Lactobacillus mucilaginosus HNGD-DF3, microbial inoculum and application of lactobacillus mucilaginosus HNGD-DF3 in reducing blood fat and blood sugar

Abstract

The invention belongs to the technical field of microorganisms, and relates to lactobacillus mucus HNGD-DF3, a microbial agent and application of the lactobacillus mucus HNGD-DF3 to lipid and sugar reduction. The lactobacillus mucus HNGD-DF3 is separated and screened from bean curds in Zunyi City in Guizhou by the inventor, and the preservation number is CCTCC M 2025193. The fermentation lactobacillus mucus HNGD-DF3 is a microorganism which is found for the first time and is used for degrading branched chain amino acid, and the strain achieves the effect of reducing blood fat and blood sugar by reducing the content of the branched chain amino acid in a living body. Further research finds that the strain has good acid resistance, cholate resistance and potential of colonization in gastrointestinal tracts at the same time.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to a fermented mucus lactobacillus. Background Art

[0002] Branched-chain amino acids include leucine, isoleucine and valine, which play an important regulatory role in the body's metabolic pathways such as energy metabolism, inflammation and glycolysis. Epidemiological studies have found that high circulating concentrations of branched-chain amino acids in the body are positively correlated with the occurrence of chronic metabolic diseases such as obesity and diabetes. A high-branched-chain amino acid diet can increase the content of branched-chain amino acids in the blood circulation and induce obesity and diabetes, while a low-branched-chain amino acid diet can reduce the content of branched-chain amino acids in the blood circulation and improve obesity and diabetes. Branched-chain amino acids in natural foods are almost all present in proteins in a bound form, and there is currently no safe, healthy and sustainable way to reduce them. Branched-chain amino acids in food proteins are absorbed from the intestines and enter the body's circulation. In this process, microorganisms in the intestines can metabolize branched-chain amino acids and affect the content of branched-chain amino acids in the blood circulation. Lactobacillus, as a type of intestinal probiotic that is recognized as safe, has the benefits of regulating body metabolism and enhancing immunity, inhibiting bacteria, and regulating intestinal flora. It has been reported that some lactic acid bacteria have the ability to utilize branched-chain amino acids and degrading enzymes. Therefore, using lactic acid bacteria to degrade branched-chain amino acids in the intestine and reduce the content of branched-chain amino acids circulating in the body may become an effective way with great development potential and achieve many health benefits. The application with publication number CN 118086155 A discloses a fermented mucus lactobacillus with weight loss and lipid-lowering effects. The bacteria achieves weight loss and lipid-lowering by regulating short-chain fatty acids in the intestine and degrading cholesterol, but its intestinal colonization ability is unclear. In order to further explore new safe, efficient, and highly active lactic acid bacteria that can colonize in the intestine and play a role in degrading branched-chain amino acids, this research group conducted in-depth exploration. Summary of the invention

[0003] In order to solve the above technical problems, the present invention proposes a fermented mucus lactobacillus HNGD-DF3, a bacterial agent and the application thereof in reducing blood lipids and blood sugar.

[0004] The technical solution of the present invention is achieved in this way: A strain of Lactobacillus mucilaginosus ( Limosilactobacillus fermentun )HNGD-DF3, taxonomic name: Limosilactobacillus fermentun It was deposited in the China Center for Type Culture Collection on January 21, 2025, with the deposit number CCTCC M 2025193, and was named: HNGD-DF3.

[0005] The fermented mucus lactobacillus HNGD-DF3 of the present invention is obtained by separation and screening from tofu in Zunyi City, Guizhou Province, and its colony characteristics are as follows: the colony diameter is about 2-3 mm, the colony is milky white, the colony is convex, round, opaque, the surface is smooth, the colony edge is irregular, it is a Gram-positive bacterium, and the bacterial body is observed to be short rod-shaped and single through a scanning electron microscope.

[0006] The present invention studies the fermentation of mucus lactobacillus ( Limosilactobacillus fermentun )Application of HNGD-DF3 in reducing lipid and blood sugar in obese mice, requesting protection: A microbial agent comprising the above-mentioned fermented Lactobacillus mucilaginosus HNGD-DF3.

