Ginseng fermentation products rich in various ginsenosides, their preparation methods and applications

Abstract

This patent application discloses a ginseng fermentation product rich in various ginsenosides, its preparation method, and its applications. The ginseng fermentation product of this invention is obtained by fermenting a ginseng fermentation substrate using microbial strains. Addressing the issue that existing ginseng fermentation products have certain effects on whitening, anti-inflammation, and enhancing the body's immunity, but none of these effects are particularly clear or prominent, the ginseng fermentation product described herein contains anti-inflammatory rare ginsenosides Rk1, Rg5, Rk3, and Rh4 at a content of over 50% by weight, and the relative content of ginsenosides Rk3 and Rh4, which are less likely to cause drowsiness, is over 30% by weight.

Description

Technical Field

[0001] This invention relates to the field of synthetic biology, and in particular to a ginseng fermentation product rich in anti-inflammatory rare ginsenosides, its bio-fermentation preparation method, and its application. Background Technology

[0002] Ginseng, a herbaceous plant belonging to the genus Panax in the family Araliaceae, is rich in various components, including ginsenosides, polysaccharides, volatile oils, proteins, organic acids, and trace elements. The properties and pharmacological activities of different ginseng components vary. Modern pharmacological research indicates that saponins (Rb1, Rb2, Rc, Rd, etc.) are the important material basis for the anti-fatigue, anti-tumor, antioxidant, immune-enhancing, and physiological function-regulating pharmacological activities of ginseng plants. However, the overall bioavailability of ginsenosides in the human body is poor, especially in the intestines where absorption is negligible. Nevertheless, studies have found that after oral administration, ginsenosides undergo a series of biotransformations and metabolisms involving gastric acid and intestinal flora, producing rarer saponins and their aglycones that are more easily absorbed by the human body and have better bioactivity, which has attracted widespread attention from researchers.

[0003] Rare ginsenosides are a special class of ginsenosides formed by the hydrolysis of the glycosyl groups or alterations in the side chains of the parent nucleus of protopanaxadiol and protopanatriol-type ginsenosides. They are present in extremely small amounts (only one in ten thousand) in ginseng plants and are difficult to obtain through traditional extraction and separation methods, hence the name "rare ginsenosides." Rare ginsenosides are secondary derivatives of ginsenosides, commonly including Rk3, Rh4, S-Rg3, R-Rg3, Rh2, Rh1, Rk1, Rg5, etc. Compared to the abundant naturally occurring ginsenosides, rare ginsenosides exhibit relatively stronger biological activities such as anti-tumor activity, inhibition of cancer cell metastasis, protection of the central nervous system, enhancement of immunity, improvement of memory, and anti-stress.

[0004] In the research of rare ginsenoside production technology, the biotransformation method of ginseng fermentation by microorganisms is considered one of the safest and most efficient in vitro conversion methods for ginsenosides. In fact, technicians have already used one or more microorganisms to ferment ginseng fermentation substrates, thereby producing a variety of rare ginsenosides.

[0005] However, a current technical problem with these existing technologies is that microbial fermentation produces a composition of various rare ginsenosides, which may contain up to dozens of different rare ginsenosides. These rare ginsenosides have different biological effects, and some even have antagonistic effects. Therefore, such ginsenoside products are more like a "cure-all," potentially offering some benefits such as skin whitening and boosting immunity, but without any particularly clear or prominent effect in any particular area.

[0006] Regarding the physiological effects of rare ginsenosides, four ginsenosides, Rk1, Rg5, Rk3, and Rh4, are considered to have anti-inflammatory effects. Ginsenosides Rk1 and Rg5 also have the function of inhibiting nerve excitation and helping sleep. Large intake may cause drowsiness in the ingested person. Summary of the Invention

[0007] In view of the above-mentioned technical problems existing in the prior art, the purpose of the present invention is to provide a ginseng fermentation product rich in rare ginsenosides with enhanced anti-inflammatory effects and less likely to cause drowsiness, its preparation method and application.

[0008] Through literature review, the inventors discovered that among rare ginsenosides, ginsenosides Rk1, Rg5, Rk3, and Rh4 possess anti-inflammatory properties. Based on this, the inventors believe that increasing the content of these ginsenosides in ginseng fermentation products would enable the preparation of ginseng fermentation products with enhanced anti-inflammatory effects.

