A method for preparing a sophora fruit ferment, the sophora fruit ferment prepared by the method, and applications thereof

Abstract

The application discloses a preparation method of a sophora japonica seed ferment, the sophora japonica seed ferment prepared by the method and application of the sophora japonica seed ferment, and belongs to the technical field of fermentation products. The sophora japonica seed ferment is prepared by the following steps: crushing and screening sophora japonica seeds, uniformly mixing sophora japonica seed powder and water to obtain a sophora japonica seed mixture, adding an enzyme preparation into the sophora japonica seed mixture, water-bath extracting to obtain a sophora japonica seed extract, adding a carbon source into the sophora japonica seed extract, sterilizing to obtain a fermentation culture medium, and adding a bacterial agent into the fermentation culture medium to ferment to obtain the sophora japonica seed ferment. The raw material sophora japonica seed and the bacterial strain used in the application are both of natural sources and have good biocompatibility. Meanwhile, the sophora japonica seed ferment is prepared by using a green fermentation method, and no organic solvent is needed in the whole process, so that the green and sustainable development concept is met.

Description

Technical Field

[0001] This invention belongs to the field of bio-fermentation application technology, specifically relating to a method for preparing Sophora japonica bud fermentation product and the obtained Sophora japonica bud fermentation product and its application. Background Technology

[0002] Many plant resources have been proven by modern science to possess various health benefits, including antioxidant, anti-inflammatory, anti-aging, and hair loss prevention properties. Leveraging the abundance of plant resources and combining them with modern technology to develop products with clearly defined efficacy has become a development trend. Naturally derived plant ingredients with antioxidant properties show promising development prospects.

[0003] However, the macromolecules in plants are difficult for the body to absorb and utilize, resulting in low absorption and utilization rates of active ingredients, which limits the application of plant extracts. Sophora japonica buds, the unopened flower buds of the legume Sophora japonica, are distributed throughout China and are rich in polysaccharides, flavonoids, volatile oils, and other active compounds. The abundant flavonoids in Sophora japonica buds possess excellent antioxidant and antibacterial effects; however, most flavonoids exist in glycoside form, resulting in lower activity and bioavailability.

[0004] Microbial fermentation is a promising biomodification technology for natural products. Extracellular enzymes produced by microorganisms, such as proteases, cellulases, glycosidases, and pectinases, can biotransform components in plants, such as polyphenols and flavonoids, thereby improving the bioavailability of plant extracts. Lactobacillus plantarum fermentation products can enhance the efficacy and bioavailability of plants. Due to the rich enzyme system produced by microorganisms, fermentation facilitates the dissolution of active ingredients in plants. Furthermore, enzymes can lead complex chemical reactions within the plant system, breaking down large molecules into smaller molecules that are easily absorbed by the body, and can also produce active ingredients, thus improving efficacy. Summary of the Invention

[0005] The purpose of this section is to outline some aspects of the embodiments of the present invention and to briefly describe some preferred embodiments.

[0006] In view of the problems existing in the above and / or prior art, the present invention is proposed.

[0007] Therefore, the purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing Sophora japonica fermented products.

[0008] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a method for preparing Sophora japonica fermented product, comprising, The Sophora japonica flowers are crushed and sieved. The resulting Sophora japonica flower powder is mixed with water to obtain a Sophora japonica flower mixture. Add enzyme preparation to the mixture, and extract in a water bath to obtain Sophora japonica extract; A carbon source was added to the Sophora japonica flower extract, and the mixture was sterilized to obtain a fermentation medium. Add microbial agents to the fermentation medium and ferment to obtain Sophora japonica bud fermentation product; The bacterial agent is Lactobacillus plantarum ( Lactobacillus plantarum FYS, deposited at the China Center for Type Culture Collection, with a suggested classification name. Lactobacillus plantarum FYS, accession number CCTCC NO:M 20252595, accession date November 19, 2025.

[0009] In a preferred embodiment of the preparation method described in this invention, the Sophora japonica flowers are crushed and sieved, wherein the particle size of the sieve is 80-100 mesh.

[0010] In a preferred embodiment of the preparation method described in this invention, the ratio of the Sophora japonica powder to water is 1g:(10~30)mL.

