Lactobacillus plantarum VB165 strain and use thereof

Abstract

The Lactobacillus plantarum VB165 strain, which was collected in the China General Microbiological Culture Collection Center on 28 September 2022 under preservation number CGMCC NO. 25839. When simulating in intestinal environment, the strain can inhibit the activity of α-glucosidase, can degrade bile salts, inhibits the activity of primary pathogenic gut bacteria, and possesses good tolerance to gastric acid and choline; said strain can be applied in the preparation of products to control blood glucose.

Description

Lactobacillus plantarum VB165 and its application

[0001] Priority information

[0002] This application claims priority to Chinese patent application number 202310921834.5, filed with the Patent Office of China on July 26, 2023, entitled “A Lactobacillus plantarum VB165 and its application”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to the technical field of microorganisms, and in particular to Lactobacillus plantarum VB165 and application thereof in hypoglycemic products. Background Art

[0004] Lactobacillus plantarum is a type of lactic acid bacteria with an optimal growth temperature of 30-35°C. It is either anaerobic or facultatively anaerobic and is a homofermentative lactic acid bacterium. As a member of the Lactobacillus genus, Lactobacillus plantarum has been widely used in food production, including kimchi, winemaking, fermented yogurt, fermented juices, synbiotic preparations, and probiotics. Furthermore, due to its high viable cell count and ability to produce lactobacilli, a biological preservative, Lactobacillus plantarum is used in aquaculture, animal husbandry, and other industries.

[0005] Lactobacillus plantarum is widely distributed in nature, found in the intestines of humans and insects, dairy products, fruits, and vegetables, and has a strong ability to metabolize sugars. As a beneficial intestinal microorganism, Lactobacillus plantarum can occupy and colonize the gastrointestinal tract through competitive inhibition, inhibiting invasion of the gastrointestinal tract by pathogenic bacteria, regulating the balance of intestinal flora, and also has immunomodulatory functions, such as regulating the proliferation of spleen cells and influencing the Th1 / Th2 cell balance. Lactobacillus plantarum is widely distributed in the intestines of adults, and the human body has immune tolerance to it, and it does not cause immune rejection effects. The U.S. Food and Drug Administration (FDA) and the European Food Safety Authority (EFSA) have recognized Lactobacillus plantarum as safe, and Chinese regulations have also approved Lactobacillus plantarum for use in food and health foods.

[0006] The incidence of diabetes is rapidly increasing worldwide. Most patients with T2DM in China experience elevated postprandial blood glucose (PPG). Elevated PPG is directly correlated with the development of diabetic complications, and controlling PPG is a key strategy for managing the onset and progression of diabetic complications. α-glucosidase inhibitors significantly control PPG by inhibiting α-glucosidase in the intestinal mucosa, slowing the breakdown of starch into glucose and reducing glucose absorption in the small intestine. Currently, the most commonly used α-glucosidase inhibitors in clinical practice are acarbose and voglibose, which are often associated with gastrointestinal adverse reactions such as abdominal discomfort, flatulence, increased flatulence, and diarrhea.

[0007] Hyperlipidemia can easily lead to cardiovascular and cerebrovascular diseases, such as myocardial infarction and coronary artery disease, and can easily trigger acute pancreatitis. Bile salts play a vital role in fat digestion. They reduce the surface tension of fat, emulsifying it into very fine chylomicrons. This increases the contact area between fat and enzymes, facilitating fat breakdown and absorption. Bile acids, primarily synthesized in the liver, are a crucial pathway in cholesterol metabolism. One-third of cholesterol catabolism occurs through bile acid synthesis. Bile salts are sodium or potassium salts formed by the binding of bile acids with glycine or taurine. Consuming bile salts not only reduces fat absorption but also promotes the metabolism of cholesterol into bile acids, thereby increasing the breakdown of cholesterol.

