Probiotic fermentation product powder capable of clearing intestines and protecting liver and preparation method of probiotic fermentation product powder

Abstract

The invention provides probiotic yeast powder capable of clearing intestines and protecting liver and a preparation method thereof, relates to the field of microbial fermentation application, and aims to solve the problems of gastrointestinal diseases and liver injury caused by factors such as irregular dietary habits, increase of life pressure and frequent drinking in modern life. By combining the traditional Chinese medicinal materials with the modern microbial fermentation technology, the probiotic fermented product with the effects of clearing intestinal toxin, protecting liver and kidney, nourishing stomach and spleen, and tonifying middle-Jiao and Qi is prepared. The specific preparation method comprises the following steps: mixing medicinal and edible materials such as radix puerariae, haws, white hyacinth beans, poria cocos and lotus leaves with defatted soybean meal, red date powder, corn starch and water, performing high-temperature sterilization to obtain a fermentation culture medium, then inoculating kluyveromyces marxianus K11, Witzia ciliaris JS518 and Lactobacillus paracasei JS12, and performing alternate fermentation, and finally, adding a microbial protective agent and a stabilizer, uniformly mixing, and freeze-drying to obtain the product.

Description

Technical Field

[0001] The invention relates to the field of microbial fermentation, and in particular to a probiotic fermentation powder capable of clearing intestines and protecting liver and a preparation method thereof. Background Art

[0002] Traditional Chinese medicine has a long history of regulating the intestines and protecting the liver, but relying solely on Chinese herbal ingredients often fails to achieve the desired effect, and Chinese herbal decoction is time-consuming and laborious. Chinese herbal decoction liquid is not conducive to preservation and has a poor taste. Modern microbial fermentation technology provides a new idea for improving the efficacy of Chinese herbal medicines. Probiotic fermentation can enhance the biological activity and absorption efficiency of Chinese herbal ingredients. Probiotics can not only improve the balance of intestinal flora, but also help enhance immune function and promote digestion. Combining traditional Chinese herbal medicines with probiotic fermentation technology is expected to develop a new product for clearing the intestines and protecting the liver. However, the following key issues need to be addressed: how to optimize the growth and metabolic activity of probiotics, increase the content and bioavailability of active ingredients in fermented products; how to effectively protect probiotics from external environmental interference and prolong the uniformity and stability of the product. Summary of the invention

[0003] To achieve the above objectives, the present invention is implemented by the following technical scheme: A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1, weighing kudzu root, hawthorn, white dung bean, Poria cocos and lotus leaf, mixing kudzu root, hawthorn, white dung bean, Poria cocos and lotus leaf in proportion, pulverizing and sieving through a 100-mesh sieve to obtain a Chinese medicine powder, adding defatted soybean powder, red date powder, corn starch and water to the Chinese medicine powder in proportion, mixing evenly, and sterilizing at high temperature to obtain a fermentation medium; S2: Inoculate Kluyveromyces marxianus K11 into the fermentation medium for aerobic fermentation, control the fermentation temperature to 26°C to 40°C and ferment for 1 to 2 hours; inoculate Weizmannella coagulans JS518 into the fermentation medium for microaerobic fermentation, control the fermentation temperature to 30°C to 37°C and ferment for 2 to 4 hours; inoculate Lactobacillus paracasei JS12 into the culture medium for microaerobic fermentation, control the fermentation temperature to 35°C to 40°C and fermentation time to 2 to 3 hours; after completing the inoculation of all strains, 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 repeat 4 times to obtain a probiotic fermentation product; 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.

[0004] 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.

[0005] The traditional Chinese medicine pharmacology of Pueraria root, hawthorn, white hyacinth bean, Poria cocos and lotus leaf is as follows: 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.

[0006] 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.

[0007] 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.

[0008] 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.

[0009] Lotus leaf: "Southern Yunnan Materia Medica": It can clear wind and heat from the head and eyes, stop dizziness, clear phlegm, relieve gas, stop vomiting, and relieve headaches. Lotus leaf has been regarded as a good medicine for weight loss in China since ancient times. Its chemical components mainly include nelumboide, citric acid, malic acid, gluconic acid, oxalic acid, succinic acid and other alkaline components with anti-mitotic effects. The nelumboide in lotus leaf tea contains a variety of effective lipogenic alkaloids, which can effectively decompose the fat in the body and strongly excrete it from the body. Nelumboide can be densely distributed on the intestinal wall of the human body, forming a layer of fat isolation membrane to prevent fat absorption and prevent fat accumulation.

