A kind of Lactobacillus plantarum with lipid-lowering function and its application

Abstract

The present invention discloses a plant lactobacillus with lipid-lowering function and its application, which belongs to the field of microbial application. The strain has been deposited in the Guangdong Provincial Microbial Culture Collection Center on January 14, 2022, and the deposit number is GDMCC No: 62211. The present invention isolates plant lactobacillus SDJ09 from sour beans. The bacterium has the ability to lower cholesterol and inhibit pancreatic lipase, has strong adhesion and excellent prebiotic properties (acid and bile salt resistance, in vitro simulated gastrointestinal fluid resistance, antibacterial ability) and safety, and the cell-free extract, metabolites and inactivated bacteria of the bacterium can reduce lipid accumulation in 3T3-L1 adipocytes. The characteristics of plant lactobacillus SDJ09 give it a wide range of application prospects, and it can be applied to medicines and animal feeds with lipid-lowering functions, providing a new way for the prevention and treatment of obesity in the body.

Description

Technical Field

[0001] The invention relates to the field of microbial applications, and in particular to a plant lactobacillus with lipid-lowering function and an application thereof. Background Art

[0002] With the increasing incidence of obesity, obesity has become a global public problem, which leads to a decline in the quality of life of obese people and even affects life expectancy. Obesity is also closely related to the occurrence of metabolic syndrome such as non-alcoholic fatty liver disease, type 2 diabetes, hyperlipidemia and atherosclerosis. The treatment of obesity is currently very challenging. The most common drug on the market is orlistat, but it still has side effects such as oily stools and gastrointestinal discomfort. Therefore, the development of more effective and safe anti-obesity drugs has become a research hotspot.

[0003] Probiotics are called living microorganisms that can have beneficial effects on the body in a certain number. Lactobacillus and Bifidobacterium are the most common species in probiotic preparations. Due to their various beneficial effects on the health of the body, they have been used to prevent and treat various medical diseases and are the focus of the next generation of probiotic research. Studies have shown that lactic acid bacteria can maintain the dynamic balance of the body's intestinal flora by colonizing the intestines, enhance the body's immunity, and have beneficial effects such as anti-inflammatory, anti-tumor, hypoglycemic and lowering serum cholesterol. As early as the 1970s, scholars discovered that lactobacillus can affect the metabolism of cholesterol in the human body, and subsequent related studies have confirmed the cholesterol-lowering effect of lactic acid bacteria.

[0004] It has been confirmed in many studies that lactic acid bacteria have a beneficial effect on reducing obesity, and its anti-obesity effect is different not only between strains, but also between inactivated bacteria and live bacteria, and between single bacteria and mixed bacteria. The research on anti-obesity effect is not limited to the strain level, but also extends to their by-products, including probiotic metabolites, functional proteins, cell lysates and short-chain fatty acids. Inactivated probiotics have the characteristics of wider application, more stable and safer. Their application is no longer limited to low-temperature yogurt, and can also be used in foods under high-temperature processes. In order to expand the scope of application of probiotics, the present invention specially screens and provides a new strain of live bacteria and its metabolites, cell-free extracts and inactivated bacteria that all have probiotic properties. Summary of the invention

[0005] The purpose of the present invention is to provide a plant lactobacillus with lipid-lowering function and an application thereof to solve the problems existing in the above-mentioned prior art. The live bacteria, cell-free extracts, metabolites and inactivated bacteria of the strain can all reduce lipid accumulation in fat cells, have broad application prospects, and provide a new approach for the prevention and treatment of obesity.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a plant lactobacillus (Lactobacillus plantarum) SDJ09 with lipid-lowering function, which has been deposited in Guangdong Provincial Microbiological Culture Collection Center on January 14, 2022, with a deposit number of GDMCC No: 62211.

[0008] The invention also provides a preparation comprising the Lactobacillus plantarum.

[0009] Furthermore, it contains one or more of the live bacteria, inactivated bacteria, metabolites and cell-free extracts of the plant lactobacillus.

