CN112438354A

CN112438354A - Plant fermentation liquor and preparation method and application thereof

Info

Publication number: CN112438354A
Application number: CN202011342693.4A
Authority: CN
Inventors: 夏越; 马明磊; 张锡科; 夏超凡; 王艳; 唐元凤
Original assignee: Shandong Sunflower Bioengineering Co ltd
Current assignee: Shandong Sunflower Bioengineering Co ltd
Priority date: 2020-11-25
Filing date: 2020-11-25
Publication date: 2021-03-05

Abstract

The invention relates to a plant fermentation liquid and a preparation method and application thereof, belonging to the technical field of fermented beverages. The feed comprises the following raw materials in parts by weight: 60-80 parts of purified water, 5-10 parts of carrot juice, 1-5 parts of dried rehmannia root extract, 1-5 parts of Chinese yam extract, 1-5 parts of dogwood extract, 1-3 parts of rhizoma alismatis extract, 1-3 parts of poria cocos extract, 1-3 parts of moutan bark extract, 0.1-1 part of cassia twig extract, 0.1-1 part of radix aconiti lateralis preparata extract, 1-5 parts of walnut juice, 1-5 parts of passion fruit juice, 2-3 parts of L-arabinose, 2-3 parts of honey, 1-5 parts of resistant dextrin, 1-2 parts of xylitol and 0.1-0.3 part of edible essence. The raw materials are firstly subjected to aerobic fermentation and then anaerobic fermentation to obtain plant fermentation liquor. According to the invention, the probiotics are used for fermenting the traditional Chinese medicine, so that the utilization rate of the medicinal materials is improved, the medicine effect is improved, and the taste of the product is better.

Description

Plant fermentation liquor and preparation method and application thereof

Technical Field

The invention belongs to the technical field of fermented beverages, and particularly relates to a plant fermentation liquid, and a preparation method and application thereof.

Background

Nowadays, the present society has great competitive pressure, great mental stress, tiredness, and many people have the symptoms of mental disorder, soreness and weakness of waist and knees, weakness of limbs, alopecia, amnesia, dull and lusterless complexion, which are all the manifestations of kidney deficiency. Kidney deficiency is usually caused by long-term accumulation, and can not be used for tonifying with the herbs of tonifying in the extreme so as to obtain the result promptly. The traditional Chinese medicine considers that the kidney has the functions of storing essence, governing water, governing bone, generating marrow, receiving qi and the like, particularly the function of storing essence of the kidney is closely related to the growth, development, reproduction and the like of human, so the kidney is called as the "congenital foundation".

At present, kidney tonifying products on the market have traditional kidney tonifying and body strengthening tablets, kidney tonifying and blood nourishing soup, five-seed kidney tonifying pills, kidney qi pills and other classic traditional Chinese medicine formulas, but the utilization rate of medicinal materials is low, and the taste is not good. Researches show that a certain synergistic effect exists between probiotics and traditional Chinese medicines, on one hand, the probiotics can promote the absorption and utilization of the traditional Chinese medicines, and a plurality of traditional Chinese medicines can be selectively absorbed by intestinal tracts after being processed and modified by microbial flora in digestive tracts; on the other hand, the effective chemical components of alkaloid, glycosides, volatile oil and tannin contained in the traditional Chinese medicine can sweep out obstacle for the proliferation of probiotics; the oil, resin, sugar, protein, pigment and the like provide nutrition for the probiotics, and some of the probiotics even directly promote the proliferation of microorganisms.

The prior patent CN109045105A relates to a kidney-tonifying traditional Chinese medicine composition for both food and medicine, which can be used as a medicine and food for nursing the body in life, but has poor taste and low utilization rate of medicinal materials; patent CN107019211A relates to a fermented composition of plant ferment with kidney tonifying and strengthening functions, which is suitable for industrial production and home production of various plant ferments, but further processed; patent CN107468806A relates to a kidney-tonifying traditional Chinese medicine composition, decoction and freeze-dried powder thereof, which can tonify kidney and strengthen yang and have stronger pertinence to applicable population. Aiming at the defects, the applicant focuses on research to improve the utilization rate of medicinal materials and the drug effect, so that the product has better taste.

Disclosure of Invention

Aiming at the problems of low utilization rate of medicinal materials of kidney tonifying products, poor taste and the like in the prior art, the invention provides plant fermentation liquor and a preparation method and application thereof, so as to solve the problems. According to the invention, probiotics are used for fermenting traditional Chinese medicines, so that the utilization rate of medicinal materials is improved, the drug effect is improved, and the taste of the product is better.

