A kind of Pediococcus pentosaceus ZKPp0412, Pueraria root fermentation liquid and its application in preparing alcohol sobering liver protection product

Abstract

The present invention relates to the field of microbial fermentation technology, and in particular to a Pediococcus pentosaceus ZKPp0412, Pueraria lobata fermentation liquid and their application in preparing alcohol sobering and liver protecting products. Pediococcus pentosaceus )ZKPp0412 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on June 21, 2023, with the deposit number CGMCC No.27674, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The Pueraria lobata fermentation liquid prepared by fermenting the Pueraria lobata extract by this strain has a lower pH and a higher number of viable bacteria, and can promote the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase in vitro. In vivo experiments have verified that the Pueraria lobata fermentation liquid has the effect of sobering up and sobering up drunk mice, and can effectively inhibit alcohol-induced liver enlargement / liver damage.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fermentation, and in particular to Pediococcus pentosaceus ZKPp0412, kudzu root fermentation liquid and applications thereof in preparing alcohol sobering and liver protecting products. Background Art

[0002] Long-term excessive drinking will have adverse effects on the human liver, stomach, kidneys, etc. After being absorbed by the intestines, ethanol is transported to the liver through the inferior vena cava. About 90% of the ethanol is mainly metabolized in the liver, and the other 10% of the ethanol is directly excreted from the body through breathing and urine. In the liver, ethanol is first converted into acetaldehyde in the presence of catalysts alcohol dehydrogenase (ADH) and cytochrome P4502E1 (CYP2E1), and then converted into acetic acid through the action of acetaldehyde dehydrogenase (ALDH), and finally generates carbon dioxide and water through the tricarboxylic acid cycle. The complete metabolism of ethanol mainly requires the catalysis of two enzymes ADH and ALDH. Therefore, increasing the activity of ADH and ALDH can accelerate the metabolism of alcohol and achieve the purpose of sobering up.

[0003] Alcohol is mainly metabolized by the liver, so excessive drinking will put a heavy burden on the liver, characterized by fatty liver in the early stage, and then develop into alcoholic hepatitis, fibrosis, cirrhosis, and even liver cancer. Studies have found that about 2 million people die from liver disease every year worldwide. Therefore, it is of great significance to find substances or methods to effectively reduce the harm of alcohol.

[0004] Pueraria root is the dried root of the leguminous plant Pueraria lobata. It was first included in the "Shennong Herbal Classic" and listed as a medium-grade product. It is said that it "can relieve thirst, fever, vomiting, various arthralgias, raise yin qi, and relieve various poisons". Pueraria root has a long history of being able to relieve alcohol. The "Food Therapy Materia Medica" records: "Steaming Pueraria root can eliminate alcohol poisoning." The alcohol-relieving effect of Pueraria root is clearly recorded in traditional medical works such as "Ben Jing", "Yao Xing Lun", "Ben Cao Shi Yi", and "Qian Jin Fang". For drunk people, take 30-60g of Pueraria root, decoct in water, and the alcohol-relieving effect is very good. According to the 2010 edition of the "Pharmacopoeia of the People's Republic of China", Pueraria root is cool in nature, sweet and spicy in taste, and has the effects of relieving muscles and fever, promoting body fluid, clearing rashes, and raising yang to stop diarrhea. In actual application, the effective ingredients in Pueraria root water are limited, the alcohol-relieving effect is relatively weak, and improper use is prone to side effects.

[0005] Traditional Chinese medicine fermentation is a processing method based on the action of microorganisms under appropriate temperature, water activity and humidity conditions. It can improve its original properties and / or produce new effects, and expand its application range to meet clinical needs. Traditional Chinese medicine fermentation has many advantages: (1) reducing the toxicity of traditional Chinese medicine; (2) improving the efficacy of traditional Chinese medicine; (3) generating new active ingredients; (4) saving traditional Chinese medicine resources. Pediococcus pentosaceus belongs to the Streptococcus family and the Pediococcus genus, a Gram-positive bacterium. Pediococcus pentosaceus can ferment a variety of sugar sources, producing acid without gas, and has good fermentation ability. It is often used as a fermentation agent for various vegetables, dairy products and meat products. It can metabolize and produce a variety of natural antibacterial substances such as organic acids, bacteriocins, hydrogen peroxide and diacetyl, which have a great promoting effect on human health. Pediococcus pentosaceus has excellent fermentation ability, but there is currently little information on its application in fermented traditional Chinese medicines with medicinal and edible properties and in alcohol-relieving and liver-protecting products. Summary of the invention

[0006] Aiming at the problem that the existing kudzu root water has limited alcohol sobering effect and cannot effectively alleviate the liver damage caused by alcohol, the present invention provides a Pediococcus pentosaceus ZKPp0412, kudzu root fermentation liquid and application thereof in preparing alcohol sobering and liver protecting products.

