Composition helpful for improving intestinal flora and application thereof

Abstract

The invention relates to a composition helpful for improving intestinal flora and application thereof, and belongs to the technical field of health care products. The composition helpful for improving intestinal flora comprises the following components: seville orange flower powder, Chinese torreya seed powder, fructus piperis longi powder and celery seed powder, and the preparation method of the composition comprises the following steps: mixing the seville orange flower powder, the Chinese torreya seed powder, the fructus piperis longi powder and the celery seed powder, and then fermenting by using a fermentation strain to obtain the composition. Wherein the fermentation strain is bifidobacterium pseudocatenum with the preservation number of CGMCC (China General Microbiological Culture Collection Center) NO.28075. The composition prepared from the components and the preparation method has the effect of remarkably improving intestinal flora.

Description

Technical Field

[0001] This invention relates to the field of health product technology, and in particular to a composition that helps improve gut microbiota and its application. Background Technology

[0002] The gut microbiota, a complex micro-ecosystem within the human body, is closely related to the host's health. Gut microbiota dysbiosis is associated with the development and progression of various diseases, such as constipation, diarrhea, weakened immunity, and metabolic disorders. Therefore, maintaining or improving gut microbiota balance has become one of the important directions in health product research and development.

[0003] Currently, the mainstream methods for improving gut microbiota mainly include directly supplementing with probiotics (such as Bifidobacterium and Lactobacillus) or using prebiotics (such as oligosaccharides) to promote the growth of beneficial bacteria. However, these methods have certain limitations. For example, directly supplemented probiotics may be largely inactivated by stomach acid and bile before reaching the intestines, resulting in less than ideal colonization and efficacy. While single prebiotics can promote gut microbiota growth, their target sites are relatively broad, and their selectivity for the proliferation of specific beneficial bacteria (such as Bifidobacterium) is not strong enough, sometimes resulting in less significant effects.

[0004] In addition, some existing technologies utilize traditional Chinese medicine or natural plant ingredients to regulate gut microbiota. However, these technologies often have several problems: First, most solutions only involve simple mixing or traditional water or alcohol extraction, resulting in limited release and bioavailability of active ingredients; second, research on specific combinations of raw materials and their synergistic effects with specific probiotic fermentation is insufficient, and there is a lack of scientifically validated specific compositions that can significantly increase the number of core beneficial bacteria such as Bifidobacterium and Lactobacillus while inhibiting harmful bacteria (such as Clostridium perfringens); finally, the strains used in existing technologies may have limited efficacy or be unsuitable for fermentation of specific raw material substrates, leading to poor efficacy of the final product.

[0005] Therefore, there is an urgent need in the field to develop a composition that can overcome the aforementioned deficiencies. This composition should be able to produce a synergistic effect through a specific combination of raw materials and a specific probiotic fermentation process, thereby more effectively and specifically improving the gut microbiota structure, promoting intestinal motility, and possessing good safety. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a composition and its application that helps improve the gut microbiota.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a composition that helps improve the gut microbiota, comprising the following components in parts by weight: 5-8 parts of bitter orange pollen, 3-5 parts of torreya nut powder, 0.5-1.5 parts of long pepper powder, and 10-15 parts of celery seed powder; the composition is prepared by mixing bitter orange pollen, torreya nut powder, long pepper powder, and celery seed powder, and then fermenting them with fermentation bacteria to obtain the composition; The fermentation strain is Bifidobacterium pseudobulb, with the preservation number CGMCCNO.28075.

[0008] The present invention, through the selection of specific components and preparation methods, produces a composition that significantly improves the intestinal flora.

[0009] Preferably, the method for preparing the composition that helps improve gut microbiota includes the following steps: S1: Mix all the raw materials in the composition evenly to obtain a mixed powder: S2: Prepare a 20-30 wt% mixture of powders using deionized water. S3: Sterilize the mixture, cool to room temperature, inoculate with fermentation starter, and ferment for 20-30 hours to obtain the fermentation broth: S4: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0010] Preferably, in step S3, the inoculum volume of the fermentation strain accounts for 8-10 v / v of the mixed liquid volume.

