Kudzuvine root fermentation product as well as preparation method and application thereof

Abstract

The invention relates to the technical field of fermentation engineering, in particular to a radix puerariae fermentation product and a preparation method and application thereof.The radix puerariae fermentation product is obtained by fermenting bifidobacterium adolescentis, saccharomyces cerevisiae and compound lactic acid bacteria and has the effects of controlling oil, repairing skin barriers and the like.

Description

Technical Field

[0001] This disclosure relates to the technical field of fermentation engineering, and in particular to a kudzu root fermentation product, its preparation method, and its uses. Background Technology

[0002] Kudzu root is the root of the legume plant *Pueraria lobata* (also known as wild kudzu). Pueraria lobata The dried root of *Puerariae Lobatae Radix* (Willd.) Ohwi, whose medicinal Latin name is *Puerariae Lobatae Radix*. This plant is also considered by some taxonomic literature to be... Pueraria montana variant Pueraria montana var. lobata However, according to legal standards such as the Chinese Pharmacopoeia, the Latin name of its original plant is still... Pueraria lobata (Willd.) Ohwi is the standard. Kudzu root is a traditional Chinese medicine that is both food and medicine. Its chemical composition is complex, mainly containing isoflavones, triterpenoids, and saponins. Among them, isoflavones such as puerarin, daidzin, daidzein, and genistein are its main pharmacologically active components. Puerarin, in particular, as a characteristic component of kudzu root, has shown significant pharmacological effects in dilating coronary arteries, improving microcirculation, and antiarrhythmic activity.

[0003] In recent years, with the increasingly widespread application of natural plant ingredients in the cosmetics industry, kudzu root extract has attracted attention due to its excellent skincare potential. Studies have shown that isoflavones in kudzu root have a structure similar to human estrogen, which can gently regulate skin metabolism and reduce melanin production by inhibiting tyrosinase activity, thus achieving a whitening effect. In addition, kudzu root extract also exhibits antioxidant, anti-inflammatory, and anti-photoaging bioactivities, reducing UV-induced fibroblast damage and increasing collagen expression levels, making it suitable for anti-wrinkle and soothing skincare applications. Currently, kudzu root is mostly used in cosmetics in the form of traditional extracts, and the effectiveness of its active ingredients mainly relies on inherent components such as puerarin.

[0004] Fermentation engineering, as an important biotechnology, has opened up new avenues for the high-value utilization of plant resources. Studies have shown that through microbial fermentation of plant materials, various hydrolytic enzymes such as cellulase and protease produced by microorganisms during metabolism can degrade plant cell wall structures, promoting the release of intracellular active substances. Simultaneously, microbial metabolism can convert macromolecules in the raw materials into more easily absorbed small-molecule active ingredients, and even generate new active compounds through biotransformation. Different fermentation substrates and fermentation strains have a significant impact on the composition of active ingredients in the final product. For example, lactic acid bacteria fermentation can increase the hydrolysis degree of plant proteins and release peptides with antioxidant or antihypertensive activities (Gao Ning, Zan Lixia, He Junting, et al. Research progress on microbial fermentation preparation and bioactivity of plant-derived bioactive peptides [J]. Food Industry Technology, 2025, 46(22): 468-478.); certain yeast fermentation can promote the enrichment of phenolic substances (such as gallic acid, catechin, quercetin, etc.) and enhance the antioxidant and anti-inflammatory effects of the products (Alessandro Stringari, et al. Revitalizing Brewers' Spent Grains and Enriching With Biogenic Compounds Through the Fermentation of Fructophilic Lactic Acid Bacteria and Yeasts [J]. Microbial biotechnology. Volume 18, Issue 6. 2025.). Therefore, by screening suitable fermentation conditions, the bioactive component profile of plant fermentation products can be directionally regulated, thereby obtaining fermentation products with specific effects.

[0005] Although fermentation engineering has made significant progress in the development of plant resources, research on fermentation of kudzu root, a traditional Chinese medicine resource, remains relatively limited. Current reports on kudzu root mainly focus on optimizing the extraction process of its natural products and evaluating their pharmacological activity. However, in-depth exploration of the impact of microorganisms on the transformation of kudzu root's active ingredients through microbial fermentation, and the application potential of fermentation products in the skincare field, is still lacking. Given that kudzu root is rich in flavonoids and other active precursors, appropriate fermentation technology holds the promise of releasing or generating new active ingredients, further enhancing its application value in the cosmetics field. Therefore, developing a kudzu root fermentation product with a novel spectrum of active ingredients and clarifying its preparation methods and uses is of significant research importance and has broad application prospects. Summary of the Invention

[0006] This disclosure relates to a kudzu root fermentation product, its preparation method, and its uses. The kudzu root fermentation product and compositions containing the kudzu root fermentation product have effects such as skin care or mucous membrane care.

[0007] On the one hand, this disclosure provides a kudzu root fermentation product, the high-performance liquid chromatography (HPLC) chromatogram of which includes at least three characteristic peaks, with the characteristic peak of puerarin as a reference peak. The relative retention times of the three characteristic peaks and the reference peak are 0.49~0.50, 1.00, and 1.36~1.38, respectively. The HPLC conditions are as follows: Chromatographic column: 4.6*250mm, 5μm, packed with octadecylsilane-bonded silica gel; Column temperature: 30℃; Injection volume: 10 μL; Detection wavelength: 250nm; Mobile phase A: 0.1% aqueous phosphoric acid solution; Mobile phase B: methanol; Flow rate: 1 mL / min; Elution gradient: 0~15min: 75%A, 25%B; 15~30min: 75%→55%A, 25%→45%B; 30~35min: 55%→10%A, 45%→90%B; 35~36min: 10%→75%A, 90%→25%B; 36~45min: 75%A, 25%B.

[0008] In some embodiments, the chromatographic column is an AQ-C18 column.

[0009] In some embodiments, the chromatographic column is an Ultimate AQ-C18 column or a column with equivalent performance.

[0010] In some implementations, the relative peak areas of the three characteristic peaks and the reference peak, in order of retention time, are 1.80%~4.34%, 100%, and 4.18%~21.03%, respectively.