[0007] In a first aspect, the present invention provides a Pediococcus pentosaceus ZKPp0412, Pediococcus pentosaceus ( Pediococcus pentosaceus ) ZKPp0412 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on June 21, 2023, with the deposit number CGMCC No.27674, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0008] In a second aspect, the present invention provides a kudzu root fermentation liquid, which is obtained by fermenting the kudzu root extract with the above-mentioned Pediococcus pentosaceus ZKPp0412.

[0009] Furthermore, the preparation method specifically comprises the following steps:

[0010] (1) preparing a kudzu root extract, adding yeast extract powder to the kudzu root extract, and sterilizing;

[0011] (2) Inoculating Pediococcus pentosaceus ZKPp0412 into MRS liquid culture medium, culturing and activating the culture medium to the third generation, and preparing a suspension of Pediococcus pentosaceus ZKPp0412;

[0012] (3) Inoculate the bacterial suspension of Pediococcus pentosaceus ZKPp0412 into the sterilized Pueraria extract, shake well, and ferment to obtain Pueraria fermentation liquid.

[0013] Furthermore, in step (1), the preparation method of Pueraria extract is as follows:

[0014] Take kudzu root and add 10 times the amount of water to soak for 1 hour, and collect the soaking water; put the soaked kudzu root into a traditional Chinese medicine cooking pot, add the soaking water, first boil it over high heat, and then simmer it over low heat for 90 minutes; filter and separate the liquid, and cook and concentrate it to a crude drug content of 1.0 g / mL to obtain a kudzu root extract.

[0015] Furthermore, in step (1), the amount of yeast extract powder is 4% to 6% by mass volume.

[0016] Furthermore, in step (1), the sterilization temperature is 121° C. and the sterilization time is 15 min.

[0017] Furthermore, in step (3), the inoculation amount of the Pediococcus pentosaceus ZKPp0412 bacterial suspension is 2% to 4% by mass.

[0018] Furthermore, in step (3), the fermentation culture conditions are: culture in a constant temperature incubator at 35-37°C for 24-48 hours.

[0019] In a third aspect, the present invention also provides an application of the Pueraria lobata fermented liquid in the preparation of alcohol sobering and liver protecting products.

[0020] Furthermore, the alcohol sobering and liver protecting product is a food, a health product, a medicine or a medical raw material.

[0021] Furthermore, alcohol sobering and liver protecting products are products that inhibit liver enlargement / liver damage caused by alcohol.

[0022] The beneficial effects of the present invention are:

[0023] 1. The Pediococcus pentosaceus ZKPp0412 provided by the present invention has a good fermentation effect on Pueraria lobata extract. The Pueraria lobata fermentation liquid obtained by fermentation has a lower pH and a higher number of viable bacteria, and can promote the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase in vitro.

[0024] 2. The Pueraria root fermentation liquid provided by the present invention has the effect of sobering up and sobering up drunk mice through in vivo experiments, and can effectively inhibit liver enlargement / liver damage caused by alcohol. The results show that the Pueraria root fermentation liquid prepared by fermenting the Pueraria root extract by Pediococcus pentosaceus ZKPp0412 can significantly reduce the drunkenness rate ( P <0.05), significantly shortened sleep time ( P <0.05); significantly reduced liver index ( P <0.05), significantly reduced AST and ALT activities ( P <0.05). BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0026] Figure 1 This is a colony morphology diagram of strain ZKPp0412 in Example 1 of the present application.

[0027] Figure 2 This is a microscopic examination result of strain ZKPp0412 after Gram staining in Example 1 of the present application. DETAILED DESCRIPTION

[0028] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0029] Example 1 Strain Isolation, Purification and Identification

[0030] 1. Sample source: Naturally fermented fruit wine from Aksu region, Xinjiang Uygur Autonomous Region in December 2012.

