A probiotic ferment powder capable of clearing the intestines and protecting the liver and its preparation method

Abstract

The present invention provides a probiotic ferment powder capable of clearing the intestines and protecting the liver, as well as a preparation method thereof, which relates to the field of microbial fermentation applications and aims to solve the problems of gastrointestinal diseases and liver damage caused by factors such as irregular eating habits, increased life stress, and frequent alcohol consumption in modern life. By combining traditional Chinese medicinal materials and modern microbial fermentation technology, the present invention prepares a probiotic ferment with the effects of clearing intestinal toxins, protecting the liver and kidneys, nourishing the stomach and spleen, and replenishing qi. The specific preparation method includes mixing medicated and edible homologous materials such as kudzu root, hawthorn, white hyacinth bean, poria cocos, and lotus leaf with defatted soybean powder, red date powder, corn starch, and water, obtaining a fermentation medium through high-temperature sterilization, then inoculating Kluyveromyces marxianus K11, Weissella coagulans JS518, and Lactobacillus paracasei JS12 for alternating fermentation, and finally adding a microbial protectant and a stabilizer, mixing evenly and freeze-drying to obtain the product.

Description

Technical Field

[0001] The present invention relates to the field of microbial fermentation, and particularly to a probiotic ferment powder capable of clearing the intestine and protecting the liver and a preparation method thereof. Background Art

[0002] Traditional Chinese medicine has a long history in regulating the intestine and protecting the liver. However, simply relying on traditional Chinese medicine ingredients often fails to achieve the ideal effect, and the decoction of traditional Chinese medicine is time-consuming and laborious. The decoction of traditional Chinese medicine is not conducive to preservation and has a poor taste. Modern microbial fermentation technology provides a new idea for improving the efficacy of traditional Chinese medicine. Through probiotic fermentation, the biological activity and absorption efficiency of traditional Chinese medicine ingredients can be enhanced. Probiotics can not only improve the balance of intestinal flora, but also help enhance immune function and promote digestion. Combining traditional Chinese medicine materials with probiotic fermentation technology is expected to develop a new product for clearing the intestine and protecting the liver. However, the following key problems need to be solved: how to optimize the growth and metabolic activities of probiotics, improve the content and bioavailability of active ingredients in the ferment; how to effectively protect probiotics from external environmental interference and extend the uniformity and stability of the product. Summary of the Invention

[0003] To achieve the above object, the present invention is realized through the following technical solutions: A preparation method of a probiotic ferment powder capable of clearing the intestine and protecting the liver, comprising the following steps:

[0004] S1. Weigh Pueraria lobata, Crataegus pinnatifida, Dolichos lablab, Poria cocos and Lotus leaf, mix Pueraria lobata, Crataegus pinnatifida, Dolichos lablab, Poria cocos and Lotus leaf evenly according to a ratio, pulverize and then screen through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soybean powder, red date powder, corn starch and water to the traditional Chinese medicine powder according to a ratio, mix evenly, and then perform high-temperature sterilization to obtain a fermentation medium;

[0005] S2: Inoculate Kluyveromyces marxianus K11 into the fermentation medium for aerobic fermentation, control the fermentation temperature at 26°C to 40°C and ferment for 1 to 2 hours; inoculate Weizmannia coagulans JS518 into the fermentation medium for microaerobic fermentation, control the fermentation temperature at 30°C to 37°C and ferment for 2 to 4 hours; inoculate Lactobacillus paracasei JS12 into the medium for microaerobic fermentation, control the fermentation temperature at 35°C to 40°C and the fermentation time at 2 to 3 hours; after all the strains are inoculated, perform alternating fermentation according to the cycle of aerobic fermentation for 1 hour, microaerobic fermentation for 3 hours, and anaerobic fermentation for 1 hour, and cycle 4 times to obtain a probiotic ferment;

[0006] S3: The probiotic fermentation product prepared in S2 is crushed to ensure that the microbial protective agent can be mixed more evenly, and the crushed probiotic fermentation product is preliminarily mixed with the pre-configured microbial protective agent. These ingredients have antioxidant and moisturizing effects, which can effectively protect the probiotics from interference from the external environment and extend the shelf life of the product; an emulsifier is added to the mixture, and a high-speed mixer is used for high-speed stirring. The stirring speed is 1200-2000rpm, and the stirring time is 10-15min. The emulsifier, the microbial protective agent and the water in the probiotic fermentation product form a microemulsified solution to wrap the probiotic fermentation product particles; then a stabilizer is added and a low-speed mixer is used for sufficient stirring. The stirring speed is 500-800rpm, and the stirring time is 20-30min. The stabilizer can form a gel network structure to prevent the probiotic fermentation product particles from aggregating and precipitating during the freeze-drying process, thereby improving the uniformity of the product. Finally, freeze-drying is performed to obtain a probiotic fermentation product powder that can cleanse the intestines and protect the liver.

