amount of Monascus is 0.5% of the weight of the plant lactobacillus fermentation product, and then aerobically ferment at 28° C. for 5 days to obtain the Monascus fermentation product.

[0050] Step 4, Bacillus subtilis fermentation: After the Monascus fermentation product is sterilized again, Bacillus subtilis is inoculated, and the inoculation amount of Bacillus subtilis is 0.3% of the weight of the Monascus fermentation product, followed by aerobic fermentation at 40° C. for 20 hours to obtain a composite fermentation product.

[0051] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0052] Comparative Example 2: A method for preparing a composite plant fermentation liquid, which is different from Example 1 in that the fermentation order of the strains in the fermentation stage is inconsistent, and comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 4:2:0.5:0.5:0.5:1:1, and then beat them to obtain slurry.

[0053] Step 1-2: Add water to adjust the water content of the slurry to 90%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0054] Step 2, Monascus fermentation: inoculating Monascus in the fermentation substrate, the inoculation amount of Monascus being 0.5% of the weight of the fermentation substrate, followed by aerobic fermentation at 28° C. for 5 days to obtain a Monascus fermentation product.

[0055] Step 3, plant lactobacillus fermentation: sterilize the Monascus fermentation product again, and then inoculate plant lactobacillus with the plant lactobacillus at an inoculation amount of 2% of the weight of the Monascus fermentation product, followed by sealed fermentation at 35° C. for 24 hours to obtain the plant lactobacillus fermentation product.

[0056] Step 4, Bacillus subtilis fermentation: sterilize the plant lactobacillus fermentation product again, inoculate Bacillus subtilis, the inoculation amount of Bacillus subtilis is 0.3% of the weight of the plant lactobacillus fermentation product, then aerobically ferment at 40° C. for 20 hours to obtain a composite fermentation product.

[0057] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0058] Comparative Example 3: A method for preparing a composite plant fermentation liquid, which is different from Example 1 in that the fermentation order of the strains in the fermentation stage is inconsistent, and comprises the following steps: Step 1, prepare the fermentation substrate: Step 1-1: chop polygonatum, schizonepeta tenuifolia, wolfberry, licorice, ginseng, pomegranate with peel removed and mulberry leaves respectively, and then mix them in a mass ratio of 4:2:0.5:0.5:0.5:1:1, and then beat them to obtain slurry.

[0059] Step 1-2: Add water to adjust the water content of the slurry to 90%, and sterilize it by high pressure at 121° C. for 15 minutes to obtain a fermentation substrate.

[0060] Step 2, Monascus fermentation: inoculating Monascus in the fermentation substrate, the inoculation amount of Monascus being 0.5% of the weight of the fermentation substrate, followed by aerobic fermentation at 28° C. for 5 days to obtain a Monascus fermentation product.

[0061] Step 3, Bacillus subtilis fermentation: After the Monascus fermentation product is sterilized again, Bacillus subtilis is inoculated, and the inoculation amount of Bacillus subtilis is 0.3% of the weight of the Monascus fermentation product, followed by aerobic fermentation at 40° C. for 20 hours to obtain the Bacillus subtilis fermentation product.

[0062] Step 4, fermentation with Bacillus plantarum: sterilize the Bacillus subtilis fermentation product again, and then inoculate with Bacillus plantarum. The inoculation amount of Bacillus plantarum is 2% of the weight of the Bacillus subtilis fermentation product. Then, the product is sealed and fermented at 35° C. for 24 hours to obtain a composite fermentation product.

[0063] Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

[0064] Comparative Example 4: A method for preparing a composite plant fermentation liquid, which is different from Example 1 in that the fermentation substrate does not contain pomegranates with peels removed.

[0065] Comparative Example 5: A method for preparing a composite plant fermentation liquid, which is different from Example 1 in that the fermentation substrate does not contain mulberry leaves.

[0066] Experiment on the effect of relieving physical fatigue: Test samples: the composite plant fermentation liquid (test group) and distilled water (control group) of the above embodiments and comparative examples.

