Propionibacterium acidipropionici BL8 with functions of reducing blood sugar and resisting aging and application thereof

Abstract

The present application relates to the field of microorganisms, and provides a propionibacterium acidifaciens BL8 with functions of reducing blood sugar and resisting aging, and application thereof, the propionibacterium acidifaciens BL8 is preserved in China General Microbiological Culture Collection Center on December 20, 2024, and the preservation number is CGMCC No.33134.The propionibacterium acidifaciens BL8 of the present application can inhibit DPP4 activity, promote INS-1 insulin secretion, inhibit acetylcholinesterase activity and superior cellulase activity.Can reduce blood sugar;Improve memory, prevent and relieve Alzheimer's disease;Promote cellulose absorption, improve constipation, and have high research and application value in reducing blood sugar and resisting aging.

Description

Technical Field

[0001] This invention relates to the field of microbiology, and in particular to a strain of Propionibacterium BL8 with hypoglycemic and anti-aging functions and its applications. Background Technology

[0002] Propionibacterium acidogeneticum is Propionibacterium A genus of microorganisms. Polymorphic bacilli, often club-shaped with rounded or pointed ends; some cells are spherical, forked, or branched, but not filamentous. Cells are single, paired, or in short chains, appearing in V or Y shapes, or arranged in squares. Gram-positive, non-motile, and non-sporogenic. Facultative anaerobic, with varying degrees of aerobic tolerance; most strains can grow in slightly oxygen-deficient air. They can ferment glucose to produce propionic acid, acetic acid, and CO2. Primarily found in cheese and dairy products.

[0003] Currently, there are few reports on *Propionibacterium oxytocinum*, and even fewer studies on its probiotic properties and metabolic mechanisms, which to some extent hinders its development and utilization. Therefore, it is necessary to increase the exploration of the characteristics of *Propionibacterium oxytocinum*, determine its efficacy based on strain function and probiotic metabolites, and clarify its application prospects. In conclusion, the research and application of *Propionibacterium oxytocinum* has broad development potential. Summary of the Invention

[0004] This invention aims to at least partially solve at least one of the technical problems existing in the prior art. To this end, this invention provides *Propionibacterium acidogeneticum*, fermentation broth, bacterial suspension, food, pharmaceutical, health product, feed, or additive, and their applications. The novel *Propionibacterium acidogeneticum* strain screened by this invention exhibits the ability to inhibit DPP4 activity, promote INS-1 insulin secretion, inhibit acetylcholinesterase activity, and possess superior cellulase activity, thus lowering blood sugar; improving memory, preventing and alleviating Alzheimer's disease; promoting cellulose absorption, and improving constipation. It has high research and application value in lowering blood sugar and anti-aging.

[0005] Therefore, in a first aspect of the present invention, the present invention provides a type of Propionibacterium acid-producing bacteria (… Propionibacterium acidipropionici BL8. According to an embodiment of the present invention, the accession number of the Propionibacterium BL8 is CGMCC No. 33134.

[0006] Preservation information:

[0007] Strain name: Propionibacterium acidogeneticum ( Propionibacterium acidipropionici )

[0008] Deposit date: December 20, 2024

[0009] Preservation Institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee

[0010] Accession number: CGMCC No. 33134

[0011] In a second aspect, the present invention provides a fermentation broth. According to an embodiment of the present invention, the fermentation broth comprises the aforementioned Propionibacterium BL8.

[0012] In a third aspect, the present invention provides a bacterial suspension. According to an embodiment of the present invention, the bacterial suspension comprises the aforementioned Propionibacterium BL8.

[0013] In a fourth aspect, the present invention provides a composition. According to embodiments of the present invention, the composition comprises at least one of the following: Propionibacterium BL8 as described in the first aspect, the fermentation broth as described in the second aspect, and the bacterial suspension as described in the third aspect.