[0007] Application of the above-mentioned fermented mucus lactobacillus HNGD-DF3 or the above-mentioned microbial agent.

[0008] Furthermore, the above application is lipid reduction and / or blood sugar reduction. The above fermented Lactobacillus mucilaginosus HNGD-DF3 reduces lipids by slowing down the accumulation of white fat. The above reduces the content of branched-chain amino acids in the blood of living organisms, and the above application is achieved through the intestinal colonization ability of the fermented Lactobacillus mucilaginosus HNGD-DF3.

[0009] In addition, the invention also requests protection for the use of fermented mucus lactobacillus HNGD-DF3 in the preparation of weight loss products, sugar control products or products for the treatment of hyperlipidemia.

[0010] The present invention has the following beneficial effects: 1. The present invention screened a strain of fermented mucus lactobacillus (Lactobacillus mucosa) from tofu in Zunyi City, Guizhou Province, which has high branched-chain amino acid transaminase activity and can efficiently degrade branched-chain amino acids. Limosilactobacillus fermentun ), the bacteria not only has good acid resistance and bile salt tolerance, but also has the potential to inhibit bacteria and colonize the gastrointestinal tract. The fermented mucus lactobacillus HNGD-DF3 can efficiently degrade branched-chain amino acids. Further research found that the strain also has good acid resistance, bile salt tolerance and the potential to colonize the gastrointestinal tract. In addition, this is the first time that a method has been discovered and used to degrade branched-chain amino acids by fermenting mucus lactobacillus, and the strain is used to reduce the content of branched-chain amino acids in the body, achieving lipid-lowering and blood sugar-lowering effects.

[0011] 2. The present invention provides a method for reducing the level of branched-chain amino acids in the body by lactic acid bacteria to achieve the purpose of reducing blood lipids and blood sugar. The bacteria can achieve a 46.31% branched-chain amino acid degradation effect when cultured in a fermentation medium for 24 hours. Further studies have found that it has high branched-chain amino acid transaminase activity, can colonize in the intestine and degrade branched-chain amino acids during the digestion and absorption of food in the intestine, thereby reducing the abnormally elevated branched-chain amino acid content in the body and restoring it to normal levels, achieving the purpose of reducing blood lipids and blood sugar. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0013] Figure 1 This is the morphological diagram of strain HNGD-DF3.

[0014] Figure 2 Phylogenetic tree of strain HNGD-DF3 based on 16S rDNA gene sequence.

[0015] Figure 3 This is the growth curve of strain HNGD-DF3 in MRS liquid medium.

[0016] Figure 4 This is the effect of strain HNGD-DF3 on the body weight of mice.

[0017] Figure 5 This is the effect of strain HNGD-DF3 on glucose tolerance in mice.

[0018] Figure 6 This is the effect of strain HNGD-DF3 on mouse adipose tissue.

[0019] Figure 7 This is the effect of strain HNGD-DF3 on the content of branched-chain amino acids in mouse blood.

[0020] Figure 8 This is the effect of strain HNGD-DF3 on the levels of four blood lipids in mice.

[0021] Fig. 9 This is the effect of strain HNGD-DF3 on lipid content and hepatocytes in the liver. DETAILED DESCRIPTION

[0022] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] Unless otherwise specified, the experimental methods used in the following experimental examples are all conventional methods; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0024] Example 1: Strain screening and identification (1) Isolation and purification of strains 10 g of tofu from Zunyi, Guizhou was added to 100 mL of sterile PBS buffer, shaken at 37°C, 150 r / min for 2 h, then 10 mL of liquid was taken for gradient dilution with PBS buffer, and the diluted liquid was evenly spread on MRS solid selective medium containing 0.4 g / L bromocresol purple, and statically cultured at 37°C for 72 h. Plates with dispersed colonies and diverse colony morphology were selected to pick strains with different forms that could produce yellow transparent circles, and streaked and purified until there were no foreign bacteria. Then, strains with positive Gram staining and negative catalase reaction obtained by Gram staining and catalase experiments were inoculated into MRS liquid medium, and after static culture for 24 h, centrifuged at 8000 rpm for 15 min, and the supernatant was taken to determine the changes in branched-chain amino acid content. Results Compared with the uninoculated MRS solution, the leucine content, isoleucine content, valine content, and total branched-chain amino acid content of the solution inoculated with HNGD-DF3 strain decreased by 45.71%, 44.42%, 48.71%, and 46.31%, respectively.