[0011] Preferably, in step S3, the fermentation temperature is 37-40℃, the fermentation pH is 6-8, and the fermentation environment is anaerobic fermentation.

[0012] In a second aspect, the present invention provides the use of the composition described in the first aspect that helps improve gut microbiota in the preparation of health products that help improve gut microbiota.

[0013] Preferably, the dosage form of the health product includes any one of liquid, pill, powder, tablet, and ointment.

[0014] Thirdly, the present invention provides an oral liquid that helps improve gut microbiota, the oral liquid comprising the composition described in the first aspect that helps improve gut microbiota.

[0015] Preferably, the oral liquid also includes sweeteners, preservatives, and drinking water.

[0016] Preferably, the sweetener is an acceptable sweetener for health products.

[0017] Preferably, the preservative is an acceptable preservative for health products.

[0018] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The activation effect of fermentation process on raw materials Microbial degradation and transformation: During anaerobic fermentation, *Bifidobacterium pseudomicrobium* (CGMCC NO.28075), as the fermentation strain, can secrete various enzymes (such as cellulase and protease) to degrade macromolecules such as cellulose, polysaccharides, and proteins in the raw materials into smaller active ingredients (such as oligosaccharides, short-chain fatty acids, and amino acids). These smaller molecules are more easily utilized by the gut microbiota, acting as prebiotics to directly promote the growth of *Bifidobacterium* and *Lactobacillus*.

[0019] Metabolic product generation: During fermentation, the microbial strain produces beneficial metabolites (such as short-chain fatty acids like lactic acid and acetic acid). These products can lower the intestinal pH, inhibit the growth of harmful bacteria (such as Clostridium perfringens), and provide energy for intestinal epithelial cells, thereby enhancing the intestinal barrier function.

[0020] 2. Synergistic mechanism between specific strains and raw materials Strain specificity: *Bifidobacterium pseudosporidis* (CGMCC NO.28075) possesses unique strain characteristics, enabling it to efficiently utilize components in raw materials (such as flavonoids in *Fructus peruviana* pollen and dietary fiber in celery seeds) and convert them into compounds with probiotic effects. Compared to other strains, this strain exhibits higher compatibility with raw materials, resulting in a stronger synergistic effect.

[0021] Prebiotic support: The raw materials, such as bitter orange pollen and torreya powder, are rich in polysaccharides and dietary fiber, which can act as prebiotics to selectively stimulate the growth and metabolism of Bifidobacterium pseudomicrobium, further amplifying the probiotic effects of its fermentation products.

[0022] 3. Mechanisms of gut microbiota regulation Beneficial bacteria proliferation: The small molecular components after fermentation can directly serve as a nutrient substrate for Bifidobacteria and Lactobacillus, promoting their colonization and proliferation, and optimizing the intestinal flora structure.

[0023] Harmful bacteria inhibition: Metabolites such as short-chain fatty acids can destroy the cell membranes of harmful bacteria, inhibit their growth, and at the same time reduce the attachment space of harmful bacteria through competitive exclusion.

[0024] 4. Mechanism of promoting intestinal peristalsis Neuroendocrine regulation: The active ingredients produced by fermentation can stimulate intestinal endocrine cells to release gastrointestinal hormones, activate the intestinal nervous system, and enhance intestinal peristalsis.

[0025] Anti-inflammatory effect: The anti-inflammatory components (such as flavonoids) in the composition can reduce intestinal inflammation and improve intestinal motility, thereby relieving constipation.

[0026] Through the above-described mechanism, the composition of the present invention not only significantly improves the balance of intestinal flora, but also effectively promotes intestinal peristalsis, achieving dual health benefits. Detailed Implementation

[0027] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0028] The raw materials used in this invention are partly sourced from the following sources: The pollen from Bitter Orange, Torreya grandis powder, Piper longum powder, and celery seed powder were all purchased from Baoji Liupanyun Biotechnology Co., Ltd. Before use, all the above raw materials will be dried to constant weight in an oven, further ground, passed through an 80-mesh sieve to obtain the corresponding raw material powder, and then sealed and stored away from light.