[0008] Summary of the Invention

[0009] The present invention provides a strain of Lactobacillus plantarum VB165, isolated from adult feces. The strain was deposited with the General Microbiology Center of the China General Culture Collection Administration on September 28, 2022, under the deposit number CGMCC No. 25839. Using adult feces as the isolation source, the strain obtained has higher safety and is more suitable for the production of health products.

[0010] The present invention provides a Lactobacillus plantarum that can effectively inhibit the activity of α-glucosidase. By utilizing its intestinal colonization and strict anaerobic characteristics, it can inhibit the activity of α-glucosidase in the intestine, degrade bile salts, and inhibit major intestinal pathogens without causing adverse reactions. It can bring many medical and health benefits in controlling the increase of host PPG, improving hyperlipidemia and cholesterol, and antagonizing intestinal infections.

[0011] The Lactobacillus plantarum provided by the present invention is in the form of living cells, for example, the strain is in the form of bacterial culture fluid.

[0012] The Lactobacillus plantarum provided by the present invention is in the form of non-viable cells, for example, the strain is in the form of freeze-dried powder.

[0013] The plant lactobacillus provided by the present invention can be used to prepare one or more of medicines, health products, foods, beverages, additives, and feeds for controlling or lowering blood sugar, and preferably controls or lowers blood sugar by inhibiting α-glucosidase activity.

[0014] The plant lactobacillus provided by the present invention can be used to prepare one or more of medicines, health products, foods, beverages, additives and feeds for degrading bile salts.

[0015] The plant lactobacillus provided by the present invention can be used to prepare one or more of medicines, health products, foods, beverages, additives and feeds for inhibiting the activity of intestinal pathogens.

[0016] The present invention provides a composition comprising the above Lactobacillus plantarum. The composition may comprise additional probiotic materials and may further comprise prebiotic materials.

[0017] The composition provided by the present invention further comprises an ingestible carrier, which can be a pharmaceutically acceptable carrier, such as a capsule, tablet or powder; or a food, such as acidified milk, yogurt, frozen yogurt, milk powder, condensed milk, cheese sauce, condiment or beverage.

[0018] The plant lactobacillus provided by the present invention can be subjected to anaerobic fermentation in a nutrient medium containing an assimilable carbon source and / or nitrogen source.

[0019] In the preferred technical solution, the assimilable carbon source is selected from starch, maltodextrin, glucose, sucrose, lactose, maltose, industrial molasses, glycerol, soybean oil, sorbitol, mannitol or any combination thereof; preferably glucose.

[0020] In the preferred technical solution, the assimilable nitrogen source is selected from yeast extract, yeast powder, yeast paste, soy lecithin, soybean cake powder, cottonseed cake powder, peanut cake powder, gluten powder, corn steep liquor, soybean meal, peptone, urea, ammonium salt, or any combination thereof; preferably, yeast extract.

[0021] In the preferred technical solution, the nutrient medium further includes an inorganic salt, which is selected from one or a combination of trisodium citrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ammonium sulfate, calcium carbonate, ferrous sulfate, zinc sulfate, copper sulfate, sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, and manganese sulfate; preferably dipotassium hydrogen phosphate and magnesium sulfate.

[0022] The present invention provides the use of the aforementioned Lactobacillus plantarum or the aforementioned composition in preventing, alleviating, and / or treating individual hyperglycemia. As previously mentioned, Lactobacillus plantarum VB165 can effectively inhibit the activity of α-glucosidase in the intestinal tract and has good tolerance to gastric acid and choline. Thus, it can be used to lower blood sugar and further effectively prevent, alleviate, and / or treat hyperglycemia and related diseases caused by it.

[0023] The present invention provides a method for preventing, alleviating and / or treating hyperglycemia in an individual, comprising administering the aforementioned Lactobacillus plantarum or the aforementioned composition to the individual. As previously mentioned, Lactobacillus plantarum VB165 can effectively inhibit the activity of α-glucosidase in the intestinal tract and has good tolerance to gastric acid and choline. Thus, it can be used to lower blood sugar. The method according to an embodiment of the present invention can effectively prevent, alleviate and / or treat hyperglycemia and related diseases caused by it.