[0010] 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.

[0011] The oxygen intake of the aerobic fermentation is 0.5-1 vvm, and the aerobic fermentation temperature is controlled at 26°C-40°C; the oxygen intake of the microaerobic fermentation is 0.02-0.1 vvm, and the aerobic fermentation temperature is controlled at 30°C-37°C; no oxygen is introduced into the anaerobic fermentation, and the anaerobic fermentation temperature is controlled at 35°C-40°C; for example, the oxygen intake of the aerobic fermentation is set to 1 vvm, which refers to the ventilation volume / fermentation tank volume / minute. For a 50L fermentation tank, the ventilation volume per minute is 50L.

[0012] 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.

[0013] The emulsifier is lecithin or polysorbate.

[0014] 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.

[0015] The present invention provides a probiotic fermentation powder capable of clearing intestines and protecting liver and a preparation method thereof, which has the following beneficial effects: 1. Alternating fermentation with Kluyveromyces marxianus K11, Weizmannella coagulans JS518 and Lactobacillus paracasei JS12. This unique fermentation cycle helps to increase the content and bioavailability of active ingredients in the fermentation.

[0016] 2. The intestinal cleansing and liver protecting probiotic fermentation powder prepared by the present invention forms a microemulsified solution by adding a microbial protective agent and an emulsifier to encapsulate the probiotic fermentation particles, effectively protecting the probiotics from interference from the external environment, extending the shelf life of the product, and improving the uniformity and stability of the product. DETAILED DESCRIPTION

[0017] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0018] Kluyveromyces marxianus in the following examples ( Kluyveromyces marxianus ) The deposit number of K11 is CGMCC No.26477.

[0019] The Lactobacillus paracasei in the following examples ( Lacticaseibacillus paracasei )The deposit number of JS12 is CGMCC No.27469.

[0020] In the following examples, Weizmannella coagulans ( Weizmannia coagulans )The deposit number of JS518 is CGMCC No.27471.

[0021] Embodiment 1:

[0022] A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1: Weigh 24 parts of kudzu root, 17 parts of hawthorn, 17 parts of white dung bean, 7 parts of poria, 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 kudzu root, hawthorn, white dung bean, poria and lotus leaf in proportion, grind and sieve through a 100-mesh sieve to obtain Chinese medicine powder, add defatted soy flour, red date powder, corn starch and water to the Chinese medicine powder in proportion, mix well, and sterilize at high temperature to obtain fermentation medium; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 1.5 hours, and 0.75vvm oxygen was provided; Weizmannella coagulans JS518 was inoculated, and the inoculation amount of Weizmannella coagulans JS518 was 1.2×10 8 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 2.8 hours, and 0.1vvm oxygen was provided; Lactobacillus paracasei JS12 was inoculated, and the inoculation amount of Lactobacillus paracasei JS12 was 2.2×108 CFU / g fermentation medium, microaerobic fermentation for 2.2 hours at 35°C-40°C, providing an oxygen input of 0.1vvm; after completing the inoculation of all strains, fermentation was carried out in accordance with the cycle of aerobic fermentation for 1 hour, oxygen input of 1vvm; microaerobic fermentation for 3 hours, oxygen input of 0.04vvm, and anaerobic fermentation for 1 hour, and the cycle was repeated 4 times 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 a microbial protective agent consisting of 10 parts of trehalose, 12 parts of glycerol, and 3 parts of whey protein, with a total weight of 25 parts, and 5 parts of emulsifier are added to the mixture, and a high-speed stirrer is used for high-speed stirring, the stirring speed is 1200-2000rpm, and the stirring time is 10-15min; then a stabilizer consisting of 4 parts of cyclodextrin, 9 parts of gelatin, and 13 parts of xanthan gum is added, and a low-speed stirrer is used for sufficient stirring, the stirring speed is 500-800rpm, and the stirring time is 20-30min. Finally, the probiotic fermentation product powder that can cleanse the intestine and protect the liver is obtained by freeze-drying.