[0029] Bifidobacterium pseudosporidis-1: accession number CGMCCNO.28075; Bifidobacterium pseudosporidis-2: accession number CCTCCNO.M 2025538; Before use, the above-mentioned *Bifidobacterium pseudobulb-1* or *Bifidobacterium pseudobulb-2* should be cultured in a suitable culture medium for *Bifidobacterium pseudobulb* until the viable count concentration reaches 1×10⁻⁶. 8 The fermentation starter culture at CFU / mL is ready for use.

[0030] Unless otherwise specified, all other materials and reagents used in the examples are commercially available.

[0031] The "room temperature" mentioned in the preparation steps is 25°C.

[0032] The raw materials and preparation method of the composition that helps improve gut microbiota are as follows: Composition 1 It is made from the following raw materials: pollen from each generation: 7 parts; Torreya nut flour: 4 parts; Piper longum powder: 1 part; Celery seed powder: 12 servings; Preparation method: S1: Mix all the raw materials in the composition evenly to obtain a mixed powder: S2: Prepare a 25wt% mixture of powders using deionized water. S3: Sterilize the mixture, cool to room temperature, inoculate with fermentation starter, and ferment for 24 hours to obtain the fermentation broth: S4: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0033] The fermentation strain was *Bifidobacterium pseudobulbarum*-1, with accession number CGMCCNO.28075, and a viable count of 1×10⁻⁶. 8 CFU / mL.

[0034] In step S3, the inoculum volume of the fermentation strain accounts for 9v / v of the mixed liquid volume; the fermentation temperature is 37℃, the fermentation pH is 7, and the fermentation environment is anaerobic fermentation.

[0035] Composition 2 It is made from the following raw materials: pollen from perennial flowers: 5 parts; Torreya nut flour: 3 parts; Piper longum powder: 0.5 parts; Celery seed powder: 10 portions; Preparation method: S1: Mix all the raw materials in the composition evenly to obtain a mixed powder: S2: Prepare a 20wt% mixture of powders using deionized water. S3: Sterilize the mixture, cool to room temperature, inoculate with fermentation starter, and ferment for 20 hours to obtain the fermentation broth: S4: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0036] The fermentation strain was *Bifidobacterium pseudobulbarum*-1, with accession number CGMCCNO.28075, and a viable count of 1×10⁻⁶. 8 CFU / mL.

[0037] The inoculum amount of the fermentation strain is 8v / v of the mixed liquid volume; the fermentation temperature is 39℃, the fermentation pH is 6.5, and the fermentation environment is anaerobic fermentation.

[0038] Composition 3 It is made from the following raw materials: Pollen from each generation: 8 parts; Torreya nut flour: 5 parts; Piper longum powder: 1.5 parts; Celery seed powder: 15 servings; Preparation method: S1: Mix all the raw materials in the composition evenly to obtain a mixed powder: S2: Prepare a 30wt% mixture of powders using deionized water. S3: Sterilize the mixture, cool to room temperature, inoculate with fermentation starter, and ferment for 30 hours to obtain the fermentation broth: S4: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0039] The fermentation strain was *Bifidobacterium pseudobulbarum*-1, with accession number CGMCCNO.28075, and a viable count of 1×10⁻⁶. 8 CFU / mL.

[0040] The inoculum amount of the fermentation strain is 10v / v of the mixed liquid volume; the fermentation temperature is 40℃, the fermentation pH is 7.5, and the fermentation environment is anaerobic fermentation.

[0041] Composition ① It is made from the following raw materials: pollen from each generation: 7 parts; Torreya nut flour: 4 parts; Piper longum powder: 1 part; Celery seed powder: 12 servings; Unlike composition 1, composition ① is prepared by an ethanol reflux method, with the specific steps as follows: A1: Mix all the raw materials in the composition evenly to obtain a mixed powder: A2: Prepare a mixture of powders into a solution using a 60wt% ethanol-water solution, wherein the ratio of powder to solution is 1g / 20mL. A3: Heat the mixture to 60°C and reflux for 2 hours, then filter to obtain the extract; A4: Freeze-dry the extract to obtain the composition.