[0067] Detection method: 270 clean-grade male Kunming mice provided by the Henan Provincial Experimental Animal Center were selected, weighing 18-22g, and divided into three groups: weight-bearing swimming test group, serum urea determination test group, and liver glycogen determination test group, with 90 mice in each group. Each group was divided into 9 small groups, with 10 mice in each group, and the above test samples were used respectively. The dosage of the test sample was 0.12mL / kg.bw, and it was diluted with water to 200mL when used, and gavage was performed once a day. After continuous use for 30 days, the effect of relieving physical fatigue was tested.

[0068] Weighted swimming test: 30 minutes after the last administration of the test substance to the mice, a lead sheet of 5% of the body weight was placed on the base of the mouse's tail and placed in a swimming box to swim. The water depth was not less than 30 cm and the water temperature was 25℃±1.0℃. The time from the start of swimming to the death of the mouse was recorded as the weighted swimming time of the mouse (seconds). If the swimming time of the experimental group was significantly longer than that of the control group, and the difference was significant (P<0.05), it can be determined that the experimental group has the effect of prolonging the weighted swimming time of the mice.

[0069] Serum urea determination test: 30 minutes after the last administration of the test substance to mice, swim without weight in water at 30°C for 90 minutes, rest, and immediately extract eyeballs and collect 0.5mL of blood 60 minutes later. After standing at 4°C for 3 hours, centrifuge at 2000rpm for 15 minutes, and take serum to determine serum urea using an automatic biochemical analyzer. If the serum urea content of the test 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 urea production in fatigue mice.

[0070] 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.

[0071] The above test results are averaged and are shown in Table 1.

[0072] Table 1:

[0073] 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.

[0074] 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.

[0075] 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.

[0076] 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 method for preparing a composite plant fermentation liquid, characterized in that: The following steps are involved: Step 1, preparing a fermentation substrate: mixing and slurrying polygonatum, smilax glabra, wolfberry, liquorice, ginseng, pomegranates with peels removed, and mulberry leaves, adjusting the water content of the slurry to 90% to 100%, and sterilizing to obtain a fermentation substrate; Step 2, Bacillus subtilis fermentation: inoculating Bacillus subtilis in a fermentation substrate, and aerobically fermenting at 40-42° C. for 20-24 hours to obtain a Bacillus subtilis fermentation product; Step 3, Monascus fermentation: sterilize the Bacillus subtilis fermentation product again, inoculate Monascus, and ferment aerobically at 28-32° C. for 5-6 days to obtain a Monascus fermentation product; Step 4, fermentation with Bacillus plantarum: sterilize the Monascus fermentation product again, inoculate Bacillus plantarum, and perform closed fermentation at 35-37° C. for 24-28 hours to obtain a composite fermentation product; Step 5: Filter the composite fermentation product, take the filtrate and sterilize it to obtain the composite plant fermentation liquid.

2. The method for preparing a composite plant fermentation liquid according to claim 1, characterized in that: The mass ratio of polygonatum, smilax china, wolfberry, liquorice, ginseng, pomegranate without peel and mulberry leaf is (4-6): (2-4): (0.5-1): (0.5-1): (0.5-1): (1-2): (1-2).

3. The method for preparing a composite plant fermentation liquid according to claim 1, characterized in that: In step 2, the inoculation amount of Bacillus subtilis is 0.3% to 0.5% of the weight of the fermentation substrate.

4. The method for preparing a composite plant fermentation liquid according to claim 1, characterized in that: In step 3, the inoculation amount of Monascus is 0.5% to 1% of the weight of the fermentation product of Bacillus subtilis.

5. The method for preparing a composite plant fermentation liquid according to claim 1, characterized in that: In step 4, the inoculation amount of Bacillus plantarum is 2% to 3% of the weight of the Monascus fermentation product.

6. The method for preparing a composite plant fermentation liquid according to claim 1, characterized in that: Step 1 is as follows: Step 1-1: chop polygonatum, smilax china, wolfberry, liquorice, ginseng, pomegranate and mulberry leaves from which the peel is removed, respectively, and then mix and beat to obtain a slurry; Step 1-2: Add water to adjust the water content of the slurry to 90%~100%, sterilize, and obtain the fermentation substrate.

7. A composite plant fermentation liquid, characterized in that: The composite plant fermentation liquid is prepared by the preparation method of the composite plant fermentation liquid according to any one of claims 1 to 6.

8. Use of the composite plant fermentation liquid as claimed in claim 7 in preparing an anti-fatigue composition.

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