[0031] 2. Strain isolation: dilute the collected samples with physiological saline to 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 times, and 100 µL of the dilution was evenly applied on the MRS-CaCO 3 In solid culture medium, invert and culture at 37℃ for 24~48h, and observe the growth of colonies. After the colonies are formed, pick single colonies with calcium rings and perform three-zone streaking three times, and repeat streaking separation many times until the colonies are purified. Inoculate the purified strains into MRS liquid culture medium for culture, take the culture fluid of different strains and 50% sterile glycerol at a ratio of 1:1 (v / v), and store them in a -80℃ refrigerator.

[0032] 3. Identification of strains

[0033] (1) Colony morphology identification

[0034] ① Culture the isolated strain on the enrichment medium plate for 48 hours, and observe and record the colony morphology and color. Figure 1 As shown, the colonies are round, milky white, smooth, slightly raised, with neat edges and opaque.

[0035] ② Gram staining: Pick the colonies on the plate for smearing, fixation, crystal violet primary staining, mordanting, decolorization, washing, safranin counterstaining, drying, and microscopic examination. The microscopic examination results of strain ZKPp0412 are shown in Figure 2 .

[0036] (2) 16S rDNA sequence determination

[0037] ① Extraction of genomic DNA of strains: Take the culture medium of the isolated strain, centrifuge at 5000r / min for 10min, discard the supernatant after centrifugation, collect the pure culture bacteria to be tested, and use a DNA extraction kit (purchased from Beijing Tiangen Biochemical Technology Co., Ltd.) to extract the genomic DNA of the strain, and operate according to the instructions.

[0038] ②PCR amplification

[0039] Based on the conserved region of Lactobacillus 16S rDNA sequence, bacterial universal primers 27F / 1492R were used.

[0040] The primer sequences are as follows:

[0041] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID No. 1);

[0042] 1492R: 5'-GGTTACCTTGTTACGACTT-3' (SEQ ID No. 2).

[0043] The PCR amplification conditions were as follows: pre-denaturation at 94°C for 4 min; denaturation at 94°C for 30 s; annealing at 55°C for 35 s; extension at 72°C for 90 s; after 32 cycles, the final extension was at 72°C for 5 min.

[0044] The PCR product was identified by 1.0% agarose gel electrophoresis, and the amplified product was sequenced. The sequencing work was completed by Beijing Qingke Biotechnology Co., Ltd. Finally, the 16S rDNA sequence of strain ZKPp0412 was obtained as follows:

[0045]

[0046] The sequencing results were compared with the NCBI database by BLAST and a phylogenetic tree was constructed. It was determined that strain ZKPp0412 was a Pediococcus pentosaceus ( Pediococcus pentosaceus ). Therefore, the strain ZKPp0412 was named Pediococcus pentosaceus ZKPp0412, and was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 21, 2023, and was classified and named: Pediococcus pentosaceus Pediococcus pentosaceus, Deposit number: CGMCC No.27674, deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0047] Example 2 Preparation of Pueraria Fermentation Broth

[0048] 1. Reagents and materials: MRS plate culture medium, MRS liquid culture medium, yeast extract powder were all purchased from Beijing Aoboxing Biotechnology Co., Ltd.; Pueraria root was purchased from Henan Yanhuang Pharmacy.

[0049] 2. Preparation method

[0050] (1) Preparation of Pueraria root extract

[0051] Take kudzu root and add 10 times water to soak for 1 hour, and collect the soaking water; put the soaked kudzu root into a traditional Chinese medicine cooking pot, add the soaking water, boil it with high heat first, and then simmer it with low heat for 90 minutes. During the decocting process, stir to prevent the pot from sticking, but avoid opening the lid frequently to prevent the loss of volatile components; filter and separate the liquid, cook and concentrate it to a crude drug content of 1.0 g / mL, and prepare the kudzu root extract.

[0052] The pH of the Pueraria root extract was measured to be 5.12.

[0053] (2) Preparation of Pueraria lobata fermentation liquid

[0054] Pueraria lobata fermentation liquid 1: Add 6% (g / mL) yeast extract powder to the Pueraria lobata extract, sterilize at 121℃ for 15min, and set aside. Inoculate the preserved Pediococcus pentosaceus ZKPp0412 into MRS liquid culture medium, culture at 37℃ for 24h, activate and pass to the third generation, and prepare bacterial suspension. Inoculate the bacterial suspension into the sterilized Pueraria lobata extract at an inoculation amount of 2% (g / g), shake well, and place in a 37℃ constant temperature incubator for 24h to prepare Pueraria lobata fermentation liquid 1.

