Bacillus lactis capable of transforming ginsenoside and cassia glycoside and application of the bacillus lactis in reducing fat and relieving fatty liver

Abstract

The application discloses a plant lactobacillus gasseri strain capable of transforming ginsenoside and cassia glycoside, and application of the strain in lipid reduction and alleviation of fatty liver, and belongs to the fields of microbial technology, medicine and functional food. The core feature of the strain is that the strain can not only transform ginsenoside Rb1 into ginsenoside Rd with higher activity, but also transform cassia glycoside into cassia glycoside base, thereby improving the efficacy. The animal experiment proves that the strain and the corresponding substrate are combined to play a better role in lipid reduction and liver protection: the serum total cholesterol and triglyceride levels of high-fat diet mice can be effectively reduced, and the liver weight, liver lipid deposition and liver tissue fatty degeneration can be reduced and improved. The strain provides a new core strain and technical scheme for developing microecological preparations for preventing and / or assisting in treating hyperlipidemia, non-alcoholic fatty liver and other metabolic diseases.

Description

Technical Field

[0001] This invention relates to a strain of *Lactobacillus plantarum* that can convert both ginsenosides and cassiasides, and its application in lowering lipids and alleviating fatty liver, belonging to the fields of microbial technology, medicine, and functional foods. Background Technology

[0002] Nonalcoholic fatty liver disease (NAFLD) is closely related to lipid metabolism disorders and has become one of the most common chronic liver diseases. Currently, there is a lack of safe and effective specific drugs. Although chemically synthesized lipid-lowering drugs such as statins can regulate blood lipids, their direct effect on improving hepatic steatosis is limited, and long-term use may pose side effects. Therefore, developing safe and effective intervention strategies derived from natural products is of great significance.

[0003] Ginseng and cassia seed are both traditional medicinal and edible ingredients with a long history of use. They are often used together in traditional Chinese medicine formulas to achieve synergistic effects. Their main active components are ginsenosides (such as Rb1) and cassia seed glycosides, respectively. However, the bioavailability of the original molecules of these glycosides is generally low. They need to be metabolized in the body into their aglycone forms (such as ginsenoside Rd and cassia seed glycoside) before they can be efficiently absorbed and exert their physiological activities.

[0004] Currently, various methods exist for converting these glycosides, each with its limitations. Chemical hydrolysis involves harsh conditions that can easily damage the active ingredients; enzymatic hydrolysis is costly, and each enzyme is typically only effective against specific glycosidic bonds. The human gut microbiota is considered the main pathway for in vivo conversion, but the significant differences in individual microbial composition lead to considerable uncertainty regarding the metabolic capacity of these two glycosides and the ultimate health benefits among different populations, making it difficult to consistently control the efficacy of compound products.

[0005] The transformation of glycosides by microorganisms depends on the activity of their glycoside hydrolases. Studies have found that the glycoside hydrolases contained in microorganisms have particularly strong specificity. Glycoside hydrolases usually only specifically hydrolyze specific chemical structures. Therefore, existing plant lactobacilli with glycoside hydrolases can often only transform one or a similar type of structure.

[0006] However, a microorganism capable of converting not only triterpenoid saponins but also pyranone compounds (cassia glycosides) is currently lacking. Therefore, this invention urgently seeks to discover a strain that can effectively hydrolyze glycosides to generate the corresponding aglycone forms and enhance their activity when developing products containing ginseng and / or cassia seeds. Summary of the Invention

[0007] The present invention aims to overcome the shortcomings of the low bioavailability of existing ginsenoside Rb1 and cassiaside, and to provide a strain of *Lactobacillus plantarum* CCFM1488 that can simultaneously and efficiently convert ginsenosides and cassiaside.

[0008] The first technical solution provided by this invention is a strain of *Lactobacillus plantarum* (… Lactiplantibacillus plantarum CCFM1488 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66135.

[0009] The second technical solution provided by the present invention is a microbial preparation containing *Lactobacillus plantarum* CCFM1488 as described in the first technical solution.

[0010] In some embodiments, the concentration of *Lactobacillus plantarum* CCFM1488 in the microbial preparation is at least 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0011] In some embodiments, the concentration of *Lactobacillus plantarum* CCFM1488 in the microbial preparation is at least 1 × 10⁻⁶. 9 CFU / mL or 1×10 9 CFU / g.

