Lactobacillus plantarum capable of lubricating intestines and passing stool and application thereof

Abstract

This invention discloses a strain of *Lactiplantibacillus plantarum* ZSHT-001 that can lubricate the intestines and relieve constipation, and its applications, belonging to the field of biotechnology. The *Lactiplantibacillus plantarum* ZSHT-001 was isolated from kimchi. The preservation number of this *Lactiplantibacillus plantarum* is GDMCC No: 66147. The fermentation product of *Lactiplantibacillus plantarum* ZSHT-001 provided by this invention has excellent antibacterial properties, good acid and bile salt resistance, and a high ability to promote gastrointestinal motility, and has the potential to relieve constipation. It is of great significance for the development of drugs, health products, and foods for the prevention or treatment of constipation.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a strain of *Lactobacillus plantarum* that can lubricate the intestines and promote bowel movements, and its applications. Technical Background

[0002] Constipation has become a prevalent digestive disorder in modern society. The symptoms such as bloating and abdominal pain caused by difficulty in defecation and hard stools significantly impact patients' quality of life. Long-term constipation not only leads to the accumulation of toxins and dysbiosis in the intestines, but may also induce organic lesions such as hemorrhoids and intestinal obstruction, creating a vicious cycle. While commonly used chemical laxatives can quickly relieve symptoms, long-term use can easily lead to drug dependence problems such as electrolyte imbalance and decreased intestinal nerve sensitivity. Traditional dietary fiber supplements have limitations such as a long onset of action and significant individual differences in response, making it difficult to meet patients' needs for safe and effective treatments.

[0003] Probiotic therapy has gained attention as an alternative intervention, with its mechanism of action—promoting metabolism by regulating gut microbiota—offering a natural safety advantage. However, the functional performance of conventional *Lactobacillus plantarum* strains in promoting intestinal motility and stimulating enteroneurin activity varies considerably, and most strains have insufficient levels of functional components such as short-chain fatty acids and bioactive peptides in their metabolites, failing to effectively synergistically improve intestinal motility and inhibit the excessive proliferation of pathogenic bacteria. More importantly, current probiotic fermentation processes generally use standardized culture media, lacking precise regulation of constipation-related metabolic pathways, resulting in a limited variety of organic acids produced by the strains. This makes it difficult to overcome the technical bottleneck of slow onset and insignificant effects in traditional probiotic preparations. Based on these issues, screening for a specific strain with constipation-relieving effects and its suitable culture system has become an important direction for solving the problem of constipation intervention. Summary of the Invention

[0004] The purpose of this invention is to provide a strain of *Lactobacillus plantarum* that can lubricate the intestines and promote bowel movements, and its applications. The fermentation product obtained by using this *Lactobacillus plantarum* as a fermentation strain has excellent antibacterial properties, good acid and bile salt resistance, and a high ability to promote gastrointestinal motility, and also has the potential to relieve constipation.

[0005] This invention isolated and screened a strain ZSHT-001 from kimchi. Physicochemical analysis and 16S rRNA gene alignment identified the strain as *Lactobacillus plantarum*, and named it *Lactobacillus plantarum* ZSHT-001. This *Lactobacillus plantarum* strain was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 14, 2025, with accession number GDMCC No: 66147.

[0006] In vitro experiments have shown that the fermentation products obtained from this Lactobacillus plantarum as a fermentation strain can significantly inhibit harmful intestinal bacteria and relieve constipation.

[0007] Preferably, the preparation of the fermentation product includes the following steps:

[0008] Step 1: Dilute the activated ZSHT-001 bacterial suspension with sterile water to a concentration of 1*10. 7 -1*10 8 CFU / mL bacterial culture solution;

[0009] Step 2: Inoculate the bacterial culture solution into the liquid culture medium at an inoculation rate of 1-3 v / v%, ferment for 24-36 h, filter, and obtain the fermentation product filtrate;

[0010] Step 3: Freeze-dry the fermentation product filtrate to obtain the fermentation product.

