Plant lactobacillus ccfm1486 transformed with cassia glycoside for enhancing lipid-lowering and weight loss and relieving liver lipid accumulation and application thereof

Abstract

The application discloses a plant lactobacillus CCFM1486 transformed with siania glycoside, which can enhance lipid-lowering, weight loss and relieve liver lipid accumulation, and application thereof. The strain is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No: 66133. The core of the application is that the strain exhibits a significant synergistic effect when combined with siania glycoside: not only can it efficiently generate siania glycoside, but also can significantly inhibit weight gain caused by high-fat diet, reduce serum total cholesterol and triglyceride levels, and target to reduce liver lipid deposition and improve liver tissue fatty degeneration, and the effect is better than that of using siania glycoside alone. The live bacteria, fermented siania glycoside and combined preparation prepared based on the strain can be used for developing innovative products for preventing and / or assisting in treating obesity, hyperlipidemia and non-alcoholic fatty liver.

Description

Technical Field

[0001] This invention relates to a strain of *Lactobacillus plantarum* CCFM1486 that transforms cassiaside to enhance its lipid-lowering, weight-loss, and liver lipid accumulation effects, and its applications, belonging to the field of microbial technology and prevention and treatment of metabolic diseases. Background Technology

[0002] Obesity and its complications, such as hyperlipidemia and non-alcoholic fatty liver disease (NAFLD), have become a major global public health challenge. NAFLD is essentially the excessive deposition of lipids in the liver (hepatic steatosis), and is closely related to obesity and lipid metabolism disorders. It can further develop into hepatitis and liver fibrosis, seriously endangering health. Currently, lifestyle interventions are the basic approach, but long-term adherence is poor; while drug treatments often involve side effects or rebound effects after discontinuation. Therefore, developing safe, effective, and naturally derived intervention strategies has an urgent clinical need and broad market prospects.

[0003] Cassia seed, a traditional food and medicine ingredient, has had its lipid-lowering activity widely confirmed. Cassia glycoside is considered one of its main characteristic active ingredients. However, as a glycoside compound, cassia glycoside has low bioavailability in its original form. It needs to be metabolized and hydrolyzed by intestinal flora to remove the glycosyl group and be converted into cassia aglycone before it can be efficiently absorbed and exert its core physiological function of regulating lipid metabolism. This conversion process has become a key bottleneck restricting the efficacy of cassia seed. Due to the significant differences in individual intestinal flora composition, there is considerable uncertainty regarding the metabolic capacity of cassia glycoside and the final health benefits obtained in different populations.

[0004] Utilizing probiotic fermentation to enhance the bioactivity and efficacy of traditional Chinese medicine components is an effective strategy. Currently, studies have reported some microorganisms capable of converting glycosides, or some probiotics with functions in regulating lipid metabolism or weight management. However, existing technologies have significant limitations: the glucosidases contained in microorganisms have strong site specificity, meaning that other microorganisms capable of deglycosides may not be able to deglycosides from cassiaside; secondly, most studies either focus solely on conversion efficiency or only evaluate common lipid-lowering or weight-loss functions. There is a particular lack of specialized strains capable of specifically converting cassiasides and, on this basis, synergistically enhancing their comprehensive advantages in the three interrelated key dimensions of "lowering blood lipids," "controlling weight gain," and "reducing hepatic lipid deposition."

[0005] Lactobacillus plantarum is a recognized safe edible probiotic, but so far no specific Lactobacillus plantarum has been reported to achieve multiple enhanced interventions for obesity, hyperlipidemia and hepatic steatosis through synergistic effects with cassiaside.

[0006] Therefore, there is an urgent need in this field for a specific strain capable of efficiently converting cassiaside and significantly enhancing its efficacy in lowering lipids, reducing weight, and alleviating hepatic lipid accumulation. Such a strain would be of great significance for developing next-generation functional foods, probiotics, or drugs targeting obesity and related metabolic syndromes. Summary of the Invention

[0007] This invention aims to overcome the shortcomings of existing technologies in addressing the interrelated metabolic problems of obesity, hyperlipidemia, and liver lipid accumulation, which involve single intervention methods and insufficient synergistic effects. It provides a strain of *Lactobacillus plantarum* capable of converting cassiaside and synergistically enhancing its efficacy in lowering lipids, reducing weight, and alleviating liver lipid accumulation. Lactiplantibacillus plantarum CCFM1486. ​​Based on the unique functions of this strain, this invention further provides a complete solution from strain to product.

