Weissmannia coagrane capable of inhibiting the activity of α-glucosidase and / or α-amylase and its applications

Abstract

Provided are Weissmannia coagulans capable of inhibiting the activity of α-glucosidase and / or α-amylase, and applications thereof. The Weissmannia coagulans SA9 strain was deposited with the Guangdong Provincial Center of Microorganisms and Seeds Preservation on June 6, 2022, under accession number GDMCC NO: 62517. Research has shown that the SA9 strain has high acid-producing and DPPH free radical-scavenging abilities, resulting in a high inhibitory effect on α-glucosidase, a key enzyme responsible for elevated blood glucose levels, achieving over 80% of the efficacy of the diabetes drug acarbose. At the same time, the strain also has excellent inhibitory ability against α-amylase, with its fermentation broth exhibiting 100% inhibition of this enzyme, achieving efficacy comparable to acarbose but safer and without toxic side effects. The SA9 strain may be used as an efficient strain for blood sugar control, alleviating diabetes and providing beneficial effects in the prevention and treatment of diabetes.

Description

[Technical Field]

[0001] The present invention relates to the technical field of microorganisms, and more particularly to Weissmannia coagulans capable of inhibiting the activity of α-glucosidase and / or α-amylase, and applications thereof. [Background technology]

[0002] Diabetes mellitus (Diabetes mellitus) is an endocrine disorder caused by genetic and acquired environmental factors. It is characterized by numerous complications, a long-term and complex illness, and significantly reduces patients' quality of life and places a heavy economic burden on families and society. Diabetes has become the third most common chronic disease affecting human health worldwide, making effective prevention and treatment of diabetes urgently needed. Research has shown that the highly active digestive enzyme α-glucosidase promotes carbohydrate digestion and absorption in the intestine, increasing plasma glucose levels. Inhibiting α-glucosidase activity can slow carbohydrate absorption and increase insulin secretion, thereby maintaining low postprandial plasma glucose levels in diabetic patients. Therefore, inhibiting α-glucosidase activity can alleviate and treat diabetes, and most commercially available antidiabetic drugs are inhibitors of this enzyme. Alternatively, α-amylase can be inhibited to block the conversion of starch to sugar, thereby lowering plasma glucose levels and controlling blood glucose levels. For example, acarbose acts as an inhibitor of both α-glucosidase and α-amylase.

[0003] In recent years, as the scientific community has deepened its research on probiotics, they have increasingly played an important role in the prevention and treatment of human diseases, demonstrating great potential. Probiotics refer to microorganisms that, when ingested in sufficient amounts, improve the health of the host. Numerous studies have shown that probiotics, prebiotics, or synbiotics can significantly alleviate the clinical symptoms of diabetic patients. According to literature reports, the supernatants of Lactobacillus GG and Bifidobacterium F-35 both exhibited excellent inhibitory effects on α-glucosidase activity, with inhibition rates reaching 29.41% and 21.82%, respectively. Chen et al. also found that Lactobacillus CCFM0528 improved blood glucose tolerance in diabetic mice, lowering blood glucose levels by approximately 43%. Furthermore, Bacillus subtilis natto has been shown to have excellent blood glucose-lowering properties. Compared to chemotherapy, probiotics are safer and have fewer toxic side effects, making biological therapy more attractive.

[0004] Weizmannia coagulans, formerly known as Bacillus coagulans, has been renamed and is one of the most popular probiotics currently being studied. Numerous studies have reported that this strain has beneficial effects in preventing and treating various human diseases, including non-alcoholic fatty liver disease, constipation, and irritable bowel syndrome. According to existing research reports, the cell-free extract of Bacillus coagulans JA845 has an inhibitory rate of 39.51% against α-glucosidase activity. Similarly, Bacillus coagulans VHProbi C08 from Qingdao Weilan Biotechnology Co., Ltd. also has excellent blood glucose lowering effect. After 6 weeks of intragastric administration, the fasting blood glucose levels of mice in the Bacillus coagulans pre-treatment group were reduced by 46.46% compared to the positive control group, but the effect needs to be improved. Currently, there is no Weissmannia coagulans that can inhibit the activities of both α-glucosidase and α-amylase. In order to improve the inhibitory efficiency, it is urgent to further develop Weissmannia coagulans with better effects and functions, which is of great significance for the production of products for the treatment or auxiliary treatment of diabetes and blood glucose control. Summary of the Invention [Means for solving the problem]

[0005] The technical problem to be solved by the present invention is to overcome the drawbacks and shortcomings of the above problems and to provide Weissmannia coagulans capable of inhibiting the activity of α-glucosidase and / or α-amylase, and applications thereof.