[0011] The kudzu root fermentation product disclosed herein contains multiple active ingredients. Besides common kudzu root active ingredients such as puerarin, it also contains at least two special post-fermentative active ingredients produced through fermentation. Under the high-performance liquid chromatography (HPLC) conditions disclosed herein, using the characteristic peak of puerarin as a reference peak, the relative retention times of the characteristic peaks of these two special post-fermentative active ingredients compared to the reference peak were 0.49–0.50 and 1.36–1.38, respectively, with relative peak areas of 1.80%–4.34% and 4.18%–21.03%, respectively. HPLC analysis confirms that the kudzu root fermentation product disclosed herein differs significantly in composition from conventional kudzu root extracts, representing a novel mixture. Efficacy experiments have verified that the kudzu root fermentation product disclosed herein can promote cell migration, repair the skin barrier, and promote re-epithelialization or healing of the skin or mucous membranes. Compared to conventional kudzu root extracts, the kudzu root fermentation product disclosed herein can further inhibit skin sebum synthesis or secretion, achieving superior oil-controlling effects.

[0012] Those skilled in the art will understand that, under stable preparation conditions, the types of chemical components contained in kudzu and the relative proportions of these components are relatively fixed. The relative retention times of characteristic peaks represent the compositional relationship of the chemical components, and the relative peak areas represent the relative proportions of their content. These specific peak spectral characteristics are unique and reproducible; therefore, by measuring kudzu extract or kudzu fermentation products under specific chromatographic conditions, it is possible to effectively identify whether they belong to the same substance as the kudzu fermentation products disclosed herein.

[0013] On the other hand, this disclosure provides a method for preparing kudzu fermentation products, comprising the following steps: pretreating kudzu raw materials to obtain kudzu treated material; mixing the kudzu treated material with a carbon source and adding a zinc ion source to prepare a liquid fermentation culture medium containing zinc ions; first inoculating the liquid fermentation culture medium with Bifidobacterium adolescentis for anaerobic fermentation, and then inoculating it with Saccharomyces cerevisiae and compound lactic acid bacteria for aerobic fermentation, thereby obtaining the kudzu fermentation product.

[0014] In this disclosure, the pretreatment method for kudzu root raw materials is not particularly limited, and conventional methods in the art can be used. As an example, the pretreatment may involve physical crushing of fresh or dried kudzu root such as pressing, slicing, crushing, or grinding; enzymatic hydrolysis using amylase, cellulase, protease, pectinase, etc.; or extraction or soaking using water, alcohol, or buffer solution. The purpose of the above treatments is to break down the plant tissue of kudzu root, causing the starch, flavonoids, and other effective components to dissolve or be exposed, so that they can be utilized by microorganisms during subsequent fermentation. In some embodiments, amylase, cellulase, protease, and pectinase are used to enzymatically hydrolyze the kudzu root raw materials. In some embodiments, the amylase is selected from one or more of α-amylase, β-amylase, γ-amylase, and isoamylase, preferably α-amylase. In some embodiments, the cellulase is selected from one or more of endoglucanase, exoglucanase, and β-glucosidase, preferably a combination of endoglucanase, exoglucanase, and β-glucosidase. In some embodiments, the protease is selected from one or more of acidic proteases, neutral proteases, and alkaline proteases, preferably neutral proteases. In some embodiments, the pectinase is selected from one or more of pectin methylgalacturonase, polygalacturonase, polygalacturonate lyase, pectin esterase, and pectin lyase, preferably a combination of pectin methylgalacturonase, polygalacturonase, polygalacturonate lyase, pectin esterase, and pectin lyase.

[0015] In this disclosure, the method for preparing the liquid fermentation medium is well known to those skilled in the art. As an example, the kudzu root treatment obtained in the aforementioned steps (such as kudzu root juice, kudzu root enzymatic hydrolysate, kudzu root aqueous extract, kudzu root powder, etc.) can be mixed with basic culture medium components such as carbon and nitrogen sources in proportions suitable for the growth of the fermentation strain. Simultaneously, a zinc ion source is added, and pH adjustment, volume adjustment, and sterilization are performed as needed to obtain a liquid fermentation medium containing zinc ions. In this disclosure, a zinc ion source refers to a source capable of providing zinc ions (Zn) in the liquid culture medium. 2+ The zinc ion source includes, but is not limited to, inorganic and organic zinc salts. In some embodiments, the zinc ion source is selected from one or more of zinc sulfate, zinc chloride, zinc nitrate, zinc acetate, zinc citrate, and zinc gluconate. The aforementioned zinc ion sources are typically added during the preparation of the culture medium in the form of a solid powder or a pre-prepared solution. In some embodiments, the concentration of zinc ions in the liquid fermentation medium is 1–10 mmol / L. In some embodiments, the concentration of zinc ions in the liquid fermentation medium is 1–5 mmol / L. In some embodiments, the concentration of zinc ions in the liquid fermentation medium is 3–5 mmol / L.

[0016] The fermentation strain can be inoculated using conventional methods known in the prior art, such as direct inoculation, liquid inoculation, and solid inoculation.

[0017] As a demonstration, the following steps can be used to inoculate brewer's yeast: - The activated seeds were inoculated into YM liquid medium and cultured in a shaker at 28°C and 150 rpm for 16 h to obtain seed solution; - Inoculate the seed culture into the liquid fermentation medium at an inoculation rate of 2% (v / v).

[0018] As an example, the following steps can be taken to inoculate with Bifidobacterium adolescentis: - Inoculate the activated seeds onto TPY medium, place them in an anaerobic culture bag with an anaerobic gas generator, seal the anaerobic culture bag, and culture in a shaker at 37°C and 150 rpm for 24 hours to obtain the seed solution. - Inoculate the seed culture into the liquid fermentation medium at an inoculation rate of 2% (v / v).

[0019] As an example, the following steps can be used to inoculate compound lactic acid bacteria: - Inoculate the bacterial powder into MRS liquid medium and culture it in a shaker at 37°C and 150 rpm for 16 h to obtain the seed culture; - Inoculate the seed culture into the liquid fermentation medium at an inoculation rate of 2% (v / v).

[0020] The term "complex lactic acid bacteria" refers to a mixed bacterial agent composed of multiple lactic acid bacteria strains with different oxygen tolerance characteristics. Based on their differences in oxygen requirements and responses, the complex lactic acid bacteria include two main categories: strictly anaerobic lactic acid bacteria and facultative anaerobic lactic acid bacteria. Strictly anaerobic lactic acid bacteria are strains that cannot grow or whose growth is significantly inhibited in the presence of oxygen, including but not limited to *Bifidobacterium* spp. Bifidobacterium ) species, such as Bifidobacterium adolescentis ( Bifidobacterium adolescentis Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium breve Bifidobacterium breve ) and Bifidobacterium infantis ( Bifidobacterium infantis Facultative anaerobic lactic acid bacteria refer to strains that can grow under both aerobic and anaerobic conditions, but prefer microaerobic or anaerobic environments, including but not limited to the genus *Lactobacillus*. Lactobacillus ) strains, such as Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus acidophilus ( Lactobacillus acidophilus Lactobacillus casei ( Lactobacillus casei ) and Lactobacillus rhamnosus ( Lactobacillus rhamnosus ); and Lactococcus spp. ( Lactococcus ) strains, such as Lactococcus lactis ( Lactococcus lactis ).

