Plant lactobacillus CCFM1487 and application of lipid-lowering and fatty liver-relieving metaplasma prepared by the plant lactobacillus CCFM1487 and transformed cassia glycoside

Abstract

This invention discloses a strain of *Lactobacillus plantarum* CCFM1487 and its prepared metagener for lowering lipids and alleviating fatty liver, as well as its application in converting cassiaside. It belongs to the fields of microbial technology and food and health product technology. This strain can efficiently convert cassiaside into cassiaside aglycone. The core of this invention lies in providing a metagener prepared from this strain through fermentation and inactivation treatment. Experiments in a high-fat diet mouse model have demonstrated that it significantly reduces serum total cholesterol and triglyceride levels, and can alleviate hepatic lipid deposition and improve hepatic steatosis. Based on this metagener, foods, medicines, or health products can be developed for the prevention and / or adjuvant treatment of metabolic disorders such as hyperlipidemia and non-alcoholic fatty liver disease, possessing significant application value.

Description

Technical Field

[0001] This invention relates to a strain of Lactobacillus plantarum CCFM1487 and its preparation of lipid-lowering and fatty liver-relieving metagenin and the application of transformed cassia glycosides, belonging to the fields of microbial technology, functional food and pharmaceutical technology. Background Technology

[0002] Nonalcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease worldwide. It is closely related to lipid metabolism disorders, and currently lacks safe and effective specific drugs. Lifestyle interventions are fundamental, but long-term adherence is poor; while lipid-lowering drugs such as statins can improve blood lipids, their direct effect on improving hepatic steatosis is limited, and long-term use may pose side effects. Therefore, developing safe and effective adjunctive intervention strategies derived from natural products has significant clinical and social implications.

[0003] Cassia seed, a traditional food and medicine ingredient, has attracted much attention for its lipid-lowering and liver-protecting activities. Cassia glycoside is one of its key active ingredients, but its bioavailability is low, requiring conversion into cassia aglycone in the intestines to fully exert its effects. Individual differences in gut microbiota lead to highly unstable conversion efficiency and ultimately unstable health benefits, limiting the application effects of cassia seed products.

[0004] Postbiotics, namely inactivated probiotic cells and their metabolites, have shown great potential in the field of microecological intervention in recent years due to their advantages such as high safety, good stability, and ease of storage and transportation. Preparing postbiotics rich in active aglycones (such as cassia seed) through in vitro fermentation of medicinal and edible raw materials (such as cassia seed) by specific probiotics is an ideal strategy to improve their efficacy stability and bioavailability.

[0005] Currently, although some studies have reported on microorganisms capable of converting glycosides, or some probiotics / postbiotics with general lipid-lowering functions, existing technologies have significant limitations: First, the glucosidase contained in microorganisms has strong site specificity, and other microorganisms capable of deglycosides may not be able to deglycosides from cassiaside; second, most studies only focus on simple conversion efficiency or general lipid-lowering functions, lacking in-depth research and product development on the targeted protective effects on the liver, a core organ of lipid metabolism. No specific *Lactobacillus plantarum* has been reported to be used to prepare dedicated postbiotic products that possess both highly efficient conversion of cassiasides and clear lipid-lowering and fatty liver-relieving effects.

[0006] Therefore, there is an urgent need in the field for a dedicated strain that can be used to prepare such functional metabiotics, as well as metabiotic products based on this strain, to meet the market's urgent demand for efficient, stable, and safe adjuvant intervention products for fatty liver. Summary of the Invention

[0007] This invention aims to overcome the deficiency in the existing technology of lacking a microecological preparation that can simultaneously and efficiently convert cassiaside and has a clear effect on lowering blood lipids and alleviating non-alcoholic fatty liver disease, and provides a strain of Lactobacillus plantarum CCFM1487, a functional metabiotic prepared from it, and its new applications in the field of metabolic health.

[0008] The first technical solution provided by this invention is a strain of *Lactobacillus plantarum* (… Lactiplantibacillus plantarum CCFM1487 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66134. The deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0009] In one embodiment of the present invention, after culturing *Lactobacillus plantarum* CCFM1487 on MRS medium for 48 hours, the colonies are round, convex, or lenticular, slightly white, opaque, and have a smooth to mucous-like soft surface. Fermentation of cassiaside using this bacterium can increase the content of cassiaside aglycone.