[0006] An object of the present invention is to provide the Weissmannia coagulan SA9 strain.

[0007] Another object of the present invention is to provide applications of the Weissmannia coagulant SA9 strain.

[0008] It is yet another object of the present invention to provide an inhibitor of α-glucosidase and / or α-amylase.

[0009] It is yet another object of the present invention to provide a method for inhibiting α-glucosidase and / or α-amylase.

[0010] The above object of the present invention is achieved by the following technical solutions.

[0011] The present invention provides the Weizmannnia coagulans SA9 strain, which is capable of inhibiting α-glucosidase and / or α-amylase activity. The strain was deposited at the Guangdong Provincial Bacterial Species Preservation Center on June 6, 2022, under accession number GDMCC NO: 62517. The SA9 strain was isolated from mulberry fermentation residue and has high acid-producing and DPPH free radical scavenging abilities. The SA9 strain exhibits a high inhibitory effect on α-glucosidase, a key enzyme responsible for elevated blood glucose levels, achieving over 80% of the efficacy of the diabetes treatment acarbose. The SA9 bacterial suspension (fermentation broth) exhibited an 85% inhibition rate, the somatic cell metabolites inhibited 96%, and the cell-free extract inhibited 87%. At the same time, the SA9 strain also has excellent inhibitory ability against α-amylase, with the fermentation broth's α-amylase inhibition rate reaching 100%, equivalent to that of acarbose, but safer than acarbose and without toxic side effects. In other words, the SA9 strain can efficiently inhibit α-glucosidase and α-amylase, and by inhibiting the activity of α-glucosidase and α-amylase, it can block the conversion and absorption of sugar, contributing to blood sugar control and the alleviation of diabetes.

[0012] Therefore, the present invention provides the use of Weissmannia coagulan SA9 strain and / or its fermentation broth in the inhibition of α-glucosidase and / or α-amylase, the manufacture of inhibitors of α-glucosidase and / or α-amylase, and the manufacture of products for the alleviation or treatment of diabetes or the control of blood glucose levels.

[0013] Preferably, the product is a food product, a dietary supplement, or a pharmaceutical product.

[0014] The present invention provides an inhibitor of α-glucosidase and / or α-amylase, comprising Weissmannia coagulan SA9 strain and / or a fermentation broth thereof.

[0015] Preferably, the fermentation broth is a fermentation supernatant, a cell-free extract, and / or somatic cell metabolites.

[0016] Preferably, the OD of the fermentation broth is 0.5 to 1.0.

[0017] Preferably, the SA9 strain is inoculated into a medium at an inoculum amount of 1 to 5%, and cultured with shaking at 35 to 40°C and 120 to 200 r / min for 28 to 48 hours to obtain the fermentation broth.

[0018] More preferably, the SA9 strain is inoculated into a medium at an inoculum amount of 1%, and cultured with shaking at 37°C and 180 r / min for 48 hours to obtain the fermentation broth.

[0019] The present invention provides a method for inhibiting α-glucosidase and / or α-amylase, in which a sample is treated with Weissmannia coagulan SA9 strain and / or a fermentation broth thereof.

[0020] The present invention has the following beneficial effects:

[0021] The present invention provides the Weizmannia coagulans SA9 strain, which is capable of inhibiting α-glucosidase and / or α-amylase activity. The SA9 strain has high acid-producing and DPPH free radical scavenging abilities, and exhibits a high inhibitory effect against α-glucosidase, a key enzyme responsible for elevated blood glucose levels, achieving over 80% of the efficacy of the antidiabetic drug acarbose. The SA9 bacterial suspension (fermentation broth) exhibited an 85% α-glucosidase inhibition rate, the somatic cell metabolites an 96% inhibition rate, and the cell-free extract an 87% inhibition rate. At the same time, the SA9 strain also exhibited excellent α-amylase inhibition, with the fermentation broth's α-amylase inhibition rate reaching 100%, achieving efficacy comparable to that of acarbose, but with greater safety and no toxic side effects.