A plant fermentation liquid comprises the following raw materials in parts by weight: 60-80 parts of purified water, 5-10 parts of carrot juice, 1-5 parts of dried rehmannia root extract, 1-5 parts of Chinese yam extract, 1-5 parts of dogwood extract, 1-3 parts of rhizoma alismatis extract, 1-3 parts of poria cocos extract, 1-3 parts of moutan bark extract, 0.1-1 part of cassia twig extract, 0.1-1 part of radix aconiti lateralis preparata extract, 1-5 parts of walnut juice, 1-5 parts of passion fruit juice, 2-3 parts of L-arabinose, 2-3 parts of honey, 1-5 parts of resistant dextrin, 1-2 parts of xylitol and 0.1-0.3 part of edible essence.

Preferably, the feed comprises the following raw materials in parts by weight: 70 parts of purified water, 5 parts of carrot juice, 4 parts of dried rehmannia root extract, 2 parts of Chinese yam extract, 2 parts of dogwood extract, 1.5 parts of rhizoma alismatis extract, 1.5 parts of poria cocos extract, 1.5 parts of moutan bark extract, 0.5 part of cassia twig extract, 0.5 part of processed monkshood extract, 5 parts of walnut juice, 5 parts of passion fruit juice, 2 parts of L-arabinose, 2 parts of honey, 5 parts of resistant dextrin, 2 parts of xylitol and 0.3 part of edible essence.

A preparation method of plant fermentation liquor comprises the following steps:

(1) squeezing carrot, passion fruit and walnut respectively for standby, then respectively crushing dried rehmannia root, Chinese yam, dogwood, rhizoma alismatis, poria cocos, tree peony bark, cassia twig and processed monkshood, adding water 1-3 cm higher than the medicinal powder, and cooking to obtain extracts of various medicines. The aim of the step is to extract the effective medicinal components in the traditional Chinese medicine.

(2) Mixing the carrot juice, passion fruit juice, walnut juice, dried rehmannia root extract, Chinese yam extract, dogwood fruit extract, rhizoma alismatis extract, poria cocos extract, moutan bark extract, cassia twig extract and radix aconiti praeparata extract prepared in the step (1) with L-arabinose, honey, resistant dextrin, xylitol and edible essence according to parts by weight to obtain mixed slurry;

(3) conveying the mixed slurry prepared in the step (2) to a sterilization container, performing high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30-40 ℃ to obtain a sterilized solution;

(4) preparing an aerobic probiotic strain into a leaven I according to the proportion that the weight part of the aerobic probiotic strain is 0.001-0.01, and the total viable count is higher than 100 hundred million/g;

(5) inoculating the starter I prepared in the step (4) into the sterilized solution prepared in the step (3), and obtaining a fermentation broth I through a fermentation process;

(6) preparing an anaerobic probiotic strain into a leavening agent II according to the proportion that the weight part of the anaerobic probiotic strain is 0.001-0.01 and the total viable count is higher than 100 hundred million/g;

(7) after the fermentation in the step (5) is finished, inoculating the leavening agent II prepared in the step (6) into the fermentation liquor I prepared in the step (5), and obtaining fermentation liquor II through the fermentation process;

(8) and (5) after the fermentation in the step (7) is finished, conveying the prepared fermentation liquor II into a high-pressure spraying wall breaking machine, setting the pressure to be 20-60 MPa, and performing pasteurization on the fermentation liquor II after wall breaking to obtain the plant fermentation liquor. The beverage of the invention is liquid and can be directly drunk.

Preferably, the aerobic probiotics in the step (4) is saccharomyces cerevisiae.

Preferably, the inoculation amount of the leavening agent I in the step (5) is 500-1000 ten thousand cfu/ml, the fermentation temperature is 37 ℃, and the fermentation time is 24 h.

Preferably, the anaerobic probiotics in step (6) at least comprise bifidobacterium lactis, and one or more of lactobacillus casei, lactobacillus plantarum and bifidobacterium longum can be used on the basis of the anaerobic probiotics.

The Bifidobacterium lactis is Bifidobacterium animalis subsp.lactis SF-B-20, is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.20924, the preservation date is 2020, 10 and 20 days, and the address of a preservation institution: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.