[0042] Composition ② Unlike composition 1, composition ② uses a different fermentation strain. The fermentation strain used in composition ② is Bifidobacterium pseudosporidis-2, with the preservation number CCTCCNO.M 2025538. The other raw materials, raw material mass fractions, preparation steps, preparation parameters, and viable cell count are the same as those in composition 1.

[0043] Composition ③ Unlike composition 1, the fermentation raw materials lack Fermented pollen. The missing mass fraction is made up by Torreya grandis powder, Piper longum powder, and celery seed powder in a mass ratio of 4:1:12. The other raw materials, raw material mass fractions, preparation steps, preparation parameters, and viable cell count are the same as those in composition 1.

[0044] Composition ④: Unlike composition 1, the fermentation raw materials lack torreya powder. The missing mass is made up by mandarin orange pollen, long pepper powder, and celery seed powder in a mass ratio of 7:1:12. The other raw materials, raw material mass, preparation steps, preparation parameters, and viable bacteria count are the same as those in composition 1.

[0045] Composition ⑤: Unlike Composition 1, the fermentation raw materials lack Piper longum powder. The missing mass is made up by Citrus reticulata pollen, Torreya grandis powder, and celery seed powder in a mass ratio of 7:4:12. The other raw materials, raw material mass, preparation steps, preparation parameters, and viable cell count are the same as those in Composition 1.

[0046] Composition ⑥: Unlike composition 1, the fermentation raw materials lack celery seed powder. The missing mass is made up by the mass ratio of mango pollen, torreya seed powder, and long pepper powder in a ratio of 7:4:1. The other raw materials, mass fractions of raw materials, preparation steps, preparation parameters, and viable cell count are the same as those of composition 1.

[0047] Composition ⑦: Unlike composition 1, composition 7 lacks the raw material powder and preparation steps. The raw materials and preparation steps for composition 7 are as follows: Raw material: MRS liquid culture medium (Beina Biotechnology, No.: BNCC363097); Preparation steps: S1: Prepare MRS liquid culture medium according to the MRS liquid culture medium instructions; S2: Sterilize the MRS liquid culture medium, cool it to room temperature, inoculate it with the fermentation starter, and ferment for 24 hours to obtain the fermentation broth: S3: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0048] The fermentation strain was *Bifidobacterium pseudobulbarum*-1, with accession number CGMCCNO.28075, and a viable count of 1×10⁻⁶. 8 CFU / mL.

[0049] In step S2, the inoculum size of the fermentation strain is 9v / v of the volume of the MRS liquid culture medium; the fermentation temperature is 37℃, the fermentation pH is 7, and the fermentation environment is anaerobic fermentation.

[0050] Components and preparation of oral liquids that help improve gut microbiota Oral liquid 1 Composed of the following components by mass percentage: Composition 1: 30 wt% Sweetener: 5 wt%; Preservative: 0.02 wt%; Drinking water replenished to 100 wt%; Preparation method: Mix the composition, sweetener, preservative and drinking water evenly, sterilize and fill; The sweetener is white sugar, and the preservative is potassium sorbate.

[0051] Oral liquid 2 Composed of the following components by mass percentage: Composition 1: 50 wt% Sweetener: 5 wt%; Preservative: 0.02 wt%; Drinking water replenished to 100 wt%; Preparation method: Mix the composition, sweetener, preservative and drinking water evenly, sterilize and fill; The sweetener is white sugar, and the preservative is potassium sorbate.

[0052] Oral liquid 3 Composed of the following components by mass percentage: Composition 1: 10 wt% Sweetener: 5 wt%; Preservative: 0.02 wt%; Drinking water replenished to 100 wt%; Preparation method: Mix the composition, sweetener, preservative and drinking water evenly, sterilize and fill; The sweetener is white sugar, and the preservative is potassium sorbate.