[0011] Preferably, the liquid culture medium is MRS liquid culture medium;

[0012] Furthermore, the liquid culture medium is a specific liquid culture medium, which comprises the following parts by weight of raw materials:

[0013] 3-6 parts okra extract;

[0014] 3-5 portions of wheat peptides;

[0015] 2-4 parts inulin;

[0016] 4-6 parts of fructooligosaccharides;

[0017] 1-3 parts polydextrose;

[0018] 2-4 parts arabinose;

[0019] 40-50 portions of sterile deionized water.

[0020] The second objective of this invention is to provide the application of Lactobacillus plantarum ZSHT-001 in the preparation of food or health products that have the effect of relieving constipation.

[0021] A third objective of this invention is to provide the application of Lactobacillus plantarum ZSHT-001 in the preparation of food or health products that have the effect of regulating intestinal flora.

[0022] Beneficial effects of this invention:

[0023] This invention provides a strain of *Lactobacillus plantarum* ZSHT-001, with the preservation number GDMCC No: 66147. Using this strain and fermentation on MRS medium or a medium prepared from okra extract, wheat peptides, inulin, fructooligosaccharides, polydextrose, and arabinose, the resulting fermentation product exhibits good constipation-relieving effects and significantly inhibits the growth of pathogenic bacteria in the gastrointestinal tract, particularly against *Escherichia coli*, *Staphylococcus aureus*, *Salmonella typhimurium*, and *Listeria monocytogenes*. It can be used to prepare products for preventing or improving constipation.

[0024] Preservation of biological materials

[0025] A strain of Lactiplantibacillus plantarum, ZSHT-001, was deposited on April 14, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No. 66147), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Detailed Implementation

[0026] The present invention will be further described below with reference to specific embodiments. It should be noted that these descriptions of embodiments are for the purpose of aiding understanding the present invention, but do not constitute a limitation thereof. Furthermore, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0027] The sources of some of the raw materials for this invention are as follows:

[0028] Okra extract: purchased from Zhongbai Xingye Food Technology (Beijing) Co., Ltd.;

[0029] Wheat peptides: purchased from Zhejiang Pingtailong Biotechnology Co., Ltd.

[0030] Inulin: Purchased from Zhongbai Xingye Food Technology (Beijing) Co., Ltd.;

[0031] Fructooligosaccharides: purchased from Quantum Hi-Tech (Guangdong) Biotechnology Co., Ltd.;

[0032] Polydextrose: purchased from Shandong Bailong Chuangyuan Biotechnology Co., Ltd.;

[0033] Arabic sugar: purchased from Zhejiang Huakang Pharmaceutical Co., Ltd.;

[0034] Unless otherwise specified, all other ingredients are commercially available.

[0035] Example 1: Isolation, Screening and Identification of Strains

[0036] 1. Discovery and isolation of strains

[0037] Materials: Pickled vegetables, purchased from Shandong Nongyu Food Co., Ltd. Take 5g of pickled vegetable leaves and homogenize them in 10mL of sterile physiological saline to obtain pickled vegetable homogenate.

[0038] Add 5 mL of kimchi homogenate to 45 mL of sterile physiological saline and mix well. Then, use physiological saline to serially dilute to 10⁻⁶. -5 10 -6 10 -7 The bacteria were spread onto MRS solid medium, with three replicates for each gradient. The plates were then incubated at 37°C for 1 day. Colonies showing significant differences were selected and streaked onto MRS solid medium for isolation. After incubation at 37°C for 1 day, the streaking was repeated three times. Single colonies were then selected for hydrogen peroxide titration. A positive catalase test was indicated by the formation of bubbles on a slide containing 5% hydrogen peroxide, while a negative test was indicated by the absence of bubbles. Strains with negative catalase tests were suspected probiotic strains. Six suspected probiotic strains were obtained using this method.

[0039] 2. Screening of the antibacterial activity of suspected probiotic strains against Listeria monocytogenes and Salmonella typhimurium.

[0040] Experimental Methods: The six suspected probiotic strains obtained from screening were labeled as No. 1-6. Strains 1-6 were cultured in MRS liquid culture at 37℃ for 1 day to obtain bacterial suspensions. The bacterial suspensions were then diluted to 10⁻⁶ with sterile deionized water. 8 The bacterial suspension was obtained at CFU / mL, and the inhibition rate of the bacterial suspension against Listeria monocytogenes and Salmonella typhimurium was determined by the filter paper disc method (three replicates were performed for each strain 1-6). The volume of bacterial suspension was 100 μL.