[0010] The second technical solution provided by the present invention is a microbial preparation containing the *Lactobacillus plantarum* CCFM1487 described in the first technical solution.

[0011] In some embodiments, the concentration of *Lactobacillus plantarum* CCFM1487 in the microbial preparation is at least 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0012] In some embodiments, the concentration of *Lactobacillus plantarum* CCFM1487 in the microbial preparation is at least 1 × 10⁻⁶. 9 CFU / mL or 1×10 9 CFU / g.

[0013] The third technical solution provided by the present invention is a synbiotic preparation containing live *Lactobacillus plantarum* CCFM1487 and cassiaside as described in the first technical solution.

[0014] 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* CCFM1487 described in the first technical solution or the microbial preparation described in the second technical solution after fermentation in a fermentation system containing cassiaside, followed by heat treatment and high-pressure homogenization.

[0015] The fifth technical solution provided by the present invention is the application of the *Lactobacillus plantarum* CCFM1487 described in the first technical solution or the microbial preparation described in the second technical solution in the preparation of products for converting cassiaside.

[0016] In some embodiments, the product is a pharmaceutical product.

[0017] The sixth technical solution provided by the present invention is a method for converting cassiaside, which uses the *Lactobacillus plantarum* CCFM1487 described in the first technical solution or the microbial preparation described in the second technical solution to ferment a substrate containing cassiaside.

[0018] In some embodiments, the fermentation is carried out at 30-40°C for 48-72 hours; the concentration of cassiaside is 0.5-1 mg / mL.

[0019] The seventh technical solution provided by this invention is the application of *Lactobacillus plantarum* CCFM1487 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.

[0020] In some implementations, the application includes at least one of the following functions: (a) Reduces individual serum total cholesterol and / or triglyceride levels; (b) Reduce individual liver weight and liver index; (c) Improves steatosis in individual liver tissue.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: ① Clear Efficacy: The core advantage of this invention lies in the fact that the metabiotic has a definite lipid-lowering and liver-protective effect. Animal experiments have confirmed that the metabiotic can significantly reduce serum total cholesterol and triglyceride levels in mice on a high-fat diet, and can effectively reduce liver weight, reduce lipid deposition in liver tissue, and improve hepatocyte steatosis. Biochemical indicators and histopathological studies have demonstrated its efficacy in alleviating non-alcoholic fatty liver disease.

[0022] ② Clear Efficacy Basis: This post-biotic is prepared by fermentation of cassiaside in a cassiaside culture medium using strain CCFM1487. This process efficiently converts cassiaside into cassia aglycone, which has higher bioavailability. Therefore, this post-biotic is a standardized functional product rich in cassia aglycone, ensuring the stability and reliability of product efficacy from the source and overcoming the problem of instability in efficacy caused by individual differences in gut microbiota.

[0023] ③ Postbiotics offer significant advantages: Compared to live bacteria preparations, this postbiotic has higher safety, better stability, and a longer shelf life, making it easier to apply and store in various products. Its effects do not depend on the colonization of the strain in the intestines, resulting in a more direct and rapid efficacy.

[0024] ④ Broad application prospects: Based on this functional metabolite, various product forms such as health foods, special dietary foods, medicines and even functional beverages can be developed to regulate lipid metabolism and help protect liver health, providing a new, safe and effective microecological intervention solution for people with metabolic syndrome.

[0025] Preservation of biological materials Lactobacillus plantarum ( Lactiplantibacillus plantarum CCFM1487, 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: 66134, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0026] Figure 1 The high-performance liquid chromatogram of cassiaside fermented by Lactobacillus plantarum CCFM1487.

[0027] Figure 2 Effects of post-biotics prepared from *Lactobacillus plantarum* CCFM1487 and its fermented cassiaside on serum lipid levels in mice on a high-fat diet; (A) serum triglyceride (TG) levels; (B) serum total cholesterol (TC) levels; (C) serum low-density lipoprotein cholesterol (LDL-C) levels; (D) serum high-density lipoprotein cholesterol (HDL-C) levels; (E) the ratio of LDL-C to HDL-C in serum.