[0024] The present invention provides the use of the aforementioned Lactobacillus plantarum or the aforementioned composition in preventing, alleviating, and / or treating individual hypercholesterolemia. As previously mentioned, Lactobacillus plantarum VB165 can effectively degrade bile salts in the intestinal tract and has good tolerance to gastric acid and choline. Therefore, this strain and its related products can effectively prevent, alleviate, and / or treat hypercholesterolemia and related diseases caused by it.

[0025] The present invention provides a method for preventing, alleviating and / or treating high bile salts in an individual, comprising administering the aforementioned Lactobacillus plantarum or the aforementioned composition to the individual. As previously mentioned, Lactobacillus plantarum VB165 can effectively degrade bile salts in the intestinal tract and has good tolerance to gastric acid and choline. Thus, it can be used to reduce bile salts. The method according to an embodiment of the present invention can effectively prevent, alleviate and / or treat high bile salts and related diseases caused by it, and improve high blood fat and cholesterol.

[0026] The present invention provides the use of the aforementioned Lactobacillus plantarum or the aforementioned composition in preventing, alleviating, and / or treating diseases related to intestinal pathogen infection. As mentioned above, Lactobacillus plantarum VB165 can inhibit the activity of major pathogens in the intestine and has good tolerance to gastric acid and choline. Therefore, this strain and its related products can effectively prevent, alleviate, and / or treat high bile salts and related diseases caused by intestinal pathogen infection.

[0027] The present invention provides a method for preventing, alleviating and / or treating related diseases caused by intestinal pathogen infection in an individual, comprising administering the aforementioned Lactobacillus plantarum or the aforementioned composition to the individual. As mentioned above, Lactobacillus plantarum VB165 can inhibit the activity of major pathogens in the intestine and has good tolerance to gastric acid and choline. Thus, the method according to an embodiment of the present invention can effectively prevent, alleviate and / or treat intestinal pathogen infection and related diseases caused thereby.

[0028] The beneficial effects of the present invention are at least:

[0029] The Lactobacillus plantarum VB165 provided by the present invention has a strong inhibitory effect on α-glucosidase, and can control or lower blood sugar through this pathway; it can also degrade bile salts, and improve the state of hyperlipidemia and hypercholesterolemia through this pathway; and it has strong ability to inhibit major intestinal pathogens and be acid and bile salt resistant, and can be used as a probiotic in the preparation of blood sugar lowering or control products.

[0030] Collection information:

[0031] Strain name: Lactobacillus plantarum VB165

[0032] Date of deposit: September 28, 2022

[0033] Depository: General Microbiology Center of China Culture Collection Administration

[0034] Accession number: CGMCC NO.25839 BRIEF DESCRIPTION OF THE DRAWINGS

[0035] FIG1 shows the results of the inhibitory ability test on α-glucosidase activity of acarbose, blank culture medium, Lactobacillus rhamnosus LGG and Lactobacillus plantarum VB165 provided by the present invention. DETAILED DESCRIPTION

[0036] The following examples are illustrative and are intended only to explain the present invention and are not to be construed as limiting the present invention. Where specific techniques or conditions are not specified in the examples, the methods were performed according to those described in the literature in the art or in accordance with the product instructions. All reagents or instruments used, for which the manufacturer is not specified, are commercially available conventional products.

[0037] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0038] Unless otherwise specified, the materials and reagents used in the following examples are all common commercial products and can be purchased on the market.

[0039] The MAGEN bacterial genomic DNA extraction kit was purchased from Shanghai Maigen Biotechnology Co., Ltd. Phusion high-fidelity DNA polymerase was purchased from Thermo Scientific.

[0040] The present invention will be further described below by way of examples, which are not intended to further limit the present invention. Those skilled in the art will appreciate that any equivalent substitutions or corresponding improvements made to the present invention are still within the scope of protection of the present invention.