[0009] Through literature review, the inventors discovered that among rare ginsenosides, ginsenosides Rk1, Rg5, Rk3, and Rh4 possess anti-inflammatory properties, while ginsenosides Rk1 and Rg5 can also inhibit nerve excitation and promote sleep. Therefore, it can be concluded that if the total content of ginsenosides Rk1, Rg5, Rk3, and Rh4 in ginseng fermentation products can be increased, while simultaneously increasing the relative content of ginsenosides Rk3 and Rh4 among these four ginsenosides, then a ginseng fermentation product with enhanced anti-inflammatory effects and less likely to induce drowsiness can be prepared.

[0010] However, the existing technical challenge is that the production of rare ginsenosides from the conversion of natural ginsenosides using microorganisms (probiotics) may be related to the conversion pathway. Among the four rare ginsenosides Rk1, Rg5, Rk3, and Rh4, ginsenoside Rg5 is more easily generated. Moreover, while a certain amount of ginsenosides Rk3 and Rh4 are generated, a larger amount of ginsenoside Rg5 is also generated. This results in either a low total content of the four ginsenosides Rk1, Rg5, Rk3, and Rh4 in existing ginseng fermentation products, or a low relative content of ginsenosides Rk3 and Rh4. Consequently, the ginseng fermentation products are unlikely to fully exert their anti-inflammatory effects while also being less likely to induce drowsiness.

[0011] Furthermore, through in-depth research, the inventors discovered that fermenting ginseng substrate with a specific combination of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, and Bifidobacterium animalis can produce ginseng fermentation products with high total content of four ginsenosides: Rk1, Rg5, Rk3, and Rh4, and also with high relative content of ginsenosides Rk3 and Rh4, thus completing this invention.

[0012] The technical solution of the present invention includes:

[0013] 1. A ginseng fermentation product, obtained by fermenting a ginseng fermentation substrate using a microbial strain, comprising ginsenosides Rk1, Rg5, Rk3, and Rh4, satisfying the following:

[0014] The total content of ginsenosides Rk1, Rg5, Rk3, and Rh4 is ≥50% by weight, with the total ginsenoside content in the ginseng fermentation product being 100% by weight; and

[0015] Of the ginsenosides Rk1, Rg5, Rk3 and Rh4, the total content of ginsenosides Rk3 and Rh4 is ≥30% by weight, preferably ≥35% by weight, with the total amount of ginsenosides Rk1, Rg5, Rk3 and Rh4 being 100% by weight.

[0016] The ginseng fermentation product can be liquid, solid, or a mixture of solid and liquid, such as fermented ginseng liquid or fermented ginseng powder.

[0017] 2. A microbial composition comprising *Lactobacillus plantarum*, *Lactobacillus casei*, *Lactobacillus paracasei*, and *Bifidobacterium animalis*; wherein,

[0018] The Lactobacillus plantarum mentioned is Lactobacillus plantarum GBFG 61, which is deposited at the China General Microbiological Culture Collection Center with accession number GGMCCNO: 31583 and deposit date of August 14, 2024.

[0019] The Lactobacillus paracasei mentioned is GBFG 34, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number GGMCC NO: 31581 and deposit date of August 14, 2024.

[0020] The Bifidobacterium animalis mentioned is Bifidobacterium animalis GBFG 52, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 31582 and deposit date of August 14, 2024.

[0021] 3. The microbial composition according to item 2 can be used to ferment ginseng fermentation substrate to produce rare ginsenosides.

[0022] Here, the ginseng fermentation substrate refers to a fermentation substrate containing ginseng fines. The ginseng fines are obtained by refining ginseng, preferably using physical methods such as extrusion, cutting, or grinding. The ginseng fines can be, for example, ginseng powder. The ginseng fermentation substrate may also contain other components typically found in fermentation substrates, such as nitrogen sources, carbon sources, inorganic salts, and pH adjusters.

[0023] 4. The microbial composition according to item 2, wherein the dry weight ratio of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei and Bifidobacterium animalis is 1:0.5-5:0.5-5:0.5-5.

[0024] 5. The bacterial composition according to item 2, wherein the Lactobacillus casei BL23 is present.