[0011] In a preferred embodiment of the preparation method described in this invention, the enzyme preparation is cellulase and protease.

[0012] In a preferred embodiment of the preparation method described in this invention, the amount of cellulase added is 0.2~2wt%, and the amount of protease added is 0.2~2wt%.

[0013] As a preferred embodiment of the preparation method described in this invention, the water bath extraction time is 1-5 hours, the enzymatic hydrolysis pH is 3-7, and the temperature is 30-60°C.

[0014] In a preferred embodiment of the preparation method described in this invention, the carbon source is glucose, galactooligosaccharide, fructooligosaccharide, and inulin; and the amount of carbon source added is 1-5 wt%.

[0015] As a preferred embodiment of the preparation method of the present invention, the microbial agent further includes one or more of Lactobacillus casei and Lactobacillus rhamnosus, and the fermentation time is 12-72 hours.

[0016] Another objective of this invention is to overcome the shortcomings of the prior art and provide a fermented Sophora japonica product.

[0017] Another objective of this invention is to overcome the shortcomings of the prior art and provide an application of Sophora japonica ferment in the preparation of pharmaceuticals or skin care products with anti-inflammatory effects.

[0018] Beneficial effects of this invention: (1) The raw materials Sophora japonica and strains used in this invention are all of natural origin and have good biocompatibility. At the same time, this invention uses a green fermentation method to prepare fermentation products, and the whole process does not require any organic solvents, which is in line with the concept of green and sustainable development. (2) The fermented Sophora japonica prepared by this invention has a simple process, uses common equipment, and is suitable for industrial production; (3) The Sophora japonica fermented product prepared by this invention has excellent antioxidant effects and shows good application prospects in the fields of food, cosmetics and medicine. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort. Wherein: Figure 1 The bar chart shows the DPPH free radical scavenging rate in Examples 1, 8, and 9 of this invention, as well as Comparative Examples 1-6.

[0020] Figure 2 This invention demonstrates the inhibitory effect of the products in Examples 1, 2, and Comparative Examples 1-6 on IL-6 expression in RAW264.7 cells.

[0021] Figure 3 This demonstrates the inhibitory effect of the product obtained in Example 2 of the present invention on IL-6 expression in RAW264.7 cells at different concentrations. Detailed Implementation

[0022] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0023] Unless otherwise specified, all chemical reagents and materials used in the embodiments of this invention are obtained commercially.

[0024] The *Lactobacillus plantarum* used in the following examples (*Lactobacillus plantarum* ... Lactobacillus plantarum FYS, deposited at the China Center for Type Culture Collection, with a suggested classification name. Lactobacillus plantarum FYS, with accession number CCTCC NO: M 20252595, accession date November 19, 2025; Lactobacillus casei CICC 20262, Lactobacillus rhamnosus CICC6141, Lactobacillus plantarum CICC23941 and Lactobacillus plantarum ATCC 14917 are from the China Industrial Microbial Culture Collection Center.