[0028] Figure 3 Effects of post-biotics prepared from Lactobacillus plantarum CCFM1487 and its fermented cassiaside on liver injury indicators in mice on a high-fat diet; (A) liver weight; (B) serum alanine aminotransferase (ALT) level; (C) serum aspartate aminotransferase (AST) level.

[0029] Figure 4 Effects of post-biotics prepared from Lactobacillus plantarum CCFM1487 and its fermented cassiaside 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.

[0030] Figure 5 This is a chromatogram of the fermentation and transformation of ginsenoside Rb1 by strains that can transform ginsenosides (with glucosidase activity) in other studies.

[0031] Figure 6 This is a high-performance liquid chromatogram of cassia glycosides fermented by the comparative strain.

[0032] "*" indicates a significant difference from the Model group (p<0.05), "**" indicates p<0.01 from the Model group, and "***" indicates p<0.001 from the Model group. Detailed Implementation

[0033] 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.

[0034] Test method: HPLC analysis of the content of cassiaside and cassia aglycone in the fermentation system: (1) Sample pretreatment: 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.

[0035] (2) Preparation of standard products Accurately weigh 10.0 mg of each cassiaside standard and dissolve it in methanol, then 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.

[0036] (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 solution, and determine the content of cassiaside in the fermentation product under the above chromatographic conditions. Calculate the content using the external standard method.

[0037] 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.

[0038] 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.

[0039] 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.

[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* CCFM1487 The specific steps are as follows: 1. Screening The samples were obtained from the feces of healthy infants in Ya'an, Sichuan. 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 then 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 then transferred to MRS liquid medium for enrichment and preserved in 30% glycerol to obtain the strain, which was named CCFM1487. The typical colonies of *Lactobacillus plantarum* were round, white, and smooth.

[0043] 2. Identification The genome of strain CCFM1487 was extracted, and the 16S rDNA of strain CCFM1487 was amplified and sequenced (performed by Biotechnology Co., Ltd., and the nucleotide sequence of the amplified 16S rDNA of CCFM1487 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 CCFM1487.

[0044] 3. Preservation of microbial strains Inoculate 5 mL of Lactobacillus plantarum CCFM1487 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: 66134, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0046] Example 2: Transformation of cassiaside by Lactobacillus plantarum CCFM1487 The specific steps are as follows: (1) Activation of strain: The strain CCFM1487 in Example 1 was inoculated into MRS liquid medium and cultured at 37°C for 24 hours, and activated for 2 generations.

[0047] (2) Fermentation transformation: The activated bacterial culture was inoculated into the fermentation substrate containing cassiaside (500 mg / L) at an inoculation rate of 5% (v / v) and fermented at a constant temperature of 37℃ for 48 h. At the same time, an uninoculated fermentation substrate was set up as a blank control.

[0048] (3) Sample processing and detection: After fermentation, the fermentation broth was centrifuged at 8000 r / min for 15 min. The supernatant was filtered through a 0.22 μm filter membrane and analyzed by HPLC to quantitatively detect the remaining concentration of cassiaside and the concentration of cassia aglycone.

[0049] like Figure 1 The results showed that after *Lactobacillus plantarum* CCFM1487 converted cassiaside, its content decreased significantly, and a significant chromatographic peak was observed in the elution time of cassiaside. Therefore, it is evident that the fermentation of cassiaside using *Lactobacillus plantarum* CCFM1487 of this invention significantly increased the content of cassiaside. Based on standard comparison and external standard method using peak area, the content of cassiaside in the fermentation broth reached 2.89 mg / L.

[0050] Example 3: Preparation of *Lactobacillus plantarum* CCFM1487 and its postbiotics and verification of its lipid-lowering and fatty liver-relieving effects 1. Preparation of post-genetic agents (1) Plant Lactobacillus plantarum CCFM1487 was activated according to the method in Example 1 to prepare seed liquid.

[0051] (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.

[0052] (3) After fermentation, the fermentation broth is subjected to heat treatment at 65°C for 30 minutes to completely inactivate the bacteria, and then broken down by high-pressure homogenization at 1000 Bar for 5 cycles.