[0012] The third technical solution provided by this invention is a synbiotic preparation containing live *Lactobacillus plantarum* CCFM1488 as described in the first technical solution, ginseng extract, and / or cassiaside. After administration, this synbiotic preparation will colonize the intestines with *Lactobacillus plantarum* CCFM1488 and enhance the intestines' ability to convert ginsenoside Rb1 and / or cassiaside, thereby improving the efficacy of ginsenoside Rb1 and / or cassiaside.

[0013] The fourth technical solution provided by the present invention is an epigenetic agent, wherein the epigenetic agent is the fermentation broth of the *Lactobacillus plantarum* CCFM1488 described in the first technical solution or the microbial preparation described in the second technical solution after fermentation in a fermentation system containing ginsenosides and / or cassiasides, followed by heat treatment and high-pressure homogenization.

[0014] The fifth technical solution provided by the present invention is the application of *Lactobacillus plantarum* CCFM1488 described in the first technical solution or the microbial preparation described in the second technical solution in the preparation of products for converting ginsenosides and / or cassiasides.

[0015] In some embodiments, the product is a pharmaceutical product.

[0016] The sixth technical solution provided by the present invention is a method for converting ginsenosides and / or cassiasides, which uses *Lactobacillus plantarum* CCFM1488 as described in the first technical solution or the microbial preparation as described in the second technical solution to ferment a substrate containing ginseng extract or cassiasides.

[0017] In some embodiments, the fermentation is carried out at 30-40°C for 48-72 hours; the concentrations of ginsenoside Rb1 and cassiaside are 1-5 mg / mL and 0.5-1 mg / mL, respectively.

[0018] The seventh technical solution provided by this invention is the application of *Lactobacillus plantarum* CCFM1488 described in the first technical solution, the microbial preparation described in the second technical solution, the synbiotic preparation described in the third technical solution, or the postbiotic described in the fourth technical solution in the preparation of a drug for lowering blood lipids and / or alleviating non-alcoholic fatty liver disease.

[0019] In some implementations, the application includes at least one of the following functions: (a) Reduce serum total cholesterol (TC) and / or triglyceride (TG) levels; (b) Reduce liver weight and liver index; (c) Improves fat accumulation in liver tissue; (d) Improve liver damage.

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows: The *Lactobacillus plantarum* CCFM1488 and its applications provided by this invention have the following significant advantages and beneficial effects: ① Extensive glucosidase activity: In this invention, *Lactobacillus plantarum* CCFM1488 can not only convert the triterpenoid saponin compound ginsenoside Rb1 into ginsenoside Rd, but also convert the pyranone compound cassiaside into cassia aglycone.

[0021] ② Fermentation enhancement: This strain can convert glycosides with low activity and bioavailability into corresponding aglycone forms with higher activity and bioavailability, thereby increasing the content of rare ginsenoside Rd and cassia seed aglycone in the product and thus enhancing the product's efficacy.

[0022] ③ High conversion efficiency: Experiments have confirmed that CCFM1488 exhibits high conversion efficiency for both substrates. Based on this strain, synbiotic preparations in live form, microbial preparations after fermentation, and direct applications in various end products such as food, pharmaceuticals, and health products can be flexibly developed to meet diverse market demands.

[0023] Preservation of biological materials A strain of Lactobacillus plantarum ( Lactiplantibacillus plantarum(CCFM1488, taxonomically named) Lactiplantibacillus plantarum It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66135, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0024] Figure 1 Liquid phase diagrams before and after fermentation of a substrate rich in ginsenoside Rb1 by *Lactobacillus plantarum* CCFM1488.

[0025] Figure 2 Liquid phase diagrams before and after fermentation of a cassia glycoside-rich substrate by *Lactobacillus plantarum* CCFM1488.

[0026] Figure 3 The effects of CCFM1488 on serum lipid levels in mice on a high-fat diet were as follows: (A) serum total cholesterol (TC) content; (B) serum triglyceride (TG) content; (C) serum low-density lipoprotein cholesterol (LDL-C) content; (D) serum high-density lipoprotein cholesterol (HDL-C) content; and (E) the ratio of LDL-C to HDL-C in serum.