[0025] 6. A method for preparing a ginseng fermentation product, comprising inoculating a microbial composition into a ginseng fermentation substrate and fermenting at 30-35°C for 48-58 hours, preferably 50-55 hours, more preferably 52 hours, thereby obtaining a ginseng fermentation product containing rare ginsenosides;

[0026] The microbial composition is any one of items 2 to 5.

[0027] 7. The preparation method according to item 6 further includes sterilizing the ginseng fermentation product.

[0028] 8. The preparation method according to item 6, wherein the ginseng fermentation product is the ginseng fermentation product according to item 1.

[0029] 9. The preparation method according to item 6, wherein the ginseng fermentation product is fermented ginseng liquid or fermented ginseng powder.

[0030] Invention Effects

[0031] This invention combines *Lactobacillus plantarum* GBFG 61, *Lactobacillus paracasei* GBFG 34, and *Bifidobacterium animalis* GBFG 52 isolated by the inventors with *Lactobacillus casei* BL23 for fermentation in a ginseng fermentation substrate. This significantly increases the total content of rare ginsenosides Rk1, Rg5, Rk3, and Rh4 in the obtained ginseng fermentation product, as well as the relative content of ginsenosides Rk3 and Rh4. Consequently, the ginseng fermentation product exhibits a clear and prominent anti-inflammatory effect and is less likely to induce drowsiness. Attached Figure Description

[0032] Figure 1High-performance liquid chromatography (HPLC) chromatogram for determining the content of four rare ginsenosides in ginseng fermentation powder (Example 1)

[0033] Figure 2 High-performance liquid chromatography (HPLC) chromatograms of four rare ginsenosides in ginseng fermentation powder (Comparative Example 3)

[0034] Figure 3 Effect of ginseng fermentation powder on cell viability (image)

[0035] Figure 4 Effect of ginseng fermentation powder on NO secretion in cells (Figure)

[0036] Figure 5 Effect of ginseng fermentation powder on TNF-α content in cells (Figure)

[0037] Figure 6 Effect of ginseng fermentation powder on IL-6 levels in cells (Figure)

[0038] Figure 7 Effect of ginseng fermentation powder on IL-1β levels in cells (Figure) Detailed Implementation

[0039] The present invention will now be described in detail with reference to specific embodiments. Unless otherwise specified, the scientific terms used in this specification have the meanings commonly understood by those skilled in the art; in case of conflict, the definitions in this specification shall prevail.

[0040] Example

[0041] The *Lactobacillus plantarum* GBFG 61 strain was isolated and cultured from soil in a ginseng cultivation base in Fusong County, Baishan City, Jilin Province, and is a β-glucosidase-producing lactobacillus strain. This *Lactobacillus plantarum* strain was named GBFG 61 and is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 31583. The deposit date is August 14, 2024. The address of the depository is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences; Postcode: 100101.

[0042] The *Lactobacillus paracasei* GBFG 34 strain was isolated and cultured from the soil of a ginseng planting base in Ji'an City (county), Tonghua City, Jilin Province, and is a β-glucosidase-producing lactobacillus strain. This *Lactobacillus paracasei* strain was named GBFG 34 and is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 31581. The deposit date is August 14, 2024. The address of the depository is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences; Postcode: 100101.

[0043] The *Bifidobacterium animalis* GBFG 52 strain was isolated and cultured from the soil of a ginseng planting base in Changbai County, Changbai Korean Autonomous County, Baishan City, Jilin Province, and produced β-glucosidase. This *Bifidobacterium animalis* strain was named GBFG 52 and is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 31582. The deposit date is August 14, 2024. The address of the depository is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Postcode: 100101.

[0044] The Lactobacillus casei BL23 was purchased from Beijing Keruisibo Biotechnology Co., Ltd.

[0045] The Lactobacillus plantarum Lp90 and Lactobacillus paracasei LC37 were purchased from Microcare Probiotics (Suzhou) Co., Ltd.

[0046] The Lactobacillus casei PG-LCG139 was purchased from Shandong Pingao Pharmaceutical Co., Ltd.

[0047] The animal bifidobacterium PG-BAD260 was purchased from Shandong Pingao Pharmaceutical Co., Ltd.