[0021] This disclosure achieves complementary metabolic advantages of different fermentation strains through the sequential synergy of anaerobic and aerobic fermentation. Combined with the metabolic transformation of zinc ions by the fermentation strains, this ensures the smooth production of the kudzu fermentation product. Specifically, anaerobic fermentation is the first fermentation stage, using *Bifidobacterium adolescentis* as the fermentation strain, and fermentation is carried out under strictly anaerobic conditions. Maintaining the anaerobic environment can be achieved using conventional techniques in the art, such as degassing by introducing sterile high-purity nitrogen gas to remove dissolved oxygen from the fermentation system, thereby meeting the initial growth and metabolic needs of *Bifidobacterium adolescentis* as a strict anaerobic bacterium. Aerobic fermentation is the second fermentation stage. After completing the first fermentation stage, *Saccharomyces cerevisiae* and a compound lactic acid bacteria are introduced into the fermentation system, and the conditions are adjusted to aerobic fermentation using conventional techniques in the art (such as introducing sterile air). *Saccharomyces cerevisiae*, as a facultative anaerobic bacterium, can grow and metabolize by consuming oxygen in an aerobic environment. The compound lactic acid bacteria include strict anaerobic and facultative anaerobic bacteria, which generally prefer a microaerobic environment and can grow and metabolize under microaerobic conditions. During aerobic fermentation, *Saccharomyces cerevisiae* provides a microaerobic environment for the complex lactic acid bacteria by consuming dissolved oxygen in the culture medium, enabling the complex lactic acid bacteria to carry out normal growth and metabolic activities. After anaerobic fermentation with *Bifidobacterium adolescentis*, followed by aerobic fermentation with *Saccharomyces cerevisiae* and the complex lactic acid bacteria, a zinc-containing liquid fermentation medium was used to obtain a kudzu root fermentation product containing the aforementioned special metagenic active components.

[0022] Post-processing of the above-mentioned kudzu fermentation products is a conventional technique in this field, aimed at removing insoluble solids and fermentation cells from the fermentation products. As an example, the fermentation broth after fermentation is terminated can be centrifuged and filtered to remove insoluble solids. Furthermore, the filtrate can be pasteurized and then filtered through a 0.22 μm filter membrane to remove cells, obtaining a clear and sterile kudzu fermentation broth.

[0023] The anaerobic fermentation process described in this disclosure uses only *Bifidobacterium adolescentis* as the fermentation strain. Therefore, the fermentation conditions should be conventional conditions known in the art that are suitable for the growth of this strain and for maintaining its normal anaerobic fermentation activity. In some embodiments, during the fermentation step, the anaerobic fermentation temperature is 34-37°C, and the fermentation time is 16-24 hours.

[0024] The aerobic fermentation process described in this disclosure uses *Saccharomyces cerevisiae* and a compound lactic acid bacteria as fermentation strains. Therefore, the fermentation temperature is a conventional condition known in the art, suitable for the growth of these strains and for maintaining their normal fermentation activity. A key parameter of the aerobic fermentation process described in this disclosure is the dissolved oxygen saturation in the liquid fermentation medium. During aerobic fermentation, *Saccharomyces cerevisiae* continuously consumes dissolved oxygen in the liquid fermentation medium. Continuously introducing sterile air to maintain oxygen supply ensures the dissolved oxygen level of the medium, thereby ensuring the normal life activities of *Saccharomyces cerevisiae*. However, with the proliferation and metabolism of *Saccharomyces cerevisiae*, its oxygen consumption rate exhibits a non-linear characteristic, causing the dissolved oxygen level of the fermentation system to fluctuate dynamically within a certain range. Therefore, it is necessary to adjust the aeration rate and stirring rate to adapt to the fermentation process. By controlling the dissolved oxygen within a certain relatively saturated range, the aerobic metabolic needs of *Saccharomyces cerevisiae* can be guaranteed, while ensuring that the dissolved oxygen level remains within the microaerobic range suitable for the growth of the compound lactic acid bacteria. This dynamic balance formed by active oxygen supply control and microbial oxygen consumption disturbance constitutes the essential basis for the synergistic fermentation of *Saccharomyces cerevisiae* and compound lactic acid bacteria. In some embodiments, during the aerobic fermentation step, the fermentation temperature is 28-32°C, the fermentation time is 16-24 hours, and the dissolved oxygen saturation in the liquid fermentation medium is maintained at 70%-100%.

[0025] In some embodiments, the compound lactic acid bacteria include Bifidobacterium lactis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus plantarum.

[0026] On the other hand, this disclosure also provides a kudzu root fermentation product, which is prepared by the above-described preparation method.

[0027] For ease of application, the kudzu fermentation product disclosed herein, or the kudzu fermentation product prepared by the above-described preparation method, can be part of various types of compositions. Therefore, in another aspect, this disclosure provides a composition comprising an effective amount of the above-described kudzu fermentation product, or an effective amount of the kudzu fermentation product prepared by the above-described preparation method, and at least one additional ingredient.

[0028] In some embodiments, the added ingredients include cosmetically or pharmaceutically acceptable diluents, cosmetically or pharmaceutically acceptable adjuvants, cosmetically or pharmaceutically acceptable excipients, cosmetically or pharmaceutically acceptable solvents or carriers, or combinations of any of the above.