[0027] Figure 4 The effects of CCFM1488 on liver damage indicators in mice on a high-fat diet: (A) liver weight; (B) serum alanine aminotransferase (ALT) level; (C) serum aspartate aminotransferase (AST) level.

[0028] Figure 5 Effects of CCFM1488 on liver pathological morphology in mice on a high-fat diet; (A) Liver H&E section; (B) Liver Oil Red O stained section; Scale bar: 100 μm.

[0029] Figure 6 This is a chromatogram of the fermentation and transformation of ginsenoside Rb1 by strains that can transform ginsenosides in other studies.

[0030] Figure 7 This is a high-performance liquid chromatogram of cassia glycosides fermented by the comparative strain. Detailed Implementation

[0031] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0032] Test method: HPLC analysis of ginsenoside Rb1 and cassiaside content in fermentation system: (1) Sample pretreatment: Ginsenoside detection: After fermentation, the supernatant of the fermentation broth was collected by centrifugation. Ten times the volume of ether was added to the supernatant three times for defatting. After standing and separating the layers, the upper layer was discarded. An equal volume of water-saturated n-butanol was added to the fermentation broth, and the mixture was thoroughly mixed. The mixture was sonicated (500W, 40kHz) for 10 min, and then allowed to stand for 20 min. The upper phase was collected, and the extraction was repeated three times. The upper phases were combined and evaporated to dryness under reduced pressure at 45℃. Chromatographic grade methanol was added to dissolve the saponins on the wall, and the volume was adjusted to 5.0 ml. The mixture was then filtered through a 0.22 μm organic filter membrane.

[0033] Cassia glycoside detection: After fermentation, the supernatant of the fermentation broth was collected by centrifugation. The supernatant was extracted with dichloromethane and ethyl acetate, respectively. The dichloromethane phase and ethyl acetate phase were evaporated by vacuum centrifugation to remove organic solvents. Then, chromatographic grade methanol was added for redissolution. Finally, the sample was filtered through a 0.22 μm organic filter membrane and then tested.

[0034] (2) Preparation of standard products Accurately weigh 10.0 mg each of ginsenoside and cassiaside standards, dissolve in methanol, and dilute to volume. Prepare a 1.0 mg / mL standard stock solution using methanol, and dilute it to concentrations of 20, 40, 80, 160, and 320 g / mL, respectively. Filter the solutions through a 0.22 μm filter membrane, and take 10 μL samples for HPLC analysis. Plot three standard curves with peak area (mAU) on the ordinate and cassiaside concentration (g / mL) on the abscissa.

[0035] (3) Sample testing Take 10 mL of the pretreated fermentation product (sample to be tested), filter it through a 0.45 μm membrane, and then dilute it to 100 mL with analytical grade 50% methanol solution. Inject the reference solution and the sample test solution, and determine the contents of ginsenoside Rb1 and cassiaside in the fermentation product under the above chromatographic conditions. Calculate the contents using the external standard method.

[0036] The chromatographic conditions were as follows: Column: X Bridge RC18 (250×4.6mm, 5μm); Mobile phase: Water (pH 2.85 + 0.6% (v / v) acetic acid): Methanol = 19:81 (v / v); Detector: Ultraviolet detector (UV) 254nm; Injection volume: 10μL; Column temperature: 35℃; Elution conditions: Flow rate 1.0mL / min.

[0037] Raw materials used in the examples: MRS liquid culture medium: yeast extract 5.0 g / L, beef extract 10.0 g / L, peptone 10.0 g / L, glucose 20.0 g / L, anhydrous sodium acetate 2.0 g / L, diammonium citrate 2.0 g / L, dipotassium hydrogen phosphate 2.6 g / L, manganese sulfate monohydrate 0.25 g / L, magnesium sulfate heptahydrate 0.5 g / L, and Tween-80 1 mL / L, pH 6.2–6.4.

[0038] MRS solid medium: yeast extract 5.0 g / L, beef extract 10.0 g / L, peptone 10.0 g / L, glucose 20.0 g / L, anhydrous sodium acetate 2.0 g / L, diammonium citrate 2.0 g / L, dipotassium hydrogen phosphate 2.6 g / L, manganese sulfate monohydrate 0.25 g / L, magnesium sulfate heptahydrate 0.5 g / L, Tween-80 1 mL / L, and agar 20.0 g / L, pH 6.2–6.4.