[0041] Preparation of culture dishes for inhibited bacteria: Prepare culture dishes containing 10... 8 CFU / mL, take 1 mL and add it to the sterilized MRS solid medium, spread it evenly, and obtain the culture dish of inhibited bacteria.

[0042] 3. Screening for the tolerance of suspected probiotic strains to artificial gastric juice and bile salts.

[0043] Preparation of artificial gastric fluid: Prepare a 0.5 wt% sodium chloride solution, add 0.3 wt% pepsin, dissolve thoroughly, adjust the pH to 1.5 with 1 mol / L HCl, and filter with a 0.22 μm microporous membrane for sterilization before use.

[0044] Bile salt preparation: Add bile salt to MRS liquid medium to prepare MRS medium containing 0.6% bile salt. After complete dissolution, filter with a 0.22μm microporous membrane for sterilization and use.

[0045] Experimental method: The labeled strains 1-6 were cultured in MRS liquid culture medium at 37℃ for 1 day to obtain bacterial suspensions. The bacterial suspensions were then diluted to 10 with sterile deionized water. 8 CFU / mL was used to obtain a bacterial suspension. 0.5 mL of the bacterial suspension was inoculated into 4.5 mL of prepared artificial gastric juice or bile salt solution and mixed well to obtain a bacterial artificial gastric juice or bile salt solution containing 1 v / v%. The bacterial artificial gastric juice or bile salt solution was incubated at 37°C. At the same time, the number of viable bacteria was detected by taking digestive fluid at 0 h and 3 h. The survival rate of the strain in artificial gastric juice and bile salt solution was calculated according to the strain survival rate formula (3 replicates were performed for each strain 1-6).

[0046] Strain survival rate = (N1 / N0) * 100%

[0047] Where N1 represents the number of viable bacteria after 3 hours of treatment, and N0 represents the number of viable bacteria after 0 hours.

[0048] The results of the antibacterial ability of the suspected probiotic strains are shown in Table 1 below.

[0049] Table 1. Detection of antibacterial ability, acid resistance, and bile salt resistance of suspected probiotic strains.

[0050]

[0051]

[0052] The results in Table 1 show that strain 3 has good antibacterial properties against both Listeria monocytogenes and Salmonella typhimurium, and strain 3 also has good tolerance to artificial gastric juice and bile salts. Therefore, strain 3 was selected as the optimal strain.

[0053] Strain morphological identification

[0054] Morphological identification of strain No. 3 revealed that it is a Gram-positive bacterium with white, smooth, raised, straight rods in short chains. It is non-spore-forming, non-flagellated, and exhibits homofermentation. It does not produce gas during glucose fermentation, does not reduce nitrite, does not produce indole, is negative for hydrogen sulfide production, and is negative for coagulase. Under a regular microscope, the strain appears rod-shaped.

[0055] molecular biological identification of strains

[0056] Strain number 3, obtained through screening, was identified using molecular biological identification methods. Sequencing revealed its 16SRNA as shown in SEQ ID NO. 1. BLAST alignment and phylogenetic tree construction were performed using this sequencing result against the NCBI database. The results showed that the strain sequence was highly homologous to the *Lactiplantibacillus plantarum* gene sequence, with a homology of 100%. Based on its morphological characteristics, it was identified as belonging to *Lactiplantibacillus plantarum*.

[0057] The strain was named Lactiplantibacillus plantarum ZSHT-001 and deposited on April 14, 2025, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, China, postal code 510070, with accession number GDMCC No:66147.

[0058] Application example: Preparation of fermentation products

[0059] A fermentation product for preventing or improving constipation, the preparation method of which includes the following steps:

[0060] Application Example 1:

[0061] Step 1: Dilute the activated ZSHT-001 bacterial suspension with sterile water to a concentration of 1×10⁻⁶. 7 CFU / mL bacterial culture solution;

[0062] Step 2: Inoculate the bacterial culture into MRS liquid medium at an inoculum rate of 2 v / v%, ferment for 30 h, filter, and obtain fermentation product filtrate;

[0063] Step 3: Freeze-dry the fermentation product filtrate to obtain the fermentation product.