[0025] (2) Branched-chain amino acid transaminase activity test 2,4-Dinitrophenylhydrazine colorimetric reagent: Accurately weigh 50 mg of 2,4-dinitrophenylhydrazine and dissolve it in 1 mol / L HCl to make up to 100 mL.

[0026] After HNDG-DF3 was inoculated in MRS liquid medium and cultured at 37℃ for 24 h, 5 mL of fermentation broth was taken as crude enzyme solution, 0.5 mL of cell wall breaking agent (0.2 mmol / L hexadecyltrimethylammonium bromide solution) was added and kept at 37℃ for 5 min, 5 mL of substrate solution (10 mmol / L pyruvate, 10 mmol / L L-amino acid, 0.1 mmol / L 5'-pyridoxal phosphate, pH8.5) which had been kept at 37℃ for 5 min was added, and after oscillation reaction (150 r / min) at 37℃ for 1 h, 0.5 mL of 10% trichloroacetic acid solution was added and fully shaken to terminate the reaction. Centrifuge at 5000 r / min for 10 min, take 2 mL of supernatant and add 2 mL of 2,4-dinitrophenylhydrazine colorimetric agent to determine the concentration of pyruvate, and calculate the enzyme activity of branched-chain amino acid transaminase according to the reduction of pyruvate.

[0027] Definition of enzyme activity: At 37°C, the amount of enzyme required to convert 1 μmol of pyruvate per hour is defined as one unit of enzyme activity (U).

[0028] Enzyme activity calculation formula: Enzyme activity (U / mL) = (C1-C2)×11×1000 / (5×88.06); Where: C1: dilute initial concentration of substrate; C2: dilution conversion concentration of substrate; 11: total volume after the reaction is completed; 5: Volume of crude enzyme solution; 88.06: The molar mass of pyruvate.

[0029] Lactobacillus mucilaginosus fermentation Limosilactobacillus fermentun ) Substrate selectivity of HNGD-DF3 (3) Morphological identification After the strain HNGD-DF3 stored at -80℃ was activated for 3 generations, it was streaked on MRS solid medium and cultured at 37℃ for 48 h to observe the colony morphology. Then the colonies were picked for Gram staining and the cell morphology was observed under a microscope.

[0030] Colony morphology: The colony diameter is about 2-3 mm, the colony is milky white, the colony is convex, round, opaque, the surface is smooth, and the colony edge is irregular; Microscopic morphology: Gram staining is positive, the cells are short rod-shaped; Morphology under electron microscope: The bacteria are short rod-shaped, single ( Figure 1 ).

[0031] (4) 16S rRNA identification After activation for three generations, the cells were inoculated into MRS liquid medium and cultured at 37°C for 18 h. The bacteria were collected and the genomic DNA of strain HNGD-DF3 was extracted using the Ezup column bacterial genomic DNA extraction kit (Sangong). PCR amplification was performed using the genomic DNA of strain HNGD-DF3 as a template and 16S rRNA universal primers 27F and 1492R.

[0032] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-GGTTACCTTGTTACGACTT-3'.

[0033] The PCR amplification program was as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 55°C for 45 s, and extension at 72°C for 60 s, for 30 cycles; and extension at 72°C for 10 min.

[0034] The purity and size of the amplified product were detected by electrophoresis, and the PCR product with the correct amplified length was sent to Shanghai Bioengineering for sequencing. The sequencing sequence is shown below.

