Fermented lactobacillus mucus A27-1, traditional Chinese medicine fermentation liquor and application

Abstract

The application provides a fermented lactobacillus mucus A27-1, a traditional Chinese medicine fermentation liquor and application, and belongs to the technical field of biological medicine and microbial fermentation. The preservation number of the fermented lactobacillus mucus A27-1 is CGMCC No. 37341. The traditional Chinese medicine compound liquid is fermented by using the fermented lactobacillus mucus A27-1, and white Candida albicans, Staphylococcus aureus and Propionibacterium acnes can be significantly inhibited; the obtained traditional Chinese medicine fermentation liquor can further improve the ability of the traditional Chinese medicine compound liquid to inhibit white Candida albicans, Staphylococcus aureus and Propionibacterium acnes, the ability to resist skin inflammation and the ability to resist oxidation, and therefore can be used for preparing a skin pathogenic bacteria inhibiting drug or a skin care product.

Description

Technical Field

[0001] This invention belongs to the fields of biomedicine and microbial fermentation technology, and particularly relates to a fermentation of Lactobacillus mucinus A27-1, a fermentation broth for traditional Chinese medicine and its application. Background Technology

[0002] The skin is the body's primary organ and the first line of defense against external environmental influences. A close relationship exists between the human skin microbiome and skin diseases. Research has found that skin diseases may be related to changes and imbalances in the microbiome. Some bacteria (such as Staphylococcus aureus and Propionibacterium acnes) and / or fungi (such as Candida albicans) can invade and cause infection after disrupting the skin barrier, leading to skin inflammation. Candida albicans, an opportunistic pathogen, proliferates rapidly when the skin is moist, resistance is lowered, and the microbiome is imbalanced, causing infections such as itchy skin, thrush, and fungal vaginitis. Staphylococcus aureus, a Gram-positive coccus, is widely found in the air, on the skin, in the nasal cavity, and on surfaces. It is a common pathogen that can enter through broken skin or hair follicles, causing various skin infections, including folliculitis, boils, carbuncles, impetigo, and cellulitis. In severe cases, it can lead to systemic infection. Propionibacterium acnes is a pathogen that causes acne. Problems such as excessive sebum secretion, blockage of the pilosebaceous duct, abnormal proliferation of Propionibacterium acnes, and changes in hormone levels can lead to skin conditions like blackheads, pimples, pustules, and cystic pimples, commonly referred to as acne bacteria. Therefore, inhibiting skin pathogens is of great significance for the prevention and treatment of skin diseases.

[0003] The scalp is an important part of human skin. The scalp has a unique micro-ecological environment (hair, hair follicles, sweat glands, and sebaceous glands, with relatively high humidity), creating favorable conditions for microbial colonization. However, an unbalanced scalp micro-ecological environment can lead to a series of problems such as hair loss and dandruff. For example, studies have found that the diversity of scalp flora is significantly reduced in people with hair loss, including a higher proportion of Propionibacterium acnes (…). Propionibacterium acnes The abundance of these cells was significantly higher than in the normal population. This indicates that improving the unbalanced scalp micro-ecosystem and maintaining scalp health remains a pressing technical challenge.

[0004] Probiotics are beneficial bacteria for humans and animals. They utilize inorganic salts, uric acid, and urea excreted by the skin's sweat glands as growth factors and nitrogen sources, thus achieving a skin-cleansing effect. Probiotics have a potential role in maintaining skin health. However, current research on the effects of probiotics on skin health, especially on scalp health, remains limited. Summary of the Invention

[0005] Therefore, the purpose of this invention is to provide a fermented Lactobacillus mucinus A27-1, a traditional Chinese medicine fermentation broth, and its application.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a fermenting Lactobacillus mucinus A27-1, the preservation number of which is CGMCC No. 37341.

[0007] This invention provides a traditional Chinese medicine fermentation liquid, which is obtained by fermenting a traditional Chinese medicine compound liquid using the above-mentioned fermented Lactobacillus myxobolus A27-1.

[0008] Preferably, the fermentation method is to culture the food in a shaker at 35~40℃ and 150~200rpm for 22~26h; The culture medium obtained after fermentation also includes centrifugation, collection of supernatant, filtration of supernatant, and obtaining filtrate.

[0009] Preferably, the traditional Chinese medicine compound liquid comprises the following raw materials in parts by weight: Prepared Polygonum multiflorum 80-150 parts, Sapindus mukorossi 80-150 parts, Artemisia argyi 60-120 parts, Angelica sinensis 60-120 parts, Ganoderma lucidum 60-120 parts, Gleditsia sinensis 60-120 parts, Mentha haplocalyx 60-120 parts, Sophora flavescens 60-120 parts, Smilax glabra 60-120 parts, Astragalus membranaceus 60-120 parts, Rehmannia glutinosa 60-120 parts, Ligustrum lucidum 60-120 parts, Clematis chinensis 60-120 parts, Carthamus tinctorius 60-120 parts, Platycladus orientalis 60-120 parts, Psoralea corylifolia 40-100 parts, Eclipta prostrata 150-200 parts, and tea seed cake 60-120 parts.

[0010] Preferably, the preparation method of the traditional Chinese medicine compound liquid includes: The prepared Polygonum multiflorum, Sapindus mukorossi, Artemisia argyi, Angelica sinensis, Ganoderma lucidum, Gleditsia sinensis, Mentha haplocalyx, Sophora flavescens, Smilax glabra, Astragalus membranaceus, Rehmannia glutinosa, Ligustrum lucidum, Clematis chinensis, Carthamus tinctorius, Platycladus orientalis, Psoralea corylifolia, Eclipta prostrata, and tea seed cake are pulverized, sieved, and mixed to obtain a mixture. The mixture is then mixed with water, soaked, and boiled over high heat while stirring. It is then simmered over low heat, filtered, and the filtrate is collected to obtain a traditional Chinese medicine compound liquid.