[0053] Oral liquid 4 Composed of the following components by mass percentage: Composition 1: 70 wt% Sweetener: 5 wt%; Preservative: 0.02 wt%; Drinking water replenished to 100 wt%; Preparation method: Mix the composition, sweetener, preservative and drinking water evenly, sterilize and fill; The sweetener is white sugar, and the preservative is potassium sorbate.

[0054] Efficacy verification: Test 1: Tests that help regulate gut microbiota Experimental methods: The experiment was conducted in accordance with the methods described in Chapter 15, "Testing methods for foods that help regulate intestinal flora," of the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)."

[0055] Test samples: Compositions 1-3, Compositions ①-⑦; The compositions were prepared into test samples at a concentration of 25 wt% using drinking water.

[0056] Sample preparation: Test animals: 18±1g male BALB / c mice, 15 mice per group, one sample per group.

[0057] Test sample administration period: 28 days; dosage: once daily, 0.3 mL / 10 g (body weight) each time.

[0058] Negative control group: physiological saline; administration time: 28 days; dosage: same as the test sample.

[0059] Experimental steps: Before administering the test samples, 0.1g of fecal matter was aseptically collected from the anus of mice and serially diluted 10-fold. Appropriate dilutions were then inoculated onto various culture media (prepared according to the culture media and methods for intestinal flora testing described in Appendix Table 15, "Test Methods for Regulating Intestinal Flora," of the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)"). After incubation, colonies were counted, and the number of bacteria per gram of wet stool was calculated. The logarithm was then statistically analyzed. Twenty-four hours after the last administration of the test samples, rectal feces were collected in the same manner as before the experiment to detect intestinal flora, using the same method as above.

[0060] Observation indicators: Body weight, Bifidobacterium, Lactobacillus, Enterococcus, Enterobacter, Clostridium perfringens.

[0061] Judgment criteria: The test sample can be considered positive in animal experiments if any of the following conditions are met.

[0062] 1. The number of Bifidobacterium and / or Lactobacillus colonies in feces increased significantly, the number of Clostridium perfringens colonies decreased or did not increase, and the number of Enterobacteriaceae and / or Enterococci colonies did not change significantly.

[0063] 2. The number of Bifidobacterium and / or Lactobacillus colonies in feces increased significantly, the number of Clostridium perfringens colonies decreased or did not increase, and the number of Enterobacteriaceae and / or Enterococci colonies increased significantly, but the increase was less than that of Bifidobacterium / Lactobacillus.

[0064] Colony change rate / % = (Number of colonies after test - Number of colonies before test) / Number of colonies before test × 100%; Average colony change rate / % = Sum of colony change rates in mice within the group / Number of mice in the group; Table 1. Effects of the composition on the intestinal flora of mice

[0065] Note: "*" indicates comparison with the negative control group, p < 0.05.

[0066] As shown in Table 1, the composition helps regulate the intestinal flora of mice. Comparing compositions 1-3 with the negative control group, the compositions provided by this invention significantly increase the number of Bifidobacterium and Lactobacillus colonies in mouse feces, inhibit the number of Clostridium perfringens colonies, and also have a certain increase in the number of Enterobacteriaceae and Enterococci, but the increase rate is far less than that of Bifidobacterium and Lactobacillus. According to the determination method recorded in the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)," compositions 1-3 are found to have the effect of regulating the intestinal flora of mice. Comparing composition 1 with compositions ① and ⑦, it can be seen that using Citrus aurantium pollen, Torreya grandis powder, and Piper longum alone... The ethanol reflux extract of a mixture of basil powder and celery seed powder, or the fermentation filtrate of *Bifidobacterium pseudobulbarbital*-1, did not have the effect of regulating mouse intestinal flora. This indicates that only the composition prepared using the raw materials, strains, and specific preparation method provided by this invention has the effect of regulating mouse intestinal flora. Comparing the results of composition 1 with compositions ②-⑥, it can be seen that when the fermentation strain or fermentation substrate in the composition is changed, the efficacy of the composition in regulating mouse intestinal flora decreases to a certain extent compared to composition 1. Comparing the results of composition 1 with composition ②, it can be seen that *Bifidobacterium pseudobulbarbital*-2 (preservation number CCTCCNO.M)... The composition obtained by fermenting a mixture of *Citrus aurantium* pollen, *Torreya grandis* powder, *Piper longum* powder, and celery seed powder using *2025538* as the fermentation strain also has a certain effect on regulating intestinal flora. However, its rate of increase in the number of *Bifidobacterium* and *Lactobacillus* colonies is much lower than that of composition 1, and its rate of inhibition of *Clostridium perfringens* colonies is also much lower than that of composition 1. This may be because even the same bacteria have different effects due to different preservation numbers. The composition obtained by using *Bifidobacterium pseudosporidis* with the preservation number specified in this invention as the fermentation strain has a significant effect on regulating intestinal flora. Comparing the results of composition 1 with those of compositions ③-⑥, it can be seen that the composition of the fermentation raw materials in this invention also has a certain impact on the efficacy of the composition in regulating intestinal flora. The absence of any one of them can reduce its efficacy.