[0010] Furthermore, the preparation method of the live bacteria is as follows:

[0011] The plant lactobacillus was inoculated into MRS liquid medium for cultivation, and then the strain concentration was adjusted to 10 8 -10 9 CFU / mL is enough.

[0012] Furthermore, the live bacteria are sterilized at high temperature to obtain inactivated bacteria; the live bacteria are ultrasonically treated and then centrifuged to obtain the supernatant to obtain a cell-free extract; the live bacteria are centrifuged to obtain the supernatant, and filtered to obtain the metabolite.

[0013] Furthermore, the inoculation amount of the plant lactobacillus is 2-3%; and the culture conditions are: culture at 35-38° C. for 24-36 hours.

[0014] Furthermore, the high temperature sterilization is specifically: sterilization at 110-150° C. for 10-30 min; the power of the ultrasonic treatment is 200-300 W, and the time is 20-30 min.

[0015] The present invention also provides an application of the plant lactobacillus or the preparation in the preparation of medicines or animal feed.

[0016] Furthermore, the plant lactobacillus or the preparation exerts a lipid-lowering effect by degrading cholesterol, inhibiting pancreatic lipase and reducing lipid accumulation in fat cells.

[0017] The present invention also provides a medicine or animal feed with lipid-lowering function, comprising the plant lactobacillus or the preparation.

[0018] The present invention discloses the following technical effects:

[0019] The invention provides a plant lactobacillus (Lactobacillus plantarum) GXSDJ09 (referred to as plant lactobacillus SDJ09) separated from sour beans, which has the ability to reduce cholesterol and inhibit pancreatic lipase, has strong adhesion and excellent probiotic properties (acid and bile salt resistance, in vitro simulated gastrointestinal fluid resistance, antibacterial ability) and safety, and the cell-free extract, metabolites and inactivated bacteria of the bacteria can all reduce lipid accumulation in 3T3-L1 adipocytes. The characteristics of plant lactobacillus SDJ09 make it have a wide range of application prospects, can be applied to medicines and animal feeds with lipid-lowering functions, and provide a new approach for the prevention and treatment of obesity in the body. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] 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 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.

[0021] Figure 1 The cholesterol standard curve and the in vitro cholesterol degradation rates of 39 lactic acid bacteria strains;

[0022] Figure 2 The gel electrophoresis diagram of the PCR product of Lactobacillus plantarum strain SDJ09 and the constructed developmental tree;

[0023] Figure 3 The growth curve and acid production curve of Lactobacillus plantarum SDJ09;

[0024] Figure 4 Oil red O staining images of 3T3-L1 adipocytes under different treatment conditions, A is the model group, B is the cell-free extract treatment group, C is the metabolite treatment group, and D is the inactivated bacterial treatment group;

[0025] Figure 5 The effects of different components of Lactobacillus plantarum SDJ09 on triglycerides in 3T3-L1 adipocytes; * indicates that the model group was significantly different from the normal group (P<0.05), and # indicates that the different treatment groups were significantly different from the model group (P<0.05). DETAILED DESCRIPTION

[0026] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0027] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0028] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0029] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0030] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0031] 1. Isolation and purification of lactic acid bacteria

[0032] A total of 10 traditional fermented foods (pickled peppers, sauerkraut, milk fan, ham, sour soup, chili sauce, sour beans, sour bamboo shoots, kimchi, and soybean sauce) were collected from four regions of Sichuan, Yunnan, Guizhou, and Guangxi as samples. 25 g of the sample was taken and placed in 225 mL of sterile saline for 30 min of shaking. Then, 200 μL of the dilution solution was diluted and spread on the MRS screening solid medium and cultured at 37 ° C for 48 h. The colony morphology was mostly centrally prominent, white in color, with obvious calcium dissolution rings around the colony, neat edges, moist and opaque surface, and accompanied by sour taste. The color and morphology of the strains suspected of lactic acid bacteria were selected on the plate. After continuous purification and culture for 3 times, a single colony was selected for catalase test and Gram staining test (experimental method reference "Screening and application of lactic acid bacteria with antioxidant function in chopped peppers"), and Gram-positive and rod-shaped under microscopy, without spores, single strain morphology and catalase-negative strains were selected for glycerol preservation and stored at -20 ° C for use. 39 strains of lactic acid bacteria were isolated and purified by the above method.