Preferably, the anaerobic probiotic is bifidobacterium lactis: lactobacillus casei: lactobacillus plantarum: bifidobacterium longum is a mixed bacterial agent with the ratio of 1:1:1: 1.

Preferably, the inoculation amount of the leavening agent II in the step (7) is 500-1000 ten thousand cfu/ml, the fermentation temperature is 37 ℃, and the fermentation time is 72 hours.

The traditional Chinese medicines adopted by the invention are as follows:

carrot: nature and taste: ping, sweet. Tonify liver, improve vision, clear heat and remove toxicity.

Dried rehmannia root: cold in nature and sweet in taste. Clear heat and cool blood, nourish yin, promote the production of body fluid.

Chinese yam: mild in nature and sweet in taste. Tonify spleen and stomach, promote the production of body fluid and nourish lung, tonify kidney and astringe essence.

Dogwood fruit: slightly warm in nature, sour and astringent in taste. Tonify liver and kidney, astringe and induce depletion.

Rhizoma alismatis: cold in nature, sweet and bland in flavor. Induce diuresis, clear damp-heat.

Tuckahoe, poria cocos: mild in nature, sweet and bland in flavor. Detoxication, diuresis, and joint movement.

Moutan bark: slightly cold in nature, bitter and pungent in flavor. Clearing heat and cooling blood, and promoting blood circulation for removing blood stasis.

Cassia twig: pungent and sweet in flavor and warm in nature. Relieving exterior syndrome, expelling pathogenic factors from muscles, warming meridians, activating collaterals, supporting yang to regulate qi, and calming the adverse-rising energy.

Processing monkshood: hot in nature, pungent and sweet in flavor and with strong toxicity. Reviving yang and rescuing from collapse, tonifying fire and supporting yang, dispelling cold and relieving pain.

Passion fruit: sweet in taste; an acid; the nature is mild. Clearing lung and moistening dryness; tranquilization and pain relief; and stopping dysentery.

Walnut: warm in nature and sweet in taste. Warming and invigorating lung and kidney, relieving asthma, eliminating phlegm, moistening intestine, and arresting seminal emission.

The plant fermentation liquid is applied to preparing the kidney tonifying drink.

The invention has the beneficial effects that:

(1) the raw materials selected by the plant fermentation liquid are all medicinal and edible raw materials, and the raw materials can be taken for a long time through a large number of clinical tests. The compatibility of the recipe is characterized in that yin qi is treated to activate yang, so that yang is transformed into fire slightly, and qi is generated by less fire, so as to warm and tonify kidney qi. The beverage of the invention warms and tonifies kidney qi, fixes essence and supplements marrow, strengthens body and enhances the resistance of human body.

(2) The invention is fermented by microorganism, has no toxic and harmful components, is rich in various probiotics and metabolites, effectively regulates the intestinal micro-ecology of human body, and promotes the absorption of functional components of traditional Chinese medicine.

(3) The invention is treated by low-temperature high-pressure wall breaking technology, the cell is completely exploded in a physical mode, the gene sequence of the cell and the activity of effective substances are effectively preserved, and the released effective substances are only one to three thousandths of microbial cells and are easier to be absorbed by human bodies.

Detailed Description

In order to make those skilled in the art 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, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

Bifidobacterium lactis SF-B-20 strain screening

(1) The strain source is as follows: feces of kidney deficiency patient

(2) Plate culture medium: 10.0g of beef extract, 5.0g of yeast extract powder, 10.0g of peptone, 20.0g of glucose and K₂HPO₄2.0g, sodium acetate 5.0g, MgSO₄·7H₂O 0.2g，MnSO₄·4H₂O0.1 g, Tween 801.0 g, ammonium citrate 2.0g, agar 15.0g, and distilled water 1000 ml.

(3) Isolation of the Strain

Ten-fold gradient dilution is carried out on the sample liquid, 100 mu L of each gradient is uniformly coated on MRS solid plate culture medium, and then the sample liquid is taken out and placed in an anaerobic tank to be anaerobically cultured overnight at 37 ℃. The next day, colonies with different shapes and sizes grow on the plate, a plurality of characteristic colonies of lactic acid bacteria are selected for the second-generation streak purification treatment, the plate is subjected to anaerobic activation for 1 day to obtain a culture, and then the strain identification is carried out.