[0041] In the culture medium used in the following examples, the percentage (%) used in the material ratio is by mass unless otherwise specified.

[0042] Example 1 Source of strains

[0043] Lactobacillus plantarum VB165 is isolated from adult feces. Fresh adult feces is added to sterile PBS buffer in a sterile container, thoroughly shaken, and then diluted and spread onto MRS solid culture plates. After 48 hours of incubation, single colonies of varying sizes and morphologies are streaked onto a fresh plate. Single colonies are then transferred to a slant surface for microscopic examination of morphology. Identification is performed after the presence of contaminants. Using adult feces as the isolation source results in a strain with enhanced safety, making it more suitable for use in the production of health products.

[0044] Example 2: Bacteria identification

[0045] The experiment was conducted according to the instructions in the Molecular Cloning Manual. After culturing the purified strain in MRS medium for one day, 1 mL of the culture was centrifuged at 10,000 rpm for 1 minute. The supernatant was discarded, and the cells were harvested for DNA extraction using the MAGEN Bacterial Genomic DNA Extraction Kit. Using the extracted DNA as a template, 16S rDNA was amplified by PCR (polymerase chain reaction) using universal primers 337F (5′-GACTCCTACGGGAGGCWGCAG-3′) and 1492R (5′-TACGGCTACCTTGTTACGACTT-3′). The PCR reaction volume was 50 μL, and the reaction system and protocol were configured according to the Phusion High-Fidelity DNA Polymerase instructions. The annealing temperature was 55°C. The PCR product was sent to Shanghai Bioengineering for sequencing.

[0046] The 16S rDNA sequence of the strain was collated and compared with the sequences of related species and genera in the GenBank database by BLAST to determine the taxonomic status of the strain. After comparison, it was found that its homology with Lactobacillus plantarum strain 1942 was as high as 99.93%, and its homology with Lactobacillus plantarum strain 1809 was as high as 99.86%. Therefore, the strain was identified as a Lactobacillus plantarum strain. The internal number of this strain is VB165. It was deposited in the General Microbiology Center of the China Culture Collection Administration on September 28, 2022. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.25839.

[0047] 16S sequencing sequence:

[0048] Example 3: Bacteria fermentation

[0049] 1. Bacteria activation

[0050] Take the glycerol tube of the original VB165 strain, thaw it at room temperature, draw 0.1 ml of the bacterial suspension and inoculate it onto the MRS solid plate, spread it evenly, put the plate into an anaerobic bag (Haibo Biological), and culture it in a 37°C incubator for 48 hours to obtain activated single colonies.

[0051] 2. Seed preparation

[0052] Take a single activated colony and scrape a loopful of it with an inoculating loop. Inoculate the loopful into a 500ml Erlenmeyer flask containing 100ml of MRS liquid medium. Wrap the flask in an anaerobic bag and incubate on a shaker at 37°C and 70 rpm for 48 hours. Ensure the OD value of the seed solution is ≥2.0. Adjust the pH of the MRS liquid medium to 7.0±0.2 before disinfection. Disinfection conditions: 121°C–123°C for 30 minutes.

[0053] 3. Fermentation preparation

[0054] Inoculate 3-5% (v / w) of the cultured seed liquid into a 500ml Erlenmeyer flask containing 100ml of liquid culture medium (MRS, artificial intestinal fluid (with enzymes, BD MRS, Coolaber), or artificial intestinal fluid plus 1% soluble starch). Wrap the flask in an anaerobic bag and incubate on a shaker at 37°C and 70 rpm for 24-48 hours. Maintain an OD value of ≥3.0. Adjust the pH of the MRS liquid culture medium to 7.0±0.2 before sterilization at 121-123°C for 30 minutes.