[0053] (4) After the inactivated and homogenized fermentation broth is frozen at -80℃, it is freeze-dried to obtain CCFM1487 post-biotic freeze-dried powder, which is stored at 4℃ for later use.

[0054] 2. Animal Experiment Design Fifty-six healthy male C57BL / 6J mice aged 6 weeks were randomly divided into 7 groups of 8 mice each: ① Control group: fed low-fat diet and administered physiological saline by gavage.

[0055] ② Model group: fed with high-fat diet and administered physiological saline by gavage.

[0056] ③ Positive control group: fed a high-fat diet and administered atorvastatin (5.2 mg / kg body weight) by gavage.

[0057] ④ Cassia glycoside group: fed with high-fat diet and administered cassia glycoside (2.25 mg / kg body weight) by gavage.

[0058] ⑤CCFM1487 group: fed a high-fat diet and administered CCFM1487 live bacteria (1×10⁻⁶) via gavage. 9 CFU / each).

[0059] ⑥ Synbiotic preparation group: fed with high-fat feed, and administered CCFM1487 live bacteria + cassia seed glycoside (2.25 mg / kg body weight) by gavage.

[0060] ⑦ Postbiotic group: fed with high-fat diet and administered CCFM1487 postbiotic by gavage (calculated as pre-fermented cassiaside, dose of 2.25 mg / kg body weight).

[0061] 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.

[0062] 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.

[0063] 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.

[0064] After fixation, dehydration, embedding, sectioning, H&E staining and Oil Red O staining, the fatty degeneration of liver tissue was observed.

[0065] 4. Results (1) Effects on blood lipid levels The results are as follows Figure 2 As shown in the figure, compared with the model group, the post-biotic group significantly reduced serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels (p<0.01), with TC, TG, and LDL-C decreasing by approximately 28%, 13%, and 26%, respectively, compared to the model group. The lipid-lowering effect of the combined preparation group (CCFM1487 live bacteria + cassia seed glycoside) was comparable, with TC, TG, and LDL-C decreasing by 24%, 22%, and 33%, respectively (p<0.01). Both were superior to the group using the same dose of cassia seed glycoside alone (TC, TG, and LDL-C decreased by approximately 15%, 10%, and 18%, respectively, p<0.05), indicating that CCFM1487 can synergistically enhance the lipid-lowering effect of cassia seed glycoside.

[0066] (2) Effects on hepatic lipid accumulation Figure 3 The results showed that, compared with the model group, both the post-suspension group and the combined-suspension preparation group significantly reduced liver weight and liver index (p<0.01), with liver weight decreasing by approximately 30% and 28%, respectively. Regarding liver function indicators, serum ALT and AST levels in the model group increased by 187% and 53% compared to the control group (p<0.001), while ALT and AST levels in the post-suspension group decreased by 26% and 30% respectively compared to the model group after intervention (p<0.001), and ALT and AST levels in the combined-suspension preparation group also decreased by 35% and 27% respectively (p<0.01), indicating that the intervention significantly reduced liver damage.

[0067] Histopathological results ( Figure 4This provides direct evidence for the above results. Numerous fat vacuoles were observed in the hepatocytes of the model group, and Oil Red O staining revealed dense red lipid droplets, with a significantly increased steatosis score. The hepatocyte morphology of the post-biotic group and the combined preparation group was significantly improved, with a significant reduction in the number and area of ​​lipid droplets, and a decrease in steatosis score of approximately 75% and 70%, respectively (p<0.001). This demonstrates that the post-biotic and combined preparations prepared from CCFM1487 and cassia seed glycosides have a significant synergistic effect in alleviating hepatic lipid deposition and improving the pathological manifestations of fatty liver.