[0039] Fermentation substrate containing ginsenoside Rb1 (g / L): ginsenoside Rb1 1 g / L, peptone 10 g / L, yeast extract 5 g / L, beef extract 10 g / L, glucose 10 g / L, anhydrous sodium acetate 2 g / L, diamine hydrogen citrate 2 g / L, K2HPO4·3H2O 2.6 g / L, MgSO4·7H2O 0.5 g / L, MnSO4·7H2O 0.25 g / L, Tween-80 1 g / L, cysteine ​​1 g / L.

[0040] Fermentation substrate containing cassiaside (g / L): cassiaside 0.5 g / L, peptone 10 g / L, yeast extract 5 g / L, beef extract 10 g / L, glucose 10 g / L, anhydrous sodium acetate 2 g / L, diamine hydrogen citrate 2 g / L, K2HPO4·3H2O 2.6 g / L, MgSO4·7H2O 0.5 g / L, MnSO4·7H2O 0.25 g / L, Tween-80 1 g / L, cysteine ​​1 g / L.

[0041] The culture medium raw materials used in the examples were all purchased from Sinopharm Chemical Reagent Co., Ltd.

[0042] Example 1: Isolation, screening, identification and preservation of *Lactobacillus plantarum* CCFM1488 The specific steps are as follows: 1. Screening The samples were obtained from the feces of healthy infants. After pretreatment, the samples were stored in 20% glycerol at -80°C. After thawing, the samples were mixed and 0.5 mL of the sample was added to 4.5 mL of physiological saline. The samples were serially diluted with physiological saline. The appropriate serial dilutions were spread on MRS solid medium and incubated at 37°C for 48 h. Typical colonies of *Lactobacillus plantarum* were picked and streaked onto MRS solid medium for purification. Single colonies were picked and transferred to MRS liquid medium for enrichment. The samples were preserved in 30% glycerol to obtain the strain, which was named CCFM1488. The typical colonies of *Lactobacillus plantarum* are round, white, and smooth.

[0043] 2. Identification The genome of strain CCFM1488 was extracted, and the 16S rDNA of strain CCFM1488 was amplified and sequenced (performed by Biotechnology Co., Ltd., and the nucleotide sequence of the amplified 16S rDNA of CCFM1488 is shown in SEQ ID NO.1). The sequence was compared with the nucleic acid sequence in NCBI, and the results showed that the strain was Lactobacillus plantarum, and it was named Lactobacillus plantarum CCFM1488.

[0044] 3. Preservation of microbial strains Inoculate 5 mL of Lactobacillus plantarum CCFM1488 into 5 mL of MRS liquid medium and incubate at 37 °C for 24 h. Take 1 mL of bacterial culture into a sterile centrifuge tube, centrifuge at 8000 r / min for 3 min, discard the upper culture medium, and resuspend the bacterial sludge in 30% glycerol solution and store at -80 °C.

[0045] 4. Preservation of bacterial strains It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66135, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0046] Example 2: Transformation of ginsenoside Rb1 by Lactobacillus plantarum CCFM1488 The specific steps are as follows: (1) The plant lactobacillus CCFM1488 in Example 1 was streaked on MRS solid medium and the plate was incubated upside down at 37°C for 48h; a single colony was picked and inoculated into 5mL of MRS liquid medium and incubated at 37°C for 48h to prepare seed liquid.

[0047] (2) Preparation of fermentation substrate containing ginsenoside Rb1: The following ingredients were added: ginsenoside Rb1 (5 mg / mL), peptone (10 g / L), yeast extract (5 g / L), beef extract (10 g / L), glucose (20 g / L), anhydrous sodium acetate (2 g / L), diamine hydrogen citrate (2 g / L), K₂HPO₄·3H₂O (2.6 g / L), MgSO₄·7H₂O (0.58 g / L), MnSO₄·7H₂O (0.25 g / L), Tween-80 (1 g / L), distilled water, and cysteine ​​hydrochloride (0.5 g / L).

[0048] In a fermentation substrate containing ginseng extract, 5% (v / v) of the seed liquid of *Lactobacillus plantarum* CCFM1488 was added, and fermentation was carried out at a constant temperature of 37°C for 48 hours to prepare the fermentation product of fermented ginsenoside Rb1.