[0055] Pueraria lobata fermentation liquid II: Add 5% (g / mL) yeast extract powder to the Pueraria lobata extract, sterilize at 121℃ for 15min, and set aside. Inoculate the preserved Pediococcus pentosaceus ZKPp0412 into MRS liquid culture medium, culture at 37℃ for 24h, activate and pass to the third generation, and prepare bacterial suspension. Inoculate the bacterial suspension into the sterilized Pueraria lobata extract at an inoculation amount of 3% (g / g), shake well, and place in a 36℃ constant temperature incubator for 36h to prepare Pueraria lobata fermentation liquid II.

[0056] Pueraria fermentation broth 3: Add 4% (g / mL) yeast extract powder to the Pueraria extract, sterilize at 121℃ for 15min, and set aside. Inoculate the preserved Pediococcus pentosaceus ZKPp0412 into MRS liquid culture medium, culture at 37℃ for 24h, activate and pass to the third generation to make a bacterial suspension. Inoculate the bacterial suspension into the sterilized Pueraria extract at an inoculation amount of 4% (g / g), shake well, and place in a 35℃ constant temperature incubator for 48h to make Pueraria fermentation broth 3.

[0057] The pH and viable bacterial count of the above Pueraria lobata fermentation broths 1, 2 and 3 were measured, and the results were as follows:

[0058] Table 1 pH and viable bacterial count of Pueraria lobata fermentation broth

[0059]

[0060] As shown in Table 1, the pH of the Pueraria lobata fermentation broth prepared according to the above method was between 3.2 and 3.5, and the number of viable bacteria was ≥1.25×10 9 CFU / mL, the fermentation effect is better.

[0061] Example 3 In vitro activation of alcohol dehydrogenase and acetaldehyde dehydrogenase by Pueraria lobata fermentation broth

[0062] 1. Reagent preparation:

[0063] (1) Sample solution 1: Pueraria extract prepared in Example 2, to which a suspension of Pediococcus pentosaceus ZKPp0412 was added, so that the number of viable bacteria in the mixed solution was 1.25×10 9 CFU / mL;

[0064] (2) Sample solution 2: Pueraria lobata fermentation solution 1 prepared in Example 2.

[0065] (3) Sample solution 3: Pueraria lobata fermentation solution 2 prepared in Example 2.

[0066] (4) Sample solution 4: Pueraria lobata fermentation solution 3 prepared in Example 2.

[0067] 2. Detection Method

[0068] (1) Method for detecting the activation rate of alcohol dehydrogenase: 120 μL of sodium pyrophosphate buffer (pH 8.8), 80 μL of 27 mmol / L oxidized coenzyme I (NAD+), and 40 μL of 11.5% ethanol were added to a 96-well ELISA plate in sequence, and then 50 μL of sample solution 1 or sample solution 2 were added (the control group was added with the same volume of distilled water); mixed, incubated at 25°C for 5 min, taken out and immediately added 10 μL of 0.5 U / mL alcohol dehydrogenase, mixed and measured the absorbance at a wavelength of 340 nm, reading once every 10 s, and measured continuously for 5 min. Plot A340 nm against time, measure the enzyme activity by the rate of change of absorbance per minute (△A340 nm / min), and calculate the activation rate according to the following formula.

[0069]

[0070] (2) Method for detecting the activation rate of acetaldehyde dehydrogenase: add 120 μL of 100 mmol / L sodium pyrophosphate buffer (pH=9.5) and 100 μL of 27 mmol / L oxidized coenzyme I (NAD+) to a 96-well ELISA plate; add 10 μL of 1 mmol / L acetaldehyde and 20 μL of 10 mmol / L pyrazole, and 30 μL of sample solution (add the same volume of distilled water to the control group); mix well, incubate at 30°C for 5 min, take out and immediately add 15 μL of 1.5 U / mL ALDH and shake well, measure the absorbance at a wavelength of 340 nm, read once every 1 min, and measure continuously for 10 min. Plot A340 nm against time, plot A340 nm against time, measure the enzyme activity by the rate of change of absorbance per minute (△A340 nm / min), and calculate the activation rate according to the formula in (1).

[0071] Table 2 In vitro activation rates of alcohol dehydrogenase and acetaldehyde dehydrogenase

[0072]

[0073] It can be seen from Table 2 that, compared with the mixture of Pediococcus pentosaceus ZKPp0412 and Pueraria extract, the Pueraria fermentation liquid provided by the present invention has a more significant activation effect on alcohol dehydrogenase and acetaldehyde dehydrogenase in vitro.

