Dampness-eliminating dried orange peel probiotic fermented afterbiogen and application thereof

Abstract

The invention discloses dried orange peel probiotic fermentation post-biogen with a dampness clearing effect and application thereof, and belongs to the field of biological medicine. The dried orange peel probiotic fermented prebiotics are prepared by performing probiotic fermentation on dried orange peel twice and then sterilizing; the primary probiotic fermentation is lactobacillus reuteri fermentation; the secondary probiotic fermentation is bifidobacterium adolescentis fermentation. Mouse experiments prove that the dried orange peel probiotics prepared by the invention have no toxic or side effect and good safety. Clinical tests verify that the dried tangerine peel probiotic fermented prebiotics prepared by the invention have the effect of clearing damp, and can obviously relieve loose thin or constipation, dry mouth, halitosis, excessive phlegm, cough, greasy hair, obesity, snoring, spot and acne formation, leucorrhea, peculiar smell and other clinical manifestations of heavy dampness, so that the invention provides a new strategy for clearing damp.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to a tangerine peel probiotic fermented postbiotic with dehumidifying effect and application thereof. Background Art

[0002] Heavy dampness refers to dampness as a heavy, turbid, and substance evil, belonging to the yin. When dampness causes disease, clinical manifestations characterized by a sense of heaviness often occur. For example, the head and body feel heavy, and the limbs feel sore. When dampness attacks the skin, it blocks the clear yang, and the clear yang does not rise, the head feels heavy as if wrapped. When dampness blocks the joints, the yang cannot be spread, and the skin becomes numb and the joints are painful. Although dampness and heat can be divided into external dampness and internal dampness, they affect each other when they cause disease, and they are combined with internal and external evils. The spleen can transport and transform water and dampness. The spleen likes dryness and hates dampness. If the spleen yang is invigorated, the spleen is healthy, and the function of transporting water and dampness is normal, dampness is not easy to cause disease. On the contrary, when dampness is too heavy and causes illness, patients often experience symptoms such as dizziness and heaviness, sore limbs, heavy and painful body, poor joint flexion and extension, chest depression, abdominal distension, nausea and vomiting, loss of appetite, loose stools, and thick and greasy tongue coating. The invasion of cold and dampness causes the spleen to fail to transport and transform, and water and dampness are retained inside the body, which accumulates with the cold in the meridians and organs, and is prone to obesity caused by cold and dampness. Such patients are mainly manifested by abdominal distension, bloating, greasy mouth, no thirst, little appetite, poor appetite, bowel sounds and diarrhea, or edema of the limbs, pale and fat tongue, greasy white tongue coating, and weak pulse.

[0003] Most of the existing medicines for removing dampness from the body and products such as dampness-removing health tea are cold and can easily damage the stomach and spleen. The spleen and stomach are the source of the biochemical transformation of qi and blood, and the spleen is responsible for transportation and transformation. If the spleen yang is damaged, the biochemical transformation of qi and blood will be blocked, and the circulation of water will stop, which will lead to edema or obesity. Therefore, the existing products cannot completely remove dampness and are prone to recurrence. Moreover, most of the drugs for draining and removing dampness are dry, and long-term use can easily damage the body's body fluids and is not suitable for long-term use, which makes it difficult to remove dampness. It is urgent to propose a new strategy for removing dampness. Summary of the invention

[0004] The purpose of the present invention is to provide a tangerine peel probiotic fermentation postbiotic with dehumidifying effect and its application, so as to solve the problems existing in the above-mentioned prior art. The tangerine peel probiotic fermentation postbiotic provided by the present invention has a significant effect on dehumidifying, and provides a new idea for dehumidifying.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a tangerine peel probiotic fermentation postbiotic, wherein the tangerine peel probiotic fermentation postbiotic is prepared by sterilizing the tangerine peel after two probiotic fermentations;

[0007] The first probiotic fermentation is Lactobacillus reuteri fermentation; the second probiotic fermentation is Bifidobacterium adolescentis fermentation.