[0025] Example 1 (1) Pulverize the dried Sophora japonica flowers for 3 minutes, pass them through a 100-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:15 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 5-6, add 0.4% cellulase (3500U / g) and 0.2% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 50℃ for 2 hours to obtain Sophora japonica flower extract; (3) Add 3% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 2% (CFU / mL), fermented at 37℃ for 24 h, and the fermentation broth was collected by centrifugation at 5000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0026] Example 2 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0027] Example 3 (1) Pulverize the dried Sophora japonica flowers for 5 minutes, pass them through a 100-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:25 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 7-8, add 0.6% cellulase (3500U / g) and 0.4% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 40℃ for 4 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9The inoculum was 4% (CFU / mL), fermented at 37℃ for 48 h, and the fermentation broth was collected by centrifugation at 7000 rpm for 5 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0028] Example 4 (1) Pulverize the dried Sophora japonica flowers for 2 minutes and pass them through an 80-mesh sieve to obtain Sophora japonica flower powder. Add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:15 to obtain a Sophora japonica flower mixture. (2) Adjust the pH of the Sophora japonica flower mixture to 5-6, add 0.8% cellulase (3500U / g) and 0.5% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 40℃ for 2 hours to obtain Sophora japonica flower extract; (3) Add 3% glucose to the Sophora japonica extract, and then sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 4% (CFU / mL), fermented at 37℃ for 24 h, and the fermentation broth was collected by centrifugation at 7000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0029] Example 5 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through a 100-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:25 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 5-6, add 0.5% cellulase (3500U / g) and 0.5% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 35℃ for 3h to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract, and then sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 5% (CFU / mL), fermented at 37℃ for 48 hours, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 minutes. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0030] Example 6 (1) Pulverize the dried Sophora japonica flowers for 5 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 5-6, add 1.2% cellulase (3500U / g) and 0.2% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 35℃ for 2 hours to obtain Sophora japonica flower extract; (3) Add 1% glucose to the Sophora japonica extract, and then sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 6% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 8000 rpm for 5 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0031] Example 7 (1) Pulverize the dried Sophora japonica flowers for 3 minutes, pass them through a 100-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:15 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 1% cellulase (3500U / g) and 0.4% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 60℃ for 2 hours to obtain Sophora japonica flower extract; (3) Add 4% glucose to the Sophora japonica extract, and then sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 7% (CFU / mL), fermented at 37℃ for 24 hours, and the fermentation broth was collected by centrifugation at 8000 rpm for 5 minutes. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0032] Example 8 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus casei CICC 20262 (5.0 × 10⁻⁶). 9The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0033] Example 9 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus rhamnosus CICC 6141 (5.0 × 10⁻⁶). 9 The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0034] Comparative Example 1 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, extract in a water bath at 45℃ for 3h, collect the extract, centrifuge at 6000rpm for 10min to take the supernatant, filter through a 0.22μm filter membrane to obtain the Sophora japonica flower fermentation broth, and obtain the unfermented Sophora japonica flower extract; Comparative Example 2 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7 and extract it in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0035] Comparative Example 3 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum CICC 23941 (5.0 × 10⁻⁶). 9 The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0036] Comparative Example 4 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Adjust the pH of the Sophora japonica flower mixture to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica flower mixture, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica flower extract; (3) Add 2% glucose to the Sophora japonica extract and sterilize at 121℃ for 20 min to obtain the fermentation medium; (4) Inoculate the fermentation medium with Lactobacillus plantarum ATCC 14917 (5.0 × 10⁻⁶). 9 The inoculum was 3% (CFU / mL), fermented at 37℃ for 36 h, and the fermentation broth was collected by centrifugation at 6000 rpm for 10 min. The supernatant was then filtered through a 0.22 μm filter membrane to obtain the Sophora japonica fermentation broth.

[0037] Comparative Example 5 (1) Take Lactobacillus plantarum CCTCC No: M20252595 and inoculate it into a shaker tube containing 6 mL of MRS medium. Incubate at 37℃ and 200 rpm for 24 h to obtain Lactobacillus plantarum liquid; wherein the OD value of the Lactobacillus plantarum liquid is between 2.3 and 2.7. (2) Prepare MRS culture medium and sterilize at 121℃ for 20 min; (3) Inoculate MRS medium with Lactobacillus plantarum (5.0 × 10⁻⁶) 9The bacterial culture was inoculated at 3% CFU / mL, with an inoculum rate of 3%. The culture was incubated at 37°C for 36 hours. The bacterial culture was collected, centrifuged at 6000 rpm for 10 minutes, and the supernatant was collected and filtered through a 0.22 μm filter membrane to obtain the Lactobacillus plantarum bacterial culture.

[0038] Comparative Example 6 (1) Pulverize the dried Sophora japonica flowers for 4 minutes, pass them through an 80-mesh sieve to obtain Sophora japonica flower powder, add purified water to the Sophora japonica flower powder at a material-to-liquid ratio of 1:20 to obtain Sophora japonica flower mixture; (2) Add 2% glucose to the Sophora japonica flower mixture and sterilize at 121℃ for 20 min to obtain the fermentation medium; (3) Inoculate the fermentation medium with Lactobacillus plantarum CCTCC No: M20252595 (5.0×10⁻⁶) 9 CFU / mL), inoculation rate 3%, fermentation at 37℃ for 36h, collection of fermentation broth centrifuged at 6000rpm for 10min, supernatant was collected and filtered through a 0.22μm filter membrane to obtain Sophora japonica fermentation broth; (4) Adjust the pH of the Sophora japonica fermentation broth to 6-7, add 0.5% cellulase (3500U / g) and 0.3% protease (120000U / g) to the Sophora japonica fermentation broth, and extract in a water bath at 45℃ for 3 hours to obtain Sophora japonica fermentation extract.