[0074] Example 4 Effects of Pueraria lobata fermented liquid on sobering up and sobering up drunken mice

[0075] (1) Experimental groups and drug administration

[0076] A. Experimental groups: 40 male KM mice, with an average age of 6-7 weeks and a body weight of 18-22 g, were randomly divided into a control group, a model group, a Pueraria extract group, and a Pueraria fermented liquid group according to their body weight, with 10 mice in each group.

[0077] B. Experimental administration: After one week of adaptive feeding, the control group and the model group were gavaged with 0.2 mL of sterile saline every day, the Pueraria extract group was gavaged with 0.2 mL of sample solution 1 in Example 3 every day, and the Pueraria fermented liquid group was gavaged with 0.2 mL of sample solution 2 in Example 3 every day, for 2 consecutive weeks. 30 minutes after the last gavage, the mice in the model group, the Pueraria extract group, and the Pueraria fermented liquid group were gavaged with 5 mL / kg of 56% alcohol, and the blank group was gavaged with the same amount of saline. The time of disappearance of righting reflex, recovery time, and sleep time of the mice were observed and recorded.

[0078] (2) Righting reflex test

[0079] After mice were gavaged with 56% alcohol by volume, they were observed for about 20 minutes. When the mice were swaying or slow in walking, they were turned over so that their backs were facing down. If the mice could not turn over to their backs up on their own within 30 seconds and their righting reflex disappeared, they were considered drunk. When the mice could turn over to their backs up on their own and their righting reflex appeared, they were considered sober. The number of mice with righting reflex disappearance, righting reflex disappearance time, and sleep time were observed and recorded. For the righting reflex disappearance time, mice that were not drunk were recorded as 0 minutes, and mice that were not sober after 7 hours were recorded as 420 minutes. For sleep time, sleep time = righting reflex recovery time - righting reflex disappearance time.

[0080] Table 3 The sobering effect of Pueraria lobata fermented liquid on mice with acute alcohol poisoning

[0081]

[0082] Note: * indicates comparison with the model group. P <0.05; ** indicates comparison with the model group, P <0.01.

[0083] As can be seen from Table 3, the Pueraria extract containing Pediococcus pentosaceus ZKPp0412 can significantly reduce the drunkenness rate of mice ( P <0.05), significantly shortened sleep time ( P <0.05); and the Pueraria fermented liquid provided by the present invention can significantly reduce the drunkenness rate ( P <0.01), significantly shortened sleep time ( P <0.01), and the hangover-relieving effect was significantly better than that of Pueraria extract containing Pediococcus pentosaceus ZKPp0412.

[0084] Example 5 Preventive effect of Pueraria lobata fermented liquid on alcoholic liver damage in mice

[0085] (1) Experimental groups and drug administration

[0086] A. Experimental groups: 40 male KM mice, with an average age of 6-7 weeks and a body weight of 18-22 g, were randomly divided into a control group, a model group, a Pueraria extract group, and a Pueraria fermented liquid group according to their body weight, with 10 mice in each group.

[0087] B. Experimental administration: After one week of adaptive feeding, the control group and the model group were gavaged with 0.2 mL of sterile saline every day, the Pueraria extract group was gavaged with 0.2 mL of sample solution 1 in Example 3 every day; the Pueraria fermented liquid group was gavaged with 0.2 mL of sample solution 2 in Example 3 every day for 2 consecutive weeks. After one week of gavage treatment, the mice in the model group, the Pueraria extract group, and the Pueraria fermented liquid group were gavaged with 5 mL / kg of 56% alcohol by volume for one consecutive week.

[0088] (2) Sample collection and index testing

[0089] After fasting for 16 h, blood was collected from the ocular vein of the mice, and the liver of the mice was completely removed and rinsed with saline to remove the accumulated blood. The liver weight was weighed after wiping and stored at -80°C for subsequent experiments.

[0090] A. Liver index: The liver index was calculated according to the following formula. The results are shown in Table 4.

[0091]

[0092] Table 4 Liver index of mice in each group

[0093]

[0094] Note: Different letters in the table represent significant differences. P <0.05.