[0022] The SA9 strain provided by the present invention inhibits the activity of α-glucosidase and α-amylase, thereby preventing the conversion and absorption of sugars, thereby facilitating blood sugar control, alleviating diabetes, and having beneficial effects on the prevention and treatment of diabetes. Weissmannia coagulans is also included in the national edible probiotics list. This strain is highly safe, has no toxic side effects, and is highly stress-resistant. Weissmannia coagulans SA9 of the present invention can be used in fermented foods, dietary supplements, pharmaceuticals, etc., providing a new option for diabetic patients' daily blood sugar control. [Brief explanation of the drawings]

[0023] [Figure 1] FIG. 1 shows the results of pre-screening to measure the acid-producing ability of different strains. [Figure 2] FIG. 2 shows the results of pre-screening measurements of the ability of different bacterial strains to scavenge DPPH free radicals. [Figure 3] Figure 3 shows the developmental tree of the strain system. [Figure 4] FIG. 4 shows the morphology of Weissmannia coagulans. [Figure 5] FIG. 5 shows the inhibition rate of α-glucosidase activity by Weissmannia coagulan. [Figure 6] FIG. 6 shows the results of an experiment on the inhibition of α-amylase activity by Weissmannia coagulan. [Figure 7] FIG. 7 shows the results of an inhibition experiment using a dose gradient of the fermentation supernatant of Weissmannia coagulans (from left to right: undiluted solution, 4-fold diluted solution, and 8-fold diluted solution). [Figure 8] FIG. 8 shows the results of an inhibition experiment using a dose gradient of Weissmannia coagulans fermentation broth (from left to right: undiluted solution, 4-fold diluted solution, and 8-fold diluted solution). DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention will now be further described in conjunction with the accompanying drawings and specific examples, which are not intended to limit the invention in any way. The reagents, methods, and equipment used in the present invention are conventional in the art unless otherwise specified.

[0025] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0026] The media used in the following examples are as follows:

[0027] MRS broth medium containing 10 g of tryptone, 10 g of beef extract, 4 g of yeast extract powder, 20 g of glucose, 2 g of dipotassium hydrogen phosphate (anhydrous), 2 g of triammonium citrate (anhydrous), 5 g of sodium acetate trihydrate, 0.2 g of magnesium sulfate (including heptahydrate), 0.05 g of manganese sulfate (including tetrahydrate), and 1 g of Tween-80, with a final pH of 6.8±0.2. [Example]

[0028] Strain screening and identification (1) Isolation and purification of strains Fresh commercially available mulberry bark is washed with water, placed in a fermentation tank, and sugar solutions of different concentrations are added for natural fermentation. After fermentation is complete, an appropriate amount of fermentation residue is weighed and added to an Erlenmeyer flask containing 100 mL of MRS liquid medium. The mixture is then cultured in a shaker at 37°C for 48 hours. 1 mL of the mixed bacterial solution is then drawn out and serially diluted with sterilized water to a concentration of 10. -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 Five serial dilutions of the above were spread onto MRS plates. Different single colonies were then randomly selected from the plates and streaked onto solid MRS plates for purification. The resulting pure cultures were then cultured at 37°C for 48 hours. The resulting pure cultures were rinsed with 20% glycerol, blotted, placed in glycerol tubes, and stored in a refrigerator at -20°C. (2) Molecular biological identification of bacteria After purification, bacterial DNA was extracted using SDS and conserved bacterial sequences were amplified using universal primers 27f (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492r (5'-TACCTTGTTACGACTT-3'). The amplified products were then sent to a sequencing company for sequencing. The resulting sequences were combined and compared with the NCBI database, yielding 22 Weissmannia coagulans strains. After prescreening to measure acid production and DPPH free radical scavenging abilities, the results are shown in Figures 1 and 2. Five strains of Bacillus coagulans with superior performance were identified and named SA19, SA9, SA12, SA4, and SA6, respectively. A tree diagram of the five strains is shown in Figure 3. The results showed that the 16S rDNA sequence of the strain of the present invention showed high homology with Weizmannia coagulans in GenBank (Note: The original Bacillus coagulans was renamed Weizmannia coagulans, and the strain was named and deposited using the latest Latin name in the present invention). (3) Morphological observation of bacteria The purified SA9 strain was inoculated into MRS liquid medium and cultured at 37°C for 48 hours. A small amount of the bacterial solution was taken and the bacterial morphology was observed using Gram staining. The results are shown in Figure 4. The bacteria were long and rod-shaped, and Gram staining was positive, revealing a purple color. Based on the results of the molecular biological identification and bacterial staining observation described above, the SA9 strain isolated by the present invention, which has high acid-producing ability and DPPH free radical scavenging ability, was identified as Weizmanniacoagulans and was preserved at the Guangdong Provincial Microorganism Species Preservation Center on June 6, 2022, with the deposit number GDMCC NO: 62517 and the deposit address being 5th floor, Building 59, Dayuan, No. 100, Xianlie Middle Road, Guangzhou City. [Example]