[0064] Application Example 2:

[0065] Step 1: Dilute the activated ZSHT-001 bacterial suspension with sterile water to a concentration of 1×10⁻⁶. 7 CFU / mL bacterial culture solution;

[0066] Step 2: Inoculate the bacterial culture solution into a specific liquid culture medium at an inoculation rate of 2 v / v%, ferment for 30 h, filter, and obtain the fermentation product filtrate;

[0067] Step 3: Freeze-dry the fermentation product filtrate to obtain the fermentation product;

[0068] The specific liquid culture medium mentioned in step 2 includes the following parts by weight of raw materials:

[0069] 5 parts okra extract;

[0070] 4 portions of wheat peptides;

[0071] 3 parts inulin;

[0072] 5 parts of fructooligosaccharides;

[0073] Two portions of polydextrose;

[0074] 3 portions of arabinose;

[0075] 45 portions of sterile deionized water.

[0076] Application Example 3:

[0077] Unlike Application Example 2, the specific liquid culture medium described in step 2 comprises the following parts by weight of raw materials:

[0078] 3 parts okra extract;

[0079] 5 portions of wheat peptides;

[0080] 2 parts inulin;

[0081] 6 parts of fructooligosaccharides;

[0082] 1 part polydextrose;

[0083] Two portions of arabinose;

[0084] 40 portions of sterile deionized water.

[0085] Application Example 4:

[0086] Unlike Application Example 2, the specific liquid culture medium described in step 2 comprises the following parts by weight of raw materials:

[0087] Okra extract 6 parts;

[0088] 3 portions of wheat peptides;

[0089] 4 parts inulin;

[0090] 4 parts of fructooligosaccharides;

[0091] 3 parts polydextrose;

[0092] 4 portions of arabinose;

[0093] 50 portions of sterile deionized water.

[0094] Comparative application example 1:

[0095] Unlike Application Example 1, Lactobacillus plantarum ZJUF SYS1 was used instead of Lactobacillus plantarum ZSHT-001. The preservation number of Lactobacillus plantarum is CCTCC NO:M2022662. The remaining parameters and steps are the same as in Application Example 2.

[0096] Compare with application example 2:

[0097] Unlike Application Example 1, Lactobacillus plantarum La-10 was used instead of Lactobacillus plantarum ZSHT-001. The preservation number of Lactobacillus plantarum is GDMCC NO: 63605. The remaining parameters and steps are the same as in Application Example 2.

[0098] Compare with example 3:

[0099] Unlike Application Example 2, the specific liquid culture medium does not contain wheat peptides. The missing mass fractions are made up with okra extract, inulin, fructooligosaccharides, polydextrose, arabinose, and sterile deionized water in a mass ratio of 5:3:5:2:3:45. The remaining parameters and steps are the same as in Application Example 2.

[0100] Compare with application example 4:

[0101] Unlike Application Example 2, the specific liquid culture medium does not contain okra extract. The missing mass fractions are made up with wheat peptides, inulin, fructooligosaccharides, polydextrose, arabinose, and sterile deionized water in a mass ratio of 4:3:5:2:3:45. The remaining parameters and steps are the same as in Application Example 2.

[0102] Efficacy verification:

[0103] 1) Validation of the efficacy of Lactobacillus plantarum fermentation products on constipation model mice

[0104] Test sample preparation: Take Application Examples 1-4 and Comparative Application Examples 1-4 and prepare them in a 15 wt% mixture using sterile deionized water, seal and set aside for use.

[0105] Two hundred SPF-grade male Kunming mice, weighing 20±2g, were selected and acclimatized for one week, during which they were allowed free access to food and water.