[0029] In some embodiments, the adjuvant includes: analgesics, agents that inhibit PAR-2 ​​activity, agents that regulate PGC-1α synthesis, agents that regulate PPARγ activity, agents that increase or decrease triglyceride content in adipocytes, agents that stimulate or delay adipocyte differentiation, lipolytic agents or agents that stimulate lipolysis, lipolytic agents, lipogenic agents, inhibitors of acetylcholine receptor aggregation, agents that inhibit muscle contraction, anticholinergic agents, elastase inhibitors, matrix metalloproteinase inhibitors, melanin synthesis stimulators or inhibitors, whitening agents or bleaching agents, pigmentation promoters, self-tanning agents, anti-aging agents, NO-synthesizers, 5α-reductase inhibitors, inhibitors of lysyl hydroxylase and / or prolyl hydroxylase, antioxidants, and free radical scavengers. Agents and / or anti-air pollution agents, active carbonyl scavengers, anti-glycation agents, antihistamines, antiviral agents, antiparasitic agents, emulsifiers, emollients, organic solvents, liquid propellants, water-retaining substances, alpha-hydroxy acids, beta-hydroxy acids, humectants, epidermal hydrolases, vitamins, amino acids, proteins, pigments, dyes, biopolymers, gelling polymers, thickeners, surfactants, softeners, adhesives, preservatives, anti-wrinkle agents, agents that reduce or treat under-eye bags, keratolytic agents, antimicrobial agents, agents that stimulate the synthesis of dermal or epidermal macromolecules and / or inhibit or prevent their degradation, agents that stimulate elastin synthesis, agents that stimulate core proteoglycan synthesis, agents that stimulate laminin synthesis, and agents that stimulate defensin synthesis. Agents, agents stimulating chaperone protein synthesis, agents stimulating cAMP synthesis, agents stimulating hyaluronic acid synthesis, agents stimulating fibronectin synthesis, agents stimulating deacetylase synthesis, agents stimulating lipid and stratum corneum component synthesis, ceramides, fatty acids, agents inhibiting elastin degradation, agents inhibiting serine proteases, agents stimulating fibroblast proliferation, agents stimulating keratinocyte proliferation, agents stimulating adipocyte proliferation, agents stimulating melanocyte proliferation, agents stimulating keratinocyte differentiation, agents inhibiting acetylcholinesterase, skin relaxants, agents stimulating glycosaminoglycan synthesis, anti-hyperkeratosis agents, comedolytic agents, anti-psoriasis agents, anti-eczema agents, DNA repair agents, DNA protectants, stabilizers, antipruritic agents, for the treatment and / or protection of skin. Agents for sensitive skin, including hardening agents, firming agents, reconstructing agents, anti-stretch mark agents, agents that regulate sebum production, antiperspirants, agents that stimulate healing, agents that assist healing, agents that stimulate re-epithelialization, agents that assist re-epithelialization, cytokines, sedatives, anti-inflammatory agents, agents that act on capillary circulation and / or microcirculation, agents that stimulate angiogenesis, agents that inhibit vascular permeability, venous tension agents, agents that act on cell metabolism, agents for improving dermal-epidermal junction, agents that induce hair growth, agents that inhibit or delay hair growth, fragrances, chelating agents, plant extracts, essential oils, marine extracts, agents derived from bio-fermentation processes, inorganic salts, cell extracts, sunscreens, and organic or inorganic photoprotective agents or mixtures thereof that effectively resist UVA and / or UVB rays.

[0030] The effective amount of the active fermentation products of this disclosure to be applied, as well as their dosage, will depend on many factors, including age, the user's condition, the severity of the condition, the route of application, and the frequency of application.

[0031] "Effective amount" means an amount of the kudzu fermentation product of this disclosure that is non-toxic but sufficient to provide the desired effect. The effective concentration of the kudzu fermentation product of this disclosure is used in the compositions of this disclosure to obtain the desired effect. In some embodiments, the concentration is between 0.00000001% (by weight) and 20% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.000001% (by weight) and 15% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.0001% (by weight) and 10% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.0001% (by weight) and 5% (by weight) relative to the total weight of the composition.

[0032] In another aspect of this disclosure, the use of the above-described kudzu fermentation product or the kudzu fermentation product prepared by the above-described preparation method in the preparation of compositions for skin or mucous membrane care is provided.

[0033] In another aspect of this disclosure, the use of the above-mentioned kudzu fermentation product or the kudzu fermentation product prepared by the above-mentioned preparation method in the preparation of compositions for repair or oil control is provided.

[0034] In another aspect of this disclosure, the use of the above-mentioned kudzu fermentation product or the kudzu fermentation product prepared by the above-mentioned preparation method in the preparation of compositions for promoting re-epithelialization or healing of skin or mucous membranes, or in the preparation of compositions for repairing the skin barrier, or in the preparation of compositions for promoting cell migration, or in the preparation of compositions for reducing skin sebum synthesis or secretion.

[0035] In this disclosure, the term "skin" should be understood as comprising its multiple layers, from the uppermost layer or stratum corneum to the lowermost layer or subcutaneous tissue, including both ends. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and / or adipocytes. In this disclosure, the term "skin" includes the scalp.

[0036] This disclosure has the following advantages and effects: 1. The kudzu root fermentation product disclosed herein can effectively inhibit oil secretion and promote cell migration, and has the effects of oil control and repair.

[0037] 2. This disclosure achieves complementary metabolic advantages of different fermentation strains through the sequential synergy of anaerobic and aerobic fermentation. Combined with the metabolic transformation of zinc ions by the fermentation strains, it ensures the stable production of kudzu fermentation products with unique active ingredients disclosed in this disclosure.

[0038] 3. The fermentation process disclosed herein can produce kudzu root fermentation products in just 2 days, which has the advantages of short fermentation time, fast output and low cost. Attached Figure Description

[0039] Figure 1 This is an HPLC chromatogram overlay of kudzu root fermentation product A from Example 1 of this disclosure, kudzu root extract from Comparative Example 3, and puerarin. From top to bottom, they are puerarin, kudzu root extract, and kudzu root fermentation product A. Detailed Implementation

[0040] To make the objects, features, and advantages of this disclosure more apparent and understandable, the disclosure will be further described in detail below with reference to the accompanying drawings and embodiments. Obviously, the described embodiments are only some, not all, of the embodiments of this disclosure. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without inventive effort are within the scope of protection of the appended claims.

[0041] The kudzu (Puerariae Lobatae Radix) used in this disclosure is produced in Luofu Mountain, Sichuan.

[0042] The cellulase used in this disclosure was purchased from Henan Nayue Biotechnology Co., Ltd., and its enzyme activity is ≥100000U / g.

[0043] The pectinase used in this disclosure was purchased from Shandong Longket Enzyme Preparation Co., Ltd., with an enzyme activity ≥30000U / g.

[0044] The α-amylase used in this disclosure was purchased from Shanghai Maclean Biotechnology Co., Ltd., with an enzyme activity ≥40000U / g.