[0039] Performance testing 1. Determination of DPPH free radical scavenging capacity: The samples obtained in Examples 1-9 and Comparative Examples 1-6 were diluted 10-fold with deionized water. 100 μL of each sample was added to a 96-well plate, and 100 μL of 0.2 mM DPPH-anhydrous ethanol solution was added to each well. The mixture was stirred and reacted at room temperature in the dark for 30 min to obtain the sample solution. The absorbance at 517 nm was measured (denoted as A). For the blank control group, anhydrous ethanol was used instead of the DPPH-anhydrous ethanol solution, and the absorbance at 517 nm was measured after the reaction (denoted as B). For the control blank group, deionized water was used instead of the sample solution, and the absorbance at 517 nm was measured after the reaction (denoted as D). The DPPH free radical scavenging rate was calculated using the following formula, and the detection results are shown in Table 1.

[0040] DPPH free radical scavenging rate (%) = [1 - (AB) / (CD)] × 100% 2. ABTS+ scavenging ability determination: 10 μL of the samples obtained in Examples 1-9 and Comparative Examples 1-6 were added to 96-well plates, followed by 190 μL of 0.7 mM ABTS solution. The mixtures were thoroughly mixed and reacted at room temperature in the dark for 15 min to obtain the sample solution. The absorbance at 734 nm was measured (denoted as A). The blank group used deionized water instead of the sample solution, and the absorbance at 734 nm was measured after the reaction (denoted as B). ABTS + The clearance rate was calculated using the following formula, and the test results are shown in Table 2.

[0041] ABTS + Clearance rate (%) = [1 - A / B] × 100% Table 1: DPPH radical scavenging rate and ABTS radical scavenging rate of Examples 1-9 and Comparative Examples 1-6

[0042] like Figure 1 As shown in Table 1, in Examples 1-9, the DPPH and ABTS radical scavenging rates both exceeded 90%, while in Comparative Example 1, the DPPH radical scavenging rate was only about 80%, and the ABTS radical scavenging rate was about 81%, indicating that fermentation significantly improved the antioxidant properties of Sophora japonica buds. In Comparative Example 2, the DPPH radical scavenging rate was only 77%, and the ABTS radical scavenging rate was about 83%, indicating that the enzymatic hydrolysis process plays an important role in improving the antioxidant capacity of the fermentation broth. Meanwhile, the DPPH and ABTS radical scavenging rates in Comparative Examples 3 and 4 were both only about 80%, comparable to the antioxidant properties in Comparative Example 1, indicating that not all strains can improve the antioxidant properties of Sophora japonica buds through fermentation. In Comparative Example 5, the DPPH and ABTS radical scavenging rates were approximately 63% and 67%, respectively, indicating that the addition of Sophora japonica buds significantly increased the DPPH radical scavenging capacity of Lactobacillus metabolites. In Comparative Example 6, the DPPH and ABTS radical scavenging rates were comparable to those in Comparative Example 1, indicating that fermentation followed by enzymatic hydrolysis does not increase the antioxidant properties of Sophora japonica buds.

[0043] 3. Anti-inflammatory capacity test: RAW 264.7 cells were cultured in DMEM containing 10% fetal bovine serum and 1% penicillin-streptomycin. Cells in the logarithmic growth phase were collected, and the cell suspension concentration was adjusted to 1×10⁻⁶. 5Seeds / mL were inoculated into 24-well plates with 500 μL of the culture medium and cultured for 24 h. The original culture medium was removed, and the plates were washed twice with PBS. 500 μL of LPS solution containing 1 μg / mL was added. DMEM medium solution of different samples (samples in Examples 1-2 and Comparative Examples 1-6) with the same concentration (0.25 mg / mL) or DMEM medium solution of the same sample (Example 2) with different concentrations (0.125, 0.25, 0.5 mg / mL) was added and incubated for 24 h.