[0048] The ginseng was purchased from a Chinese medicinal herb market, while other raw materials and reagents were obtained through commercial channels.

[0049] Example 1: Preparation of Fermented Ginseng Powder with Enhanced Anti-inflammatory Effects Rich in Rare Ginsenosides

[0050] Ginseng powder was mixed with 70% ethanol at a mass ratio of 1:20 to obtain ginseng aqueous extract, which was then sterilized at 121℃ for 15 min to obtain ginseng extract solution.

[0051] The Lactobacillus plantarum GBFG 61, Lactobacillus casei BL23, Lactobacillus paracasei GBFG 34, and Bifidobacterium animalis GBFG 52 were mixed in a dry weight ratio of 1:2:3:4 to prepare a bacterial culture composition, which was then activated.

[0052] The activated bacterial culture composition was inoculated into the ginseng extract at 1% by weight, and fermented at 30℃ to 35℃ for 52 hours to obtain fermented ginseng liquid.

[0053] The fermented ginseng liquid was filtered and centrifuged, and the resulting supernatant was freeze-dried under vacuum to prepare fermented ginseng powder 1.

[0054] Example 2

[0055] Fermentation was carried out as in Example 1 to prepare fermented ginseng powder 2, except that: Lactobacillus plantarum GBFG 61, Lactobacillus casei BL23, Lactobacillus paracasei GBFG 34, and Bifidobacterium animalis GBFG 52 were mixed in a dry weight ratio of 1:0.5:0.5:0.5 to prepare a bacterial culture composition.

[0056] Example 3

[0057] Fermentation was carried out as in Example 1 to prepare fermented ginseng powder 3, except that: Lactobacillus plantarum GBFG 61, Lactobacillus casei BL23, Lactobacillus paracasei GBFG 34, and Bifidobacterium animalis GBFG 52 were mixed in a dry weight ratio of 1:5:5:5 to prepare a bacterial culture composition.

[0058] Comparative Example 1

[0059] Fermentation was carried out as in Example 1 to prepare comparative fermented ginseng powder 1, except that Lactobacillus plantarum GBFG 61 was replaced with Lactobacillus plantarum Lp90.

[0060] Comparative Example 2

[0061] Fermentation was carried out as in Example 1 to prepare comparative fermented ginseng powder 2, the difference being that Lactobacillus casei BL23 was replaced with Lactobacillus casei PG-LCG139.

[0062] Comparative Example 3

[0063] Fermentation was carried out as in Example 1 to prepare comparative fermented ginseng powder 3, except that Lactobacillus paracasei GBFG 34 was replaced with Lactobacillus paracasei LC37.

[0064] Comparative Example 4

[0065] Fermentation was carried out as in Example 1 to prepare comparative fermented ginseng powder 4, the difference being that the animal bifidobacterium GBFG 52 was replaced with animal bifidobacterium PG-BAD260.

[0066] Example 4: Determination of the content of rare ginsenosides with anti-inflammatory effects

[0067] Fermented ginseng powders prepared in Examples 1-3 and Comparative Examples 1-4 were dissolved in methanol to prepare solutions of 80 mg / mL. 1 mL of each solution was filtered through a 0.45 μm microporous membrane to obtain a clear liquid of fermented ginseng powder. The total ginsenoside content and the content of various rare ginsenosides were then determined. The total content (wt%) of ginsenosides Rk1, Rg5, Rk3, and Rh4, with the total ginsenoside content as 100 wt%, and the relative content (wt%) of ginsenosides Rk3 and Rh4, with the total amount of ginsenosides Rk1, Rg5, Rk3, and Rh4 as 100 wt%, were calculated. The results are shown in Table 1.

[0068] The determination method for total ginsenosides was performed according to the method for content determination under the ginsenosides section of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China. The results are expressed as ginsenoside Re(C 48 H 82 O 18 )count.

[0069] The detection method for rare ginsenosides shall be in accordance with the method for determining the content of rare ginsenosides in T / CNHFA 001-2021.

[0070] As an example, the high-performance liquid chromatograms of the contents of ginsenosides Rk3, Rh4, Rk1, and Rg5 in the fermented ginseng powder prepared in Example 1 and Comparative Example 3 are shown below. Figure 1 and Figure 2 As shown.