[0007] The mass proportions of the kudzu root, hawthorn, white hyacinth bean, poria, lotus leaf, defatted soy flour, red date powder, corn starch, water, microbial protective agent, emulsifier and stabilizer are 20-25 parts of kudzu root, 16-18 parts of hawthorn, 15-18 parts of white hyacinth bean, 5-10 parts of poria, 20-30 parts of lotus leaf, 20-30 parts of defatted soy flour, 15-26 parts of red date powder, 15-24 parts of corn starch, 90-150 parts of water, 20-40 parts of microbial protective agent, 5-8 parts of emulsifier and 18-35 parts of stabilizer.

[0008] The traditional Chinese medicine pharmacology of Pueraria root, hawthorn, white hyacinth bean, Poria cocos and lotus leaf is as follows:

[0009] Pueraria root: According to Compendium of Materia Medica, Pueraria root is cool in nature, flat in qi, sweet in taste, and has the effects of clearing away heat, reducing internal heat, and detoxifying. Modern medical research shows that puerarin, an isoflavone compound in Pueraria root, has a certain effect on hypertension, hyperlipidemia, hyperglycemia, and cardiovascular and cerebrovascular diseases.

[0010] Hawthorn: It tastes sour, sweet, and slightly warm in nature. It enters the spleen, stomach, and liver meridians. It has the effects of eliminating food accumulation and removing stagnation and blood stasis. It is mainly used to treat food stagnation, abdominal distension and pain, diarrhea, dysmenorrhea due to blood stasis, amenorrhea, postpartum abdominal pain, lochia retention, hernia or testicular swelling and pain, and hyperlipidemia.

[0011] White lentil: According to Compendium of Materia Medica, white lentil can nourish the five internal organs and treat vomiting. Long-term use can prevent hair from turning gray, relieve wind and treat leucorrhea in women. It can detoxify alcohol, dolphin fish, and all kinds of plant poisons. It can stop diarrhea, relieve heat, warm the spleen and stomach, remove dampness and heat, and stop thirst.

[0012] Poria: It has a light and penetrating smell, its nature is upward, it produces body fluids, opens the pores, nourishes the water source and descends, and promotes urination.

[0013] Lotus leaf: "Compendium of Materia Medica in Yunnan": It clears the wind-heat in the upper head, stops dizziness, clears phlegm, relieves qi, stops vomiting, and relieves headache and stuffiness. Since ancient times in China, lotus leaves have been regarded as excellent medicines for slimming. Its chemical components mainly include nuciferine, citric acid, malic acid, gluconic acid, oxalic acid, succinic acid and other alkaline components with anti-mitotic effects. The nuciferine in lotus leaf tea contains a variety of effective lipid-lowering alkaloids, which can effectively decompose the fat in the body and strongly excrete it out of the body. Nuciferine can strongly cover the intestinal wall of the human body, forming a layer of fat isolation membrane to prevent fat absorption and prevent fat accumulation.

[0014] The Kluyveromyces marxianus K11, Weissella coagulans JS518 and Lactobacillus paracasei JS12 are respectively inoculated into the fermentation medium after activation. The inoculation amount of Kluyveromyces marxianus K11 is 10 7 ~10 9 CFU / g of the fermentation medium, the inoculation amount of Weissella coagulans JS518 is 10 7 ~10 9 CFU / g of the fermentation medium, the inoculation amount of Lactobacillus paracasei JS12 is 10 7 ~10 9 CFU / g of the fermentation medium.

[0015] The oxygen input amount for the aerobic fermentation is 0.5 - 1 vvm, and the aerobic fermentation temperature is controlled at 26°C - 40°C; the oxygen input amount for the micro-aerobic fermentation is 0.02 - 0.1 vvm, and the aerobic fermentation temperature is controlled at 30°C - 37°C; no oxygen is input for the anaerobic fermentation, and the anaerobic fermentation temperature is controlled at 35°C - 40°C; for example, the oxygen input amount for the aerobic fermentation set at 1 vvm refers to the ventilation volume / fermentation tank volume / minute. For a 50L fermentation tank, the ventilation volume per minute is 50L.

[0016] The microbial protectant is composed of 6 - 10 parts by mass of trehalose, 12 - 25 parts of glycerol, and 3 - 5 parts of whey protein.

[0017] The emulsifier is lecithin or polysorbate.

[0018] The stabilizer is composed of 3 - 8 parts by mass of cyclodextrin, 6 - 11 parts of gelatin, and 9 - 18 parts of xanthan gum.

[0019] The present invention provides a probiotic ferment powder capable of clearing the intestines and protecting the liver and its preparation method, which has the following beneficial effects:

[0020] 1. Alternating fermentation is carried out using Kluyveromyces marxianus K11, Weissella coagulans JS518 and Lactobacillus paracasei JS12. This unique fermentation cycle helps to improve the content and bioavailability of active ingredients in the ferment.

[0021] 2. The probiotic ferment powder for intestinal cleansing and liver protection prepared by the present invention forms a microemulsion solution by adding a microbial protectant and an emulsifier to wrap the probiotic ferment particles, effectively protecting the probiotics from external environmental interference, extending the shelf life of the product, and improving the uniformity and stability of the product. Detailed implementation manners

[0022] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0023] Kluyveromyces marxianus ( Kluyveromyces marxianus ) K11 in the following embodiments has a deposit number of CGMCC No. 26477.

[0024] Lactobacillus paracasei ( Lacticaseibacillus paracasei ) JS12 in the following embodiments has a deposit number of CGMCC No. 27469.