[0008] The present invention also provides a method for preparing the tangerine peel probiotic fermentation postbiotics, comprising the following steps:

[0009] Ultrafine powder is obtained by ultrafine grinding of dried tangerine peel, soaked in water, and ground to obtain slurry;

[0010] The slurry is inoculated with Lactobacillus reuteri liquid, and the first fermentation is carried out, and the slurry is sterilized; then the slurry is inoculated with Bifidobacterium adolescentis liquid, and the second fermentation is carried out, and the slurry is sterilized, and the supernatant is obtained by centrifugation and vacuum freeze drying to obtain tangerine peel probiotic fermentation postbiotics;

[0011] The sterilization conditions are 95° C., 40 min.

[0012] Furthermore, the preservation number of the Lactobacillus reuteri is CICC 6123, and the preservation number of the Bifidobacterium adolescentis is CICC 6178.

[0013] Furthermore, the inoculation amount of the Lactobacillus reuteri bacterial solution is 3%, and the inoculation amount of the Bifidobacterium adolescentis bacterial solution is 7%.

[0014] Furthermore, the concentration of the Lactobacillus reuteri liquid is 5×10 9 cfu / mL, the concentration of the Bifidobacterium adolescentis bacterial solution was 2×10 8 cfu / mL.

[0015] Furthermore, the volume ratio of the ultrafine powder to water is 1:10.

[0016] Furthermore, the conditions of the first fermentation are pH 7 and fermentation at 39° C. for 25 hours.

[0017] Furthermore, the second fermentation was performed under the conditions of pH 7 and 35° C. for 34 hours.

[0018] The present invention also provides the use of the tangerine peel probiotic fermentation postbiotics in the preparation of dehumidifying products.

[0019] The present invention discloses the following technical effects:

[0020] The tangerine peel probiotic fermentation postbiotics of the present invention is tangerine peel fermented by Lactobacillus reuteri and then fermented by Bifidobacterium adolescentis. Mouse experiments have confirmed that there is no toxic side effect and good safety. And clinical trials have verified that the tangerine peel probiotic fermentation postbiotics prepared by the present invention has a dampness-removing effect, which can significantly alleviate the clinical manifestations of heavy moisture such as loose stools or constipation, dry mouth and bad breath, cough with sputum, greasy hair, obesity, snoring, spots and acne, and leucorrhea odor. It can be seen that the present invention provides a new strategy for dampness removal. DETAILED DESCRIPTION

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0023] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0024] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0025] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0026] The Lactobacillus reuteri used in the present invention has a preservation number of CICC 6123 and the Bifidobacterium adolensentis has a preservation number of CICC 6178, both of which are purchased from the China Industrial Microbiological Culture Collection Center.

[0027] The public can purchase the above strains from the above depository institutions. The specific research is as follows.

[0028] Example 1

[0029] Lactobacillus reuteri was inoculated into NA medium and cultured anaerobically at 30°C for 18 hours to obtain activated Lactobacillus reuteri. Then, 10 wt% of the inoculum was inoculated into MRS medium and cultured anaerobically at 30°C for 48 hours to obtain a viable cell count of 5×109 cfu / mL of Lactobacillus reuteri liquid; inoculate Bifidobacterium adolescentis into TPY medium, and culture anaerobically at 37°C for 18h to obtain activated Bifidobacterium adolescentis. Then inoculate into MRS medium at a 5wt% inoculation amount, and culture anaerobically at 37°C for 48h to obtain a viable cell count of 2×10 8 cfu / mL of Bifidobacterium adolescentis bacterial solution.

[0030] Take tangerine peel and grind it into ultrafine powder, add 10 times the volume of pure water to soak and grind to obtain slurry;

[0031] The slurry was inoculated with Lactobacillus reuteri liquid at an inoculum rate of 3%, and the first fermentation (pH=7, fermentation at 39°C for 25 hours) was carried out, and the mixture was sterilized at 95°C for 40 minutes. Then, the slurry was inoculated with Bifidobacterium adolescentis liquid at an inoculum rate of 7%, and the second fermentation (pH=7, fermentation at 35°C for 34 hours) was carried out, and the mixture was sterilized at 95°C for 40 minutes. The supernatant was collected by centrifugation and vacuum freeze-dried to obtain tangerine peel probiotic fermentation postbiotics.