[0106] After one week of rearing, 100 mice weighing 18–22 g were randomly selected as experimental animals and randomly divided into 10 groups of 10 mice each: a blank control group, a model control group, application examples 1–4, and control application examples 1–4. Except for the blank control group, all other groups were administered loperamide hydrochloride (5 mg / kg bw) by gavage twice daily for 7 consecutive days to induce a functional constipation model in mice. The blank control group was administered an equal volume of physiological saline by gavage daily. Subsequently, experimental samples (application examples 1–4 and control application examples 1–4) were administered to the experimental groups by gavage daily at a volume of 0.2 mL / 10 g body weight. During this period, the blank control group and the model group were administered an equal volume of physiological saline by gavage. During the experiment, all mice had free access to food and water and were fed continuously for 14 days. After the 14th day, each group of mice was fasted but allowed free access to water for 12 hours. Then, each group underwent a final gavage. One hour after the gavage, each group was given 0.1 mL / 10 g body weight of activated charcoal suspension. Twenty minutes after the end of the gavage, the mice were euthanized by cervical dislocation. The small intestine was removed from the abdomen, and the distance the charcoal was propelled and the total length of the small intestine were measured. The propulsion rate (%) was calculated. The experimental results are average values ​​rounded to two decimal places, as shown in Table 1.

[0107] The activated carbon suspension was prepared by using deionized water and gum arabic to form a 5% (w / v) activated carbon suspension from activated carbon powder.

[0108] Propulsion rate % = Carbon powder propulsion distance / Total length of small intestine * 100%.

[0109] The higher the propagation rate, the better the effect of relieving constipation.

[0110] Table 2 Verification of Constipation Relief Efficacy

[0111] Group Propulsion rate / % Blank group 70.36％±4.14* Model control group 36.54％±2.49 Application Example 1 63.43％±3.25* Application Example 2 74.37％±2.32* Application Example 3 70.62％±4.17* Application Example 4 72.31％±3.45* Comparative Application Example 1 50.51％±6.31* Comparative Application Example 2 47.34％±4.36* Comparative Application Example 3 65.36％±5.49* Comparative Application Example 4 67.93％±3.27*

[0112] Note: "*" indicates that p < 0.05 compared with the model control group.

[0113] As shown in Table 1, the small intestinal propulsion rate was significantly lower in the model control group compared with the blank group, indicating that the constipation model was successfully established.

[0114] The results of Application Example 1 and the model control group show that Lactobacillus plantarum ZSHT-001 provided by the present invention has a certain effect in relieving constipation.

[0115] The results of Application Examples 1-2 show that the fermentation product of Lactobacillus plantarum ZSHT-001 provided by the present invention, fermented using the special culture medium specified by the present invention, has a better effect in relieving constipation.

[0116] A comparison of the results of Application Example 1 and Comparative Application Examples 1-2 shows that, compared with other Lactobacillus plantarum with similar effects, the Lactobacillus plantarum ZSHT-001 provided by this invention has a better effect in relieving constipation.

[0117] A comparison of the results of Application Example 2 and Comparative Application Examples 3-4 shows that changes in the composition of the culture medium have a certain impact on the small intestinal propulsion rate. The fermentation products cultured using the special culture medium specified in this invention have a better effect on relieving constipation.

[0118] 2) Verification of the inhibitory efficacy of Lactobacillus plantarum ZSHT-001 fermentation products against Escherichia coli and Staphylococcus aureus.

[0119] Test sample preparation: Take Application Example 1-2 and Comparative Application Example 1-2 and prepare a 15wt% mixture using sterile deionized water, seal and set aside for use.

[0120] Experimental method: The antibacterial ability against different harmful bacteria was determined using the filter paper disc method. *Escherichia coli* and *Staphylococcus aureus* were inoculated separately into LB liquid medium and cultured at 37°C for 24 hours on a shaker. The bacterial suspension was then diluted to 10⁻⁶. 8 CFU / mL, 0.1 mL of the diluted bacterial suspension was evenly spread onto a nutrient agar plate. Two sterile filter paper discs (5 mm in diameter) were then placed on the plate. For the experimental group, 10 μL of the test sample was added to the filter paper disc, while for the control group, an equal volume of sterile deionized water was added. The plates were incubated at 37℃ for 24 h, and the diameter of the inhibition zone was observed and measured. The experiment was performed in triplicate, and the results are expressed as mean ± standard deviation, as shown in Table 2.

[0121] Table 3 Antibacterial test of fermentation products of Lactobacillus plantarum ZSHT-001

[0122]

[0123] The results showed that Lactobacillus plantarum ZSHT-001 had a strong inhibitory effect on two common pathogenic bacteria, Escherichia coli and Staphylococcus aureus, with the best effect on Escherichia coli and the second best effect on Staphylococcus aureus.