[0044] The level of the pro-inflammatory cytokine IL-6 in macrophage supernatant was measured according to the guidelines of a commercially available ELISA kit (Signalway Antibody, USA). In this assay, IL-6, as a typical cellular inflammatory cytokine, clearly represents the intensity of cellular inflammation. The lower the IL-6 expression level, the weaker the cellular inflammation level. If the product can significantly inhibit the expression of IL-6 in cells, it indicates that the product has a significant anti-inflammatory effect.

[0045] like Figure 2 and Figure 3 As shown, the Sophora japonica fermentation broth prepared in Examples 1-2 significantly inhibited LPS-induced inflammatory responses, and the inhibitory effect was significantly better than that obtained from other comparative examples. Notably, the expression level of IL-6 showed a typical negative correlation with the concentration of the Sophora japonica fermentation broth prepared in Example 2; that is, as the concentration of the Sophora japonica fermentation broth increased, the level of IL-6 measured in LPS-induced RAW264.7 cells gradually decreased. This indicates that the samples prepared in these examples have significant anti-inflammatory effects.

[0046] 4. Sensory evaluation: Forty people were randomly selected and scored according to the sensory evaluation criteria shown in the table below. The scores are shown in Table 3. Table 2 Sensory evaluation indicators and scores

[0047] Table 3: Sensory evaluation of Examples 1-9 and Comparative Examples 1-6

[0048] As shown in Table 3, the properties of Sophora japonica flowers changed significantly after fermentation with Lactobacillus, and consumers were more satisfied with their color, smell, and viscosity.

[0049] This invention is the first to utilize Lactobacillus fermentation of Sophora japonica extract. Through enzymatic hydrolysis and Lactobacillus fermentation, the components in the Sophora japonica extract are biotransformed, improving its DPPH and ABTS free radical scavenging rates. Therefore, the Sophora japonica fermentation broth is a green product with good antioxidant capacity and excellent biological activity, making it a promising new cosmetic ingredient.

[0050] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the present invention.

Claims

1. A method for preparing a fermented product of Sophora japonica buds, characterized in that: include, The Sophora japonica flowers are crushed and sieved. The resulting Sophora japonica flower powder is mixed with water to obtain a Sophora japonica flower mixture. Add enzyme preparation to the mixture, and extract in a water bath to obtain Sophora japonica extract; A carbon source was added to the Sophora japonica flower extract, and the mixture was sterilized to obtain a fermentation medium. Add microbial agents to the fermentation medium and ferment to obtain Sophora japonica bud fermentation product; The bacterial agent is Lactobacillus plantarum ( Lactobacillus plantarum FYS, deposited at the China Center for Type Culture Collection, with a suggested classification name. Lactobacillus plantarum FYS, accession number CCTCC NO: M20252595, accession date November 19, 2025.

2. The preparation method according to claim 1, characterized in that: The process of crushing and sieving the Sophora japonica flowers involves using a sieve with a particle size of 80-100 mesh.

3. The preparation method according to claim 1 or 2, characterized in that: The ratio of the sophora japonica powder to water is 1g:(10~30)mL.

4. The preparation method according to claim 3, characterized in that: The enzyme preparations are cellulase and protease.

5. The preparation method according to claim 4, characterized in that: The amount of cellulase added is 0.2~2wt%, and the amount of protease added is 0.2~2wt%.

6. The preparation method according to claim 1, characterized in that: The water bath extraction time is 1-5 hours, the enzymatic hydrolysis pH is 5-8, and the temperature is 30-60℃.

7. The preparation method according to claim 1 or 6, characterized in that: The carbon source is glucose, galactooligosaccharide, fructooligosaccharide, and inulin; the amount of carbon source added is 1-5 wt%.

8. The preparation method according to claim 1, characterized in that: The microbial agent also includes one or more of Lactobacillus casei CICC20262 and Lactobacillus rhamnosus CICC6141, and the fermentation time is 12-72 hours.

9. The preparation method according to any one of claims 1 to 8 yields the Sophora japonica fermented product.

10. The use of the Sophora japonica ferment as described in claim 9 in the preparation of pharmaceuticals or skin care products with anti-inflammatory effects.

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