[0049] (3) Detect the content of ginsenoside Rb1 in the fermentation broth. The reference solution and the test solution were prepared according to the above method for processing ginsenoside fermentation broth, and the determination was performed under chromatographic conditions. The HPLC chromatogram is shown below. Figure 1 As shown.

[0050] The results showed that after fermentation, the content of ginsenoside Rb1 in the fermentation medium decreased significantly, and a chromatographic peak of ginsenoside Rd appeared. The standard curve confirmed that the content of ginsenoside Rd increased by 5 mg / L after fermentation. Therefore, the *Lactobacillus plantarum* CCFM1488 of this invention can effectively convert ginsenoside Rb1 into ginsenoside Rd.

[0051] Example 3: Transformation of cassiaside by Lactobacillus plantarum CCFM1488 The specific steps are as follows: (1) The plant lactobacillus CCFM1488 in Example 1 was streaked on MRS solid medium and the plate was incubated upside down at 37°C for 48h; a single colony was picked and inoculated into 5mL of MRS liquid medium and incubated at 37°C for 48h to prepare seed liquid.

[0052] (2) Preparation of fermentation substrate containing cassiaside: Cassia glycoside 1 mg / mL, peptone 10 g / L, yeast extract 5 g / L, beef extract 10 g / L, glucose 20 g / L, anhydrous sodium acetate 2 g / L, diamine hydrogen citrate 2 g / L, K₂HPO₄·3H₂O 2.6 g / L, MgSO₄·7H₂O 0.58 g / L, MnSO₄·7H₂O 0.25 g / L, Tween-80 1 g / L, distilled water, cysteine ​​hydrochloride 0.5 g / L.

[0053] Add 5% (v / v) of the bacterial culture of *Lactobacillus plantarum* CCFM1488 to the fermentation substrate containing cassiaside, and ferment at a constant temperature of 37°C for 48 hours.

[0054] (3) After fermentation, centrifuge at 8000 r / min for 15 min, take the fermentation supernatant to obtain the fermentation product of fermented cassia glycosides, and store it at 4℃.

[0055] (4) Prepare the reference solution and the sample test solution according to the above method for processing cassiaside fermentation broth, and determine the chromatogram according to the chromatographic conditions. Figure 2 As shown.

[0056] The results showed that the content of cassiaside was significantly reduced after fermentation with *Lactobacillus plantarum* CCFM1488, and a significant chromatographic peak was found in the elution time of cassiaside. After fermentation of cassiaside with *Lactobacillus plantarum* CCFM1488 of the present invention, the content of cassiaside was significantly increased. The standard curve showed that the amount of cassiaside generated was 2.87 mg / L.

[0057] Example 4: Verification of the lipid-lowering and fatty liver-relieving effects of Lactobacillus plantarum CCFM1488 and its prepared fermented cassiaside. 1. Preparation of cassia glycosides by fermentation with CCFM1488 (1) Plant Lactobacillus plantarum CCFM1488 was activated according to the method in Example 1 to prepare seed liquid.

[0058] (2) In a fermentation substrate containing cassiaside (concentration of 500 mg / L), 5% (v / v) of seed liquid was added and fermented at a constant temperature of 37°C for 48 hours.

[0059] (3) After fermentation, the fermentation liquid is subjected to heat treatment at 65°C for 30 minutes to completely inactivate the bacteria.

[0060] (4) After freezing the inactivated fermentation broth at -80℃, freeze-dry it to obtain CCFM1488 fermented cassia glycoside freeze-dried powder, which is stored at 4℃ for later use.

[0061] 2. Animal Experiment Design Fifty-six healthy 6-week-old SPF-grade male C57BL / 6J mice were purchased from Vital River Pharmaceuticals (Shanghai) Co., Ltd., and randomly divided into 7 groups of 8 mice each: ① Control group: fed low-fat diet and administered physiological saline by gavage.

[0062] ② Model group: fed with high-fat diet and administered physiological saline by gavage.

[0063] ③ Positive control group: fed a high-fat diet and administered atorvastatin (5.2 mg / kg body weight) by gavage.

[0064] ④ Cassia glycoside group: fed with high-fat diet and administered cassia glycoside (2.25 mg / kg body weight) by gavage.