[0025] Weizmannia coagulans ( Weizmannia coagulans ) JS518 in the following embodiments has a deposit number of CGMCC No. 27471.

[0026] Example 1:

[0027] A preparation method of a probiotic ferment powder for intestinal cleansing and liver protection includes the following steps:

[0028] S1: Weigh 24 parts of kudzu root, 17 parts of hawthorn, 17 parts of white hyacinth bean, 7 parts of poria cocos, 25 parts of lotus leaf, 25 parts of defatted soy flour, 21 parts of red date powder, 20 parts of corn starch and 120 parts of water. Mix the kudzu root, hawthorn, white hyacinth bean, poria cocos and lotus leaf evenly according to the proportion, crush them and sieve through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soy flour, red date powder, corn starch and water to the traditional Chinese medicine powder according to the proportion, mix evenly, and perform high-temperature sterilization to obtain a fermentation medium;

[0029] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g of the fermentation medium, and perform aerobic fermentation at 26°C to 40°C for 1.5 hours, providing an oxygen inlet volume of 0.75 vvm; inoculate Weizmannia coagulans JS518, and the inoculation amount of Weizmannia coagulans JS518 is 1.2 × 10 8CFU / g of the fermentation medium, carry out microaerobic fermentation at 30°C to 37°C for 2.8 hours, and provide an oxygen inlet volume of 0.1 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.2×10 8 CFU / g of the fermentation medium, carry out microaerobic fermentation at 35°C to 40°C for 2.2 hours, and provide an oxygen inlet volume of 0.1 vvm; after inoculating all the strains, carry out aerobic fermentation for 1 hour with an oxygen inlet volume of 1 vvm; carry out microaerobic fermentation for 3 hours with an oxygen inlet volume of 0.04 vvm, and carry out alternating fermentation according to the cycle of anaerobic fermentation for 1 hour, and cycle 4 times to obtain the probiotic fermentate;

[0030] S3: Crush the probiotic fermentate prepared in S2, and preliminarily mix the crushed probiotic fermentate with a microbial protectant composed of 10 parts of trehalose, 12 parts of glycerol, and 3 parts of whey protein, with a total weight of 25 parts. Add 5 parts of an emulsifier to the mixture and use a high-speed blender for high-speed stirring, with a stirring speed of 1200 - 2000 rpm and a stirring time of 10 - 15 min; then add a stabilizer composed of 4 parts of cyclodextrin, 9 parts of gelatin, and 13 parts of xanthan gum, with a total weight of 26 parts, and use a low-speed blender for sufficient stirring, with a stirring speed of 500 - 800 rpm and a stirring time of 20 - 30 min, and finally obtain the probiotic fermentate powder that can clear the intestines and protect the liver by freeze-drying.

[0031] Example 2:

[0032] A preparation method of a probiotic fermentate powder that can clear the intestines and protect the liver, comprising the following steps:

[0033] S1: Weigh 22 parts of kudzu root, 17 parts of hawthorn, 16 parts of white hyacinth bean, 8 parts of poria cocos, 28 parts of lotus leaf, 28 parts of defatted soy flour, 23 parts of red date powder, 18 parts of corn starch, and 135 parts of water. Mix the kudzu root, hawthorn, white hyacinth bean, poria cocos, and lotus leaf evenly according to the ratio, crush them, and sieve them through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soy flour, red date powder, corn starch, and water to the traditional Chinese medicine powder according to the ratio, mix evenly, and perform high-temperature sterilization to obtain the fermentation medium;

[0034] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 2.7 ×10 7 CFU / g of the fermentation medium, carry out aerobic fermentation at 26°C to 40°C for 1.6 hours, and provide an oxygen inlet volume of 0.92 vm; inoculate Weissella confusa JS518, and the inoculation amount of Weissella confusa JS518 is 3.2×10 8CFU / g of the fermentation medium, perform microaerobic fermentation at 30°C to 37°C for 3 hours, and provide an oxygen inlet volume of 0.08 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 5.1×10 8 CFU / g of the fermentation medium, perform microaerobic fermentation at 35°C to 40°C for 2.5 hours, and provide an oxygen inlet volume of 0.04 vvm; after inoculating all the strains, perform aerobic fermentation for 1 hour with an oxygen inlet volume of 0.75 vvm; perform microaerobic fermentation for 3 hours with an oxygen inlet volume of 0.05 vvm, and perform anaerobic fermentation for 1 hour in an alternating fermentation cycle, and cycle 4 times to obtain the probiotic fermentate;

[0035] S3: Crush the probiotic fermentate prepared in S2, and preliminarily mix the crushed probiotic fermentate with a microbial protectant composed of 7 parts of trehalose, 25 parts of glycerol, and 3 parts of whey protein, with a total weight of 35 parts. Add 6 parts of an emulsifier to the mixture and use a high-speed blender to perform high-speed stirring at a stirring speed of 1200 - 2000 rpm for 10 - 15 min; then add a stabilizer composed of 4 parts of cyclodextrin, 6 parts of gelatin, and 17 parts of xanthan gum, with a total weight of 27 parts, and use a low-speed blender to perform sufficient stirring at a stirring speed of 500 - 800 rpm for 20 - 30 min, and finally obtain a probiotic fermentate powder that can clear the intestines and protect the liver by freeze-drying.