[0023] Embodiment 2:

[0024] A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1: Weigh 22 parts of kudzu root, 17 parts of hawthorn, 16 parts of white dung bean, 8 parts of poria, 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 kudzu root, hawthorn, white dung bean, poria and lotus leaf in proportion, grind and sieve through a 100-mesh sieve to obtain Chinese medicine powder, add defatted soy flour, red date powder, corn starch and water to the Chinese medicine powder in proportion, mix well, and sterilize at high temperature to obtain fermentation medium; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 2.7 × 10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 1.6 hours, providing 0.92vm oxygen input; inoculated with Weizmannella coagulans JS518, the inoculation amount of Weizmannella coagulans JS518 was 3.2×10 8 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 3 hours, and 0.08vvm oxygen was provided; Lactobacillus paracasei JS12 was inoculated, and the inoculation amount of Lactobacillus paracasei JS12 was 5.1×10 8CFU / g fermentation medium, microaerobic fermentation for 2.5 hours at 35°C-40°C, providing an oxygen input of 0.04vvm; after completing the inoculation of all strains, fermentation was carried out alternately according to the cycle of aerobic fermentation for 1 hour, oxygen input of 0.75vvm; microaerobic fermentation for 3 hours, oxygen input of 0.05vvm, and anaerobic fermentation for 1 hour, and the cycle was repeated 4 times 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 a microbial protective agent consisting of 7 parts of trehalose, 25 parts of glycerol, and 3 parts of whey protein, with a total weight of 35 parts, and 6 parts of emulsifier are added to the mixture, and a high-speed stirrer is used for high-speed stirring, the stirring speed is 1200-2000rpm, and the stirring time is 10-15min; then a stabilizer consisting of 4 parts of cyclodextrin, 6 parts of gelatin, and 17 parts of xanthan gum is added, with a total weight of 27 parts, and a low-speed stirrer is used for sufficient stirring, the stirring speed is 500-800rpm, and the stirring time is 20-30min. Finally, the probiotic fermentation product powder that can cleanse the intestine and protect the liver is obtained by freeze-drying.

[0025] Embodiment 3:

[0026] A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1: Weigh 25 parts of kudzu root, 16 parts of hawthorn, 18 parts of white dung bean, 9 parts of poria, 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 kudzu root, hawthorn, white dung bean, poria and lotus leaf in proportion, grind and sieve through a 100-mesh sieve to obtain Chinese medicine powder, add defatted soy flour, red date powder, corn starch and water to the Chinese medicine powder in proportion, mix well, and sterilize at high temperature to obtain fermentation medium; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 7.2×10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 2 hours, and 0.50vm oxygen was provided; Weizmannella coagulans JS518 was inoculated, and the inoculation amount of Weizmannella coagulans JS518 was 1.1×10 8 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 2.5 hours, and 0.05vvm oxygen was introduced; Lactobacillus paracasei JS12 was inoculated, and the inoculation amount of Lactobacillus paracasei JS12 was 5.1×10 8CFU / g fermentation medium, microaerobic fermentation for 2.5 hours at 35°C-40°C, with an oxygen input of 0.02vvm; after completing the inoculation of all strains, aerobic fermentation for 1 hour, with an oxygen input of 1.0vvm; microaerobic fermentation for 3 hours, with an oxygen input of 0.1vvm, and anaerobic fermentation for 1 hour, alternate fermentation, cycle 4 times, and obtain probiotic fermentation product; S3: The probiotic fermentation product prepared in S2 is crushed, and the crushed probiotic fermentation product is preliminarily mixed with a microbial protective agent consisting of 6 parts of trehalose, 20 parts of glycerol, and 5 parts of whey protein, with a total weight of 31 parts, and 8 parts of emulsifier are added to the mixture, and a high-speed stirrer is used for high-speed stirring, the stirring speed is 1200-2000rpm, and the stirring time is 10-15min; then a stabilizer consisting of 8 parts of cyclodextrin, 11 parts of gelatin, and 9 parts of xanthan gum is added, with a total weight of 28 parts, and a low-speed stirrer is used for sufficient stirring, the stirring speed is 500-800rpm, and the stirring time is 20-30min. Finally, freeze-drying is performed to obtain a probiotic fermentation product powder that can cleanse the intestines and protect the liver.