[0008] The first technical solution provided by this invention is a strain of *Lactobacillus plantarum* (… Lactiplantibacillus plantarum The strain CCFM1486, which can convert cassiaside into cassia aglycone and is used for synergistic intervention in obesity, hyperlipidemia and liver lipid accumulation, was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66133.

[0009] The second technical solution provided by the present invention is a microbial preparation containing *Lactobacillus plantarum* CCFM1486 as described in the first technical solution.

[0010] In some embodiments, the concentration of *Lactobacillus plantarum* CCFM1486 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* CCFM1486 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 the present invention is a synbiotic preparation containing live Lactobacillus plantarum CCFM1486 and cassiaside as described in the first technical solution.

[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* CCFM1486 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.

[0014] In some embodiments, the inactivation conditions are heat treatment at 60–70°C for 25–35 minutes; and the high-pressure homogenization conditions are 300–1500 Bar.

[0015] The fifth technical solution provided by this invention is the application of *Lactobacillus plantarum* CCFM1486 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 or health product.

[0017] The sixth technical solution provided by the present invention is a method for converting cassiaside, which uses the *Lactobacillus plantarum* CCFM1486 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 concentration of cassiaside in the fermentation substrate containing cassiaside is 500-1000 mg / L; the inoculum amount is 2-8% (v / v); and the fermentation is carried out at a constant temperature of 30-40℃ for 36-72 hours.

[0019] The seventh technical solution provided by this invention is the application of *Lactobacillus plantarum* CCFM1486 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) Inhibit weight gain or promote weight loss under a high-fat diet; (b) Reduce serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels; (c) Reduce liver weight and lipid accumulation in the liver; (d) Improves fatty degeneration of liver tissue.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: ① Multiple Efficacy: The core advantage of this invention lies in its synergistic intervention capability. Animal experiments have confirmed that the combined use of CCFM1486 and cassia seed glycosides can not only significantly inhibit weight gain caused by a high-fat diet, but also effectively reduce key blood lipid indicators such as serum total cholesterol and triglycerides, and target the reduction of liver weight and lipid deposition within the liver. This simultaneous improvement effect on "general obesity," "dyslipidemia," and "liver lipid accumulation" demonstrates its unique advantages of multi-target and comprehensive regulation, which is superior to single-function intervention strategies.

[0022] ② Clear Mechanism: This invention not only confirms the apparent improvement in body weight, blood lipids, and liver lipids, but also reveals part of its mechanism of action at the molecular level. The formulation can effectively regulate the expression of key genes in liver lipid metabolism (such as FASN, ATGL, MGL, HSL), thereby exerting a comprehensive metabolic regulatory effect by inhibiting fat synthesis and promoting fat breakdown.

[0023] ③ Wide range of applications: The strains and preparations described are derived from recognized safe probiotics and traditional food-medicine homologous raw materials. Animal experiments have shown that they do not cause liver damage (ALT and AST levels are normal), demonstrating high biocompatibility and suitability for long-term use. Based on this strain, various forms of products can be developed, including probiotics, fermented cassia glycosides, and synbiotic preparations, applicable to multiple fields such as functional foods, health products, and pharmaceuticals. The production process is mature and easily scaled up.

[0024] Products with enhanced lipid-lowering effects can be produced using Lactobacillus plantarum CCFM1486 and cassiaside. The production process uses cassiaside as a raw material to enhance its efficacy. This process has the advantages of being safe, efficient, low-cost, and having a mild reaction, making it suitable for large-scale industrial production.

[0025] Preservation of biological materials Lactobacillus plantarum ( Lactiplantibacillus plantarum CCFM1486, taxonomically named Lactiplantibacillus plantarum It was deposited on April 11, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO: 66133, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0026] Figure 1 High-performance liquid chromatography (HPLC) chromatograms of *Lactobacillus plantarum* CCFM1486 before and after fermentation.