(4) Determination of 16S rRNA sequence

The 16S rRNA amplification primers used were 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' and 1492R: 5'-GGTTACCTTGTTACGACTT-3' are provided. The product was commissioned for Sanger sequencing in a qualified third party laboratory. The sequence was compared to the 16 srna gene in the GenBank database using the BLAsT algorithm. BLAST analysis showed that the strain SF-B-20 is very close to Bifidobacterium lactis (99% relativity).

The result of the detection

CACCTTAGACGGCTCCCCCCACAAGGGTCGGGCCACCGGCTTCGGGTGCTACCCACTTTCATGACTTGACGGGCGGTGTGTACAAGGCCCGGGAACGCATTCACCGCGGCGTTGCTGATCCGCGATTACTAGCGACTCCGCCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGACCGGTTTTCAGCGATCCGCCCCACGTCACCGTGTCGCACCGCGTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACGTAAGGGGCATGATGATCTGACGTCATCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCCCACATGAGTTCCCGGCATCACCCGCTGGCAACATGCGGCGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTGAACCGGCCCCGAAGGGAAACCGTGTCTCCACGGCGATCCGGCACATGTCAAGCCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGATGCTTAACGCGTTGGCTCCGACACGGGACCCGTGGAAAGGGCCCCACATCCAGCATCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTGACGGCCCAGAGACCTGCCTTCGCCATTGGTGTTCTTCCCGATATCTACACATTCCACCGTTACACCGGGAATTCCAGTCTCCCCTACCGCACTCCAGCCCGCCCGTACCCGGCGCAGATCCACCGTTAGGCGATGGACTTTCACACCGGACGCGACGAACCGCCTACGAGCCCTTTACGCCCAATAAATCCGGATAACGCTCGCACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCGAACAATCCACTCAACACGGCCGAAACCGTGCCTTGCCCTTGAACAAAAGCGGTTTACAACCCGAAGGCCTCCATCCCGCACGCGGCGTCGCTGCATCAGGCTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTATCTCAGTCCCAATGTGGCCGGTCACCCTCTCAGGCCGGCTACCCGTCAACGCCTTGGTGGGCCATCACCCCGCCAACAAGCTGATAGGACGCGACCCCATCCCATGCCGCAAAAGCATTTCCCACCCCACCATGCGATGGAGCGGAGCATCCGGTATTACCACCCGTTTCCAGGAGCTATTCCGGTGCACAGGGCAGGTTGGTCACGCATTACTCACCCGTTCGCCACTCTCACCC

The strain is preserved in the common microorganism center of the national microorganism culture preservation and management committee, the preservation number is CGMCC No.20924, and the preservation date is 10 months and 21 days in 2020. The address of the depository institution: the microbial research institute of western road 1 institute of Chinese academy of sciences, north jing, chaoyang district, zip code: 100101, telephone: 86-10-64807355.

Example 2

Preparation of plant fermentation broth

(1) Squeezing carrot, passion fruit and walnut respectively for standby, crushing dried rehmannia root, Chinese yam, dogwood, rhizoma alismatis, poria cocos, tree peony bark, cassia twig and processed monkshood, and adding water 1-3 cm higher than the medicinal powder respectively for cooking to prepare extracts of various medicines;

(2) mixing 5 parts of carrot juice, 5 parts of passion fruit juice, 5 parts of walnut juice, 4 parts of rehmanniae extract, 2 parts of yam extract, 2 parts of dogwood extract, 1.5 parts of rhizoma alismatis extract, 1.5 parts of poria cocos extract, 1.5 parts of moutan bark extract, 0.5 part of cassia twig extract, 0.5 part of rhizoma typhonii extract, 2 parts of L-arabinose, 2 parts of honey, 5 parts of resistant dextrin, 2 parts of xylitol and 0.3 part of edible essence according to parts by weight to obtain mixed slurry;

(4) preparing a saccharomyces cerevisiae strain (a commercially available strain) into a leavening agent I according to the proportion that the weight part of the saccharomyces cerevisiae strain is 0.001 part and the total viable count is higher than 100 hundred million/g;

(5) inoculating the starter I prepared in the step (4) into the sterilized solution prepared in the step (3), wherein the inoculation amount is 1000 ten thousand cfu/ml, the fermentation temperature is 37 ℃, and the fermentation time is 24 hours, so as to obtain fermentation liquor I;