[0027] Embodiment 4:

[0028] A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1: Weigh 23 parts of kudzu root, 17 parts of hawthorn, 16 parts of white dung bean, 7 parts of poria, 24 parts of lotus leaf, 24 parts of defatted soy flour, 20 parts of red date powder, 20 parts of corn starch and 125 parts of water. Mix kudzu root, hawthorn, white dung bean, poria and lotus leaf in proportion, grind and sieve through a 100-mesh sieve to obtain Chinese medicine powder, add defatted soy flour, red date powder, corn starch and water to the Chinese medicine powder in proportion, mix well, and sterilize at high temperature to obtain fermentation medium; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 6.1 × 10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 1.5 hours, and 0.75vm oxygen was introduced; Weizmannella coagulans JS518 was inoculated, and the inoculation amount of Weizmannella coagulans JS518 was 1.05×10 8 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 2.8 hours, and 0.05vvm oxygen was introduced; Lactobacillus paracasei JS12 was inoculated, and the inoculation amount of Lactobacillus paracasei JS12 was 2.18×10 8CFU / g fermentation medium, microaerobic fermentation for 2.1 hours at 35°C-40°C, with an oxygen input of 0.03vvm; after completing the inoculation of all strains, aerobic fermentation for 1 hour, with an oxygen input of 0.60vvm; microaerobic fermentation for 3 hours, with an oxygen input of 0.03vvm, and anaerobic fermentation for 1 hour, alternate fermentation, cycle 4 times, and obtain probiotic fermentation product; S3: The probiotic fermentation product prepared in S2 is pulverized, and the pulverized probiotic fermentation product is preliminarily mixed with a microbial protective agent consisting of 6 parts of trehalose, 13 parts of glycerol, and 4 parts of whey protein, with a total weight of 23 parts, and 7 parts of emulsifier are added to the mixture, and a high-speed stirrer is used for high-speed stirring, the stirring speed is 1200-2000rpm, and the stirring time is 10-15min; then a stabilizer consisting of 5 parts of cyclodextrin, 9 parts of gelatin, and 18 parts of xanthan gum is added, with a total weight of 32 parts, and a low-speed stirrer is used for sufficient stirring, the stirring speed is 500-800rpm, and the stirring time is 20-30min. Finally, the probiotic fermentation product powder that can cleanse the intestine and protect the liver is obtained by freeze-drying.

[0029] Embodiment 5:

[0030] A method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver comprises the following steps: S1: Weigh 24 parts of kudzu root, 16 parts of hawthorn, 18 parts of white dung bean, 6 parts of poria, 27 parts of lotus leaf, 27 parts of defatted soy flour, 25 parts of red date powder, 18 parts of corn starch and 135 parts of water. Mix kudzu root, hawthorn, white dung bean, poria and lotus leaf in proportion, grind and sieve through a 100-mesh sieve to obtain Chinese medicine powder, add defatted soy flour, red date powder, corn starch and water to the Chinese medicine powder in proportion, mix well, and sterilize at high temperature to obtain fermentation medium; S2: Inoculate Kluyveromyces marxianus K11, the inoculation amount of Kluyveromyces marxianus K11 is 7.9×10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 1 hour, and 1.0vm oxygen was provided; Weizmannella coagulans JS518 was inoculated, and the inoculation amount of Weizmannella coagulans JS518 was 8.0×10 7 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 2.7 hours, providing 0.07vvm oxygen flow; inoculation of Lactobacillus paracasei JS12, the inoculation amount of Lactobacillus paracasei JS12 was 2.05×10 8CFU / g fermentation medium, microaerobic fermentation for 2.1 hours at 35°C-40°C, providing an oxygen intake of 0.06vvm; after completing the inoculation of all strains, aerobic fermentation for 1 hour, oxygen intake of 0.90vvm; microaerobic fermentation for 3 hours, oxygen intake of 0.07vvm, and anaerobic fermentation for 1 hour were performed alternately, and the cycle was repeated 4 times 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 a microbial protective agent consisting of 7 parts of trehalose, 20 parts of glycerol, and 4 parts of whey protein, with a total weight of 33 parts, and 7 parts of emulsifier are added to the mixture, and a high-speed stirrer is used for high-speed stirring, the stirring speed is 1200-2000rpm, and the stirring time is 10-15min; then a stabilizer consisting of 5 parts of cyclodextrin, 6 parts of gelatin, and 10 parts of xanthan gum is added, with a total weight of 21 parts, and a low-speed stirrer is used for sufficient stirring, the stirring speed is 500-800rpm, and the stirring time is 20-30min. Finally, the probiotic fermentation product powder that can cleanse the intestine and protect the liver is obtained by freeze-drying.