[0027] Figure 2 The effects of CCFM1486 combined with cassiaside on obesity characteristics in mice; (A) pre-dissection body weight; (B) weight gain; (C) liver index; (D) epididymal fat index.

[0028] Figure 3 Effects of CCFM1486 combined with cassia seed glycosides on blood lipid levels in mice; serum (A) total cholesterol; (B) triglycerides; (C) low-density lipoprotein cholesterol; (D) high-density lipoprotein cholesterol; (E) low-density lipoprotein cholesterol / high-density lipoprotein cholesterol.

[0029] Figure 4 Effects of CCFM1486 combined with cassiaside on liver injury and liver pathological morphology in mice; serum (A) aspartate aminotransferase; (B) alanine aminotransferase; (C) liver H&E section; (D) liver oil red O stained section; scale bar: 100 μm.

[0030] Figure 5 The effects of CCFM1486 combined with cassiaside on the expression of lipid metabolism genes in mouse liver; relative expression levels of liver (A) fatty acid synthase; (B) triglyceride lipase; (C) monoacylglycerol lipase; (D) hormone-sensitive lipase.

[0031] Figure 6 This is a chromatogram of the fermentation and transformation of ginsenoside Rb1 by strains that can transform ginsenosides (with glucosidase activity) in other studies.

[0032] Figure 7 This is a high-performance liquid chromatogram of cassia glycosides fermented by the comparative strain.

[0033] "*" 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

[0034] 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.

[0035] 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.

[0036] (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.

[0037] (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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] The culture medium raw materials used in the examples were all purchased from Sinopharm Chemical Reagent Co., Ltd.

[0043] Example 1: Isolation, screening, identification and preservation of *Lactobacillus plantarum* CCFM1486 The specific steps are as follows: 1. Screening The samples were obtained from the feces of healthy infants in Changsha, Hunan Province. 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 and then spread on modified MRS solid medium. The samples were incubated at 37°C for 48 h. Typical colonies of *Lactobacillus plantarum* were picked and streaked onto modified MRS solid medium for purification. Single colonies were transferred to modified MRS liquid medium for enrichment and preserved in 30% glycerol. The resulting strain was named CCFM1486. ​​The typical colonies of *Lactobacillus plantarum* were white, raised, with smooth edges, short and thick cells, and were rod-shaped.

[0044] 2. Identification The genome of strain CCFM1486 was extracted, and the 16S rDNA of strain CCFM1486 was amplified and sequenced (performed by Biotechnology Co., Ltd., and the nucleotide sequence of the amplified 16S rDNA of CCFM1486 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 CCFM1486.

[0045] 3. Preservation of microbial strains Inoculate 5 mL of modified MRS liquid medium with Lactobacillus plantarum CCFM1486 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.

[0046] 4. Preservation of bacterial strains Lactobacillus plantarum CCFM1486 was deposited on April 11, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO: 66133, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0047] Example 2: Transformation of cassiaside by Lactobacillus plantarum CCFM1486 The specific steps are as follows: (1) The plant lactobacillus CCFM1486 in Example 1 was streaked on a modified MRS solid medium and the plate was incubated upside down at 37°C for 48 h; a single colony was picked and inoculated into 5 mL of modified MRS liquid medium and incubated at 37°C for 48 h to prepare seed culture.

[0048] (2) The activated bacterial solution was inoculated into the fermentation substrate containing cassia glycoside (500 mg / L) at an inoculation rate of 5% (v / v) and fermented at a constant temperature of 37°C for 48 hours.

[0049] (3) Sample preparation: The fermentation broth was centrifuged at 8000 r / min for 15 min, and the supernatant was taken and filtered through a 0.22 μm filter membrane for HPLC analysis. The fermentation substrate without bacterial inoculation was used as a blank control.

[0050] The results are as follows Figure 1 As shown, compared with the blank control, after 48 hours of fermentation with CCFM1486, the peak area of ​​cassiaside in the fermentation broth was significantly reduced. Simultaneously, a new and significant peak appeared at the elution position of cassia aglycone. Based on standard comparison and external standard method using peak area, the content of cassia aglycone in the fermentation broth reached 2.47 mg / L.

[0051] Example 3: Effects of synbiotic preparations made from *Lactobacillus plantarum* CCFM1486 and fermented cassiaside on mice on a high-fat diet 1. 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.