[0033] 2. Screening of Lactobacillus plantarum SDJ09

[0034] 2.1 Screening of cholesterol-lowering lactic acid bacteria

[0035] Drawing of cholesterol standard curve: Take 0.1, 0.2, 0.3, 0.4, 0.5, 0.6mL of cholesterol stock solution in clean test tubes respectively, evaporate the liquid with nitrogen at 60℃, add o-phthalaldehyde and concentrated sulfuric acid successively, mix well, let stand and measure the absorbance at 550nm, and draw the standard curve ( Figure 1 (top middle picture).

[0036] After seed MRS culture medium was used for the 39 strains, they were inoculated into MRS culture medium containing cholesterol at a 3% inoculation rate. After constant temperature culture at 37°C for 36 hours, the culture was centrifuged at 8000r / min for 10 minutes. The cholesterol degradation ability was determined by the o-phthalaldehyde method. The operation steps were as follows: take 1mL of supernatant in a test tube, add 4mL of 95% ethanol and 3mL of 33% KOH, bathe in a 60°C water bath for 15 minutes, add 5mL of n-hexane and 4mL of distilled water, shake well and let stand to separate layers, take 3mL of the upper layer liquid in a small test tube, evaporate the reagent with nitrogen at 60°C, add 5mL of o-phthalaldehyde and 3mL of concentrated sulfuric acid, mix well, detect the absorbance at a wavelength of 550nm, calculate the cholesterol removal rate according to the formula, and repeat the experiment five times to calculate the average value.

[0037] Cholesterol degradation rate = (blank group OD 550 - Experimental group OD 550 ) / blank group OD 550 x100%.

[0038] The results of the cholesterol degradation test of 39 lactic acid bacteria are shown in Figure 1 In the lower middle figure, the five lactic acid bacteria strains SDJ07, SDJ09, SS08, SDJ13 and SS05 with higher cholesterol-lowering ability were involved in the following experiments.

[0039] 2.2 Determination of pancreatic lipase inhibition rate

[0040] The operation steps of the pancreatic lipase inhibition rate of the five strains were as follows: the strains were inoculated with 3% of MRS medium and cultured for 18 h, washed twice with PBS at pH = 7.2, and then resuspended in PBS to adjust the OD 600 To 0.5±0.02(10 8 -10 9CFU / mL). Prepare Tris-Hcl buffer with pH=8.5, add 12.5μL 5mM 4-nitrooctanoic acid phenyl ester (NPC) to 500μL Tris-Hcl buffer and mix with 125μL bacterial solution, then add 12.5μL pancreatic lipase (5mg / mL Tris-Hcl buffer) and mix for 2min, and incubate the mixture at 37°C for 30min, take 150μL in a 96-well plate, and measure the absorbance at 412nm wavelength (n=3). The sample blank is dimethyl sulfoxide (DMSO) instead of NPC, the control sample is only NPC without lactic acid bacteria, and the blank control is DMSO instead of NPC but without lactic acid bacteria. Calculate the inhibition rate according to the following formula:

[0041] Pancreatic lipase inhibition rate (%) = 1-((C-C0) / (N-N0)) × 100%

[0042] In the formula: C0 represents the OD value of the sample blank group at a wavelength of 412nm, C represents the OD value of the sample at a wavelength of 412nm, N0 represents the OD value of the control blank group at a wavelength of 412nm, and N represents the OD value of the control sample at a wavelength of 412nm. The experimental results are shown in Table 1.