[0032] Comparative Example 1

[0033] According to the operation described in Example 1, Lactobacillus reuteri was prepared to have a viable count of 5×10 9 cfu / mL of Lactobacillus reuteri liquid;

[0034] Take tangerine peel and grind it into ultrafine powder, add 10 times the volume of pure water to soak and grind to obtain slurry;

[0035] The slurry was inoculated with Lactobacillus reuteri at an inoculum rate of 3%, and fermented (pH=7, 39°C for 25 hours), sterilized at 95°C for 40 minutes, centrifuged to obtain the supernatant, and vacuum freeze-dried to obtain tangerine peel probiotic fermentation postbiotics.

[0036] Comparative Example 2

[0037] According to the operation described in Example 1, Bifidobacterium adolescentis was prepared with a viable count of 2×10 8 cfu / mL of Bifidobacterium adolescentis bacterial solution;

[0038] Take tangerine peel and grind it into ultrafine powder, add 10 times the volume of pure water to soak and grind to obtain slurry;

[0039] The slurry was inoculated with 7% of Bifidobacterium adolescentis liquid for fermentation (pH=7, 35°C for 34 hours), sterilized at 95°C for 40 minutes, centrifuged to obtain the supernatant, and vacuum freeze-dried to obtain tangerine peel probiotic fermentation postbiotics.

[0040] Effect example

[0041] 1. Toxicity verification experiment

[0042] 1. Experimental animals and groups

[0043] ICR mice weighing 18-22g, half male and half female. Mice were raised in the laboratory by sex, at room temperature of 20℃-26℃, relative humidity of 40-70%, and natural light. Water and feed were freely available. After one week of feeding before the experiment, mice with normal behavior, diet, and mental state were selected for the experiment.

[0044] Take 40 mice and fast for 12 hours before administration. Divide into four groups: control group, gavage with distilled water at 0.4mL / 10g body weight; Example 1 group: use distilled water to resuspend the postbiotics prepared in Example 1, the mass concentration is 10%, and gavage is 0.4mL / 10g body weight; Comparative Example 1 group: use distilled water to resuspend the postbiotics prepared in Comparative Example 1, the mass concentration is 10%, and gavage is 0.4mL / 10g body weight; Comparative Example 2 group: use distilled water to resuspend the postbiotics prepared in Comparative Example 2, the mass concentration is 10%, and gavage is 0.4mL / 10g body weight. Gavage once a day for one week. Observe and record the behavioral activities, status, diet, stool, urine, fur color, secretions and death of each group of mice in detail.

[0045] 2. Statistics

[0046] The data were analyzed by one-way analysis of variance using SPSS11.0 software.

[0047] 3. Results

[0048] 3.1 General

[0049] After intragastric administration, the mice in each group showed no abnormal physical signs or appearance, moved freely, had normal eating, drinking, and feces, had smooth skin, and had clean mouth, nose, eyes, and anus without secretions.

[0050] 3.2 Weight changes

[0051] After 7 days of intragastric administration, all groups of mice showed a trend of weight gain. Before administration, the weights of mice in each group were: control group (20.29 ± 0.51) g, Example 1 group (20.31 ± 0.42) g, comparative example 1 group (20.31 ± 0.40) g, comparative example 2 group (20.30 ± 0.48) g. After administration, the weights of mice in each group were: control group (24.69 ± 1.31) g, Example 1 group (24.36 ± 1.34) g, comparative example 1 group (24.42 ± 1.29) g, comparative example 2 group (24.45 ± 1.37) g. The weights of the groups were compared, P> 0.05, and the difference was not statistically significant.

[0052] 3.3 Macroscopic observation of major organ anatomy

[0053] All 40 mice in the experiment survived without obvious poisoning reactions. Dissection findings: no abnormal fluid was found in the chest and abdominal cavities, no flatulence was found in the intestines, no abnormal color or morphology was found in important organs such as the heart, liver, spleen, lungs, kidneys, and brain, no bleeding spots or other pathological changes, the gastric mucosa was ruddy in color, without bleeding spots or ulcers, and no abnormalities were found in the autopsy.