[0029] α-Glucosidase activity inhibition experiment by Weissmannia coagran SA9 1. Solution Preparation 1 U / mL α-glucosidase solution: 1 mg of α-glucosidase was accurately weighed, and 33 mL of PBS solution was added to prepare a 1 U / mL α-glucosidase solution. 5 mol / L p-nitrophenyl-α-D-glucopyranoside (PNPG): 0.015 g of PNPG was accurately weighed and placed in a 50 mL volumetric flask, and PBS solution was added to prepare a 5 mol / L PNPG solution. 0.1 mol / L Na2CO3 solution: 0.53 g of anhydrous Na2CO3 was accurately weighed, dissolved in distilled water, and the solution was adjusted to a constant volume of 50 mL. 2. Preparation of Different Weissmannian Coagulans (1) Strain activation: The SA9 strain isolated and identified in Example 1 was taken out of the refrigerator at -20°C, inoculated into a test tube at a 1% inoculum dose, and activated for two generations before use. (2) Preparation of fermentation broth (bacterial suspension): The fermentation broth of the activated Weissmannia coagulant SA9 strain was inoculated into 100 mL of liquid medium at an inoculum amount of 1%, and cultured with shaking at 37°C and 180 r / min for 48 hours to obtain a fermentation broth. (3) Preparation of fermentation supernatant: The fermentation broth prepared above was centrifuged at 4°C and 6000 r / min for 10 min, and the supernatant was collected and filtered through a 0.22 μm water microfiltration membrane to obtain a sample of the fermentation supernatant, which was then refrigerated at 4°C. (4) Preparation of bacterial cell fraction (CFS): The fermentation broth prepared above was centrifuged at 4°C and 4000 r / min for 15 min to collect the bacteria. After washing twice with 0.1 mol / L sterile PBS (pH = 6.8), the bacteria were resuspended in PBS and the concentration of the bacterial solution was measured by OD 600 The pH was adjusted to 1.0, and the mixture was shaken at 37°C and 180 r / min for 24 hours. The mixture was then centrifuged at 4°C and 6000 r / min for 10 minutes to collect the supernatant, which was then filtered through a 0.22 μm water microfiltration membrane to obtain the bacterial cell metabolites, which were then refrigerated at 4°C. (5) Preparation of cell-free extract (CFE): The fermentation broth prepared above was centrifuged at 4°C and 4000 r / min for 15 min to collect the bacterial cells. The cells were washed twice with 0.1 mol / L sterile PBS (pH = 6.8), and then resuspended in PBS. The concentration of the bacterial solution was adjusted to OD . 600 The pH was adjusted to 1.0, 1.5 mg / mL lysozyme was added, and the mixture was incubated at 37°C for 3 hours. The mixture was then sonicated in an ice bath (power output 800 W, ultrasonic time 15 min, 2 sec on, 2 sec off). The sonicated solution was centrifuged at 4°C, 6000 r / min for 15 min, and the supernatant was collected and filtered through a 0.22 μm water microfiltration membrane to obtain a cell-free extract, which was then refrigerated at 4°C. 3. Sample Preparation (1) Sample group: 50 μL of the bacterial suspension (fermentation broth), fermentation supernatant, somatic cell metabolites, and cell-free extract prepared above were each pipetted and mixed with 100 μL of PNPG. After placing in a 37°C water bath for 10 minutes, the mixture was removed. 100 μL of α-glucosidase solution (1 U / mL) was then aspirated and reacted at 37°C for 30 minutes. The mixture was then removed and 1 mL of sodium carbonate (0.1 M) was added to stop the reaction. 100 μL of the mixture was aspirated and transferred to a 96-well plate, and the α-glucosidase activity was measured at 400 nm. (2) Sample blank group: 50 μL of the sample obtained above was removed and pipetted. This was mixed with 100 μL of PNPG and 100 μL of PBS and placed in a 37°C water bath for 10 minutes. 100 μL of α-glucosidase solution (1 U / mL) was then added and the mixture was incubated at 37°C for 40 minutes. 1 mL of sodium carbonate (0.1 M) was added to stop the reaction. 100 μL of the sample was then pipetted and transferred to a 96-well plate, and the α-glucosidase activity was measured at 400 nm. (3) Control group: 50 μL of PBS was pipetted and mixed with 100 μL of PNPG. The mixture was placed in a 37°C water bath for 10 minutes, then removed. 100 μL of α-glucosidase solution (1 U / mL) was then aspirated and reacted at 37°C for 30 minutes. 1 mL of sodium carbonate (0.1 M) was added to stop the reaction. 100 μL was aspirated and transferred to a 96-well plate, and α-glucosidase activity was measured at 400 nm. (4) Blank group: 150 μL of PBS was pipetted and mixed with 100 μL of PNPG. The mixture was placed in a 37°C water bath for 40 minutes, then removed and 1 mL of 0.1 M sodium carbonate was added to stop the reaction. 100 μL was pipetted and transferred to a 96-well plate, and α-glucosidase activity was measured at 400 nm. (5) Acarbose group: 50 μL was pipetted and mixed with 100 μL of PNPG. The mixture was placed in a 37°C water bath for 10 minutes. 100 μL of α-glucosidase solution (1 U / mL) was then pipetted and reacted at 37°C for 30 minutes. 1 mL of sodium carbonate (0.1 M) was added to stop the reaction. 100 μL was then pipetted and transferred to a 96-well plate. α-glucosidase activity was measured at 400 nm. (6) The parameters and conditions of the different experimental groups are shown in the table below. Three parallel experiments were conducted in each group. [Table 1] (7) The formula for calculating the inhibition rate of α-glucosidase is as follows:

number

[0030] α-Amylase inhibition experiment with Weissmannia coagran SA9 1. Sample Preparation (1) Experimental samples: The fermentation broth (bacterial suspension), fermentation supernatant, bacterial cell metabolites, and cell-free extract of the SA9 strain were prepared in the same manner as in Example 2 for the acarbose group and the PBS group, using lactic acid solutions with pH values ​​of 3.5 and 4.0. (2) Starch medium: 6 g of soluble starch and 3 g of agar were accurately weighed and placed in a 250 ml Erlenmeyer flask, which had been sterilized. (3) α-amylase solution (1 mg / L): 5 mg of the required α-amylase (extracted from porcine pancreas, purchased commercially) was accurately weighed and dissolved in 5 mL of PBS buffer solution, and used immediately after preparation. 2. Experimental Method A small amount of starch medium was poured into a sterile plate. Once solidified, an Oxford cup (6 mm inner diameter, 8 mm outer diameter, 10 mm height) was placed on the surface of the medium using tweezers. The plate was then filled with medium up to about two-thirds of the height of the Oxford cup. Once solidified, the Oxford cup was removed. Three Oxford cups were placed on each plate. 50 μL of each of the above samples was mixed with 50 μL of α-amylase solution and added to the wells. A blank control was prepared by mixing 50 μL of PBS with 50 μL of α-amylase solution. A negative medium control was prepared by mixing 50 μL of MRS with 50 μL of α-amylase solution. An acidity negative control was prepared by mixing 50 μL of lactic acid with 50 μL of α-amylase solution. The plates were then placed in a 37°C incubator and incubated for 24 hours. The plates were then removed, stained with diluted iodine solution, and observed. The size of the clear zone was measured using a vernier caliper. 3, results The inhibitory effects of different sample groups on α-amylase are shown in Figure 6. The acarbose-positive group showed 100% inhibition of α-amylase, with no clear zone formed. The bacterial suspension group also showed no clear zone. The diameter of the clear zone in the fermentation supernatant was 14.5 mm. The pH 4.0 lactic acid-negative control group showed a clear zone diameter of 19.5 mm. The MRS control group showed a clear zone diameter of 20.4 mm. The PBS blank control group showed a clear zone diameter of 20.6 mm. These results indicate that lactic acid and MRS components have no inhibitory effect on α-amylase, meaning that the lactic acid and medium components in the fermentation broth do not affect the experimental results. The inhibitory effect of Weissmannia coagulans SA9 fermentation broth on α-amylase was similar to that of acarbose, with 100% inhibition of enzyme activity, and the fermentation supernatant showed an inhibition rate of approximately 50%. Bacterial cell metabolites and cell-free extracts had no significant effect.