(6) lactobacillus casei SF-L-12: bifidobacterium lactis SF-B-20: lactobacillus plantarum SF-L-28: 0.01 part of mixed strain of bifidobacterium longum SF-B-27: 1:1:1 by weight and the proportion that the total viable count is higher than 100 hundred million/g are prepared into a leaven II;

the lactobacillus casei SF-L-12 is preserved in the common microorganism center of the national microorganism culture preservation and management committee, the preservation number is CGMCC No.20927, and the preservation date is 10 months and 21 days in 2020. The address of the depository institution: the microbial research institute of western road 1 institute of Chinese academy of sciences, north jing, chaoyang district, zip code: 100101, telephone: 86-10-64807355;

the lactobacillus plantarum SF-L-28 is preserved in the common microorganism center of the national microorganism culture preservation and management committee, the preservation number is CGMCC No.20925, and the preservation date is 10 months and 21 days in 2020. The address of the depository institution: the microbial research institute of western road 1 institute of Chinese academy of sciences, north jing, chaoyang district, zip code: 100101, telephone: 86-10-64807355;

the bifidobacterium longum SF-B-27 is preserved in the common microorganism center of the national microorganism culture preservation and management committee, the preservation number is CGMCC No.20926, and the preservation date is 10 months and 21 days in 2020. The address of the depository institution: the microbial research institute of western road 1 institute of Chinese academy of sciences, north jing, chaoyang district, zip code: 100101, telephone: 86-10-64807355;

(7) after the fermentation in the step (5) is finished, inoculating the leavening agent II prepared in the step (6) into the fermentation liquid I prepared in the step (5), wherein the inoculation amount is 500 ten thousand cfu/ml, the fermentation temperature is 37 ℃, and the fermentation time is 72 hours, so as to obtain the fermentation liquid II;

(8) and (5) after the fermentation in the step (7) is finished, conveying the prepared fermentation liquor II into a high-pressure spraying wall breaking machine, setting the pressure to be 20MPa, and performing pasteurization on the fermentation liquor II after wall breaking to obtain the plant fermentation liquor.

Example 3

Comparative experiment on effects of plant fermentation liquid prepared in example 2 of the invention

The kidney-tonifying plant fermented beverage prepared in example 2 was compared with decoction prepared by a conventional method and fermented beverage prepared by bifidobacterium lactis HN019 on the market by the same process. The protective effect of the plant fermentation beverage and the control sample prepared by the embodiment of the invention on kidney injury is researched by preparing a kidney disease model by injecting puromycin aminonucleoside into the tail vein of an SD mouse.

Firstly, Chinese herbal medicines are decocted according to the proportion of 4 parts of dried rehmannia root, 2 parts of Chinese yam, 2 parts of dogwood, 1.5 parts of oriental waterplantain rhizome, 1.5 parts of tuckahoe, 1.5 parts of tree peony bark, 0.5 part of cassia twig and 0.5 part of processed aconite by the tradition to obtain decoction as a traditional Chinese medicine.

And then, the plant fermented beverage obtained by replacing the bifidobacterium lactis SF-B-20 with the bifidobacterium lactis HN019 by the same implementation method of the invention is used as a comparison group.

1. Experimental methods

1. SD mice were randomly divided into 5 groups: control group, model group, traditional Chinese medicine group, experimental group and comparison group, 10 of them are used. Wherein, the control group is not treated, and distilled water with the same volume is infused every day after the experiment begins; the other groups were made into renal disease models by tail vein injection of puromycin aminonucleosides. The model group was not treated with drugs after modeling, and each mouse was drenched with distilled water every day; the Chinese herbal medicine group drenches Chinese herbal medicine decoction (200mg/kg) to each mouse every day within 14 days after modeling; experimental groups the fermented plant drink prepared in example 2 (200mg/kg) was drenched to each mouse daily for 14 days after modeling; the control group drenched bifidobacterium lactis HN019 plant fermented drink (200mg/kg) to each mouse every day within 14 days after modeling; keep the other living environments of the mice consistent.

2. Urine protein creatinine ratio detection 24 hour urine from mice was collected on experiment day 13, protein quantification and urine creatinine were measured, and the urine protein creatinine ratio was calculated.

3. Carrying out biochemical index detection on each group of mice 14 days after modeling, injecting 33mg/100g chloral hydrate into the abdominal cavity of an anesthetized mouse, opening the abdominal cavity under aseptic condition, collecting blood from the abdominal aorta, standing for 2h and then centrifuging. And detecting serum creatinine, urea nitrogen, uric acid, albumin, total cholesterol and triglyceride according to the steps of the kit specification.