[0068] Comparative Example 1: Effect of Microbial Strains on Cassia glycoside Conversion The specific implementation method is the same as in Example 2, except that *Lactobacillus plantarum* CCFM1487 is replaced with *Lactobacillus plantarum* CCFM1274 (accession number GDMCC No. 62798, described in the patent application text with application (patent) number CN202311184003.0); *Lactobacillus plantarum* CCFM1366 (accession number GDMCC No. 63998, described in the patent application text with publication number CN118374396A); *Pediococcus acidilactici* CCFM1364 (accession number GDMCC No: 63996, described in the patent text with publication number CN118360201A); *Bifidobacterium breve* FHNX48M6; and *Bifidobacterium animalis*. *Bifidobacterium longum* subsp. *lactis* SCYA1M1, *Bifidobacterium longum* subsp. *longum* FGSYC28M5, *Lactobacillus gasseri* FGSZY12L1, and *Lactobacillus mucosa* Fynlj51M3 were obtained from the Food Microbiology Culture Collection Center of Jiangnan University. Changes in cassiasides in each component before and after fermentation and the presence of cassia aglycone formation were detected.

[0069] The results are as follows Figure 6 As shown, the chromatograms of the comparative strains showed no significant chromatographic peaks at 32 minutes, indicating that these strains did not have the ability to convert cassiaside into cassia aglycone. This proves that the ability to convert cassiaside into cassia aglycone is strain-specific, and not all *Lactobacillus plantarum* or *Lactobacillus* have the ability to convert cassiaside into cassia aglycone.

[0070] Comparative Example 2: Lactobacillus plantarum CCFM1274 (accession number GDMCC No. 62798) was selected, as described in the article "Enhancing the anti-fatigue effect of ginsenoside extract by..." Bifidobacterium animalis subsp. lactis CCFM1274 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.

[0071] First, a fermentation substrate containing ginsenoside Rb1 was prepared, with the following specific components: 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, and cysteine ​​1 g / L.

[0072] Fermentation product of fermented ginsenoside Rb1 was prepared by adding 5% (v / v) of seed liquid of *Lactobacillus plantarum* CCFM1274 to a fermentation substrate containing ginseng extract and fermenting at 37°C for 48 hours.

[0073] The method is the same as in Example 2, except that Bacillus plantarum CCFM1487 is replaced with Bacillus plantarum CCFM1274, and the fermentation substrate containing cassiaside is replaced with the fermentation substrate containing ginsenoside Rb1.

[0074] from Figure 5 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 glucosidase activity to convert ginsenoside Rb1, and can convert ginsenoside Rb1 to ginsenoside Rd. Figure 6 The results showed that *Lactobacillus plantarum* CCFM1274 did not exhibit significant chromatographic peaks at 32 minutes after fermentation, indicating that *Lactobacillus plantarum* CCFM1274 lacks the ability to convert cassiaside into cassia aglycone. This demonstrates that although *Lactobacillus plantarum* CCFM1274 possesses glucosidase activity, the glucosidase contained in this strain cannot act on the glycosides of cassiaside, highlighting the specific specificity of the glucosidase for cassiaside conversion.

[0075] 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 CCFM1487, characterized in that, It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66134.

2. A microbial preparation, characterized in that, Contains the *Lactobacillus plantarum* CCFM1487 as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The concentration of *Lactobacillus plantarum* CCFM1487 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* CCFM1487 as described in claim 1 and cassiaside.

5. An epigenetic agent, characterized in that, The post-biotic is the fermentation broth of *Lactobacillus plantarum* CCFM1487 as described in claim 1 or the microbial preparation as described in any one of claims 2 to 3, after fermentation in a fermentation system containing cassiaside and subsequent heat treatment and high-pressure homogenization.

6. The use of the *Lactobacillus plantarum* CCFM1487 of claim 1 or the microbial preparation of any one of claims 2-3 in the preparation of products for converting cassiaside.

7. A method for converting cassiaside, characterized in that, The substrate containing cassiaside is fermented using the *Lactobacillus plantarum* CCFM1487 as described in claim 1 or the microbial preparation as described in any one of claims 2 to 3.

8. The method according to claim 7, characterized in that, The fermentation is carried out at 30-40°C for 48-72 hours; the concentration of cassiaside is 0.5-1 mg / mL.

9. The use of *Lactobacillus plantarum* CCFM1487 as described in claim 1, the microbial preparation as described in any one of claims 2-3, 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) Reduces individual serum total cholesterol and / or triglyceride levels; (b) Reduce individual liver weight and liver index; (c) Improves steatosis in individual liver tissue.

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