[0065] ⑤CCFM1488 group: fed a high-fat diet and administered CCFM1488 live bacteria (1×10⁻⁶) via gavage. 9 CFU / each).

[0066] ⑥ Fermented cassia glycoside group: fed with high-fat diet, and administered CCFM1488 fermented cassia glycoside by gavage (calculated as cassia glycoside before fermentation, the dose was 2.25 mg / kg body weight).

[0067] ⑦ Synbiotic preparation group: fed with high-fat feed, and gavaged with CCFM1488 live bacteria + cassia seed glycoside (live bacteria 1×10^9 CFU / animal, cassia seed glycoside dosage 2.25 mg / kg body weight).

[0068] The experiment lasted 12 weeks: one week of acclimatization feeding, eight weeks of high-fat feeding modeling, followed by a four-week intervention. All mice had free access to food and water during this period.

[0069] 3. Indicator Testing After the intervention, blood and tissue samples were collected from mice. Intact liver tissue was sampled and washed with phosphate-buffered saline (PBS), blotted dry on filter paper, and wet weight was measured. The liver index was then calculated.

[0070] The collected whole blood samples were centrifuged (3000×g, 10min) after standing for 30min to obtain serum. TC, TG, LDL-C, HDL-C, ALT, and AST were detected using a fully automated biochemical analyzer.

[0071] After fixation, dehydration, embedding, sectioning, H&E staining and Oil Red O staining, the fatty degeneration of liver tissue was observed.

[0072] 4. Results (1) Effects on blood lipid levels The results are as follows Figure 3The results show that the combination of CCFM1488 and cassia seed glycosides has a clear lipid-lowering effect, and CCFM1488 can enhance the efficacy of cassia seed glycosides. Compared with the model group, the synbiotic preparation of CCFM1488 and cassia seed glycosides and fermented cassia seed glycosides significantly reduced serum TC, TG, and LDL-C levels (P<0.05). The synbiotic preparation reduced these levels by 20.4%, 23.9%, and 25.9%, respectively, while fermented cassia seed glycosides reduced them by 17.2%, 16.2%, and 20.1%, respectively. These effects were superior to those of using the same dose of cassia seed glycosides alone (which reduced these levels by 9.4%, 10.7%, and 10.3%, respectively), indicating that the cassia seed glycosides generated after in vitro directed conversion have a better lipid-lowering effect than cassia seed glycosides. Furthermore, CCFM1488 in the synbiotic preparation can colonize the intestine, thereby enhancing the directed conversion of cassia seed glycosides in the intestine and thus improving the efficacy of cassia seed glycosides.

[0073] (2) Effects on hepatic lipid accumulation Figure 4 The results showed that, compared with the model group, the combined preparation of CCFM1488 and cassia seed glycoside and fermented cassia seed glycoside significantly reduced liver weight (P<0.01) and serum ALT and AST levels (P<0.05). The combined preparation reduced these levels by 28.3%, 28.4%, and 29.9%, respectively, while fermented cassia seed glycoside reduced them by 24.4%, 28.6%, and 26.5%, respectively. Moreover, the effects were better than those of the group using the same dose of cassia seed glycoside alone (which reduced these levels by 14.6%, 16.1%, and 17.4%, respectively).

[0074] Histopathological results ( Figure 5 This provides direct evidence for the above results. The model group showed numerous fat vacuoles and red lipid droplets in the hepatocytes, while the synbiotic preparation of CCFM1488 and cassia seed glycoside, and the fermented cassia seed glycoside group showed significantly improved hepatocyte morphology and a significant reduction in lipid droplets. This demonstrates that the combination of CCFM1488 and cassia seed glycoside has a significant "relieving fatty liver" effect. Furthermore, CCFM1488 enhances the efficacy of cassia seed glycoside through in vitro fermentation or in vivo colonization to improve intestinal transformation capacity.

[0075] Comparative Example 1 The specific implementation method is the same as in Examples 2 and 3, except that *Lactobacillus plantarum* CCFM1488 is replaced with *Lactobacillus plantarum* CCFM1274 (accession number GDMCC No. 62798, described in the article "Enhancing the anti-fatigue effect of ginsenoside extract by..."). Bifidobacterium animalis subsp. lactisCCFM1274 fermentation through biotransformation of Rb1, Rb2, and Rc to Rd. (DOI: https: / / doi.org / 10.1016 / j.fbio.2024.105251) This study investigated whether strains capable of transforming ginsenosides also possess the ability to transform cassiasides.