[0014] In a fifth aspect of the invention, the use of the Propionibacterium BL8, the fermentation broth of the second aspect, the bacterial suspension of the third aspect, or the composition of the fourth aspect in the preparation of a medicine for the prevention, relief, treatment, or adjunctive treatment of type 2 diabetes.

[0015] In a sixth aspect of the invention, the use of the Propionibacterium BL8, the fermentation broth of the second aspect, the bacterial suspension of the third aspect, or the composition of the fourth aspect in the preparation of health products that help improve memory, and / or in the preparation of medicines for the prevention and relief of Alzheimer's disease.

[0016] In a seventh aspect of the invention, the use of the Propionibacterium BL8, the fermentation broth of the second aspect, the bacterial suspension of the third aspect, or the composition of the fourth aspect in the preparation of a medicine for improving constipation.

[0017] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 Figure: BL8 degradation results on cellulose. Detailed Implementation

[0020] The embodiments of the present invention are described in detail below, with examples of these embodiments shown in the accompanying drawings. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all commercially available conventional products.

[0021] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "multiple" or "a plurality of" means at least two, two types, such as two, two, three, three, etc., unless otherwise explicitly specified.

[0022] In this document, the terms “comprising,” “having,” or “including” are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0023] In this document, the term “optionally” generally means that an event or condition described below may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0024] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0025] To facilitate understanding of the invention, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined elsewhere in this document, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains.

[0026] In this document, the terms “treatment” and “relief” refer to the attainment of a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of disease or its symptoms, and / or therapeutic in terms of partial or complete cure of disease and / or adverse effects caused by disease. As used herein, “treatment” encompasses diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of disease in susceptible individuals who have not yet been diagnosed with the disease; (b) suppression of disease, such as inhibiting disease progression; or (c) relief of disease, such as reducing disease-related symptoms. As used herein, “treatment” encompasses any medication that administers a strain, fermentation broth, bacterial suspension, or composition to an individual to treat, cure, relieve, improve, reduce, or suppress the individual’s disease, including but not limited to administering to an individual in need a strain, fermentation broth, bacterial suspension, or composition described herein.

[0027] In this document, the term "carrier" includes any solvent, pharmaceutical stabilizer, or combination thereof, which are known to those skilled in the art. Except in cases where any conventional carrier is incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is covered.

[0028] Fermentation liquid

[0029] In some embodiments, the present invention provides a fermentation broth derived from the aforementioned Propionibacterium BL8.

[0030] It should be noted that the "fermentation broth" of the present invention refers to the solution obtained after culturing Propionibacterium acidogeneticum BL8 for a period of time, which mainly contains Propionibacterium acidogeneticum BL8 and its metabolites; or the supernatant after further treatment by centrifugation, filtration and other means, which mainly contains the metabolites of Propionibacterium acidogeneticum BL8.

[0031] bacterial suspension

[0032] In some embodiments, the present invention provides a bacterial suspension comprising the aforementioned Propionibacterium BL8.

[0033] It should be noted that the bacterial suspension can be obtained by processing the above-mentioned fermentation broth through centrifugation, resuspension and other means.

[0034] Food, beverages, pharmaceuticals, health products, animal feed, cosmetics, or additives

[0035] According to embodiments of the present invention, the aforementioned food, beverage, pharmaceutical, health product, feed, cosmetic, or additive may further include at least one of the following additional technical features:

[0036] According to embodiments of the present invention, it further includes pharmaceutically acceptable excipients or carriers, food or health product acceptable excipients or carriers, or animal feed acceptable excipients or carriers.

[0037] In this article, "acceptable in food" refers to substances or compositions that are edible for human consumption, which may be adjusted according to the food requirements of different countries.

[0038] In this article, "acceptable in health supplements" refers to substances or compositions that can be consumed by humans, which may be adjusted according to the health supplement requirements of different countries.

[0039] In this document, "pharmaceutical acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with other components of the formulation and / or the mammals to which it is treated. Preferably, "pharmaceutical acceptable" as used herein means approved by a federal regulatory agency or national government, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals, particularly in humans.