[0035] The 16S rRNA gene sequence of strain HNGD DF3 obtained by sequencing has 1477 bases and was compared by BLAST analysis at NCBI. According to the BLAST results, strain HNGD-DF3 and Lactobacillus fermentans ( Limosilactobacillus fermentun The similarity of 16S rRNA gene sequences of the two strains was 99.86%. The phylogenetic tree was constructed by MAGE11 software. Figure 2 As shown in the figure, the step size is set to 1000, and the strain is determined to be fermentative mucus Lactobacillus. Limosilactobacillus fermentun ) HNGD-DF3, and was deposited in the China Center for Type Culture Collection on January 21, 2025, with the address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, China Center for Type Culture Collection, Wuhan University, Postal Code 430072; the deposit number is CCTCC M 2025193.

[0036] Example 2: Study on strain characteristics (1) Plotting of strain growth curve The strain after activation for 3 generations was inoculated into MRS liquid medium with an inoculation loop, and the MRS liquid medium without strain was used as blank. The OD600 was measured every 2 hours, and the process was repeated three times. The growth curve was drawn with time and OD600 as the horizontal and vertical coordinates, respectively. The results are shown in Figure 2. Figure 3 As shown, under static culture conditions at 37°C, the strain was in the lag phase from 0 to 10 h, entered the exponential growth phase after 10 h, and entered the steady growth phase after about 20 h.

[0037] (2) Acid and bile salt resistance test Preparation of artificial gastric juice: A pepsin (1:2500) enzyme activity solution with a mass concentration of 9 mg / mL was prepared with 0.01 M PBS buffer, and then the pH value was adjusted to 3.0 with 1 M HCl solution and sterilized by filtration with a 0.22 μm microporous membrane.

[0038] Preparation of artificial intestinal fluid: A trypsin (1:300) enzyme activity solution with a mass concentration of 1 mg / mL was prepared with 0.01 M PBS buffer, and then 0.3% (m / v) ox bile salt was added. The pH value was adjusted to 8.0 with 1 M HCl solution and then sterilized by filtration through a 0.22 μm microporous membrane.

[0039] After the strain HNGD-DF3 was activated for 3 generations, it was inoculated into MRS liquid medium and cultured at 37°C for 18 h. 5 mL of bacterial solution was centrifuged (6000 r / min, 10 min) to collect the bacteria and washed twice with an equal volume of 0.01 M PBS buffer. The washed bacteria were cultured with an equal volume of artificial gastric juice and artificial intestinal juice for 0 and 4 h. The treated bacteria were gradient diluted with PBS buffer and the number of viable bacteria was determined by plate counting method. The plate count of the dilution gradient with a colony count between 30 and 300 was selected.

[0040] Survival rate = (number of viable bacteria after 4 h of treatment) / (number of viable bacteria after 0 h of treatment) × 100%.

[0041] Result analysis: The survival rate of strain HNGD-DF3 after being treated in artificial gastric juice and artificial intestinal juice for 4 hours is as follows: This indicates that the strain has good resistance to acid and bile salt solutions and has the potential to survive in the gastrointestinal tract.

[0042] (3) Cell hydrophobicity and self-aggregation ability test After activating the strain HNGD-DF3 for 3 generations, it was inoculated into MRS liquid medium and cultured at 37°C for 18 h. 5 mL of bacterial solution was collected by centrifugation (8000 r / min, 10 min), washed twice with an equal volume of 0.01 M PBS buffer, and then resuspended with PBS buffer and the OD600 value was adjusted to about 0.8, which was recorded as A. 0 , and prepare a lactic acid bacteria suspension.

[0043] Add 1 mL of chloroform to 3 mL of lactic acid bacteria suspension and mix thoroughly. Then let it stand at room temperature for 20 minutes to allow the organic phase and aqueous phase to separate. Remove the organic phase and measure the OD value of the aqueous phase at 600 nm using PBS buffer as a blank, which is recorded as A. 1 The experiment was repeated three times.

[0044] Cell hydrophobicity = [(A 0 -A 1 ) / A 0 ]×100%.

[0045] Take 4 mL of lactic acid bacteria suspension and incubate at room temperature for 5 h, then carefully aspirate 1 mL of the top suspension, and measure the OD value at 600 nm using PBS buffer as a blank, which is recorded as A. 2 The experiment was repeated three times.