[0067] In summary, the composition obtained using the fermentation raw materials, fermentation strains, and fermentation conditions specified in this invention has a significant effect on regulating the intestinal flora.

[0068] Test 2: Effect of the composition on promoting intestinal motility in mice Test samples: Compositions 1-3, Compositions ①-⑦; The compositions were prepared into test samples at a concentration of 25 wt% using drinking water.

[0069] Experimental animals: 18±1g male BALB / c mice, 10 mice per group, each group corresponding to one sample.

[0070] Blank control group: physiological saline.

[0071] Test sample administration time: 14 days.

[0072] Experimental steps: The experiment consisted of 10 treatment groups, 1 model control group, and a blank control group. Treatment groups received the test sample orally once daily for 14 consecutive days via gavage. The gavage volume for the treatment group was 0.3 mL / 10 g (body weight) / day. The model control group and the blank control group received the same volume of physiological saline via gavage.

[0073] Fourteen days later, the mice were fasted but allowed free access to water for 16 hours. The blank control group was given saline by gavage, while the model control group and the treatment group were given loperamide (4 mg / kg BW) by gavage. 0.5 hours after gavage, the treatment group, model control group, and blank control group were given ink (containing 5 wt% activated charcoal powder, 10 wt% gum arabic, and the remainder deionized water) by gavage, with normal drinking water and fasting. After 25 minutes, the mice in each group were immediately euthanized by cervical dislocation, the abdominal cavity was opened, the mesentery was separated, and the intestinal segment from the upper end of the pylorus to the lower end of the ileocecal junction was cut off and placed on a tray. The small intestine was gently pulled into a straight line, and the length of the intestinal segment was measured as the "total length of the small intestine". The length from the pylorus to the leading edge of the ink was measured as the "ink propulsion length". The average propulsion rate of the small intestine in each group was calculated.

[0074] Table 2 Effects of the composition on promoting intestinal motility in mice Test sample test animals / only Average propulsion rate of ink in small intestine / % Blank control group 10 63.2 Model control group 10 37.5 Composition 1 10 60.4 Composition 2 10 57.6 Composition 3 10 59.3 Composition ① 10 40.6 Composition ② 10 47.5 Composition ③ 10 42.1 Composition ④ 10 49.2 Composition ⑤ 10 51.3 Composition ⑥ 10 52.8 Composition ⑦ 10 43.7 As shown in Table 2, the composition helps promote intestinal peristalsis in mice. Comparison of the model control group and the blank control group indicates that the constipation mouse model of this invention was successfully established. Comparison of compositions 1-3 with the model control group shows that the compositions provided by this invention can significantly enhance intestinal peristalsis in constipation model mice. Comparison of compositions 1, ①, and ⑦ shows that the ethanol extract of the fermentation raw materials or the fermentation filtrate of the fermentation strains in this invention did not significantly enhance intestinal peristalsis in constipation model mice. However, the compositions obtained using the fermentation raw materials and fermentation strains specified in this invention and following the preparation steps specified in this invention have significantly enhanced intestinal peristalsis in constipation model mice. The efficacy of composition 1 in enhancing intestinal peristalsis in constipated mice indicates a significant synergistic effect between the two. A comparison of composition 1 and composition 2 shows that the bioactivity of *Bifidobacterium pseudosporidis* with different preservation numbers varies. The composition obtained using *Bifidobacterium pseudosporidis* with the preservation number specified in this invention has a significant effect on enhancing intestinal peristalsis in constipated mice. A comparison of composition 1 and compositions 3-6 shows that the ratio of fermentation raw materials in this invention has a significant impact on the efficacy of enhancing intestinal peristalsis in constipated mice. The composition obtained using fermentation raw materials with the specified types and mass ratios specified in this invention can significantly enhance the efficacy of enhancing intestinal peristalsis in constipated mice.