The experimental data are analyzed by SPSS18.0 software, and the measured data are averaged plus or minus standard deviation

The group comparisons conforming to the normal distribution were performed by one-way analysis of variance, and the abnormal distribution and the measurement data were subjected to rank test at a test level α of 0.05.

Table 1 comparison of urine protein creatinine ratios for each group of mice (mg/g,

)

group of	Number of mice	Urinary protein creatinine ratio
			Control group	9	308.09±54.89
Building module	10	2550.15±469.75^A
			Experimental group	10	625.84±321.52^B
Chinese medicine	10	723.45±140.36^B
			Comparison group	9	793.45±140.36^B

Note: a <0.05 compared to normal group; b compares P <0.05 to the modeled group.

TABLE 2 serum biochemical indices of various groups of mice

Note: a <0.05 compared to control; b <0.05 compared to the modeling group; c P <0.05 compared to experimental group.

2. Results of the experiment

1 mouse in the control group died, mainly caused death by improper operation during drug administration; the control group mice died 1, mainly under anesthesia, due to dose-failure.

Table 1 shows that the urine protein creatinine ratio of the building block group is significantly increased (P <0.05) compared to the control group; compared with the modeling group, the urine protein creatinine ratio of the traditional Chinese medicine group, the experimental group and the comparative group is obviously reduced (P is less than 0.05). The ratios of the experimental group, the traditional Chinese medicine group and the comparative group protein creatinine are not significantly different (P >0.05), but the ratio of the experimental group to the traditional Chinese medicine group and the comparative group protein creatinine is lower. The traditional Chinese medicine decoction, the plant fermented beverage and the bifidobacterium lactis HN019 plant fermented beverage can reduce the ratio of urine protein creatinine caused by puromycin amino nucleoside, and the effect of the plant fermented beverage is better.

Table 2 shows that, according to the biochemical indexes of albumin in each group, compared with the control group, the modeling group is obviously increased (P <0.05), the experimental group has no significant difference (P >0.05), and the traditional Chinese medicine group and the control group are obviously decreased (P < 0.05); compared with the modeling group, the experimental group, the traditional Chinese medicine group and the control group are obviously increased (P is less than 0.05). According to the biochemical indexes of urea nitrogen of each group, compared with a control group, the modeling group is obviously increased (P is less than 0.05), and the experimental group, the traditional Chinese medicine group and the control group have no significant difference (P is more than 0.05); compared with the modeling group, the experimental group, the traditional Chinese medicine group and the control group are obviously increased (P is less than 0.05). There was no significant difference between groups according to biochemical indices of creatinine in each group (P > 0.05). According to biochemical indexes of uric acid and total cholesterol in each group, compared with a control group, the modeling group is obviously increased (P is less than 0.05), and the experimental group, the traditional Chinese medicine group and the comparison group have no significant difference (P is more than 0.05); compared with the modeling group, the experimental group, the traditional Chinese medicine group and the comparison group are obviously reduced (P is less than 0.05). According to the biochemical indexes of triglyceride of each group, compared with a control group, the building group and the comparison group are obviously increased (P is less than 0.05), and the experimental group and the traditional Chinese medicine group have no significant difference (P is more than 0.05); compared with the modeling group, the experimental group is obviously reduced (P <0.05) and the traditional Chinese medicine group and the comparison group have no significant difference (P > 0.05). Experimental results show that the traditional Chinese medicine decoction, the plant fermented drink and the bifidobacterium lactis HN019 plant fermented drink can achieve treatment effects on reduction of albumin, increase of urea nitrogen, uric acid, total cholesterol and triglyceride in mouse serum caused by puromycin aminonucleoside, and the plant fermented drink is relatively good in treatment effect.

Example 4

Example 2 experiment of protective action of plant fermentation broth on mouse kidney

The kidney-tonifying plant fermented drink prepared by the invention is proved to have a protection effect on the kidney injury of SD mice. A nephrosis model is prepared by injecting puromycin aminonucleoside into the tail vein of an SD mouse, sirolimus is used as a positive control drug to treat, and the protective effect of the plant fermentation drink prepared by the embodiment of the invention on kidney injury is researched.