[0040] In this article, “acceptable adjuvants or carriers in animal feed” refers to substances or compositions that can be consumed by animals, and these can be adjusted according to the animal feed requirements of different countries.

[0041] In this document, the term "pharmaceutically acceptable carrier" includes any solvent, drug stabilizer, or combination thereof known to those skilled in the art. It covers the use of any conventional carrier in therapeutic or pharmaceutical compositions, except in cases where any conventional carrier is incompatible with the active ingredient.

[0042] In this document, the term "pharmaceuticalally acceptable excipient" may include any solvent suitable for the specific target dosage form. The use of any conventional excipients is also within the scope of consideration of this disclosure, except for any range of incompatibilities with the Propionibacterium BL8 disclosed herein, such as any adverse biological effects produced or harmful interactions with any other component of the pharmaceutically acceptable composition.

[0043] According to an embodiment of the present invention, at least one of the aforementioned Propionibacterium BL8, fermentation broth, and bacterial suspension is added to or inoculated into food, beverage, health product, feed, cosmetic, or additive, or added to a drug.

[0044] For example, the aforementioned food products include, but are not limited to: probiotic tablets, fermented dairy products (such as probiotic yogurt), probiotic solid beverages, probiotic milk powder, probiotic cheese, probiotic soy products, probiotic candies, probiotic fermented vegetables, etc.

[0045] For example, the aforementioned drugs include, but are not limited to, human drugs and veterinary drugs. The aforementioned veterinary drugs can be for pets, livestock, or wild animals.

[0046] For example, the aforementioned health products include, but are not limited to: health products for human use and health products for veterinary use.

[0047] It should be noted that the characteristics and advantages described above for Propionibacterium acidogeneticum BL8 also apply to this food, beverage, pharmaceutical, health product, feed, cosmetic or additive, and will not be repeated here.

[0048] Composition

[0049] According to embodiments of the present invention, the above composition may further include at least one of the following additional technical features:

[0050] According to embodiments of the present invention, the composition further includes an ingestible excipient and / or a carrier.

[0051] According to an embodiment of the present invention, the excipient includes at least one selected from adhesives, disintegrants, lubricants, flow aids, stabilizers, fillers, diluents, and sustained-release agents.

[0052] According to an embodiment of the present invention, the carrier comprises at least one selected from sugars, cellulose and its derivatives, calcium phosphates, alkaline earth metal stearates, vegetable oils, nonionic surfactants, cationic surfactants, anionic surfactants, fatty alcohols, and hydrolyzed cereal solids.

[0053] According to embodiments of the present invention, the dosage form of the composition includes at least one selected from oral liquids, powders, granules, capsules, tablets, and pills.

[0054] Example 1 Isolation and Identification of Strains

[0055] 1. Sample collection

[0056] The Propionibacterium acidogeneticus BL8 strain was isolated from breast milk of lactating women within the company by personnel from the strain technology platform and stored at -80℃.

[0057] 2. Strains Isolation

[0058] After serially diluting the sample tenfold, several appropriate serial dilutions were selected and evenly spread on MRS agar plates. The plates were then inverted and anaerobically cultured at 37°C for 48 hours. Single colonies were picked from the plates with different serial dilutions and transferred for further culture. Finally, a strain with good growth performance was obtained and named BL8.

[0059] 3. Strain identification

[0060] The colonies of strain BL8 of this invention are milky white, smooth, round, and easily picked up on MRS agar plates. The optimal growth pH is 6.0-7.0, and the optimal growth temperature is 35-37℃. 16S rDNA sequencing results show that the 16S rDNA sequence of BL8 has >99% sequence homology with known Propionibacterium acidogenetic standard strains.

[0061] Therefore, strain BL8 was identified as Propionibacterium acidogeneticum (… Propionibacterium acidipropionici The strain BL8 was deposited on December 20, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 33134.