[0055] MRS liquid medium: yeast extract 0.5%, peptone 1.0%, beef extract 1.0%, glucose 2.0%, dipotassium hydrogen phosphate 0.2%, diammonium hydrogen citrate 0.2%, sodium acetate 0.5%, magnesium sulfate 0.02%, manganese sulfate 0.005% (all percentages are by mass), Tween 80 0.1% (volume / mass ratio).

[0056] MRS solid medium: Add 1.5% agar powder to MRS liquid medium.

[0057] Artificial intestinal fluid: contains enzymes, BD MRS, and Coolaber.

[0058] Artificial intestinal fluid + starch: artificial intestinal fluid, 1% soluble starch.

[0059] Example 4: Detection of the inhibitory ability of α-glucosidase activity

[0060] In order to detect the ability of strains to produce AGI in different environments, especially intestinal environments, artificial simulated small intestinal fluid was used to simulate the intestinal state, and artificial small intestinal fluid with 1% soluble starch added was used to simulate the state of eating carbohydrates. The inhibitory activity of the fermentation supernatants of VB165 and Lactobacillus rhamnosus LGG (ATCC53103) on α-glucosidase was compared in these environments.

[0061] After 48 hours of incubation, the culture supernatant was collected by centrifugation. For the α-glucosidase activity assay, the reaction system consisted of a 100 μL reaction mixture. First, 25 μL of the substrate 4-nitrophenol-α-D-pyranoglucopyranoside (PNPG, 10 mmol / L, Solebol) and 25 μL of the sample were mixed and incubated at 37°C for 10 minutes. Then, 50 μL of α-glucosidase (40 U / L, Sigma) was added, mixed, and incubated at 37°C for 30 minutes. The reaction was then quickly terminated by the addition of 100 μL of Na₂CO₃ (0.1 mol / L). The absorbance at 405 nm was measured using a microplate reader. Each experiment was repeated in triplicate. The absorbance of the sample was calibrated using the absorbance of a sample blank control (50 μL of PBS was used instead of α-glucosidase in the system). For the negative control (no sample inhibiting α-glucosidase in the system), 25 μL PBS was used instead of the sample. For the negative blank control (no α-glucosidase activity in the system), 25 μL PBS was used instead of the sample and 50 μL PBS was used instead of α-glucosidase.

[0062] The calculation formula of α-glucosidase inhibition rate is:

[0063] α-glucosidase inhibition rate = (1-(AB) / (CD)) × 100%

[0064] Where: A is the absorbance of the sample at 405 nm; B is the absorbance of the sample blank control at 405 nm; C is the absorbance of the negative control at 405 nm; D is the absorbance of the negative blank control at 405 nm.

[0065] The results are shown in Figure 1. The inhibition rates of 10 μM acarbose on α-glucosidase were 92.3% (MRS), 97.0% (artificial intestinal fluid), and 97.0% (artificial intestinal fluid + starch); the inhibition rates of blank culture medium were 1.4% (MRS), 4.4% (artificial intestinal fluid), and 3.8% (artificial intestinal fluid + starch); the inhibition rates of Lactobacillus rhamnosus LGG were 28.1% (MRS), 8.9% (artificial intestinal fluid), and 13.1% (artificial intestinal fluid + starch); and the inhibition rates of Lactobacillus plantarum VB165 were 56.6% (MRS), 63.6% (artificial intestinal fluid), and 68.7% (artificial intestinal fluid + starch). The results showed that under conditions of simulated artificial intestinal fluid, including both with and without the addition of starch, the fermentation supernatant of Lactobacillus plantarum VB165 had a strong inhibitory effect on α-glucosidase, and was significantly higher than that of Lactobacillus rhamnosus LGG.

[0066] Example 5 Bile Salt Degradation Ability Detection

[0067] To test the ability of the strain to degrade bile salts, the following six major bile salts (source leaf organisms) were added to Lactobacillus MRS culture medium: 4 mmol / L sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), and sodium glycocholate (GC); and 2 mmol / L sodium glycochenodeoxycholate (GCDC) and sodium glycodeoxycholate (GDC). Bile salt degradation was assessed based on the consumption rate after incubation. 2 ml of bile salt-containing MRS medium was added to each well of a 48-well plate. Resuscitated strains were then added at 1% of the volume. The culture was placed in an anaerobic bag and incubated anaerobically at 37°C and 90 rpm for 24 hours. Sample processing: 1 ml of the plate medium was centrifuged, and the supernatant was collected for HPLC analysis to determine bile salt degradation ability.