[0045] The neutral protease used in this disclosure was purchased from Shanghai Maclean Biotechnology Co., Ltd., with an enzyme activity ≥50000U / g.

[0046] The Bifidobacterium adolescentis used in this disclosure ( Bifidobacterium adolescentis Purchased from Guangdong Provincial Center for Microbial Culture Collection, accession number GDMCC NO. 1.1262.

[0047] The brewing yeast used in this disclosure ( Saccharomyces cerevisiae Purchased from Guangdong Provincial Center for Microbial Culture Collection, accession number GDMCC NO. 2.243.

[0048] The compound lactic acid bacteria used in this disclosure were purchased from Shandong Zhongke Jiayi Biotechnology Co., Ltd., with product number Novum CHMO15, and consist of Bifidobacterium lactis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus plantarum.

[0049] The Bifidobacterium culture medium used in this disclosure was purchased from Qingdao Haibo Biotechnology Co., Ltd., product number HB8527-1.

[0050] Unless otherwise specified, all experimental reagents and materials used in this disclosure are commercially available. Example 1

[0051] A fermented kudzu root product is obtained through the following steps: S1. Crush kudzu root slices and pass them through a 40-mesh sieve. Mix the kudzu root powder with distilled water at a solid-liquid ratio (g / mL) of 1:20. Add α-amylase, cellulase, neutral protease, and pectinase. Place the mixture in a shaker at 50℃ and 200rpm for 3 hours for enzymatic hydrolysis to obtain the kudzu root treated product. The weight ratio of each enzyme relative to the sieved kudzu root powder is 0.1%.

[0052] S2. Inoculate the plate-activated Bifidobacterium adolescentis seed culture into Bifidobacterium liquid medium, place it in an anaerobic culture bag equipped with an anaerobic gas generator, seal the anaerobic culture bag, and culture in a shaker at 37℃ and 150 rpm for 24 h to obtain Bifidobacterium adolescentis seed culture. Inoculate the plate-activated Saccharomyces cerevisiae seed culture into YM liquid medium and culture in a shaker at 28℃ and 150 rpm for 16 h to obtain Saccharomyces cerevisiae seed culture. Inoculate the compound lactic acid bacteria powder into MRS liquid medium and culture in a shaker at 37℃ and 150 rpm for 16 h to obtain compound lactic acid bacteria seed culture.

[0053] S3. Add glucose powder and zinc sulfate heptahydrate to the kudzu root treatment material from step S1 to prepare a liquid fermentation medium containing zinc ions, and then autoclave at 121°C for 15 minutes. The concentration of glucose in the liquid fermentation medium is 10 g / L, and the concentration of zinc sulfate heptahydrate is 1 g / L.

[0054] S4. Place the sterilized liquid fermentation medium from step S3 into a fermenter, purge with sterile nitrogen to remove dissolved oxygen, inoculate with *Bifidobacterium adolescentis* seed culture, and seal the fermenter for anaerobic fermentation. The inoculation amount of *Bifidobacterium adolescentis* seed culture is 3% (v / v), the fermentation temperature is controlled at 34~37℃, and the fermentation time is 16 hours.

[0055] After fermentation in step S4 (S5), open the fermenter and introduce sterile air into the liquid fermentation medium to wash away dissolved nitrogen. Once the dissolved oxygen saturation in the liquid fermentation medium reaches 100%, inoculate with *Saccharomyces cerevisiae* seed culture and *Lactobacillus thuringiensis* seed culture for aerobic fermentation. During aerobic fermentation, continuously monitor and adjust the dissolved oxygen saturation in the liquid fermentation medium to ensure it remains within the range of 70%–100%. The inoculation amount of both *Saccharomyces cerevisiae* seed culture and *Lactobacillus thuringiensis* seed culture is 3% (v / v), the fermentation temperature is controlled at 28–32℃, and the fermentation time is 16 hours.

[0056] S6. After fermentation in step S5 is completed, the fermentation broth is centrifuged and filtered. The supernatant is pasteurized and then filtered through a 0.22μm aqueous filter membrane to obtain the kudzu fermentation product (hereinafter referred to as kudzu fermentation product A). Comparative Example 1

[0057] A fermented kudzu root product is obtained through the following steps: S1. Crush kudzu root slices and pass them through a 40-mesh sieve. Mix the kudzu root powder with distilled water at a solid-liquid ratio (g / mL) of 1:20. Add α-amylase, cellulase, neutral protease, and pectinase. Place the mixture in a shaker at 50℃ and 200rpm for 3 hours for enzymatic hydrolysis to obtain the kudzu root treated product. The weight ratio of each enzyme relative to the sieved kudzu root powder is 0.1%.

[0058] S2. Inoculate the plate-activated Bifidobacterium adolescentis seed culture into Bifidobacterium liquid medium, place it in an anaerobic culture bag equipped with an anaerobic gas generator, seal the anaerobic culture bag, and culture in a shaker at 37℃ and 150 rpm for 24 h to obtain Bifidobacterium adolescentis seed culture. Inoculate the plate-activated Saccharomyces cerevisiae seed culture into YM liquid medium and culture in a shaker at 28℃ and 150 rpm for 16 h to obtain Saccharomyces cerevisiae seed culture. Inoculate the compound lactic acid bacteria powder into MRS liquid medium and culture in a shaker at 37℃ and 150 rpm for 16 h to obtain compound lactic acid bacteria seed culture.

[0059] S3. Add glucose powder to the kudzu root treatment material from step S1 to prepare a liquid fermentation medium, and then autoclave at 121°C for 15 minutes. The concentration of glucose in the liquid fermentation medium is 10 g / L.

[0060] S4. Place the sterilized liquid fermentation medium from step S3 into a fermenter, purge with sterile nitrogen to remove dissolved oxygen, inoculate with *Bifidobacterium adolescentis* seed culture, and seal the fermenter for anaerobic fermentation. The inoculation amount of *Bifidobacterium adolescentis* seed culture is 3% (v / v), the fermentation temperature is controlled at 34~37℃, and the fermentation time is 16 hours.

[0061] After fermentation in step S4 (S5), open the fermenter and introduce sterile air into the liquid fermentation medium to wash away dissolved nitrogen. Once the dissolved oxygen saturation in the liquid fermentation medium reaches 100%, inoculate with *Saccharomyces cerevisiae* seed culture and *Lactobacillus thuringiensis* seed culture for aerobic fermentation. During aerobic fermentation, continuously monitor and adjust the dissolved oxygen saturation in the liquid fermentation medium to ensure it remains within the range of 70%–100%. The inoculation amount of both *Saccharomyces cerevisiae* seed culture and *Lactobacillus thuringiensis* seed culture is 3% (v / v), the fermentation temperature is controlled at 28–32℃, and the fermentation time is 16 hours.