[0075] Experiment 3: Acute toxicity test of oral solutions that help improve gut microbiota Test samples: oral liquid 1-4; Experimental procedure: Eighty SD rats weighing 250±5g were selected, half male and half female; Environment: temperature 23±1℃, humidity 54±2%; The test sample was administered orally to the rats once by gavage, and the dosage is shown in Table 3. The rats were fasted for 16 hours before gavage and observed for two consecutive weeks after gavage. The symptoms of poisoning and mortality were recorded as shown in Table 3. During the gavage period, the rats were fed with standard feed normally.

[0076] Table 3 Acute toxicity test of oral liquids that help improve gut microbiota Group gender quantity dose Death count Oral liquid 1 female 10 1mL / 100g / d 0 Oral liquid 1 male 10 1mL / 100g / d 0 Oral liquid 2 female 10 1mL / 100g / d 0 Oral liquid 2 male 10 1mL / 100g / d 0 Oral liquid 3 female 10 1mL / 100g / d 0 Oral liquid 3 male 10 1mL / 100g / d 0 Oral liquid 4 female 10 1mL / 100g / d 0 Oral liquid 4 male 10 1mL / 100g / d 0 The acute toxicity test results showed that no obvious signs of poisoning were observed in the rats during the test, and no animals died, indicating that the oral liquid prepared in this invention is non-toxic.

[0077] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition that helps improve gut microbiota, characterized in that, The composition comprises the following raw materials in parts by weight: Pollen from perennial flowers: 5-8 parts; Torreya nut flour: 3-5 parts; Piper longum powder: 0.5-1.5 parts; Celery seed powder: 10-15 servings; The composition is prepared by mixing mandarin orange pollen, torreya seed powder, long pepper powder, and celery seed powder, and then fermenting them with fermentation bacteria to obtain the composition. The fermentation strain is Bifidobacterium pseudosporidis, with the preservation number CGMCCNO.28075.

2. The composition according to claim 1, characterized in that, The method for preparing the composition includes the following steps: S1: Mix all the raw materials in the composition evenly to obtain a mixed powder: S2: Prepare a 20-30 wt% mixture of powders using deionized water. S3: Sterilize the mixture, cool to room temperature, inoculate with fermentation starter, and ferment for 20-30 hours to obtain the fermentation broth: S4: Sterilize the fermentation broth, filter it to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

3. The composition according to claim 2, characterized in that, In step S3, the inoculation amount of fermentation strain accounts for 8-10 v / v of the mixed liquid volume.

4. The composition according to claim 2, characterized in that, In step S3, the fermentation temperature is 37-40℃, the fermentation pH is 6.5-7.5, and the fermentation environment is anaerobic fermentation.

5. The use of the composition according to any one of claims 1-4 in the preparation of a health product that helps improve gut microbiota.

6. The application as described in claim 5, characterized in that, The dosage forms of the health products include any one of liquid, pill, powder, tablet, and ointment.

7. An oral liquid that helps improve gut microbiota, characterized in that, Includes the composition according to any one of claims 1-4.

8. The oral liquid as described in claim 7, characterized in that, The oral liquid also includes sweeteners, preservatives, and drinking water.

9. The oral liquid as described in claim 8, characterized in that, The sweetener is an acceptable sweetener for health products.

10. The oral liquid as described in claim 8, characterized in that, The preservative mentioned is an acceptable preservative for health products.

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