[0031] Comparative Example 1: Comparative Example 1 is the fermentation medium prepared in step S1 of Example 1.

[0032] Comparative Example 2: Comparative Example 2 is a constant micro-aerobic fermentation, which specifically comprises the following steps: S1: The raw material ratio, preparation method, and operating conditions of step S1 in comparative example 2 are exactly the same as those of step S1 in example 1; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g fermentation medium; inoculated with Weizmannella coagulans JS518, the inoculation amount of Weizmannella coagulans JS518 was 1.2×10 8 CFU / g fermentation medium; inoculated with Lactobacillus paracasei JS12, the inoculation amount of Lactobacillus paracasei JS12 was 2.2×10 8 CFU / g fermentation medium; after completing the inoculation of all bacterial species, microaerobic fermentation is carried out at 30°C to 37°C for 24-32 hours, and the oxygen intake of microaerobic fermentation is 0.1vvm to obtain probiotic fermentation products; S3: The raw material ratio, preparation method and operating conditions of step S3 in comparative example 2 are exactly the same as those of step S3 in example 1.

[0033] Comparative Example 3: Comparative Example 3 is an aerobic and hypoxic alternating fermentation, which specifically includes the following steps: S1: The raw material ratio, preparation method, and operating conditions of step S1 in comparative example 3 are exactly the same as those of step S1 in example 1; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g fermentation medium, aerobic fermentation was carried out at 26℃~40℃ for 1.5 hours, and 0.75vvm oxygen was provided; Weizmannella coagulans JS518 was inoculated, and the inoculation amount of Weizmannella coagulans JS518 was 1.2×10 8 CFU / g fermentation medium, microaerobic fermentation was carried out at 30℃~37℃ for 2.8 hours, and 0.25vvm oxygen was introduced; Lactobacillus paracasei JS12 was inoculated, and the inoculation amount of Lactobacillus paracasei JS12 was 2.2×10 8 CFU / g fermentation medium, microaerobic fermentation for 2.2 hours at 35°C-40°C, providing an oxygen intake of 0.4vvm; after completing the inoculation of all strains, alternating fermentation according to the cycle of aerobic fermentation for 1 hour, oxygen intake of 1vvm; microaerobic fermentation for 4 hours, oxygen intake of 0.1vvm, and repeating 4 times to obtain a probiotic fermentation product; S3: The raw material ratio, preparation method and operating conditions of step S3 in comparative example 3 are exactly the same as those of step S3 in embodiment 1.

[0034] Comparative Example 4: Comparative Example 4 is a low oxygen and anaerobic alternating fermentation, which specifically comprises the following steps: S1: The raw material ratio, preparation method, and operating conditions of step S1 in Comparative Example 4 are exactly the same as those of step S1 in Example 1; S2: Inoculate Kluyveromyces marxianus K11. The inoculation amount of Kluyveromyces marxianus K11 is 3.5 × 10 7 CFU / g fermentation medium; inoculated with Weizmannella coagulans JS518, the inoculation amount of Weizmannella coagulans JS518 was 1.2×10 8 CFU / g fermentation medium; inoculated with Lactobacillus paracasei JS12, the inoculation amount of Lactobacillus paracasei JS12 was 2.2×10 8 CFU / g fermentation medium; after completing the inoculation of all bacterial species, the fermentation was carried out in a cycle of microaerobic fermentation for 4 hours with an oxygen intake of 0.1vvm and anaerobic fermentation for 1 hour, and the cycle was repeated 4 times to obtain a probiotic fermentation product; S3: The raw material ratio, preparation method and operating conditions of step S3 in comparative example 4 are exactly the same as those of step S3 in embodiment 1.

[0035] Active ingredient detection: The probiotic fermentation powders for cleansing intestines and protecting liver prepared in Examples 1 to 5, Comparative Examples 2 to 4 and Comparative Example 1 were tested. Pueraria isoflavones, puerarin and nuciferine were determined by high performance liquid chromatography, crude polysaccharides were determined according to the national standard SN / T4260, dietary fiber content was determined by enzymatic hydrolysis, and free amino acid content was determined by ninhydrin method. The test results are shown in Table 1: Table 1 Test items Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Pueraria isoflavones (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 During the fermentation process of Kluyveromyces marxianus K11, Weizmannella coagulans JS518 and Lactobacillus paracasei JS12 with traditional Chinese medicine ingredients such as Pueraria root, hawthorn, white lentil, Poria cocos and lotus leaf, the growth and metabolic activity of probiotics can be optimized by precisely controlling the temperature, oxygen intake and time of the fermentation process, thereby significantly improving the content and bioavailability of active ingredients in the fermentation product.