[0046] Cell self-aggregation ability = (1-A 2 / A 0 )×100%.

[0047] Result analysis: The cell hydrophobicity and self-aggregation ability of strain HNGD-DF3 are as follows: Cell hydrophobicity is a prerequisite for probiotics to adhere to intestinal epithelial cells and colonize the gastrointestinal tract to exert their beneficial effects. The self-aggregation ability of cells can evaluate the ability to adhere to intestinal cells and avoid pathogen colonization.

[0048] The results showed that strain HNGD-DF3 had good hydrophobicity and self-aggregation ability, and had the potential to colonize the intestine and inhibit pathogenic bacteria.

[0049] Example 3: Effects of fermented Lactobacillus mucilaginosus HNDG-DF3 on glucose and lipid metabolism in obese mice induced by a high-fat diet (1) Animal husbandry Forty 6-week-old male C57BL / 6 mice purchased from the Animal Experimental Center of Zhengzhou University were placed under control conditions of 25°C, 55% humidity, and a 12-h light / dark cycle and fed with a normal diet and a high-fat diet (60% of calories from fat).

[0050] (2) Experimental process The mice were randomly divided into 4 groups (ND&S group, ND&B group, HF&S group and HF&B group), with 10 mice in each group. During the one-week adaptation period, they were provided with unlimited food and drinking water. After the adaptation period, the ND&B group and the HF&B group were gavaged with fermented Lactobacillus mucosus HNGD-DF3 (10 8 CFU / mL), 200 microliters / mouse / day, and the ND&S group and HF&S group were gavaged with the same volume of normal saline. During the experiment, all mice drank purified water, the ND&S group and ND&B group were fed with ordinary feed, and the HF&S group and HF&B group were fed with high-fat feed for 8 consecutive weeks. Body weight was recorded every week during the experiment. At the end of the experiment, blood was collected by eyeball method and then the mice were killed by dislocation. After dissection, tissues or organs such as abdominal fat, epididymal fat, perirenal fat, brown fat and liver were obtained.

[0051] (3) Weight record Mice were gavaged daily with fermented Lactobacillus mucilaginosus HNGD-DF3 (10 8 CFU / mL) 200 μL, and continued feeding for 9 weeks. Figure 4 As shown, the weight gain rates of mice in the ND&B group and the ND&S group were similar; compared with the ND&S group, the weight gain in the ND&B group was significantly slowed down, indicating that fermented Lactobacillus mucinus HNGD-DF3 significantly alleviated the weight gain of mice induced by a high-fat diet.

[0052] (4) Oral glucose tolerance test (OGTT) The experiment was conducted in the last week before the end of the experiment. The mice were fasted for 12-14 h overnight, but not water. A 200 mg / mL D-glucose solution was prepared with PBS. The glucose solution was intraperitoneally injected at a dose of 2 g / kg (10 μL / g) at 9:00 am the next day. The blood glucose level of the mice was tested at 0 min, 15 min, 30 min, 60 min, and 120 min. Figure 5 As shown in the figure, the glucose concentration of all animals reached a peak 15 minutes after oral glucose administration. During the OGTT, the blood glucose concentration of the ND&S group was the highest; compared with the ND&S group, the ND&B group reduced the blood glucose concentration, and the area under the curve (AUC) of the ND&B group was also significantly lower than that of the ND&S group, indicating that the fermented mucoid Lactobacillus HNGD-DF3 has a good hypoglycemic effect.

[0053] (5) Fat content and adipocyte staining After the autopsy, the inguinal fat, epididymal fat, perirenal fat, brown fat and other fat were completely removed and weighed. After weighing, part of the fat was added to the paraformaldehyde fixative for fixation. After fixation, the sections were sealed with wax and then H&E staining was performed to observe the changes in adipose tissue. Figure 6 As shown, compared with the ND&S group, the weight of inguinal fat, epididymal fat, perirenal fat and body weight in the HF&S group were significantly increased; while the ND&B group reduced the increase in fat content caused by a high-fat diet. In addition, H&E staining of epididymal fat showed that the adipocytes in the HF&B group were smaller and more densely arranged than those in the HF&S group. HE staining of brown fat showed that the number and volume of vacuoles in the HF&B group were reduced compared with those in the HF&S group. The results showed that fermented mucus lactobacillus HNGD-DF3 has the ability to slow down the accumulation of white fat caused by a high-fat diet.