[0124] Acute toxicity test:

[0125] Acute toxicity tests were conducted on the test samples to verify their safety.

[0126] Test samples: Application Examples 1-4;

[0127] Experimental methods:

[0128] Forty SPF-grade Kunming mice, weighing 20±2g each (half male and half female), were selected. 20g of the test sample was diluted with purified water to 60mL to reach the maximum permissible gavage concentration. The mice were administered the sample orally twice daily at 6-hour intervals, with each gavage volume being 0.3mL / 10g body weight, equivalent to a dose of 20.00g / kg body weight. Mice were fasted for 6 hours before the first gavage. They were observed for two weeks after gavage, and any signs of poisoning and mortality were recorded.

[0129] Experimental Results: No obvious symptoms of poisoning were observed in Kunming mice after gavage, and no deaths were observed during the 14-day observation period. At the end of the observation period, the test animals were euthanized and dissected. No obvious abnormalities were found in the major organs such as the liver, spleen, kidneys, intestines, heart, and lungs. The median lethal dose (LD50) of the fermentation product described in this invention for Kunming mice was greater than 20.00 g / kg·bw. According to the acute toxicity dose grading standard in GB15193.3-2014, it belongs to the practically non-toxic category.

[0130] A health supplement for preventing or improving constipation, the preparation method of which includes the following steps:

[0131] Health Supplement 1:

[0132] The fermentation product obtained in Application Example 1 was prepared into a 15 wt% mixture using drinking water, and then 1 wt% white sugar and 0.05 wt% vitamin C were added, mixed evenly, bottled, and sealed for storage.

[0133] Health Supplement 2:

[0134] The fermentation product obtained in Application Example 2 was prepared into a 15 wt% mixture using drinking water, and then 1 wt% white sugar and 0.05 wt% vitamin C were added, mixed evenly, bottled, and sealed for storage.

[0135] Health Supplement 3:

[0136] The fermentation product obtained in Application Example 3 was prepared into a 15 wt% mixture using drinking water, and then 1 wt% white sugar and 0.05 wt% vitamin C were added, mixed evenly, bottled, and sealed for storage.

[0137] Health Supplement 4:

[0138] The fermentation product obtained in Application Example 4 was prepared into a 15 wt% mixture using drinking water, and then 1 wt% white sugar and 0.05 wt% vitamin C were added, mixed evenly, bottled, and sealed for storage.

[0139] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. A fermentation product of *Lactobacillus plantarum* ZSHT-001, characterized in that, The method for preparing the fermentation product includes the following steps: Step 1: Dilute the activated ZSHT-001 bacterial suspension with sterile water to a concentration of 1×10⁻⁶. 7 -1×10 8 CFU / mL bacterial culture solution; Step 2: Inoculate the bacterial culture solution into the liquid culture medium at an inoculation rate of 1-3 v / v%, ferment for 24-36 h, filter, and obtain the fermentation product filtrate; Step 3: Freeze-dry the fermentation product filtrate to obtain the fermentation product; The liquid culture medium mentioned in step 2 is a specific liquid culture medium; The specific liquid culture medium mentioned in step 2 comprises the following parts by weight of raw materials: 3-6 parts okra extract; 3-5 portions of wheat peptides; 2-4 parts inulin; 4-6 parts of fructooligosaccharides; 1-3 parts polydextrose; 2-4 parts arabinose; 40-50 parts of sterile deionized water; The Lactobacillus plantarum ZSHT-001 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 14, 2025, with accession number GDMCC No:66147.

2. The use of the fermentation product according to claim 1 in the preparation of a product for preventing or improving constipation; the product is a health product.

3. A health product that helps with bowel movements, characterized in that, It contains the fermentation product as described in claim 1.

4. The use of the fermentation product of claim 1 in the preparation of health products that help regulate intestinal flora.

5. A health food for helping to regulate intestinal flora, characterized by, It contains the fermentation product as described in claim 1.

6. The health care product according to claim 3 or 5, wherein This also includes food-acceptable additives.

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