[0036] Detection of effective viable bacteria count: 1 g of the probiotic fermentation powder for clearing intestines and protecting liver prepared in Examples 1 to 5 and Comparative Examples 2 to 3 was placed in a test tube, and 9 ml of normal saline was added respectively. The mixture was shaken at 35° C. and 120 rpm for 2 h. The total number of effective viable bacteria in the probiotic fermentation powder for clearing intestines and protecting liver in different groups was counted by the plate counting method. The test results are shown in Table 2: Table 2 <![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 Comparative Example 2 0.27 1.28 0.54 Comparative Example 3 1.59 0.87 0.21 Comparative Example 4 0.12 0.26 1.53 Diarrhea regulation test: Healthy SPF Kunming mice were randomly divided into 6 groups, with 10 mice in each group, 5 males and 5 females, and kept in separate cages. The mice were adaptively raised in a barrier environment for one week, with free access to food and water, and the bedding was changed every day. The experiment began after 5 days of adaptive feeding. The experimental period was 10 days, with 1-5 days as the modeling period and 6-10 days as the recovery period. Grouping and treatment methods are detailed in Table 3; the changes in the water content of mouse feces were recorded, as detailed in Table 4: Table 3 Group Modeling Period Processing Recovery period treatment Normal group Basic feed, free feeding. Basic feed, free feeding, no treatment. Control group The basic feed was fed freely, and magnesium sulfate was prepared into a 50 mg / mL magnesium sulfate aqueous solution, and each mouse was given 0.2 mL / 10 g of magnesium sulfate aqueous solution by gavage once a day. Basic feed, free feeding, no treatment. Experimental Group 1 The basic feed was fed freely, and magnesium sulfate was prepared into a 50 mg / mL magnesium sulfate aqueous solution, and each mouse was given 0.2 mL / 10 g of magnesium sulfate aqueous solution by gavage once a day. The basic feed was fed freely, and the probiotic fermentation powder for clearing intestines and protecting liver prepared in Example 1 was added, at a dosage of 10 mg / 10 g of mouse body weight, and the probiotic fermentation powder for clearing intestines and protecting liver was given once a day. Experimental Group 2 The basic feed was fed freely, and magnesium sulfate was prepared into a 50 mg / mL magnesium sulfate aqueous solution, and each mouse was given 0.2 mL / 10 g of magnesium sulfate aqueous solution by gavage once a day. The basic feed was fed freely, and the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 2 was added, at a dosage of 10 mg / 10 g of mouse body weight, and the probiotic fermentation powder for clearing intestines and protecting liver was given once a day. Experimental Group 3 The basic feed was fed freely, and magnesium sulfate was prepared into a 50 mg / mL magnesium sulfate aqueous solution, and each mouse was given 0.2 mL / 10 g of magnesium sulfate aqueous solution by gavage once a day. The basic feed was fed freely, and the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 3 was added, at a dosage of 10 mg / 10 g of mouse body weight, and the probiotic fermentation powder for clearing intestines and protecting liver was given once a day. Experimental Group 4 The basic feed was fed freely, and magnesium sulfate was prepared into a 50 mg / mL magnesium sulfate aqueous solution, and each mouse was given 0.2 mL / 10 g of magnesium sulfate aqueous solution by gavage once a day. The basic feed was fed freely, and the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 4 was added, at a dosage of 10 mg / 10 g of mouse body weight, and the probiotic fermentation powder for clearing intestines and protecting liver was given once a day. Table 4 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% Mouse hangover test: Healthy SPF Kunming mice were randomly divided into 6 groups, with 10 mice in each group, 5 males and 5 females, and kept in separate cages. The mice were adaptively raised in a barrier environment for one week, with free access to food and water, and the bedding was changed every day. The experiment began after 7 days of adaptive feeding. The experimental period was 25 days, and the grouping and treatment methods were as follows: the liquor used in the experiment was the commercially available 52° Langjiu, as shown in Table 5 for details. The levels of alanine aminotransferase AST and aspartate aminotransferase ALT in mice of different groups are shown in Table 6 for details: Table 5 Group Treatment Normal group The mice were fed with basic feed freely, and each mouse was given 0.1 mL / 10 g of normal saline by gavage once every 12 h a day. On the 25th day, after the first administration of 0.1 mL / 10 g of normal saline, the blood was collected after fasting for 12 h. Control