[0036] Example 3:

[0037] A preparation method of a probiotic fermentate powder that can clear the intestines and protect the liver, comprising the following steps:

[0038] S1: Weigh 25 parts of kudzu root, 16 parts of hawthorn, 18 parts of white hyacinth bean, 9 parts of poria cocos, 26 parts of lotus leaf, 26 parts of defatted soy flour, 22 parts of red date powder, 22 parts of corn starch, and 130 parts of water. Mix the kudzu root, hawthorn, white hyacinth bean, poria cocos, and lotus leaf evenly in proportion, crush them, and sieve through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soy flour, red date powder, corn starch, and water to the traditional Chinese medicine powder in proportion, mix evenly, and perform high-temperature sterilization to obtain the fermentation medium;

[0039] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 7.2×10 7 CFU / g of the fermentation medium, perform aerobic fermentation at 26°C to 40°C for 2 hours, and provide an oxygen inlet volume of 0.50 vm; inoculate Weizmannia coagulans JS518, and the inoculation amount of Weizmannia coagulans JS518 is 1.1×10 8CFU / g of the fermentation medium, carry out microaerobic fermentation at 30°C to 37°C for 2.5 hours, and provide an oxygen input of 0.05 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 5.1×10 8 CFU / g of the fermentation medium, carry out microaerobic fermentation at 35°C to 40°C for 2.5 hours, with an oxygen input of 0.02 vvm; after inoculating all the strains, carry out aerobic fermentation for 1 hour, providing an oxygen input of 1.0 vvm; carry out microaerobic fermentation for 3 hours, with an oxygen input of 0.1 vvm, and carry out alternating fermentation in the cycle of anaerobic fermentation for 1 hour, and cycle 4 times to obtain the probiotic fermentate;

[0040] S3: Crush the probiotic fermentate prepared in S2, and preliminarily mix the crushed probiotic fermentate with a microbial protectant composed of 6 parts of trehalose, 20 parts of glycerol, and 5 parts of whey protein, with a total weight of 31 parts. Add 8 parts of an emulsifier to the mixture and use a high-speed blender for high-speed stirring, with a stirring speed of 1200 - 2000 rpm and a stirring time of 10 - 15 min; then add a stabilizer composed of 8 parts of cyclodextrin, 11 parts of gelatin, and 9 parts of xanthan gum, with a total weight of 28 parts, and use a low-speed blender for sufficient stirring, with a stirring speed of 500 - 800 rpm and a stirring time of 20 - 30 min, and finally obtain the probiotic fermentate powder that can clear the intestines and protect the liver by freeze-drying.

[0041] Example 4:

[0042] A preparation method of a probiotic fermentate powder that can clear the intestines and protect the liver, comprising the following steps:

[0043] S1: Weigh 23 parts of Pueraria lobata, 17 parts of Hawthorn, 16 parts of White Hyacinth Bean, 7 parts of Poria cocos, 24 parts of Lotus Leaf, 24 parts of defatted soybean powder, 20 parts of red date powder, 20 parts of corn starch, and 125 parts of water. Mix Pueraria lobata, Hawthorn, White Hyacinth Bean, Poria cocos, and Lotus Leaf evenly according to the ratio, crush them and sieve through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soybean powder, red date powder, corn starch, and water to the traditional Chinese medicine powder according to the ratio, mix evenly, and carry out high-temperature sterilization to obtain the fermentation medium;

[0044] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 6.1 ×10 7 CFU / g of the fermentation medium, carry out aerobic fermentation at 26°C to 40°C for 1.5 hours, and provide an oxygen input of 0.75 vm; inoculate Weizmannia coagulans JS518, and the inoculation amount of Weizmannia coagulans JS518 is 1.05×10 8CFU / g fermentation medium, carry out microaerobic fermentation at 30°C to 37°C for 2.8 hours, and provide an oxygen inlet volume of 0.05 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.18×10 8 CFU / g fermentation medium, carry out microaerobic fermentation at 35°C to 40°C for 2.1 hours, with an oxygen inlet volume of 0.03 vvm; after inoculating all strains, carry out aerobic fermentation for 1 hour, providing an oxygen inlet volume of 0.60 vvm; carry out microaerobic fermentation for 3 hours, with an oxygen inlet volume of 0.03 vvm, and carry out alternating fermentation in a cycle of anaerobic fermentation for 1 hour, and cycle 4 times to obtain a probiotic fermentate;

[0045] S3: Crush the probiotic fermentate prepared in S2, and preliminarily mix the crushed probiotic fermentate with a microbial protectant composed of 6 parts of trehalose, 13 parts of glycerol, and 4 parts of whey protein, with a total weight of 23 parts. Add 7 parts of emulsifier to the mixture and use a high-speed blender to carry out high-speed stirring, with a stirring speed of 1200 - 2000 rpm and a stirring time of 10 - 15 min; then add a stabilizer composed of 5 parts of cyclodextrin, 9 parts of gelatin, and 18 parts of xanthan gum, with a total weight of 32 parts, and use a low-speed blender to carry out sufficient stirring, with a stirring speed of 500 - 800 rpm and a stirring time of 20 - 30 min, and finally freeze-dry to obtain a probiotic fermentate powder that can clear the intestines and protect the liver.