[0043] Table 1 Pancreatic lipase inhibition rate of 5 lactic acid bacteria strains

[0044]

[0045]

[0046] 2.3 Adhesion test

[0047] Using Lactobacillus rhamnosus LGG as the control bacteria, HT-29 cells were cultured with 1640 complete medium containing 10% fetal bovine serum at 37°C and 5% CO2. When the cells proliferated and became fused to about 80%, they were digested with 0.25% trypsin and diluted with 1640 complete medium without double antibody to adjust the cell number to 2x10 5 cell / mL, take 0.5mL and culture in a 24-well plate until the proliferation and fusion reaches 80%, wash the cells with sterile PB, add the test solution, incubate for 2h, wash the cells with PBS, add trypsin digestion for 2min to terminate the reaction, then add 1mL0.05% TritionX-100, repeatedly blow and beat to completely lyse the cells, let stand for 5min, transfer the lysate to a sterile test tube, dilute and spread, and calculate the adhesion according to the following formula:

[0048] Adhesion = (total number of adherent bacteria / number of HT-29 cells per dish) x 100%;

[0049] The experimental results are shown in Table 2:

[0050] Table 2 Adhesion test of 6 lactic acid bacteria strains

[0051]

[0052] Combined with the above experimental results, a strain with high cholesterol-lowering ability, high pancreatic lipase inhibition rate and highest adhesion was screened out: SDJ09, which was isolated from sour beans in a market in Guangxi.

[0053] 3. 16S rRNA gene sequence analysis

[0054] The above-screened strains were identified by PCR, and the genomic DNA of the strains was extracted using a bacterial DNA extraction kit. The universal primers 27F and 1492R were used to amplify the strains. The amplification conditions were: 94°C pre-denaturation for 3 min, 94°C denaturation for 30 s, 55°C annealing for 30 s, 72°C extension for 1 min, 30 cycles, 72°C extension for 5 min, and storage at 4°C. The PCR products were analyzed by 1% agarose gel electrophoresis ( Figure 2 The 16S rRNA gene sequences were compared in NCBI, BLAST was used for homology analysis, and MEGA5.0 software was used to build a developmental tree ( Figure 2 After identification, SDJ09 was identified as Lactobacillus plantarum, named Lactobacillus plantarum GXSDJ09 (abbreviated as Lactobacillus plantarum SDJ09), and preserved in Guangdong Microbial Culture Collection Center, the preservation address is: 5th Floor, Laboratory Building, Provincial Institute of Microbiology, No. 100, Xianlie Middle Road, Guangzhou, China, the preservation date is January 14, 2022, and the preservation number is GDMCCNo: 62211.

[0055] 4. Probiotic properties of lipid-lowering lactic acid bacteria

[0056] 4.1 Growth curve and acid production curve of Lactobacillus plantarum SDJ09

[0057] The strain was inoculated into MRS liquid medium at 3% inoculum, cultured at 37°C, and samples were taken every 2 hours to measure the absorbance and pH value of the fermentation broth at 600nm, and then the growth curve and acid production curve were drawn. Figure 3 , Lactobacillus plantarum SDJ09 has excellent growth performance and acid production ability.

[0058] 4.2 Acid and bile salt resistance

[0059] A single colony was picked and cultured in MRS liquid medium for 20 h, then centrifuged at 4°C, 8000 r / min for 10 min, and then resuspended in sterile PBS to adjust the OD 600To 0.5±0.02, and then inoculated into MRS liquid culture medium with pH=2 and 0.3% ox bile salt at a 3% inoculum size, diluted and spread at 0, 2, and 3h, and the survival rate was determined by calculating the number of live bacteria on the plate before and after treatment. The results are shown in Table 3. Compared with the control bacteria LGG, Lactobacillus plantarum SDJ09 has a higher survival rate and exhibits excellent acid and bile resistance.