group The mice were fed with basic feed freely, and each mouse was given 0.1 mL / 10 g of liquor by oral gavage once every 12 hours a day. On the 25th day, after the first administration of 0.1 mL / 10 g of liquor, the mice were fasted for 12 hours and blood was collected. Experimental Group 1 The basic feed was fed freely, and each mouse was given 0.1 mL of liquor / 10 g of mouse body weight by gavage once every 12 hours a day; the probiotic fermentation powder for clearing intestines and protecting liver prepared in Example 1 was added, and the probiotic fermentation powder for clearing intestines and protecting liver was given at a dosage of 10 mg / 10 g of mouse body weight, once a day. On the 25th day, after the first administration of 0.1 mL of liquor / 10 g of mouse body weight, blood was collected after fasting for 12 hours. Experimental Group 2 The basic feed was fed freely, and each mouse was given 0.1 mL / 10 g of white wine by gavage once every 12 hours a day; the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 2 was added, and the probiotic fermentation powder for clearing intestines and protecting liver was given at a dosage of 10 mg / 10 g of mouse body weight, once a day. After the first administration of 0.1 mL / 10 g of white wine on the 25th day, blood was collected after fasting for 12 hours. Experimental Group 3 The basic feed was freely fed, and each mouse was given 0.1 mL / 10 g of white wine by gavage once every 12 hours a day; the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 3 was added, and the probiotic fermentation powder for clearing intestines and protecting liver was given at a dosage of 10 mg / 10 g of mouse body weight, once a day. After the first administration of 0.1 mL / 10 g of white wine on the 25th day, blood was collected after fasting for 12 hours. Experimental Group 4 The basic feed was fed freely, and each mouse was given 0.1 mL / 10 g of white wine by gavage, once every 12 hours a day; the probiotic fermentation powder for clearing intestines and protecting liver prepared in Comparative Example 4 was added, and the probiotic fermentation powder for clearing intestines and protecting liver was given at a dosage of 10 mg / 10 g of mouse body weight, once a day. After the first administration of 0.1 mL / 10 g of white wine on the 25th day, blood was collected after fasting for 12 hours. Table 6 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 Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that 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 capable of 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: Inoculate Kluyveromyces marxianus K11 into the fermentation medium for aerobic fermentation, control the fermentation temperature to 26°C-40°C and ferment for 1-2 hours; inoculate Weizmannella coagulans JS518 into the fermentation medium for microaerobic fermentation, control the fermentation temperature to 30°C-37°C and ferment for 2-4 hours; inoculate Lactobacillus paracasei JS12 into the culture medium for microaerobic fermentation, control the fermentation temperature to 35°C-40°C and fermentation time to 2-3 hours; after all strains are inoculated, perform alternating fermentation according to a predetermined fermentation cycle: aerobic fermentation for 1 hour, microaerobic fermentation for 3 hours, anaerobic fermentation for 1 hour, and perform 4 cycles in total 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 a pre-configured microbial protective agent, 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. Finally, freeze-drying is performed to obtain a probiotic fermentation product powder that can cleanse the intestines and protect the liver.

2. The method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver according to claim 1, characterized in that: 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.

3. The method for preparing a probiotic fermentation powder capable of 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.

4. The method for preparing a probiotic fermentation powder capable of 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.

5. The method for preparing a probiotic fermentation powder capable of 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.

6. The method for preparing a probiotic fermentation powder capable of clearing intestines and protecting liver according to claim 1, characterized in that: The emulsifier is lecithin or polysorbate.

7. The method for preparing a probiotic fermentation powder capable of clearing intestines 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.

8. A probiotic fermentation powder capable of clearing intestines and protecting liver, characterized in that: The powder is prepared by the method for preparing the probiotic fermentation product powder capable of clearing intestine and protecting liver as described in any one of claims 1 to 7.