[0054] In summary, the postbiotics prepared in the embodiments and comparative examples of the present invention have no toxic reactions and are safe.

[0055] 2. Clinical Validation

[0056] Outpatients diagnosed with heavy dampness and willing to accept intervention measures were selected, with clinical manifestations of loose stools or constipation, dry mouth and bad breath, cough with sputum, greasy hair, obesity, snoring, spots and acne, and leucorrhea with odor. Exclusion criteria: those who were found to be in a disease state through a comprehensive physical examination; healthy people; pregnant women, women preparing for pregnancy, and breastfeeding women; suspected or confirmed history of alcohol or drug abuse; known allergies to the components of the postbiotics of the present invention; patients participating in clinical trials of other drugs; and those who are unwilling to cooperate.

[0057] A total of 120 patients were clinically verified, including 50 males and 70 females, with the oldest age of 63 years old and the youngest age of 25 years old, and were randomly divided into 3 groups: Example 1 group, taking 2g of the postbiotics prepared in Example 1 of the present invention before going to bed every night; Comparative Example 1 group, taking 2g of the postbiotics prepared in Comparative Example 1 of the present invention before going to bed every night; Comparative Example 2 group, taking 2g of the postbiotics prepared in Comparative Example 2 of the present invention before going to bed every night. One month is a course of treatment, and Table 1 shows the result data of each group after taking one course of treatment.

[0058] Table 1 Comparison of clinical manifestations before and after taking (unit: person)

[0059]

[0060] As can be seen from Table 1, the tangerine peel probiotic fermentation postbiotics prepared in Example 1 of the present invention has a good dehumidifying effect and has significant therapeutic effects in clinical applications. However, due to the lack of the fermentation step of Bifidobacterium adolescentis or Lactobacillus reuteri in Comparative Examples 1 and 2, the therapeutic effects are obviously inferior to those in Example 1.

[0061] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A tangerine peel probiotic fermentation postbiotic, characterized in that: The tangerine peel probiotic fermentation postbiotics are prepared by sterilizing tangerine peel after two probiotic fermentations; The first probiotic fermentation is Lactobacillus reuteri fermentation; the second probiotic fermentation is Bifidobacterium adolescentis fermentation.

2. The method for preparing tangerine peel probiotic fermentation postbiotics as claimed in claim 1, characterized in that, The following steps are involved: Ultrafine powder is obtained by ultrafine grinding of dried tangerine peel, soaked in water, and ground to obtain slurry; The slurry is inoculated with Lactobacillus reuteri liquid, and the first fermentation is carried out, and the slurry is sterilized; then the slurry is inoculated with Bifidobacterium adolescentis liquid, and the second fermentation is carried out, and the slurry is sterilized, and the supernatant is obtained by centrifugation and vacuum freeze drying to obtain tangerine peel probiotic fermentation postbiotics; The sterilization conditions are 95° C., 40 min.

3. The preparation method according to claim 2, characterized in that: The preservation number of the Lactobacillus reuteri is CICC6123, and the preservation number of the Bifidobacterium adolescentis is CICC 6178.

4. The preparation method according to claim 2, characterized in that: The inoculation amount of the Lactobacillus reuteri bacterial solution is 3%, and the inoculation amount of the Bifidobacterium adolescentis bacterial solution is 7%.

5. The preparation method according to claim 2, characterized in that: The concentration of the Lactobacillus reuteri liquid is 5×10 9 cfu / mL, the concentration of the Bifidobacterium adolescentis bacterial solution was 2×10 8 cfu / mL.

6. The preparation method according to claim 2, characterized in that: The volume ratio of the superfine powder to water is 1:

10.

7. The preparation method according to claim 2, characterized in that: The conditions of the first fermentation were pH 7 and fermentation at 39° C. for 25 hours.

8. The preparation method according to claim 2, characterized in that: The second fermentation was carried out under the conditions of pH 7 and 35° C. for 34 hours.

9. Use of the tangerine peel probiotic fermentation postbiotics as claimed in claim 1 in the preparation of dehumidifying products.

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