[0054] (6) Determination of serum indicators Blood was collected from the mouse eyeballs and placed at 4°C for 30 min, then centrifuged at 12,000 rpm for 30 min to obtain serum. The contents of branched-chain amino acids (BCAAs), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in the serum were tested. Figure 7As shown, compared with the ND&S group, the BCAAs content in the blood of mice in the ND&B group was significantly reduced; compared with the HF&S group, the BCAAs content in the blood of mice in the HF&B group was also significantly reduced. The results showed that fermented mucus lactobacillus HNGD-DF3 can reduce the level of BCAAs in the blood of mice. After 9 weeks of high-fat diet intake, the TC, TG, and LDL-C levels in the blood of the HF&S group increased significantly, and the HDL-C level decreased significantly. In the HF&B group, oral administration of fermented mucus lactobacillus HNGD-DF3 significantly inhibited the increase of TC, TG, and LDL-C, and increased the HDL-C level ( Figure 8 ), indicating that fermented Lactobacillus mucilaginosus HNGD-DF3 could improve the increase in blood lipid levels induced by a high-fat diet.

[0055] (7) Determination of liver lipid content and liver tissue staining 0.1 g of fresh liver tissue was added to 0.9 ml of sterile saline and homogenized using a homogenizer to obtain a 10% liver homogenate. The contents of TC, TG, HDL-C, and LDL-C in the liver homogenate were determined. The results are shown in Fig. 9 As shown in the figure, compared with the ND&S group, the TC, TG, and LDL-C levels of the HF&S group increased significantly, while the HDL-C level decreased significantly; while the HF&B group significantly reduced the TC, TG, and LDL-C levels in the liver and increased the HDL-C level by oral administration of fermented Lactobacillus mucosa HNGD-DF3. The liver tissue was fixed in paraformaldehyde, sealed with wax, and then sliced ​​and stained with H&E and ORO to observe the changes in liver tissue. Fig. 9 As shown, the H&E and ORO staining results showed that fermented Lactobacillus mucinus HNGD-DF3 improved the abnormal liver lipid metabolism and liver cell abnormalities caused by a high-fat diet, and also improved lipid accumulation in the liver.

[0056] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A strain of Lactobacillus mucilaginosus HNGD-DF3, whose taxonomic name is Limosilactobacillus fermentun , deposited in the China Center for Type Culture Collection on January 21, 2025, with the deposit number CCTCC M 2025193, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province.

2. A microbial agent comprising the fermented mucilaginous Lactobacillus HNGD-DF3 according to claim 1.

3. Use of the fermented mucus lactobacillus HNGD-DF3 according to claim 1 or the microbial agent according to claim 2.

4. The use according to claim 3, characterized in that: The application is lipid-lowering and / or blood sugar-lowering.

5. The use according to claim 4, characterized in that: The fermented Lactobacillus mucilaginosus HNGD-DF3 achieves lipid reduction by slowing down the accumulation of white fat.

6. The use according to claim 3, characterized in that: The application is to reduce the content of branched-chain amino acids in the blood of a living being.

7. The use according to claim 3, characterized in that: The application is achieved through the intestinal colonization ability of fermented Lactobacillus mucilaginosus HNGD-DF3.

8. Use of the fermented mucus lactobacillus HNGD-DF3 according to claim 1 in preparing weight loss products.

9. Use of the fermented mucus lactobacillus HNGD-DF3 according to claim 1 in the preparation of a sugar-control product.

10. Use of the fermented Lactobacillus mucilaginosus HNGD-DF3 according to claim 1 in preparing a product for treating hyperlipidemia.

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