[0062] 16S rDNA sequence of Propionibacterium acidogeneticum BL8:

[0063]

[0064] Example 2 Preparation of Cellular Metabolites (CFS)

[0065] Propionibacterium acidogenetice BL8 was cultured anaerobicly in MRS medium at 37°C for 18 h, and the bacterial cells were collected by centrifugation at 3000 rpm for 10 min at 4°C. The cells were washed twice with sterile phosphate-buffered saline (PBS, pH 6.8), resuspended in PBS, and the bacterial concentration was adjusted to 1 × 10⁻⁶. 9 CFU / ml was prepared as a bacterial suspension for later use. The concentration was adjusted to 10. 9 CFU / ml of Propionibacterium acidogenetice was incubated anaerobically at 37°C for 16 h, then centrifuged at 3000 rpm for 10 min at 4°C. The supernatant was filtered through a 0.22 μm aqueous microfiltration membrane to obtain cell metabolites, which were then stored at -80°C.

[0066] Example 3: Inhibitory effect of Propionibacterium acidogeneticum BL8 on dipeptidyl peptidase-IV (DPP4)

[0067] Take a 96-well plate and add 25 μL of glycyl-prolyl-p-nitroaniline (1.6 mmol / L) and 25 μL of CFS to the reaction wells using a micropipette; incubate at 37°C for 10 min, then add 50 μL of DPP4 (10 U / L); react at 37°C for 60 min, and finally stop the reaction by adding 100 μL of sodium acetate buffer (pH 4.0, 1 mol / L). Measure the absorbance of the reaction solution at 405 nm using a microplate reader. Perform three replicates for each sample. The inhibition rate is calculated as shown in the formula:

[0068] Inhibition rate (%) = {1 - (A)} 样品 -A 样品空白 ) / (A 阳性 -A 阴性 )﹜×100

[0069] In the formula: positive is glycyl-prolyl-p-nitroaniline + Tris-HCl buffer (100 mmol / L, pH 8.0) + DPP4 + sodium acetate buffer;

[0070] The negative result is glycyl-prolyl-p-nitroaniline + Tris-HCl buffer (100 mmol / L, pH 8.0) + Tris-HCl buffer + sodium acetate buffer;

[0071] The sample consisted of glycyl-prolyl-p-nitroaniline + sample + DPP4 + sodium acetate buffer.

[0072] The sample blank consisted of glycyl-prolyl-p-nitroaniline + sample + Tris-HCl buffer (100 mmol / L, pH 8.0) + sodium acetate buffer.

[0073] Results: As shown in Table 1, compared with the no inhibitory effect of the blank medium MRS, the CFS of Propionibacterium BL8 had a high inhibitory effect on DPP4 enzyme activity, with an inhibition rate of 81.3%.

[0074] Table 1: BL8 DPP4 Inhibition Results

[0075]

[0076] Example 4: Propionibacterium BL8 promotes insulin secretion

[0077] Cell culture: INS-1 cells (rat islet cell tumor cells) were cultured in RPMI 1640 complete medium (containing 10% fetal bovine serum, 1mM sodium pyruvate, 50μM mercaptoethanol, 100... μ (g / ml streptomycin) was cultured in a 37°C, 5% CO2 incubator. Cells adhered and grew; once the cell density reached 80%-90%, they were fused and passaged.

[0078] Promote insulin secretion: Cultured INS-1 cells were inoculated at a rate of 1×10⁻⁶. 5 CFU / ml was seeded into 12-well plates at 1.5 ml / well. After incubation at 37°C and 5% CO2 for 24 h, the RPMI 1640 medium was discarded, and the cells were gently washed twice with KRBH buffer. The cells were then pre-incubated with KRBH for 2 h to sensitize them to glucose. Fresh KRBH buffer containing CFU / ml was then added, and incubation continued for 1 h. The supernatant was collected and stored at -20°C for insulin concentration assay.