[0060] Table 3 Acid and bile resistance characteristics of Lactobacillus plantarum SDJ09

[0061]

[0062] 4.3 Resistance to simulated gastrointestinal fluid in vitro

[0063] The in vitro static digestion model was used to simulate gastrointestinal fluid. 3g / L pepsin was dissolved in PBS (pH=3) to simulate gastric fluid, and 1.0g / L trypsin was dissolved in PBS (pH=8) to simulate intestinal fluid, and then treated with a 0.22μm filter membrane to sterilize. On this basis, the lactic acid bacteria in the simulated gastric fluid were cultured at 37°C for 3h, and 1mL of the mixture was added to 9mL of simulated intestinal fluid. After continuing to culture for 8h, gradient dilution and coating were performed. The survival rate was determined according to the number of viable bacteria on the plate before and after treatment. The results are shown in Table 4. The survival rate of Lactobacillus plantarum SDJ09 in simulated in vitro gastrointestinal fluid was higher than that of LGG, and it had good resistance to simulated in vitro gastrointestinal fluid.

[0064] Table 4 Resistance of Lactobacillus plantarum SDJ09 to simulated gastrointestinal fluid in vitro

[0065]

[0066]

[0067] 4.4 Antibacterial ability of strain metabolites

[0068] The antibacterial ability of the strain metabolites was determined by the Oxford cup method. The indicator strains Escherichia coli and Staphylococcus aureus were adjusted to a concentration of 10 8 CFU / mL, evenly spread on LB plate, take the culture solution after 24h of lactic acid bacteria fermentation, centrifuge at 8000r / min for 10min, take the supernatant and treat it with 0.22μm filter membrane, then take 100μL in Oxford cup, culture for 24h, measure the inhibition zone, and evaluate its antibacterial effect according to the diameter of the inhibition zone. The results are shown in Table 5. The diameter of the inhibition zone of Lactobacillus plantarum SDJ09 metabolites against Escherichia coli and Staphylococcus aureus is greater than 10mm, which has good antibacterial ability.

[0069] Table 5 Antibacterial ability of Lactobacillus plantarum metabolites

[0070]

[0071] 4.5 Antibiotic sensitivity of Lactobacillus plantarum SDJ09

[0072] The drug sensitivity test of lactic acid bacteria uses the drug sensitivity paper agar diffusion method (KB method). Fresh bacteria are centrifuged at 4°C, 8000r / min for 10 minutes, the supernatant is discarded, and the bacteria are collected. Resuspend with PBS and adjust the concentration to 10 7 CFU / mL, divide the MRS solid medium into areas and mark them, draw 200μL and apply it on the MRS medium, then put the antibiotic susceptibility sheet on the surface of the plate covered with bacteria, culture at 37℃ for 48h, observe the inhibition zone and measure the diameter of the susceptibility zone. The experimental results are shown in Table 6. Lactobacillus plantarum SDJ09 is sensitive or intermediate to 8 antibiotics and has a certain safety.

[0073] Table 6 Antibiotic sensitivity of Lactobacillus plantarum SDJ09

[0074]

[0075] 5. Effects of different components of Lactobacillus plantarum SDJ09 on lipid accumulation in 3T3-L1 adipocytes

[0076] 5.1 Sample preparation

[0077] Preparation of cell-free extract (CFE): Lactobacillus plantarum SDJ09 was cultured in MRS medium for 24 h, centrifuged at 8000 rpm for 10 min at 4 °C, washed twice with PBS, resuspended in distilled water, and the OD was adjusted. 600 to 0.5±0.02, and ultrasonically treated in an ice bath for 30 min (200 W, working 5 s, rest 5 s), and then centrifuged to obtain the supernatant for freeze-drying.

[0078] Preparation of metabolites (CM): After activation, Lactobacillus plantarum SDJ09 was inoculated in MRS liquid medium at a 2% inoculum volume and cultured at 37°C for 24 h. The cultured bacterial liquid was centrifuged at 4°C, 8000 r / min, for 10 min, and washed twice with sterile PBS. The strain concentration was adjusted to 1×10 8 CFU / mL, culture at 37℃ for 2h, then centrifuge at 4℃, 8000r / min for 10min, take the supernatant and filter it through 0.22μm filter membrane to obtain the lactic acid bacteria metabolite sample.