[0031] To further illustrate the α-amylase inhibitory effect of the fermentation broth group, a dose gradient experiment was conducted using a 2-fold dilution method on Weissmannia coagulan fermentation broth and fermentation supernatant. The results are shown in Figures 7 and 8. From left to right, the samples are undiluted, diluted 4-fold, and diluted 8-fold. The diameters of the clear zones in the supernatant group were 15.0 mm, 18.5 mm, and 20.7 mm, respectively, while those in the fermentation broth group were 11.3 mm, 12.1 mm, and 16.4 mm, respectively. As the concentration decreased, the clear zones became larger and the edges became more distinct. After an 8-fold dilution, the fermentation supernatant's α-amylase inhibition rate was nearly zero, while the fermentation broth maintained 50% inhibition even after an 8-fold dilution. This indicates that the α-amylase inhibition by Weissmannia coagulan SA9 and its fermentation products was dose-dependent.

[0032] As can be seen from the above, the Weissmannia coagulan SA9 strain provided by the present invention has excellent inhibitory effects on α-glucosidase and α-amylase, and by inhibiting the activity of α-glucosidase and α-amylase, it blocks the conversion and absorption of sugars, thereby facilitating blood sugar control, alleviating diabetes, and having beneficial effects on the prevention and treatment of diabetes, and can be used in products to lower blood sugar levels. Currently, Weissmannia coagulan is registered as an edible probiotic in many countries and regions. Its characteristics include high safety, high temperature resistance, spore formation, and processability. Weissmannia coagulan can be provided to diabetic patients in various product forms, providing more treatment options and benefiting diabetic patients in terms of blood sugar control and maintaining a healthy and good mental state. Furthermore, Weissmannia coagulan has simple fermentation conditions, is easy to industrialize, and is low-cost, making it a probiotic strain with great market potential.

[0033] The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be considered as equivalent substitutes and fall within the protection scope of the present invention.

Claims

1. A Weizmannia coagulans SA9 strain capable of inhibiting the activity of α-glucosidase and / or α-amylase, characterized in that the Weizmannia coagulans SA9 strain was deposited at the Guangdong Provincial Center of Microorganisms and Species Preservation on June 6, 2022 under deposit number GDMCC NO: 62517.

2. 2. The use of the Weissmannia coagulan SA9 strain according to claim 1 and / or its fermentation broth in inhibiting α-glucosidase and / or α-amylase.

3. 2. Use of the Weissmannia coagulan SA9 strain according to claim 1 and / or its fermentation broth in the production of an inhibitor of α-glucosidase and / or α-amylase.

4. 10. Use of the Weissmannia coagulan SA9 strain according to claim 1 and / or its fermentation broth in the manufacture of a product for alleviating or treating diabetes or controlling blood sugar levels.

5. 5. The application according to claim 4, characterized in that the product is a food product, a nutritional supplement or a pharmaceutical product.

6. An α-glucosidase and / or α-amylase inhibitor, comprising the Weissmannia coagranulosa SA9 strain according to claim 1 and / or a fermentation broth thereof.

7. The inhibitor according to claim 6, characterized in that the fermentation broth is a fermentation supernatant, a cell-free extract, and / or a bacterial cell metabolite.

8. The inhibitor according to claim 6, wherein the OD of the fermentation broth is 0.5 to 1.

0.

9. The inhibitor according to claim 6, wherein the SA9 strain is inoculated into a medium at an inoculum amount of 1 to 10%, and cultured with shaking at 35 to 40°C and 120 to 200 r / min for 28 to 48 hours to obtain the fermentation broth.

10. A method for inhibiting α-glucosidase and / or α-amylase, comprising treating a sample with the Weissmannia coagulan SA9 strain according to claim 1 and / or a fermentation broth thereof.

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