[0046] Example 5:

[0047] A preparation method of a probiotic fermentate powder that can clear the intestines and protect the liver, comprising the following steps:

[0048] S1: Weigh 24 parts of Pueraria lobata, 16 parts of Hawthorn, 18 parts of White Hyacinth Bean, 6 parts of Poria cocos, 27 parts of Lotus Leaf, 27 parts of defatted soybean powder, 25 parts of red date powder, 18 parts of corn starch and 135 parts of water. Mix Pueraria lobata, Hawthorn, White Hyacinth Bean, Poria cocos and Lotus Leaf evenly according to the ratio, crush and sieve through a 100-mesh sieve to obtain traditional Chinese medicine powder. Add defatted soybean powder, red date powder, corn starch and water to the traditional Chinese medicine powder according to the ratio, mix evenly, and carry out high-temperature sterilization to obtain a fermentation medium;

[0049] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 7.9×10 7 CFU / g fermentation medium, carry out aerobic fermentation at 26°C to 40°C for 1 hour, and provide an oxygen inlet volume of 1.0 vm; inoculate Weizmannia coagulans JS518, and the inoculation amount of Weizmannia coagulans JS518 is 8.0×10 7CFU / g fermentation medium, carry out microaerobic fermentation at 30°C to 37°C for 2.7 hours, and provide an oxygen input of 0.07 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.05×10 8 CFU / g fermentation medium, carry out microaerobic fermentation at 35°C to 40°C for 2.1 hours, and provide an oxygen input of 0.06 vvm; after all the strains are inoculated, carry out aerobic fermentation for 1 hour with an oxygen input of 0.90 vvm; carry out microaerobic fermentation for 3 hours with an oxygen input of 0.07 vvm, and carry out alternating fermentation in the cycle of anaerobic fermentation for 1 hour, and cycle 4 times to obtain the probiotic fermentate;

[0050] S3: Crush the probiotic fermentate prepared in S2, and preliminarily mix the crushed probiotic fermentate with a microbial protectant composed of 7 parts of trehalose, 20 parts of glycerol, and 4 parts of whey protein, with a total weight of 33 parts. Add 7 parts of emulsifier to the mixture and stir at high speed using a high-speed stirrer. The stirring speed is 1200 - 2000 rpm, and the stirring time is 10 - 15 min; then add a stabilizer composed of 5 parts of cyclodextrin, 6 parts of gelatin, and 10 parts of xanthan gum, with a total weight of 21 parts, and stir thoroughly using a low-speed stirrer. The stirring speed is 500 - 800 rpm, and the stirring time is 20 - 30 min. Finally, freeze-dry to obtain a probiotic fermentate powder that can clear the intestines and protect the liver.

[0051] Comparative Example 1:

[0052] Comparative Example 1 is the fermentation medium prepared in Step S1 of Example 1.

[0053] Comparative Example 2:

[0054] Comparative Example 2 is a constant microaerobic fermentation, which specifically includes the following steps:

[0055] S1: The raw material ratio, preparation method, and operating conditions in Step S1 of Comparative Example 2 are exactly the same as those in S1 of Example 1;

[0056] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 3.5 ×10 7 CFU / g fermentation medium; inoculate Weissella confusa JS518, and the inoculation amount of Weissella confusa JS518 is 1.2×10 8 CFU / g fermentation medium; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.2×10 8 CFU / g fermentation medium; after all the strains are inoculated, carry out microaerobic fermentation at 30°C to 37°C for 24 - 32 hours, and the oxygen input for microaerobic fermentation is 0.1 vvm to obtain the probiotic fermentate;

[0057] S3: The raw material ratio, preparation method, and operating conditions in step S3 of Comparative Example 2 are exactly the same as those in S3 of Example 1.

[0058] Comparative Example 3:

[0059] Comparative Example 3 is aerobic and hypoxic alternating fermentation, which specifically includes the following steps:

[0060] S1: The raw material ratio, preparation method, and operating conditions in step S1 of Comparative Example 3 are exactly the same as those in S1 of Example 1;

[0061] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g of fermentation medium, and carry out aerobic fermentation at 26°C to 40°C for 1.5 hours, providing an oxygen inlet volume of 0.75 vvm; inoculate Weissella confusa JS518, and the inoculation amount of Weissella confusa JS518 is 1.2×10 8 CFU / g of fermentation medium, and carry out microaerobic fermentation at 30°C to 37°C for 2.8 hours, providing an oxygen inlet volume of 0.25 vvm; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.2×10 8 CFU / g of fermentation medium, and carry out microaerobic fermentation at 35°C to 40°C for 2.2 hours, providing an oxygen inlet volume of 0.4 vvm; after inoculating all the strains, carry out alternating fermentation according to the cycle of aerobic fermentation for 1 hour with an oxygen inlet volume of 1 vvm; microaerobic fermentation for 4 hours with an oxygen inlet volume of 0.1 vvm, and cycle 4 times to obtain the probiotic fermented product;

[0062] S3: The raw material ratio, preparation method, and operating conditions in step S3 of Comparative Example 3 are exactly the same as those in S3 of Example 1.