[0062] S6. After fermentation in step S5 is completed, the fermentation broth is centrifuged and filtered. The supernatant is pasteurized and then filtered through a 0.22μm aqueous filter membrane to obtain the kudzu fermentation product (hereinafter referred to as kudzu fermentation product B). Comparative Example 2

[0063] A fermented kudzu root product is obtained through the following steps: S1. Crush kudzu root slices and pass them through a 40-mesh sieve. Mix the kudzu root powder with distilled water at a solid-liquid ratio (g / mL) of 1:20. Add α-amylase, cellulase, neutral protease, and pectinase. Place the mixture in a shaker at 50℃ and 200rpm for 3 hours for enzymatic hydrolysis to obtain the kudzu root treated product. The weight ratio of each enzyme relative to the sieved kudzu root powder is 0.1%.

[0064] S2. Inoculate the plate-activated Bifidobacterium adolescentis seed culture into Bifidobacterium liquid medium, place it in an anaerobic culture bag equipped with an anaerobic gas generator, seal the anaerobic culture bag, and culture in a shaker at 37℃ and 150 rpm for 24 h to obtain Bifidobacterium adolescentis seed culture. Inoculate the plate-activated Saccharomyces cerevisiae seed culture into YM liquid medium and culture in a shaker at 28℃ and 150 rpm for 16 h to obtain Saccharomyces cerevisiae seed culture. Inoculate the compound lactic acid bacteria powder into MRS liquid medium and culture in a shaker at 37℃ and 150 rpm for 16 h to obtain compound lactic acid bacteria seed culture.

[0065] S3. Add glucose powder and zinc sulfate heptahydrate to the kudzu root treatment material from step S1 to prepare a liquid fermentation medium containing zinc ions, and then autoclave at 121°C for 15 minutes. The concentration of glucose in the liquid fermentation medium is 10 g / L, and the concentration of zinc sulfate heptahydrate is 1 g / L.

[0066] S4. Place the sterilized liquid fermentation medium from step S3 into a fermenter, and introduce sterile air until the dissolved oxygen saturation in the liquid fermentation medium reaches 100%. Then, inoculate with brewer's yeast seed liquid and compound lactic acid bacteria seed liquid for aerobic fermentation. During aerobic fermentation, continuously monitor and adjust the dissolved oxygen saturation in the liquid fermentation medium to ensure that it is within the range of 70% to 100%. The inoculation amount of both brewer's yeast seed liquid and compound lactic acid bacteria seed liquid is 3% (v / v), the fermentation temperature is controlled at 28~32℃, and the fermentation time is 16 hours.

[0067] After fermentation in step S4 (S5), sterile nitrogen gas is introduced into the liquid fermentation medium to wash away dissolved oxygen. The *Bifidobacterium adolescentis* seed culture is then inoculated, and the fermenter is sealed for anaerobic fermentation. The inoculation amount of the *Bifidobacterium adolescentis* seed culture is 3% (v / v), the fermentation temperature is controlled at 34-37℃, and the fermentation time is 16 hours.

[0068] S6. After fermentation in step S5 is completed, the fermentation broth is centrifuged and filtered. The supernatant is pasteurized and then filtered through a 0.22μm aqueous filter membrane to obtain the kudzu fermentation product (hereinafter referred to as kudzu fermentation product C). Comparative Example 3

[0069] A kudzu root extract is prepared by the following steps: Pulverized kudzu root slices were passed through a 40-mesh sieve. Kudzu root powder was then mixed with distilled water at a solid-liquid ratio of 1:20 (g / mL). The mixture was refluxed at 100℃ for 2 hours, filtered, and the filtrate was filtered through a 0.22μm aqueous filter membrane to obtain kudzu root extract.

[0070] Application Example 1: Kudzu Root Fermented Extract A kudzu root fermented extract is prepared according to the following steps: Table 1. Kudzu Root Fermented Extract According to the formula in Table 1, after mixing phase A, heat to 60℃ to dissolve and stir evenly, then cool to room temperature and add phase B raw materials to obtain the final product. SC-GLUCAN HD is a commercially available product composed of water, glycerin, 1,2-hexanediol, 1,2-pentanediol, and β-glucan. Test Example 1

[0071] Using puerarin standard as a reference, high performance liquid chromatography was used to analyze pueraria fermentation product A and pueraria extract of Comparative Example 3, including the following steps: Instruments: Shimadzu high performance liquid chromatography system (model: LC-20AT), SPD-M20A diode array detector.

[0072] Preparation of the test solution: Take an appropriate amount of this product, accurately weigh it, place it in a volumetric flask, add 50% methanol to dissolve it and prepare a solution containing 0.1g per 1mL, shake well, filter, and the solution is obtained.

[0073] Preparation of puerarin reference solution: Accurately weigh about 10 mg of reference standard, place it in a 20 mL volumetric flask, add an appropriate amount of 50% methanol to dissolve it, dilute to the mark, and shake well to obtain the solution.

[0074] The chromatographic conditions are as follows: Column: Octadecylsilane-bonded silica gel as the packing material (Ultimate AQ-C18, 4.6×250mm, 5μm, or a column with equivalent performance). Column temperature: 30℃; Injection volume: 10 μL; Detection wavelength: 250nm; Mobile phase A: 0.1% aqueous phosphoric acid solution (v / v); Mobile phase B: methanol; Flow rate: 1 mL / min; Elution gradient: 0~15min: 75%A, 25%B; 15~30min: 75%→55%A, 25%→45%B; 30~35min: 55%→10%A, 45%→90%B; 35~36min: 10%→75%A, 90%→25%B; 36~45min: 75%A, 25%B.

[0075] Assay: Accurately pipette 10 μL each of the diluent (50% methanol), reference solution, and test solution into the high-performance liquid chromatograph (HPLC), and record the HPLC chromatograms, as shown below. Figure 1 As shown.