[0068] The analysis conditions were as follows: chromatographic column: Ultimate AQ-C18 (5 μm, 4.6×250 mm); flow rate: 1.0 ml / min; detection wavelength: 204 nm; column temperature: 45°C; mobile phase A: phosphate buffer, mobile phase B: acetonitrile, A:B = 56:44.

[0069] The results showed that Lactobacillus plantarum VB165 could degrade five bile salts: sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), and sodium glycodeoxycholate (GDC). Lactobacillus rhamnosus LGG could only degrade sodium glycochenodeoxycholate (GCDC) and sodium glycodeoxycholate (GDC). Therefore, Lactobacillus plantarum VB165 has a stronger bile salt degradation ability.

[0070] Table 1 Test results of the ability of strains to degrade bile salts

[0071] Example 6 Antagonistic Pathogen Ability Test

[0072] The antibacterial ability of the strains was evaluated using the double-layer plate culture method.

[0073] Preparation of the lower layer probiotics: Thaw the activated frozen tubes of different single bacteria, spot them on the MRS solid plate, put them in an anaerobic bag, and culture them anaerobically at 37°C for 48 hours. The inoculation volume of the bacteria is 2μl / spot.

[0074] Preparation of the upper pathogenic bacteria layer: The upper culture medium sterilized by high temperature is cooled to 40-50°C, and different pathogens are added as required (the final concentration of pathogens in the culture medium is about 10 6 100 mL / plate) was mixed thoroughly. 7 mL / plate of the pathogenic bacteria culture medium was added to each of the previously grown probiotic single-strain plates. After solidification, the culture was performed according to the growth conditions of different pathogens (Table 2). After the upper plate was fully grown, the presence of an inhibition zone around the probiotics was observed to determine their antibacterial activity. All pathogenic bacteria strains were purchased from Qingdao Haibo Biotechnology.

[0075] Table 2 Types of pathogenic strains and culture conditions

[0076] As shown in Table 3, Lactobacillus plantarum VB165 can inhibit all 7 common pathogens included in Table 2 and has the ability to antagonize pathogens.

[0077] Table 3 Antibacterial effect of Lactobacillus plantarum VB165

[0078] Example 7 Acid and bile salt resistance

[0079] 1 mL of a 24-hour bacterial suspension was centrifuged and the supernatant removed. The suspension was then resuspended in either artificial gastric fluid (Coolaber, pH 3.0) or 0.1% ox bile salts (Coolaber, prepared in PBS) and incubated at 37°C for 2 hours. Serial dilutions were performed and colonies were counted on plates to calculate the survival rate of the test bacteria. The results showed that Lactobacillus plantarum VB165 had a 2-hour survival rate of 99.50% and a 4-hour survival rate of 98.30% in artificial gastric fluid (pH 3.0). In addition, Lactobacillus plantarum VB165 had a 2-hour survival rate of 98.70% and a 4-hour survival rate of 95.60% in 0.1% ox bile salts, demonstrating strong acid and bile salt resistance.

[0080] Table 4 Acid and bile salt resistance of Lactobacillus plantarum VB165

[0081] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A Lactobacillus plantarum VB165, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with a deposit number of CGMCC NO.25839 and a deposit date of September 28, 2022.

2. The plant lactobacillus according to claim 1, wherein The Lactobacillus plantarum is in the form of living cells.

3. The plant lactobacillus according to claim 2, wherein The Lactobacillus plantarum is in the form of bacterial culture solution.