[0095] When the liver is damaged by alcohol, the cells will swell. At the same time, metabolic disorders are prone to occur during fat metabolism, causing fat to accumulate in liver cells, resulting in an increase in the overall mass and volume of the liver and an abnormal increase in the liver index. As can be seen from Table 4, the liver index of the model group mice was significantly higher than that of the control group ( P <0.05), indicating that drinking alcohol can cause liver enlargement; the liver index of the Pueraria extract group and the Pueraria fermentation liquid group was significantly lower than that of the model group ( P <0.05), indicating that the two groups of samples can inhibit alcohol-induced liver enlargement, and the Pueraria root fermentation liquid group is significantly lower than the Pueraria root extract group ( P <0.05), indicating that the Pueraria root fermented liquid is more effective in inhibiting alcohol-induced hepatomegaly than the unfermented Pueraria root extract.

[0096] B. Serum AST and ALT activities: Blood was collected from mouse eyeballs and centrifuged at 4°C, 3000 r / min for 10 min to obtain serum. The aspartate aminotransferase (AST) kit and alanine aminotransferase (ALT) kit (both purchased from Nanjing Jiancheng Bioengineering Institute) were used to determine AST and ALT activities. The results are shown in Table 5.

[0097] Table 5 AST and ALT activities of mice in each group

[0098]

[0099] Note: Different letters in the same column of data in the table represent significant differences. P <0.05.

[0100] Abnormal liver function caused by alcohol-mediated oxidative stress leads to the release of liver enzymes such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum, lipid peroxidation, and loss of antioxidant function. Therefore, the activity of AST and ALT enzymes can be used to measure the degree of liver function damage. As shown in Table 5, compared with the control group, the activity of ALT and AST in the model group increased significantly ( P <0.05), indicating that alcohol can cause liver damage; the ALT and AST activities in the Pueraria extract group and the Pueraria fermentation liquid group were significantly lower than those in the model group ( P <0.05), indicating that the two groups of samples can inhibit alcohol-induced liver damage, and the Pueraria root fermentation liquid group is significantly lower than the Pueraria root extract group ( P <0.05), indicating that the Pueraria root fermented liquid provided by the present invention has a more significant effect in inhibiting liver damage than the unfermented Pueraria root extract.

[0101] The above results show that fermented Pueraria root extract has a significant effect on sobering up drunken mice and has a significant preventive and protective effect on alcoholic liver damage compared with unfermented Pueraria root extract ( P <0.05).

[0102] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A Pueraria lobata fermented liquid, characterized in that: By Pediococcus pentosaceus ( Pediococcus pentosaceus )ZKPp0412 was fermented with Pueraria extract; Pediococcus pentosaceus ZKPp0412 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on June 21, 2023, with the deposit number CGMCC No.27674, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; The preparation method specifically comprises the following steps: (1) preparing a kudzu root extract, adding yeast extract powder to the kudzu root extract, and sterilizing; (2) Inoculating Pediococcus pentosaceus ZKPp0412 into MRS liquid culture medium, culturing and activating the culture medium to the third generation, and preparing a suspension of Pediococcus pentosaceus ZKPp0412; (3) inoculating the bacterial suspension of Pediococcus pentosaceus ZKPp0412 into the sterilized Pueraria extract, shaking well, and fermenting to obtain Pueraria fermentation liquid; In step (1), the preparation method of Pueraria extract is as follows: Take kudzu root and add 10 times water to soak for 1 hour, collect the soaking water; put the soaked kudzu root into a traditional Chinese medicine cooking pot, add the soaking water, boil it with high heat first, and then simmer it with low heat for 90 minutes; filter and separate the liquid, boil and concentrate it to the crude drug content of 1.0g / mL, and prepare the kudzu root extract; In step (3), the inoculation amount of the Pediococcus pentosaceus ZKPp0412 bacterial suspension is 2% to 4% by mass.

2. The Pueraria lobata fermented liquid according to claim 1, characterized in that: In step (1), the amount of yeast extract powder is 4% to 6% by mass volume.

3. The Pueraria lobata fermented liquid according to claim 1, characterized in that: In step (1), the sterilization temperature is 121°C and the sterilization time is 15 min.

4. The Pueraria lobata fermented liquid according to claim 1, characterized in that: In step (3), the fermentation culture conditions are: culture in a constant temperature incubator at 35-37°C for 24-48 hours.

5. Use of the Pueraria lobata fermented liquid as claimed in claim 1 in preparing a product for sobering up and protecting the liver.

6. The use according to claim 5, characterized in that Alcohol sobering and liver protecting products are products that inhibit liver enlargement / liver damage caused by alcohol.

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