[0079] Insulin level detection: Rat insulin detection kit (Solepro, catalog number: SEKR-0033-96T)

[0080] Results: As shown in Table 2, CFS of Propionibacterium BL8 can significantly promote insulin secretion by INS-1 cells.

[0081] Table 2: Results of BL8 promoting insulin secretion

[0082]

[0083] Example 5: Inhibitory effect of Propionibacterium BL8 on acetylcholinesterase activity

[0084] Add 2.65 ml of PBS (pH 8.0), 50 μL of acetylcholinesterase (AChE, 1 U / ml), 100 μL of dithiodinitrobenzic acid (DTNB, 15 mM), and 100 μL of sample solution (CFS) to a test tube. Preheat at 37°C for 2 min, then add 100 μL of thioacetylcholine (ATch, 15 mM). Incubate at 37°C for 20 min, then immediately add 1 ml of sodium dodecyl sulfate (SDS, 0.4%) to terminate the reaction. Measure the absorbance of the resulting solution at 412 nm immediately. The inhibition rate is calculated as shown in the formula:

[0085] Inhibition rate (%) = {A 对照 -(A 样品 -A 样品空白 )﹜ / A 对照 ×100

[0086] In the formula: the control is 2.75 ml PBS (pH 8.0) + AChE + DTNB + ATch + SDS

[0087] The sample consisted of 2.65 ml PBS (pH 8.0) + AChE + DTNB + 100 μL sample (CFS) + ATch + SDS.

[0088] The sample blank consisted of 2.65 ml PBS (pH 8.0) + AChE + DTNB + 100 μL sample background + ATch + SDS.

[0089] Results: As shown in Table 3, CFS of Propionibacterium BL8 inhibits the activity of acetylcholinesterase, with an inhibition rate of 23.33%, which can effectively prevent and alleviate Alzheimer's disease.

[0090] Table 3: Results of BL8's inhibition of acetylcholinesterase activity

[0091]

[0092] Example 6: Propionibacterium acidogeneticum BL8 can produce cellulase.

[0093] The cellulose degradation capacity of Propionibacterium acidogeneticum was identified and determined using the agar diffusion method. The cellulose medium used was MRS agar medium + 1.5% sodium carboxymethyl cellulose. During the experiment, 3 μL of Propionibacterium acidogeneticum bacterial suspension (1 × 10⁻⁶ viable cells) was added dropwise to the experimental group. 9 (CFU / ml), the control group was added with 3μL of blank MRS medium and cultured in a constant temperature anaerobic incubator at 37℃ for 2-3 days before Congo red staining.

[0094] Results: Compared with the control group treated with blank culture medium, BL8 significantly degraded cellulose and formed distinct degradation zones. Figure 1This indicates that Propionibacterium BL8 can produce cellulase, which can effectively relieve and improve constipation in people.

[0095] Example 7: Determination of acid resistance of Propionibacterium BL8

[0096] Propionibacterium acidogeneticum BL8 was inoculated into MRS broth at a 1% inoculum size and anaerobically cultured at 37°C for 18 hours. The cells were then collected by centrifugation at 3000 rpm for 10 minutes at 4°C. The collected cells were resuspended in artificial gastric fluid (pH 2.5) and the bacterial concentration was adjusted to 10-1. 9 CFU / ml, after anaerobic incubation at 37℃ for 0h, 2h, and 4h, plate counts were performed to calculate the survival rate of *Propionibacterium acidogeneticum* and assess acid tolerance. The survival rate calculation is shown in the formula:

[0097] Survival rate (%) = (Number of viable bacteria at the end of the time / Number of viable bacteria at time 0) × 100

[0098] Results: In artificial gastric fluid, the survival rate of BL8 after 4 hours was 53.5% (Table 4), indicating good acid tolerance.