[0076] from Figure 6 It can be seen that the content of ginsenoside Rd in the fermentation broth of *Lactobacillus plantarum* CCFM1274 increased significantly after fermentation, proving that *Lactobacillus plantarum* CCFM1274 has the ability to convert and generate ginsenoside Rd. Figure 7 The results showed that *Lactobacillus plantarum* CCFM1274 did not exhibit significant chromatographic peaks at 32 minutes after fermentation, indicating that *Lactobacillus plantarum* CCFM1274 lacked the ability to convert cassiaside into cassia aglycone. This demonstrates that the ability to simultaneously convert ginsenoside Rb1 into ginsenoside Rd and cassiaside into cassia aglycone is not universal, and that the strain's glucosidase activity is specific.

[0077] Comparative Example 2 The specific implementation method is the same as in Example 2, except that *Lactobacillus plantarum* CCFM1488 is replaced with *Lactobacillus plantarum* CCFM1274 (accession number GDMCC No. 62798, described in the patent application text with application (patent) number CNCN117562258A); *Lactobacillus plantarum* CCFM1366 (accession number GDMCC No. 63998, described in the patent application text with publication number CN118374396A); *Pediococcus lactis* ( Pediococcus acidilactici CCFM1364 (accession number GDMCC No: 63996, described in the patent text with publication number CN118360201A); Bifidobacterium breve ( Bifidobacterium breve FHNX48M6, Bifidobacterium animalis ( Bifidobacterium animalis subsp.lactis SCYA1M1, Bifidobacterium longum ( Bifidobacterium longum subsp.longum FGSYC28M5, Lactobacillus gasseri ( Lactobacillus gasseri FGSZY12L1, Mucosal Lactobacillus ( Limosilactobacillus mucosae Fynlj51M3 was obtained from the Food Microbiology Culture Collection Center of Jiangnan University. The study investigated changes in cassiasides in each component before and after fermentation, and whether cassiaside aglycones were generated.

[0078] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A strain of *Lactobacillus plantarum* ( Lactiplantibacillus plantarum CCFM1488, characterized in that, It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66135.

2. A microbial preparation, characterized in that, Contains the *Lactobacillus plantarum* CCFM1488 as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The concentration of *Lactobacillus plantarum* CCFM1488 in the microbial preparation is at least 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

4. A synthetic preparation, characterized in that, It contains live Lactobacillus plantarum CCFM1488 as described in claim 1, ginseng extract, and / or cassiaside.

5. An epigenetic agent, characterized in that, The post-biotic is the fermentation broth of *Lactobacillus plantarum* CCFM1488 as described in claim 1 or the microbial preparation as described in claim 2 or 3, after fermentation in a fermentation system containing ginsenosides and / or cassia seed glycosides, followed by heat treatment and high-pressure homogenization.

6. The use of the *Lactobacillus plantarum* CCFM1488 of claim 1 or the microbial preparation of claim 2 or 3 in the preparation of products for converting ginsenosides and / or cassiasides.

7. A method for converting ginsenosides and / or cassiasides, characterized in that, The substrate containing ginseng extract or cassia glycoside is fermented using the *Lactobacillus plantarum* CCFM1488 as described in claim 1 or the microbial preparation as described in claim 2 or 3.

8. The method according to claim 7, characterized in that, The fermentation is carried out at 30-40℃ for 48-72 hours; the concentrations of ginsenoside Rb1 and cassiaside are 1-5 mg / mL and 0.5-1 mg / mL, respectively.

9. The use of *Lactobacillus plantarum* CCFM1488 as described in claim 1, the microbial preparation as described in claim 2 or 3, the synbiotic preparation as described in claim 4, or the metabiotic as described in claim 5 in the preparation of a medicament for lowering blood lipids and / or alleviating non-alcoholic fatty liver disease.

10. The application according to claim 9, characterized in that, The application includes at least one of the following functions: (a) Reduce serum total cholesterol (TC) and / or triglyceride (TG) levels; (b) Reduce liver weight and liver index; (c) Improves fat accumulation in liver tissue; (d) Improve liver damage.

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