1. Experimental methods

(1) SD mice were randomly divided into 4 groups: control group, model group, sirolimus group, experimental group, 10 pieces per group. Wherein, the control group is not treated, and distilled water with the same volume is infused every day after the experiment begins; the other groups were made into renal disease models by tail vein injection of puromycin aminonucleosides. The model group was not treated with drugs after modeling, and each mouse was drenched with distilled water every day; the sirolimus group was administered sirolimus (9mg/kg) per mouse daily for 14 days after modeling; the experimental group drenches 200mg/kg of the plant fermentation drink of the invention to each mouse every day within 14 days after modeling; keep the other living environments of the mice consistent.

(2) Urine protein creatinine ratio detection 24 hour urine from mice was collected on experiment day 13, protein quantification and urine creatinine were measured, and the urine protein creatinine ratio was calculated.

(3) Carrying out biochemical index detection on each group of mice 14 days after modeling, injecting 33mg/100g chloral hydrate into the abdominal cavity of an anesthetized mouse, opening the abdominal cavity under aseptic condition, collecting blood from the abdominal aorta, standing for 2h and then centrifuging. And detecting serum creatinine, urea nitrogen, uric acid, albumin, total cholesterol and triglyceride according to the steps of the kit specification.

)

group of	Number of mice	Urinary protein creatinine ratio
			Control group	9	317.09±59.89
Building module	8	2450.25±456.25^A
			Sirolimus group	10	623.45±140.36^B
Experimental group	10	615.84±321.52^B

Note: a <0.05 compared to normal group; b <0.05 compared to model group.

TABLE 2 serum biochemical indices of various groups of mice

Note: a <0.05 compared to normal group; b <0.05 compared to model group; c < 0.05P compared to sirolimus group.

2. Results of the experiment

The control mice died 1 and the model mice died 2, mainly under anesthesia, due to dose-failure.

As shown in table 1, the urine protein creatinine ratio of the modeling group was significantly increased (P <0.05) compared to the normal group; compared with the modeling group, the ratio of the urine protein to creatinine was significantly reduced in the sirolimus group and the experimental group (P < 0.05). Sirolimus and the plant fermented drink of the invention can reduce the urinary protein creatinine ratio caused by puromycin amino nucleoside.

As shown in Table 2, compared with the normal group, the model group mice have obviously increased urea nitrogen, uric acid, total cholesterol and triglyceride, obviously reduced albumin (P <0.05), unobvious creatinine change and no statistical significance (P > 0.05); compared with a modeling group, the sirolimus group and the experimental group are obviously reduced in urea nitrogen, uric acid, total cholesterol and triglyceride, obviously increased in albumin (P <0.05), unobvious in creatinine change and free of statistical significance (P > 0.05); the albumin in the experimental group is higher than that in the sirolimus group (P <0.05), and the uric acid level in the experimental group is lower than that in the sirolimus group (P < 0.05). Experimental results show that the plant fermented drink and sirolimus have a treatment effect on the nephropathy caused by puromycin amino nucleoside.

Example 5

The plant fermented beverage has the treatment effect on patients with kidney deficiency

1. Test members: 120 patients with kidney deficiency.

Of these patients, 60 male patients and 60 female patients were aged 30 to 58 years, and the average age was (43.5 ± 5.8) years. The symptoms of mental disorder, soreness and weakness of waist and knees, weakness of limbs, dysuria, alopecia, amnesia, dull and lusterless complexion, hyposexuality, fatigue and the like can be described, and male patients also have symptoms of impotence, premature ejaculation, female patients have irregular menstruation and the like.

2. The taking method comprises the following steps: the plant fermented drink is taken 1 time a day, 10mL each time, and 1 month is 1 course of treatment.

3. And (3) evaluating the curative effect: after the traditional Chinese medicine composition is taken for one treatment course, the effect of kidney deficiency of men and women can be recovered, the sexual activity and power of men and women can be obviously improved after the traditional Chinese medicine composition is taken for the second treatment course, and the effective rate of the traditional Chinese medicine composition for the people suffering from impotence and premature ejaculation reaches more than 98%.

The clinical curative effect tracking is that the tracking curative effect evaluation is carried out on 60 patients of men and women through the standards, and after the male takes the medicine for one month, the male feels that the waist is not acid, the head is not dim, the feet are not acid, the sleep is good, and the energy is vigorous. Female patients have good sleep, ruddy complexion and vigorous energy, and the normal state of the female patients is recovered by 98 percent after taking the medicine for one month, and the effective rate is 100 percent.