[0099] Table 4: Acid Resistance Results of BL8

[0100]

[0101] Example 8: Determination of bile salt tolerance in Propionibacterium BL8

[0102] Propionibacterium acidogeneticum BL8 was inoculated into MRS broth at a 1% inoculum size and anaerobically cultured at 37°C for 18 h. The cells were then collected by centrifugation at 3000 rpm for 10 min at 4°C. The collected cells were resuspended in simulated intestinal fluid (pH 8.0) containing 0.1%, 0.2%, and 0.3% bile salts, and adjusted to a concentration of 10⁻⁶. 9 CFU / ml, after anaerobic incubation at 37℃ for 0h, 2h, and 4h, plate counts were performed to calculate the survival rate of Propionibacterium acidogeneticum and assess bile salt tolerance.

[0103] Results: In bile salt solutions with concentrations of 0.1%, 0.2%, and 0.3%, the survival rate of BL8 after 4 hours was over 90% (Table 5), demonstrating good bile salt tolerance. The survival rate was calculated as follows:

[0104] Survival rate (%) = number of viable bacteria at the end of the period / number of viable bacteria at time 0 × 100.

[0105] Table 5: Results of BL8 bile salt tolerance

[0106]

[0107] Example 9: Hydrophobicity determination of Propionibacterium BL8

[0108] Propionibacterium acidogeneticum BL8 was inoculated into MRS broth at a 1% inoculum size and cultured anaerobically at 37°C for 18 hours. The cells were collected by centrifugation at 3000 rpm for 10 minutes at 4°C. The cells were washed twice with sterile PBS (pH 8.0). The bacterial concentration was adjusted to an OD600 between 0.6 and 1. 4 ml of the adjusted bacterial solution was mixed with 0.5 ml of xylene and ethyl acetate, respectively. The mixture was vortexed for 2 minutes and then allowed to stand at room temperature for 15 minutes. The aqueous layer was carefully aspirated, and its OD value was measured at 600 nm. The hydrophobicity was calculated as shown in the following formula:

[0109] Cell surface hydrophobicity (%) = (initial OD - final OD) / initial OD × 100

[0110] Results: BL8 exhibited a hydrophobicity of 25.3% in xylene and 29% in ethyl acetate (Table 6). It belongs to the moderately hydrophobic strain.

[0111] Table 6: Hydrophobicity Results of BL8

[0112]

[0113] In the description of this specification, references to terms such as "one embodiment," "some embodiments," "embodiment," or "specific embodiment," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment is included in at least one embodiment of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment. Furthermore, the specific features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments and features described in this specification.

[0114] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A strain of Propionibacterium acidogenicum ( Propionibacterium acidipropionici BL8, characterized in that, The Propionibacterium BL8 was deposited on December 20, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 33134.

2. A fermentation broth, characterized in that, The fermentation broth is obtained by fermentation of Propionibacterium BL8 as described in claim 1.

3. A bacterial suspension, characterized in that, The bacterial suspension includes Propionibacterium BL8 as described in claim 1.

4. A composition, characterized in that, The composition comprises at least one of the following: Propionibacterium BL8 of claim 1, the fermentation broth of claim 2, and the bacterial suspension of claim 3.

5. The composition according to claim 4, characterized in that, The composition also includes excipients or carriers.

6. Use of the Propionibacterium BL8 of claim 1, the fermentation broth of claim 2, the bacterial suspension of claim 3, or the composition of claim 4 in the preparation of a medicine for the prevention, relief, treatment, or adjunctive treatment of type 2 diabetes.

7. Use of the Propionibacterium BL8 of claim 1, the fermentation broth of claim 2, the bacterial suspension of claim 3, or the composition of claim 4 in the preparation of health products that help improve memory, and / or in the preparation of medicines for the prevention and relief of Alzheimer's disease.

8. Use of the Propionibacterium BL8 of claim 1, the fermentation broth of claim 2, the bacterial suspension of claim 3, or the composition of claim 4 in the preparation of a medicine for improving constipation.

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