[0063] Comparative Example 4:

[0064] Comparative Example 4 is hypoxic and anoxic alternating fermentation, which specifically includes the following steps:

[0065] S1: The raw material ratio, preparation method, and operating conditions in step S1 of Comparative Example 4 are exactly the same as those in S1 of Example 1;

[0066] S2: Inoculate Kluyveromyces marxianus K11, and the inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g of fermentation medium; inoculate Weissella confusa JS518, and the inoculation amount of Weissella confusa JS518 is 1.2×10 8CFU / g fermentation medium; inoculate Lactobacillus paracasei JS12, and the inoculation amount of Lactobacillus paracasei JS12 is 2.2×10 8 CFU / g fermentation medium; after inoculating all the strains, carry out alternating fermentation according to the cycle of microaerobic fermentation for 4 hours with an oxygen inlet amount of 0.1 vvm and anaerobic fermentation for 1 hour, and cycle 4 times to obtain the probiotic fermentate;

[0067] S3: The raw material ratio, preparation method, and operating conditions in step S3 of Comparative Example 4 are exactly the same as those in S3 of Example 1.

[0068] Detection of active ingredients: Detect the probiotic fermentate powder capable of clearing the intestine and protecting the liver prepared in Examples 1 to 5, Comparative Examples 2 to 4, and Comparative Example 1. Puerarin isoflavones, puerarin, and nuciferine are determined by high performance liquid chromatography, crude polysaccharides are determined with reference to the national standard SN / T4260, and the dietary fiber content is determined by an enzymatic method; the free amino acid content is detected by the ninhydrin method, and the detection results are shown in Table 1:

[0069] Table 1

[0070] Test items Example 1 Example 2 Example 3 Example 4 Example 5 Control Example 1 Control Example 2 Control Example 3 Control Example 4 Puerarin isoflavone (mg / g) 0.215 0.197 0.186 0.214 0.209 0.097 0.121 0.155 0.116 Puerarin (ug / g) 0.425 0.418 0.376 0.398 0.388 0.157 0.221 0.276 0.188 Polysaccharides (mg / g) 3.438 3.360 3.756 0.315 0.298 1.246 1.735 2.193 1.495 Nuciferine (mg / g) 0.303 0.346 0.309 0.298 0.246 0.113 0.158 0.205 0.136 Dietary fiber (mg / g) 112.256 110.676 88.954 94.212 119.65 41.092 57.526 73.962 49.308 Free amino acids (mg / g) 1.863 1.456 1.597 1.439 1.480 0.691 0.966 1.242 0.828

[0071] During the fermentation process of Kluyveromyces marxianus K11, Weissella confusa JS518, and Lactobacillus paracasei JS12 with traditional Chinese medicine ingredients such as Pueraria lobata, Crataegus pinnatifida, Dolicho lablab, Poria cocos, and Nelumbo nucifera, by precisely controlling the temperature, oxygen inlet amount, and time during the fermentation process to optimize the growth and metabolic activities of probiotics, the content and bioavailability of active ingredients in the fermentate can be significantly improved.

[0072] Detection of the effective viable count: Take 1 g of the probiotic fermentate powder for clearing the intestine and protecting the liver prepared in Examples 1 to 5 and Comparative Examples 2 to 3 and place it in a test tube. Add 9 ml of normal saline respectively, shake and mix at 35 °C on a shaker at 120 rpm for 2 h, and use the plate counting method to count the total number of effective viable bacteria in the probiotic fermentate powder for clearing the intestine and protecting the liver in different groups. The detection results are shown in Table 2:

[0073] Table 2

[0074] <![CDATA[Kluyveromyces marxianus K11 (×10 8 CFU / g)]]> <![CDATA[Weizmannia coagulans JS518 (×10 8 CFU / g)]]> <![CDATA[Lactobacillus paracasei JS12 (×10 8 CFU / g)]]> Example 1 4.92 1.09 0.92 Control Example 2 0.27 1.28 0.54 Control Example 3 1.59 0.87 0.21 Control Example 4 0.12 0.26 1.53

[0075] Diarrhea regulation test: Healthy SPF-grade Kunming mice are randomly divided into 6 groups, with 10 mice in each group, 5 males and 5 females, and housed separately in cages. The mice are adaptively fed in a barrier environment for one week, with free access to food and water, and the bedding is changed daily. The experiment starts after 5 days of adaptive feeding. The experimental period is 10 days, with days 1 - 5 being the modeling period and days 6 - 10 being the recovery period. The grouping and treatment methods are shown in detail in Table 3; record the changes in the fecal water content of the mice, as shown in detail in Table 4:

[0076] Table 3

[0077] Group Treatment during modeling period Treatment during recovery period Normal group Basal diet, fed ad libitum. Basal diet, fed ad libitum, without treatment. Control group Basal diet, fed ad libitum. Prepare magnesium sulfate aqueous solution at 50 mg / mL, and give each mouse 0.2 mL / 10 g body weight of magnesium sulfate aqueous solution by gavage once a day. Basal diet, fed ad libitum, without treatment. Experimental group 1 Basal diet, fed ad libitum. Prepare magnesium sulfate aqueous solution at 50 mg / mL, and give each mouse 0.2 mL / 10 g body weight of magnesium sulfate aqueous solution by gavage once a day. Basal diet, fed ad libitum. Add the probiotic fermented powder for clearing the intestine and protecting the liver prepared in Example 1, and give the probiotic fermented powder for clearing the intestine and protecting the liver at a dose of 10 mg / 10 g body weight of the mouse once a day. Experimental group 2 Basal diet, fed ad libitum. Prepare magnesium sulfate aqueous solution at 50 mg / mL, and give each mouse 0.2 mL / 10 g body weight of magnesium sulfate aqueous solution by gavage once a day. Basal diet, fed ad libitum. Add the probiotic fermented powder for clearing the intestine and protecting the liver prepared in Control Example 2, and give the probiotic fermented powder for clearing the intestine and protecting the liver at a dose of 10 mg / 10 g body weight of the mouse once a day. Experimental group 3 Basal diet, fed ad libitum. Prepare magnesium sulfate aqueous solution at 50 mg / mL, and give each mouse 0.2 mL / 10 g body weight of magnesium sulfate aqueous solution by gavage once a day. Basal diet, fed ad libitum. Add the probiotic fermented powder for clearing the intestine and protecting the liver prepared in Control Example 3, and give the probiotic fermented powder for clearing the intestine and protecting the liver at a dose of 10 mg / 10 g body weight of the mouse once a day. Experimental group 4 Basal diet, fed ad libitum. Prepare magnesium sulfate aqueous solution at 50 mg / mL, and give each mouse 0.2 mL / 10 g body weight of magnesium sulfate aqueous solution by gavage once a day. Basal diet, fed ad libitum. Add the probiotic fermented powder for clearing the intestine and protecting the liver prepared in Control Example 4, and give the probiotic fermented powder for clearing the intestine and protecting the liver at a dose of 10 mg / 10 g body weight of the mouse once a day.

[0078] Table 4

[0079] Group Day 5 Day 6 Day 8 Day 10 Normal group 35.9% 36.4% 34.5% 35.8% Control group 67.8% 61.4% 54.3% 50.1% Experimental group 1 68.4% 52.4% 45.4% 37.1% Experimental group 2 64.2% 60.4% 54.7% 49.4% Experimental group 3 65.9% 56.7% 49.1% 46.3% Experimental group 4 66.4% 61.4% 51.6% 48.7%

[0080] Mouse anti-alcoholism test: Healthy SPF-grade Kunming mice were randomly divided into 6 groups, with 10 mice in each group (5 males and 5 females), and they were housed separately in cages. The mice were adaptively raised in a barrier environment for one week. The mice had free access to food and water, and the bedding was changed daily. The experiment started after 7 days of adaptive breeding. The experimental period was 25 days. For the grouping and treatment methods, the white liquor used in the experiment was commercially available 52° Langjiu. For details, see Table 5. For the contents of alanine aminotransferase (AST) and aspartate aminotransferase (ALT) in mice of different groups, see Table 6 for details:

[0081] Table 5

[0082] Group Treatment method Normal group The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of normal saline per 10 g of body weight once every 12 h. After the first administration of 0.1 mL of normal saline per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected. Control group The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of white liquor per 10 g of body weight once every 12 h. After the first administration of 0.1 mL of white liquor per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected. Experimental group 1 The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of white liquor per 10 g of body weight once every 12 h. The probiotic fermented powder for clearing the intestine and protecting the liver prepared in Example 1 was added, and the probiotic fermented powder for clearing the intestine and protecting the liver was administered at a dose of 10 mg per 10 g of body weight of the mouse once a day. After the first administration of 0.1 mL of white liquor per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected. Experimental group 2 The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of white liquor per 10 g of body weight once every 12 h. The probiotic fermented powder for clearing the intestine and protecting the liver prepared in Comparative Example 2 was added, and the probiotic fermented powder for clearing the intestine and protecting the liver was administered at a dose of 10 mg per 10 g of body weight of the mouse once a day. After the first administration of 0.1 mL of white liquor per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected. Experimental group 3 The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of white liquor per 10 g of body weight once every 12 h. The probiotic fermented powder for clearing the intestine and protecting the liver prepared in Comparative Example 3 was added, and the probiotic fermented powder for clearing the intestine and protecting the liver was administered at a dose of 10 mg per 10 g of body weight of the mouse once a day. After the first administration of 0.1 mL of white liquor per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected. Experimental group 4 The basal diet was provided ad libitum. Each mouse was gavaged with 0.1 mL of white liquor per 10 g of body weight once every 12 h. The probiotic fermented powder for clearing the intestine and protecting the liver prepared in Comparative Example 4 was added, and the probiotic fermented powder for clearing the intestine and protecting the liver was administered at a dose of 10 mg per 10 g of body weight of the mouse once a day. After the first administration of 0.1 mL of white liquor per 10 g of body weight on the 25th day, the mice were fasted for 12 h and then blood was collected.