[0079] Preparation of inactivated bacteria (HK): After culturing for 24 h, Lactobacillus plantarum SDJ09 was washed twice with sterile PBS and resuspended in PBS to adjust the OD 600 To 0.5±0.02, after sterilization at 121℃ for 15min, the lactic acid bacteria are heat-killed.

[0080] 5.2 Effects of different components of Lactobacillus plantarum SDJ09 on lipid accumulation in 3T3-L1 cells

[0081] The classic cocktail method was used to induce differentiation of 3T3-L1 adipocytes (refer to "Effects of Bacillus Subtilis-Fermented White Sword Bean Extract on Adipogenesis and Lipolysis of 3T3-L1 Adipocytes."). 10% fetal bovine serum DMEM culture medium containing three components of Lactobacillus plantarum SDJ09 and 10% fetal bovine serum DMEM culture medium (model group) were added to the mature adipocytes after induced differentiation and cultured for 24 hours. Normally grown 3T3-L1 cells were used as the control group (NC), the culture medium was removed, and the cells were carefully washed three times with PBS, and Oil Red O staining and triglyceride content were determined.

[0082] Oil red O staining of 3T3-L1 adipocytes under different treatment conditions Figure 4 As shown, red represents lipid droplets, purple represents cytoplasm, where A is the model group (CON), B is the cell-free extract treatment group (CFE), C is the metabolite treatment group (CM), and D is the inactivated bacteria treatment group (HK).

[0083] Determination of triglyceride content:

[0084] Add 100 μL of RIPA cell lysis buffer containing 10 μL of PMSF to each well of the treated cells, fully lyse on ice, centrifuge at 12000 r / min for 10 min, take the supernatant, and measure the triglyceride content with a triglyceride kit and the total protein with a BCA kit. Figure 5 As shown, all three fractions of Lactobacillus plantarum SDJ09 could reduce the triglyceride content in 3T3-L1 adipocytes.

[0085] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A plant lactobacillus with lipid-lowering function ( Lactobacillus plantarum )SDJ09, characterized in that, It was deposited in the Guangdong Provincial Microbiological Culture Collection Center on January 14, 2022, with the deposit number GDMCC No: 62211.

2. A preparation, characterized in that Contains Lactobacillus plantarum ( Lactobacillus plantarum ) one or more of live bacteria, inactivated bacteria, metabolites and cell-free extracts of SDJ09; the deposit number of the Lactobacillus plantarum SDJ09 is GDMCC No: 62211; The preparation method of the live bacteria is as follows: The plant lactobacillus SDJ09 was inoculated into MRS liquid medium for cultivation, and then the strain concentration was adjusted to 10 8 -10 9 CFU / mL is enough; The live bacteria are sterilized at high temperature to obtain inactivated bacteria; the live bacteria are ultrasonically treated and then centrifuged to obtain the supernatant to obtain a cell-free extract; the live bacteria are centrifuged to obtain the supernatant, and the supernatant is filtered to obtain the metabolite.

3. The preparation according to claim 2, characterized in that The inoculation amount of the Lactobacillus plantarum SDJ09 is 2-3%; the culture conditions are: culture at 35-38° C. for 24-36 h.

4. The preparation according to claim 2, characterized in that The high temperature sterilization is specifically: sterilization at 110-150° C. for 10-30 min; the power of the ultrasonic treatment is 200-300 W, and the time is 20-30 min.

5. Use of the Lactobacillus plantarum SDJ09 according to claim 1 or the preparation according to any one of claims 2 to 4 in the preparation of lipid-lowering drugs or animal feed.

6. The use according to claim 5, characterized in that: The Lactobacillus plantarum SDJ09 or the preparation exerts a lipid-lowering effect by degrading cholesterol, inhibiting pancreatic lipase and reducing lipid accumulation in fat cells.

7. A drug or animal feed having lipid-lowering function, characterized in that: A preparation comprising the Lactobacillus plantarum SDJ09 according to claim 1 or any one of claims 2 to 4.

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