[0071] Table 1. Content of rare ginsenosides with anti-inflammatory effects in fermented ginseng powder

[0072]

[0073] As shown in Table 1, when the strains of *Lactobacillus plantarum* GBFG 61, *Lactobacillus casei* BL23, *Lactobacillus paracasei* GBFG 34, and *Bifidobacterium animalis* GBFG52 of this invention were selected, the content of anti-inflammatory rare ginsenosides in the resulting fermented ginseng powder was significantly increased, and the relative contents of ginsenosides Rk3 and Rh4, which do not cause drowsiness, were also significantly increased. This suggests that the resulting fermented ginseng powder has enhanced anti-inflammatory effects and is less likely to cause drowsiness.

[0074] Example 5: Verification of the anti-inflammatory effects of various fermented ginseng powders

[0075] This experiment mainly compares the fermented ginseng powder produced in Example 1 (fermented ginseng powder 1) with the fermented ginseng powder produced in Comparative Example 3 (comparative fermented ginseng powder 3) to verify its anti-inflammatory efficacy.

[0076] 1. Cell Culture and Processing

[0077] The mouse mononuclear macrophage cell line RAW264.7 was obtained from Cybio (Shanghai) Biotechnology Co., Ltd. It was cultured in a humidified environment of 37℃, 95% air and 5% CO2, in DMEM complete medium supplemented with 10% FBS. When the cell density reached 90%, it was passaged at a volume ratio of 1:3, and cells in the logarithmic growth phase were used for subsequent experiments.

[0078] 2. Cytotoxicity assay

[0079] Cell viability was determined using the MTT assay. RAW264.7 B16 cells in logarithmic growth phase were used, and the cells were divided into two groups of 1 × 10⁶ cells per well. 4 Cells were seeded at a density in 96-well plates and incubated at 37°C with 5% CO2 for 12 h (6 replicates per group). The original culture medium was aspirated, and 100 μL of 200 μg / mL fermented ginseng powder sample solution was added to each well. The plates were then incubated for 24 h, followed by another 24 h at 37°C with 5% CO2. 10 μL of 5 mg / mL MTT was added to each well, and after 4 h, the supernatant was discarded. 150 μL of DMSO was added, and the OD value was measured at 490 nm to calculate the cell proliferation rate for each group.

[0080] 3. The impact of NO release

[0081] Logarithmic phase cells were seeded at a concentration of 1 × 10⁻⁶. 4The samples were densely inoculated into 96-well plates and incubated at 37°C with 5% CO2 for 24 h. The model and sample groups were treated with 150 μL of 1.0 μg / mL LPS, while the blank group was treated with complete culture medium. All samples were incubated for another 24 h. The sample groups were treated with 150 μL of 200 μg / mL fermented ginseng powder sample solution and incubated at 37°C with 5% CO2 for 24 h. The supernatant was then collected, and NO secretion was measured according to the NO kit instructions. The secretion levels of inflammatory factors TNF-α, IL-6, and IL-1β were detected using an ELISA kit according to the kit instructions.

[0082] 4. Results Analysis

[0083] 4.1 Effect on cell viability

[0084] RAW264.7 cells were cultured for 24 h at 400 μg / mL fermented ginseng powder. Cell viability was detected and calculated using the MTT assay. The results are as follows: Figure 3 As shown in the figure. Compared with the control group, the survival rate of RAW264.7 cells treated with 200 μg / mL fermented ginseng powder 1 and control fermented ginseng powder 3 remained basically unchanged, indicating that the two concentrations of fermented ginseng powder at this experimental concentration showed no toxicity to RAW264.7 cells and can be used for subsequent experimental studies.

[0085] 4.2 Effect of fermented ginseng powder on NO secretion in RAW264.7 cells

[0086] Nitric oxide (NO) is a signaling molecule that plays a crucial role in cell survival and death, and it exhibits various pro-inflammatory effects on macrophages. The effect of fermented ginseng powder on NO release is as follows: Figure 4 As shown in the figure, compared with the control group, the release of NO in the model group increased significantly after LPS stimulation, indicating that the model was successfully established and RAW264.7 cells showed inflammatory imbalance. Compared with the model group, the NO concentration in cells treated with 200 μg / mL fermented ginseng powder 1 was significantly reduced, while the NO concentration in cells treated with fermented ginseng powder 3 remained high. This indicates that fermented ginseng powder 1 has a strong effect in reducing the pro-inflammatory mediator NO and alleviating the LPS-induced inflammatory response in RAW264.7 cells.