Tracking clinical curative effect: according to the comprehensive evaluation of all the male and female parties through the standards, 58 male people are cured in the treatment process of 1-2 months, 60 female people with weak sexual efficacy are cured, 59 male people are cured, and the comprehensive evaluation rate is 100%.

The test result shows that the plant fermented beverage can warm and tonify kidney qi and improve the symptoms of patients with kidney deficiency; has effects of warming and invigorating kidney qi, consolidating essence, strengthening body constitution, and enhancing body resistance.

Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Sequence listing

<120> plant fermentation liquor and preparation method and application thereof

<141> 2020-11-25

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1378

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

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Claims

1. A plant fermentation liquid is characterized by comprising the following raw materials in parts by weight: 60-80 parts of purified water, 5-10 parts of carrot juice, 1-5 parts of dried rehmannia root extract, 1-5 parts of Chinese yam extract, 1-5 parts of dogwood extract, 1-3 parts of rhizoma alismatis extract, 1-3 parts of poria cocos extract, 1-3 parts of moutan bark extract, 0.1-1 part of cassia twig extract, 0.1-1 part of rhizoma typhonii extract, 1-5 parts of walnut juice, 1-5 parts of passion fruit juice, 2-3 parts of L-arabinose, 2-3 parts of honey, 1-5 parts of resistant dextrin, 1-2 parts of xylitol and 0.1-0.3 part of edible essence;

carrying out aerobic fermentation on the raw materials, and then carrying out anaerobic fermentation to obtain plant fermentation liquor;

the strains used for anaerobic fermentation at least comprise bifidobacterium lactis; the Bifidobacterium lactis is Bifidobacterium animalis subsp.lactis SF-B-20, is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.20924, the preservation date is 2020, 10 and 21 days, and the address of a preservation institution is as follows: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.

2. The plant fermentation broth of claim 1, comprising the following raw materials in parts by weight: 70 parts of purified water, 5 parts of carrot juice, 4 parts of dried rehmannia root extract, 2 parts of Chinese yam extract, 2 parts of dogwood extract, 1.5 parts of rhizoma alismatis extract, 1.5 parts of poria cocos extract, 1.5 parts of moutan bark extract, 0.5 part of cassia twig extract, 0.5 part of processed monkshood extract, 5 parts of walnut juice, 5 parts of passion fruit juice, 2 parts of L-arabinose, 2 parts of honey, 5 parts of resistant dextrin, 2 parts of xylitol and 0.3 part of edible essence.

3. The preparation method of the plant fermentation liquor is characterized by comprising the following steps:

(1) squeezing carrots, passion fruits and walnuts respectively for standby, then respectively crushing dried rehmannia roots, Chinese yams, dogwoods, rhizoma alismatis, poria cocos, tree peony barks, cassia twigs and processed monkshood, and adding water 1-3 cm higher than medicinal powder for cooking to prepare extracts of various medicines;

(8) and (5) after the fermentation in the step (7) is finished, conveying the prepared fermentation liquor II into a high-pressure spraying wall breaking machine, setting the pressure to be 20-60 MPa, and performing pasteurization on the fermentation liquor II after wall breaking to obtain the plant fermentation liquor.

4. The method according to claim 3, wherein the aerobic probiotic bacteria in step (4) is Saccharomyces cerevisiae.

5. The method according to claim 3, wherein the amount of the starter culture I inoculated in the step (5) is 500 to 1000 ten thousand cfu/ml, the fermentation temperature is 37 ℃ and the fermentation time is 24 hours.

6. The preparation method according to claim 3, wherein the anaerobic probiotics in step (6) comprise one or more of Bifidobacterium lactis, Lactobacillus casei, Lactobacillus plantarum and Bifidobacterium longum.

7. The method according to any one of claims 3 or 6, wherein the anaerobic probiotic bacteria in step (6) are Bifidobacterium lactis: lactobacillus casei: lactobacillus plantarum: bifidobacterium longum is a mixed bacterial agent with the ratio of 1:1:1: 1.

8. The preparation method according to claim 3, wherein the inoculation amount of the fermentation agent II in the step (7) is 500 to 1000 ten thousand cfu/ml, the fermentation temperature is 37 ℃, and the fermentation time is 72 hours.

9. Use of the plant fermentation broth of claim 1 in the preparation of a kidney-tonifying beverage.