4. The plant lactobacillus according to claim 1, wherein The Lactobacillus plantarum is in the form of non-living cells.

5. The plant lactobacillus according to claim 4, wherein The Lactobacillus plantarum is in the form of freeze-dried powder.

6. A use of Lactobacillus plantarum according to any one of claims 1 to 5, characterized in that: Used to prepare one or more of medicines, health products, foods, beverages, additives, and feeds for controlling or lowering blood sugar.

7. The use according to claim 6, characterized in that Control or lower blood sugar by inhibiting α-glucosidase activity.

8. A use of Lactobacillus plantarum according to any one of claims 1 to 5, characterized in that: Used for preparing one or more of medicines, health products, foods, beverages, additives, and feeds for degrading bile salts.

9. A use of Lactobacillus plantarum according to any one of claims 1 to 5, characterized in that: Used to prepare one or more of medicines, health products, foods, beverages, additives, and feeds that inhibit the activity of intestinal pathogens.

10. A composition, characterized in that The composition comprises the Lactobacillus plantarum according to any one of claims 1 to 5.

11. The composition according to claim 10, characterized in that The composition may also comprise additional probiotic material.

12. The composition according to claim 10, characterized in that The composition also comprises a prebiotic material.

13. The composition according to any one of claims 10 to 12, characterized in that The composition also includes an ingestible carrier.

14. The composition according to claim 13, characterized in that The ingestible carrier is a pharmaceutically acceptable carrier.

15. The composition according to claim 13, characterized in that The ingestible carrier is a capsule, tablet or powder.

16. The composition according to claim 13, characterized in that The ingestible carrier is a food.

17. The composition according to claim 16, characterized in that The food is acidified milk, yogurt, frozen yogurt, milk powder, concentrated milk, cheese sauce, condiment or beverage.

18. A fermentation method of Lactobacillus plantarum according to any one of claims 1 to 5, characterized in that: The fermentation method comprises carrying out anaerobic fermentation in a nutrient medium containing an assimilable carbon source and / or nitrogen source.

19. The fermentation method according to claim 18, characterized in that The assimilable carbon source is selected from starch, maltodextrin, glucose, sucrose, lactose, maltose, industrial molasses, glycerol, soybean oil, sorbitol, mannitol or any combination thereof.

20. The fermentation method according to claim 18, characterized in that The assimilable carbon source is glucose.

21. The fermentation method according to claim 18, characterized in that The assimilable nitrogen source is selected from yeast extract, yeast powder, yeast extract, soybean lecithin, soybean cake powder, cottonseed cake powder, peanut cake powder, gluten powder, corn steep liquor, soybean meal, peptone, urea, ammonium salt or any combination thereof.

22. The fermentation method according to claim 18, characterized in that The assimilable nitrogen source is yeast extract.

23. The fermentation method according to claim 18, characterized in that The nutrient medium also includes an inorganic salt, which is selected from one of trisodium citrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ammonium sulfate, calcium carbonate, ferrous sulfate, zinc sulfate, copper sulfate, sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, manganese sulfate, or any combination thereof.

24. The fermentation method according to claim 18, characterized in that The nutrient medium also includes dipotassium hydrogen phosphate and magnesium sulfate.

25. Use of the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 in preventing, alleviating and / or treating hyperglycemia in an individual.

26. A method for preventing, alleviating and / or treating hyperglycemia in an individual, characterized in that: The method comprises administering the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 to the individual.

27. Use of the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 in preventing, alleviating and / or treating high bile salts in an individual.

28. A method for preventing, alleviating and / or treating hypercholesterolemia in an individual, characterized in that: The method comprises administering the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 to the individual.

29. Use of the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 in preventing, alleviating and / or treating diseases related to intestinal pathogen infection.

30. A method for preventing, alleviating and / or treating diseases related to intestinal pathogen infection in an individual, characterized in that: The method comprises administering the Lactobacillus plantarum according to any one of claims 1 to 5 or the composition according to any one of claims 10 to 17 to the individual.

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