[0087] 4.3 Effects of fermented ginseng powder on the release of TNF-α and IL-6 in RAW264.7 cells

[0088] The effects of fermented ginseng powder on the release levels of TNF-α and IL-6 are as follows: Figure 5 , Figure 6As shown, compared with the control group, the release of TNF-α and IL-6 in the model group was significantly increased after LPS stimulation, indicating that RAW264.7 cells experienced inflammatory imbalance. Compared with the model group, the levels of TNF-α and IL-6 were significantly reduced after treatment with fermented ginseng powder 1, while the effect of fermented ginseng powder 3 on reducing the levels of TNF-α and IL-6 in cells was limited, indicating that fermented ginseng powder 1 had a better anti-inflammatory effect than fermented ginseng powder 3.

[0089] 4.4 Effect of fermented ginseng powder on IL-1β release in RAW264.7 cells

[0090] The effect of fermented ginseng powder on IL-1β release is as follows: Figure 7 As shown, compared with the blank group, the release of IL-1β in the model group was significantly increased after LPS stimulation. Compared with the model group, the IL-1β content after treatment with 200 μg / mL fermented ginseng powder 1 was significantly reduced, while the reduction effect of control fermented ginseng powder 3 on IL-1β content was limited, indicating that fermented ginseng powder 1 has a better anti-inflammatory effect than control fermented ginseng powder 3.

[0091] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. For those skilled in the art, other variations or modifications can be made without departing from the principles of the present invention, and the resulting variations are still within the scope of protection of the present invention.

[0092] Industrial applicability

[0093] This invention provides a ginseng fermentation product containing rare ginsenosides with enhanced anti-inflammatory effects, its preparation method, and its application.

Claims

1. A microbial composition for producing rare ginsenosides from ginseng fermentation substrate, comprising *Lactobacillus plantarum*, *Lactobacillus casei*, *Lactobacillus paracasei*, and *Bifidobacterium animalis*; wherein, The plant lactobacillus is *Lactobacillus plantarum* (… Lactobacillus plantarum GBFG 61 is deposited at the China General Microbiological Culture Collection Center (CGC) with accession number GGMCC NO: 31583 and deposit date of August 14, 2024. The *Lactobacillus paracasei* is *Lactobacillus paracasei* (… Lactobacillus paracasei GBFG 34 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number GGMCC NO: 31581 and deposit date of August 14, 2024. The animal bifidobacteria is animal bifidobacteria ( Bifidobacterium animalis GBFG 52, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 31582, on August 14, 2024; and The Lactobacillus casei is Lactobacillus casei ( Lactobacillus casei BL23; The dry weight ratio of *Lactobacillus plantarum*, *Lactobacillus casei*, *Lactobacillus paracasei*, and *Bifidobacterium animalis* is 1:0.5~5:0.5~5:0.5~5.

2. A method for preparing a ginseng fermentation product, comprising inoculating a microbial composition into a ginseng fermentation substrate and fermenting at 30-35°C for 50-55 hours to obtain a ginseng fermentation product containing rare ginsenosides; in, The bacterial composition is the bacterial composition according to claim 1.

3. The preparation method according to claim 2 further includes sterilizing the ginseng fermentation product.

4. The preparation method according to claim 2, wherein, The ginseng fermentation product contains ginsenosides Rk1, Rg5, Rk3, and Rh4, which satisfy the following: The total content of ginsenosides Rk1, Rg5, Rk3 and Rh4 is ≥50% by weight, with the total amount of ginsenosides in the ginseng fermentation product being 100% by weight. as well as Of the ginsenosides Rk1, Rg5, Rk3 and Rh4, the total content of ginsenosides Rk3 and Rh4 is ≥30% by weight, with the total amount of ginsenosides Rk1, Rg5, Rk3 and Rh4 being 100% by weight.

5. The preparation method according to claim 2, wherein, The ginseng fermentation product is fermented ginseng liquid or fermented ginseng powder.

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