[0052] ② Model group: fed with high-fat diet and administered physiological saline by gavage.

[0053] ③ Positive control group: fed a high-fat diet and administered atorvastatin (5.2 mg / kg body weight) by gavage.

[0054] ④ Cassia glycoside group: fed with high-fat diet and administered cassia glycoside (2.25 mg / kg body weight) by gavage.

[0055] ⑤CCFM1486 group: fed with high-fat diet and administered CCFM1486 live bacteria (1×10^9 CFU / animal) by gavage.

[0056] ⑥ Synbiotic preparation group: fed with high-fat feed, and administered CCFM1486 live bacteria (1×10^9 CFU / animal) + cassia seed glycoside (2.25 mg / kg body weight) by gavage.

[0057] ⑦ Fermented cassia glycoside group: fed with high-fat diet, and administered CCFM1486 fermented cassia glycoside by gavage (calculated as cassia glycoside before fermentation, the dose was 2.25 mg / kg body weight).

[0058] 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.

[0059] 2. Indicator Testing During the intervention period, the mice's mental state was observed daily, and their weight was accurately weighed and recorded weekly.

[0060] After the intervention, blood and tissue samples were collected from mice. Intact liver tissue and epididymal adipose tissue were sampled and washed with phosphate-buffered saline (PBS). After being blotted dry on filter paper, wet weight was measured and organ index was calculated.

[0061] 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.

[0062] After fixation, dehydration, embedding, sectioning, H&E staining and Oil Red O staining, the fatty degeneration of liver tissue was observed.

[0063] The mRNA expression levels of FASN, ATGL, MGL, and HSL genes in liver tissue were detected using real-time quantitative PCR (qPCR), with GAPDH as an internal control.

[0064] 3. Results Changes in obesity characteristics in mice after intervention, such as Figure 2 As shown; the changes in blood lipid levels in mice are as follows Figure 3 As shown; liver H&E sections and Oil Red O stained sections are as follows. Figure 4 As shown; the expression results of liver lipid metabolism-related genes are as follows. Figure 5 As shown.

[0065] (1) Effects on obesity phenotype and weight like Figure 2As shown, compared with the control group, the final body weight, weight gain, and epididymal fat mass of the model group were all significantly increased (p<0.001), indicating that the obesity model was successfully established. Compared with the model group, the fermented cassia seed glycoside group showed an inhibition of body weight gain of approximately 31.6% and a reduction in epididymal fat index of approximately 23.9% (p<0.01); the combined preparation group showed an inhibition of body weight gain of approximately 24.6% and a reduction in epididymal fat index of approximately 20.2% (p<0.05); while the weight change was not significant when using cassia seed glycoside or CCFM1486 live bacteria alone (p>0.05). This indicates that the combined use of CCFM1486 and cassia seed glycoside has a significant synergistic effect on weight loss and inhibition of fat accumulation.

[0066] (2) Effects on serum lipid levels Changes in blood lipid levels in mice as follows Figure 3 As shown in the figure, serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) in the model group increased by 326%, 36%, and 44.2%, respectively, compared with the control group (p<0.001). In the synthetic preparation group, TC, TG, and LDL-C decreased by 21%, 23%, and 20%, respectively, compared with the model group (p<0.01). The fermented cassia seed glycoside group showed a better lipid-lowering effect, with TC, TG, and LDL-C decreasing by 18%, 29%, and 35%, respectively (p<0.001), and high-density lipoprotein cholesterol (HDL-C) levels increasing (p>0.05).

[0067] (3) Effects on liver lipid accumulation and damage Depend on Figure 4 It was found that the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the model group mice were significantly higher than those in the control group (p<0.001). Compared with the model group, the fermented cassia seed glycoside group showed a 40.8% decrease in ALT and a 35.7% decrease in AST (p<0.001); the combined preparation group showed a 39.5% decrease in ALT and a 38.8% decrease in AST (p<0.001); and the cassia seed glycoside group showed a 22.5% decrease in ALT and a 24.9% decrease in AST (p<0.05).