[0076] Figure 1 The results showed that the retention time of the characteristic peak of puerarin was approximately 20 min. Both kudzu fermentation product A and kudzu extract exhibited characteristic peaks at retention times of approximately 20 min, and these peaks highly overlapped with the characteristic peak of puerarin, confirming that these peaks were indeed characteristic peaks of puerarin in kudzu fermentation product A and kudzu extract. On the other hand, kudzu fermentation product A showed UV absorption characteristic peaks at retention times of approximately 10 min and 27.5 min, while kudzu extract did not show significant UV absorption characteristic peaks at the same retention times. This indicates that compared to kudzu extract, kudzu fermentation product A contains at least two additional components, and the total chemical composition of kudzu fermentation product A is significantly different from that of kudzu extract. The substance corresponding to the characteristic peak at a retention time of approximately 10 min is designated as component A, and the substance corresponding to the characteristic peak at a retention time of approximately 27.5 min is designated as component B.

[0077] Four groups of kudzu root fermentation product A samples were prepared repeatedly according to the preparation method of Example 1, and analyzed under the above chromatographic conditions. The presence of components A and B was found in all four groups. Therefore, under the above chromatographic conditions, kudzu root fermentation product A has three common characteristic peaks. Integral analysis was performed on the characteristic peaks of components A, B, and puerarin, with the integration parameters set as follows: Half-peak width: 5 sec; Slope: 1000 µV / min; Drift: 0 µV / min; Variable parameter time: 1000 min; Minimum area: 1000 count.

[0078] Using the characteristic peak of puerarin as a reference peak, the analytical results of the high-performance liquid chromatogram of kudzu fermentation product A are shown in Table 2.

[0079] Table 2. Common characteristic peak information of high performance liquid chromatogram of kudzu root fermentation product A Table 2 shows that the relative retention times of components A and B are basically the same, but the relative peak areas of component A range from 1.80% to 4.34%, while those of component B range from 4.18% to 21.03%, exhibiting significant fluctuations. During aerobic fermentation, the dissolved oxygen saturation of the liquid fermentation medium is not a fixed value. Fluctuations in dissolved oxygen levels make the fermentation system unstable, affecting the metabolic activities of the fermentation strains and thus causing significant fluctuations in the relative contents of components A and B. This further confirms that components A and B are both new substances produced by kudzu root after fermentation.

[0080] Test Example 2: Oil Content Test 2.1 Reagents and Materials 0.25% trypsin digestion solution (prepared by mixing 0.25g trypsin with 100mL water), PBS, complete culture medium, FFA (prepared by mixing linoleic acid and palmitic acid in a molar ratio of 1:1), and Oil Red O staining kit.

[0081] 2.2 Instruments Constant temperature CO2 incubator, clean bench, microplate reader.

[0082] 2.3 Cell lines Human sebaceous gland cells SZ-95.

[0083] 2.4 Samples to be tested and grouping 2.4.1 Sample to be tested Kudzu root fermentation product A, kudzu root fermentation product B, kudzu root fermentation product C, and kudzu root extract of Comparative Example 3 were diluted with PBS and tested at concentrations of 1% (v / v) and 2% (v / v). Isotretinoin, tested at a concentration of 0.05 μM.

[0084] 2.4.2 Grouping Experimental group: Sample to be tested, FFA; Blank control group: PBS; Model groups: PBS, FFA.

[0085] 2.5 Experimental Methods Take one flask of SZ-95 cells in good exponential growth phase, add 0.25% trypsin digestion solution, digest to detach the adherent cells, and count (1~4)×10⁻⁶ cells. 5 Cells were cultured at a concentration of [number] cells / mL to prepare a cell suspension. An appropriate amount of the cell suspension was seeded into a 12-well plate containing complete culture medium and incubated in a CO2 incubator for 24 h. Except for the blank control group which received PBS, each well in the other wells was treated with FFA to achieve a final concentration of 225 μmol / L. Simultaneously with cell modeling induction, the experimental groups received the corresponding test samples, while the model group received the same amount of PBS. All wells were incubated in a CO2 incubator for 48 h. The culture medium was then discarded, and staining was performed according to the Oil Red O staining kit instructions.

[0086] Discard the solution in the well plate, add isopropanol to dissolve the Oil Red O staining solution, and measure the absorbance at 490 nm using a microplate reader. Calculate the relative content of lipid secretion from the cells based on the absorbance values.

[0087] 2.6 Experimental Results Oil Red O is a fat-soluble dye that is highly soluble in fat. Its staining principle is that Oil Red O specifically adsorbs onto neutral triglycerides, lipids, and lipoproteins in tissues and cells, thus staining the fat. Free-floxacin (FFA) is an inducer; under FFA stimulation, SZ-95 cells secrete large amounts of oil that can be stained by Oil Red O. This experiment used test samples to treat FFA-induced SZ-95 cells, and by detecting the amount of oil produced by SZ-95 cells, determined whether the test sample could inhibit the secretion of oil by human sebaceous gland cells.

[0088] The results of the effects of the test samples on lipid secretion in SZ-95 cells are shown in Table 3.

[0089] Table 3. Relative lipid secretion of SZ-95 cells (Mean±SD) Note: Compared with the blank control group ### P<0.001; compared with the model group, *** P <0.001.

[0090] Experimental results showed that, compared to the model group, isotretinoin significantly reduced sebum secretion in SZ-95 cells, proving that the experiments disclosed in this paper can effectively verify the effect of the test samples on sebum secretion in SZ-95 cells. Compared to the model group, kudzu fermentation product A, kudzu fermentation product B, kudzu fermentation product C, and kudzu extract all significantly inhibited the synthesis and secretion of sebum by sebaceous gland cells, slowing down sebum deposition. In particular, compared to kudzu extract, kudzu fermentation product A showed a more significant effect in inhibiting sebum secretion. P <0.05%, demonstrating that the kudzu fermentation product A disclosed herein has a superior effect in inhibiting sebum secretion. However, compared to kudzu extract, 1% and 2% kudzu fermentation products B and C both showed inferior effects in inhibiting sebum secretion; compared to kudzu fermentation products B and C, 1% and 2% kudzu fermentation product A both showed more significant effects in inhibiting sebum secretion. P The result (<0.01) demonstrates that fermentation of kudzu root may not necessarily enhance its ability to inhibit sebum secretion, and the effects of fermentation conditions and procedures on oil control are unpredictable. This disclosure utilizes the synergistic effect of zinc ions and a specific fermentation procedure to obtain a kudzu root fermentation product with superior oil control properties. This product can significantly inhibit skin sebum synthesis or secretion and can be used to improve problems such as excessive oil production and skin water-oil imbalance.

[0091] Test Example 3: Cell Migration Test 3.1 Reagents and Materials 0.25% trypsin digestion solution (prepared by using 0.25g trypsin with 100mL of water), complete culture medium, PBS.