[0083] Table 6

[0084] Group AST (IU / L) ALT (IU / L) Normal group 1.56±0.24 2.69±0.68 Control group 2.98±0.36 3.89±0.23 Experimental group 1 1.67±0.54 2.98±0.63 Experimental group 2 2.31±0.36 3.56±0.21 Experimental group 3 1.99±0.21 3.21±0.57 Experimental group 4 2.17±0.41 3.17±0.39

[0085] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a probiotic fermentation powder for clearing intestines and protecting liver, characterized in that: The following steps are involved: S1, Pueraria root, hawthorn, white hyacinth bean, Poria cocos and lotus leaf are mixed evenly in proportion, crushed and sieved through a 100-mesh sieve to obtain a Chinese medicine powder, defatted soybean powder, red date powder, corn starch and water are added to the Chinese medicine powder in proportion, mixed evenly, and sterilized at high temperature to obtain a fermentation medium; S2: inoculating Kluyveromyces marxianus K11 with a preservation number of CGMCC No. 26477 into a fermentation medium for aerobic fermentation, controlling the fermentation temperature to 26°C to 40°C and fermenting for 1 to 2 hours; inoculating Weizmannella coagulans JS518 with a preservation number of CGMCC No. 27471 into a fermentation medium for microaerobic fermentation, controlling the fermentation temperature to 30°C to 37°C and fermenting for 2 to 4 hours; inoculating Lactobacillus paracasei JS12 with a preservation number of CGMCC No. 27469 into a culture medium for microaerobic fermentation, controlling the fermentation temperature to 35°C to 40°C and fermenting for 2 to 3 hours; after all strains are inoculated, performing alternating fermentations according to a predetermined fermentation cycle: aerobic fermentation for 1 hour, microaerobic fermentation for 3 hours, and anaerobic fermentation for 1 hour, for a total of 4 cycles, to obtain a probiotic fermentation product; S3: the probiotic fermentation product prepared in S2 is crushed, and the crushed probiotic fermentation product is preliminarily mixed with the pre-configured microbial protective agent, an emulsifier is added to the mixture, and a high-speed stirrer is used for high-speed stirring at a stirring speed of 1200-2000 rpm for 10-15 minutes, and the emulsifier, the microbial protective agent and the water in the probiotic fermentation product form a microemulsified solution to wrap the probiotic fermentation product particles; a stabilizer is then added, and a low-speed stirrer is used for sufficient stirring at a stirring speed of 500-800 rpm for 20-30 minutes, and finally freeze-dried to obtain a probiotic fermentation product powder for clearing intestines and protecting liver; The mass proportions of the kudzu root, hawthorn, white hyacinth bean, poria, lotus leaf, defatted soy flour, red date powder, corn starch, water, microbial protective agent, emulsifier and stabilizer are 20-25 parts of kudzu root, 16-18 parts of hawthorn, 15-18 parts of white hyacinth bean, 5-10 parts of poria, 20-30 parts of lotus leaf, 20-30 parts of defatted soy flour, 15-26 parts of red date powder, 15-24 parts of corn starch, 90-150 parts of water, 20-40 parts of microbial protective agent, 5-8 parts of emulsifier and 18-35 parts of stabilizer.

2. The method for preparing a probiotic fermentation powder for clearing intestines and protecting liver according to claim 1, characterized in that: The Kluyveromyces marxianus K11, Weizmannella coagulans JS518 and Lactobacillus paracasei JS12 were activated and inoculated into the fermentation medium respectively. The inoculation amount of Kluyveromyces marxianus K11 was 10 7 ~10 9 CFU / g fermentation medium, the inoculum size of Weizmannella coagulans JS518 was 10 7 ~10 9 CFU / g fermentation medium, the inoculation amount of Lactobacillus paracasei JS12 was 10 7 ~10 9 CFU / g fermentation medium.

3. The method for preparing a probiotic fermentation powder for clearing intestines and protecting liver according to claim 1, characterized in that: The oxygen intake for aerobic fermentation is 0.5-1 vvm, and the temperature of aerobic fermentation is controlled at 26°C-40°C; the oxygen intake for microaerobic fermentation is 0.02-0.1 vvm, and the temperature of microaerobic fermentation is 30°C-37°C; no oxygen is introduced into anaerobic fermentation, and the temperature of anaerobic fermentation is controlled at 35°C-40°C.

4. The method for preparing a probiotic fermentation powder for clearing intestines and protecting liver according to claim 1, characterized in that: The microbial protective agent consists of 6 to 10 parts of trehalose, 12 to 25 parts of glycerol and 3 to 5 parts of whey protein in terms of mass.

5. The method for preparing the probiotic fermentation powder for clearing intestine and protecting liver according to claim 1, characterized in that: The emulsifier is lecithin or polysorbate.

6. The method for preparing the probiotic fermentation powder for clearing intestine and protecting liver according to claim 1, characterized in that: The stabilizer is composed of 3 to 8 parts of cyclodextrin, 6 to 11 parts of gelatin and 9 to 18 parts of xanthan gum in terms of mass.

7. A probiotic fermentation powder for clearing intestines and protecting liver, characterized in that: The powder is prepared by the method for preparing the intestinal cleansing and liver protecting probiotic fermentation product powder according to any one of claims 1 to 6.