[0068] HE staining results showed that the liver cells of the control group mice had intact and regular structures, normal lobular structures, and clearly defined hepatic cords. Compared with the control group, the liver cells of the model group mice were swollen and enlarged, with scattered fat vacuoles of varying sizes distributed within the cells, disrupted lobular structures, and disordered hepatic cord arrangement. After drug intervention, the morphology of liver cells in mice in the atorvastatin group, cassia glycoside group, CCFM1486 group, synthetic preparation group, and fermented cassia glycoside group improved to varying degrees, and the number of fat vacuoles decreased to varying degrees, with the fermented cassia glycoside group showing the best effect. Oil Red O staining results also showed the same trend as H&E staining. Compared with the control group, the liver cells of the model group mice had diffusely distributed red lipid droplets, while the accumulation of lipid droplets in the liver cells of mice in the atorvastatin group, cassia glycoside group, CCFM1486 group, synthetic preparation group, and fermented cassia glycoside group showed varying degrees of reduction, with the fermented cassia glycoside group showing the best reduction. Liver section results showed that the proportion of fat vacuoles in hepatocytes was approximately 45-50% in the model group, while it decreased to approximately 10-15% in the fermented cassia seed glycoside group and approximately 15-20% in the combined preparation group. The oil red O staining showed a consistent trend of decreasing lipid droplet area. This indicates that CCFM1486 combined with cassia seed glycoside can effectively alleviate hepatic lipid accumulation and damage. Furthermore, CCFM1486 combined with cassia seed glycoside can effectively alleviate high-fat diet-induced hepatic lipid accumulation, with fermented cassia seed glycoside showing the best effect.

[0069] (4) Effects on the expression of hepatic lipid metabolism genes Depend on Figure 5 Compared with the model group, the fermented cassia glycoside group showed a 51% decrease in fatty acid synthase expression, a 106% increase in triglyceride lipase, a 109% increase in monoacylglycerol lipase, and a 107% increase in hormone-sensitive lipase. In contrast, the combined preparation group showed a 45% decrease in fatty acid synthase, a 100% increase in triglyceride lipase, a 124% increase in monoacylglycerol lipase, and an 82% increase in hormone-sensitive lipase. This indicates that the CCFM1486-cassia glycoside preparation exerts its lipid-lowering and hepatic steatosis-alleviating effects by bidirectionally regulating the expression of key genes in hepatic lipid metabolism, inhibiting fat synthesis, and promoting fat breakdown.

[0070] In summary, this embodiment fully demonstrates that *Lactobacillus plantarum* CCFM1486 can not only convert cassiaside, but also synergistically enhance its comprehensive effects in three aspects: "lipid reduction", "weight loss" and "relieving liver lipid accumulation". Moreover, the fermented cassiaside form obtained by fermenting cassiaside with *Lactobacillus plantarum* CCFM1486 has particularly outstanding effects.

[0071] 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* CCFM1486 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 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 cassia aglycones were formed.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] The method is the same as in Example 2, except that Bacillus plantarum CCFM1486 is replaced with Bacillus plantarum CCFM1274, and the fermentation substrate containing cassiaside (500 mg / L) is replaced with a fermentation substrate containing ginsenoside Rb1.

[0077] 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 glucosidase activity to convert ginsenoside Rb1, and can convert ginsenoside Rb1 to 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 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.

[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 CCFM1486, characterized in that, It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC NO: 66133.

2. A microbial preparation, characterized in that, Contains the *Lactobacillus plantarum* CCFM1486 as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The concentration of *Lactobacillus plantarum* CCFM1486 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* CCFM1486 as described in claim 1 and cassiaside.

5. An epigenetic agent, characterized in that, The post-biotic is the fermentation broth of *Lactobacillus plantarum* CCFM1486 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* CCFM1486 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* CCFM1486 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 concentration of cassiaside in the fermentation substrate containing cassiaside is 500-1000 mg / L; the inoculum amount is 2-8% (v / v); and the fermentation is carried out at a constant temperature of 30-40℃ for 36-72 hours.

9. The use of *Lactobacillus plantarum* CCFM1486 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) Inhibit weight gain or promote weight loss under a high-fat diet; (b) Reduce serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels; (c) Reduce liver weight and lipid accumulation in the liver; (d) Improves fatty degeneration of liver tissue.

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