[0092] 3.2 Instruments Biological inverted microscope, constant temperature CO2 incubator, ultra-clean workbench.

[0093] 3.3 Cell lines Human keratinocytes (HaCaT).

[0094] 3.4 Samples to be tested and grouping 3.4.1 Sample to be tested Kudzu root fermentation product A and kudzu root extract of Comparative Example 3 were diluted with PBS, and the test concentrations were both 2% (v / v).

[0095] 3.4.2 Grouping Sample group: Samples to be tested; Blank control group: PBS.

[0096] 3.5 Experimental Methods One flask of cells in the exponential growth phase was digested with 0.25% trypsin solution to prepare a cell suspension. This suspension was then seeded into a 12-well plate containing complete culture medium and incubated in a CO2 incubator for 24 hours. The medium was changed, and two vertical lines were drawn in the center of each well using a yellow pipette tip. The cells were washed once with PBS, and photographs were taken at the positions marked above and below the horizontal lines. After photographing, complete culture medium containing 2% FBS and the test sample were added to the sample group; the blank control group was added to complete culture medium containing 2% FBS and PBS. The plates were then incubated in a CO2 incubator for 24 hours. After 24 hours of incubation, photographs were taken at the same locations using an inverted biological microscope, and the area of ​​the scratches was calculated.

[0097] Scratch healing rate (%) = (Scratch area before sample application - Scratch area after sample application) / Scratch area before sample application × 100%.

[0098] 3.6 Experimental Results Cell migration is one of the basic functions of normal cells, a physiological process of normal growth and development, and a common form of movement among living cells. Cell migration is similar to the skin wound healing process; therefore, promoting cell migration has a positive effect on skin repair.

[0099] The results of the effects of the test samples on cell migration are shown in Table 4.

[0100] Table 4. Effects of test samples on cell migration Group Scratch healing rate (Mean±SD) Blank control group 18.89%±4.01% Kudzu Fermentation Products Group A 37.73%±2.19%*** Kudzu root extract group 27.17%±6.48% Note: Compared with the blank control group, *** P <0.001.

[0101] The results showed that the kudzu root fermentation product A disclosed herein significantly promoted keratinocyte migration, exhibiting a significant effect in promoting skin or mucous membrane re-epithelialization or wound healing, and repairing the skin barrier, thereby achieving a repairing effect. Compared with the blank control group, the effect of kudzu root extract on cell migration was not statistically significant, indicating that it could not effectively promote keratinocyte migration. The above results demonstrate that the kudzu root fermentation product obtained after fermentation in this disclosure produces a new substance that can promote cell migration and has a repairing effect.

[0102] In this disclosure, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or terminal device that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or terminal device. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or terminal device that includes said element.

[0103] While specific embodiments of this disclosure have been described for illustrative purposes, various modifications or alterations can be made by those skilled in the art without departing from the spirit and scope of this disclosure. All such modifications or alterations should fall within the scope of the appended claims.

Claims

1. A fermented kudzu root product, characterized in that, Its high-performance liquid chromatography (HPLC) chromatogram includes at least three characteristic peaks, with the characteristic peak of puerarin as a reference peak. The relative retention times of the three characteristic peaks and the reference peak are 0.49~0.50, 1.00, and 1.36~1.38, respectively. The HPLC conditions are as follows: Chromatographic column: 4.6*250mm, 5μm, packed with octadecylsilane-bonded silica gel; Column temperature: 30℃; Injection volume: 10 μL; Detection wavelength: 250nm; Mobile phase A: 0.1% aqueous phosphoric acid solution; Mobile phase B: methanol; Flow rate: 1 mL / min; Elution gradient: 0~15min: 75%A, 25%B; 15~30min: 75%→55%A, 25%→45%B; 30~35min: 55%→10%A, 45%→90%B; 35~36min: 10%→75%A, 90%→25%B; 36~45min: 75%A, 25%B.

2. The kudzu root fermentation product according to claim 1, characterized in that, The chromatographic column is an AQ-C18 column, which can be an Ultimate AQ-C18 column or a column with equivalent performance.

3. The kudzu root fermentation product according to claim 1, characterized in that, According to the order of retention time, the relative peak areas of the three characteristic peaks and the reference peak are 1.80%~4.34%, 100%, and 4.18%~21.03%, respectively.

4. A method for preparing kudzu root fermentation products, characterized in that, Includes the following steps: The kudzu root raw material is pretreated to obtain kudzu root treated product; the kudzu root treated product is mixed with a carbon source and a zinc ion source is added to prepare a liquid fermentation culture medium containing zinc ions; first, Bifidobacterium adolescentis is inoculated into the liquid fermentation culture medium for anaerobic fermentation, and then Saccharomyces cerevisiae and compound lactic acid bacteria are inoculated for aerobic fermentation to obtain the kudzu root fermentation product.

5. The preparation method according to claim 4, characterized in that, In the anaerobic fermentation step, the fermentation temperature is 34~37℃ and the fermentation time is 16~24h.

6. The preparation method according to claim 4, characterized in that, In the aerobic fermentation step, the fermentation temperature is 28~32℃, the fermentation time is 16~24h, and the dissolved oxygen saturation in the liquid fermentation medium is maintained at 70%~100%.

7. The preparation method according to claim 4, characterized in that, The compound lactic acid bacteria include Bifidobacterium lactis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus plantarum.

8. A fermented product of kudzu root, characterized in that, The kudzu fermentation product is prepared by the preparation method according to any one of claims 4-7.

9. A composition, characterized in that, It contains an effective amount of the kudzu fermentation product according to any one of claims 1-3, or an effective amount of the kudzu fermentation product according to claim 8, and at least one additional ingredient.

10. Use of the kudzu fermentation product according to any one of claims 1-3 or the kudzu fermentation product according to claim 8 in the preparation of a composition for skin or mucous membrane care.

11. Use of the kudzu fermentation product according to any one of claims 1-3 or the kudzu fermentation product according to claim 8 in the preparation of compositions for repair or oil control.

12. Use of the kudzu fermentation product according to any one of claims 1-3 or the kudzu fermentation product according to claim 8 in the preparation of a composition for promoting re-epithelialization or healing of skin or mucous membranes, or in the preparation of a composition for repairing the skin barrier, or in the preparation of a composition for promoting cell migration, or in the preparation of a composition for reducing skin sebum synthesis or secretion.

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