[0011] Preferably, the mass ratio of the mixture to the volume of water is 1g:8~15mL; the soaking method is soaking at 20~25℃ for 1~3h; the stirring rate is 80~200rpm / min; the boiling time is 1.5~2.5h; the decocting is performed 1~3 times, and each decocting time is 2~4h.

[0012] This invention provides an application of the above-mentioned traditional Chinese medicine fermentation liquid in at least one of the following (1) to (7): (1) Inhibits skin pathogens; (2) Preparation of drugs to inhibit skin pathogens; (3) Preparation of antioxidants; (4) Preparation of drugs to inhibit skin inflammation; (5) Prepare skin care products; (6) Enhance the ability of the traditional Chinese medicine compound solution to inhibit skin pathogens; (7) Prepare drugs that enhance the ability of traditional Chinese medicine compound liquid to inhibit skin pathogens.

[0013] Preferably, the pathogenic bacteria include one or more of Candida albicans, Staphylococcus aureus, and Propionibacterium acnes.

[0014] This invention provides a method for increasing the content of metabolites in a traditional Chinese medicine compound solution, wherein the traditional Chinese medicine compound solution is the aforementioned traditional Chinese medicine compound solution; The method includes the step of fermenting a traditional Chinese medicine compound solution using the aforementioned fermented Lactobacillus mucinus A27-1.

[0015] Preferably, the metabolites include one or more of hexadecylsphingosine, oxymatrine, shikonin, gallic acid, (R)-red acid, D-(+)-phenyllactic acid, and azelaic acid.

[0016] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a fermented Lactobacillus mucinus A27-1, a traditional Chinese medicine fermentation broth, and its applications. The fermented traditional Chinese medicine compound broth using the Lactobacillus mucinus A27-1 significantly inhibits Candida albicans, Staphylococcus aureus, and Propionibacterium acnes. The resulting fermented traditional Chinese medicine broth further enhances the ability of the traditional Chinese medicine compound broth to inhibit Candida albicans, Staphylococcus aureus, and Propionibacterium acnes, as well as its anti-skin inflammation and antioxidant capabilities. Therefore, it can be used to prepare drugs that inhibit skin pathogens or to prepare skin care products.

[0017] Biological preservation information: The fermenting myxobacterium is named A27-1 and classified as *Lactobacillus fermentatus*. Lactobacillus fermentum It was deposited on January 9, 2026 at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 37341. Attached Figure Description

[0018] Figure 1 Morphological image of strain A27-1 (×1000x).

[0019] Figure 2 This is a phylogenetic tree diagram of strain A27-1.

[0020] Figure 3The changes in DPPH activity and superoxide anion free radical scavenging capacity of the traditional Chinese medicine fermentation broth A27-1 are shown in Figure 1. A represents the DPPH free radical scavenging capacity of the traditional Chinese medicine fermentation broth A27-1; B represents the superoxide anion free radical scavenging capacity of the traditional Chinese medicine fermentation broth A27-1. Different letters indicate significant differences between different groups (p < 0.05), while the same letter indicates no significant differences between different groups (p > 0.05).

[0021] Figure 4 The results show the antibacterial activity of traditional Chinese medicine fermentation broth A27-1 against Candida albicans. A represents the antibacterial activity of traditional Chinese medicine fermentation broth A27-1 against clinical Candida albicans 115, and B represents the antibacterial activity of traditional Chinese medicine fermentation broth A27-1 against standard Candida albicans ATCC10231. Different letters indicate significant differences between different groups (p < 0.05), while the same letter indicates no significant differences between different groups (p > 0.05).

[0022] Figure 5 The results show the ability of fermentation broth A27-1 to inhibit Staphylococcus aureus. Different letters indicate significant differences between different groups (p < 0.05), while the same letter indicates no significant differences between different groups (p > 0.05).

[0023] Figure 6 The results show the ability of fermented Chinese medicine A27-1 to inhibit Propionibacterium acnes. Different letters indicate significant differences between different groups (p < 0.05), while the same letter indicates no significant differences between different groups (p > 0.05).

[0024] Figure 7 The effect of treating traditional Chinese medicine compound solution and fermentation broth obtained from A27-1 fermentation on IL-6 mRNA expression in HaCaT cells for 1 h.

[0025] Figure 8 This is a bar chart showing the top 20 metabolites by abundance in different groups under positive ion mode.

[0026] Figure 9 The results show the screening of scalp care potential metabolites in different groups under positive ion mode.

[0027] Figure 10 This is a bar chart showing the top 20 metabolites by abundance in different groups under negative ion mode.

[0028] Figure 11 The results show the screening of potential metabolites for scalp care in different groups under negative ion mode. Detailed Implementation

[0029] This invention provides a fermenting Lactobacillus mucinus A27-1, the preservation number of which is CGMCC No. 37341.

[0030] The fermented Lactobacillus mucinus A27-1 strain of this invention is derived from traditional fermented pickled vegetables in Guizhou Province. It is a Gram-positive bacillus, and the A27-1 strain has an anti-Helicobacter pylori effect.

[0031] This invention provides a traditional Chinese medicine fermentation liquid, which is obtained by fermenting a traditional Chinese medicine compound liquid using the above-mentioned fermented Lactobacillus myxobolus A27-1.

[0032] In this invention, the fermentation method is to culture at 35-40℃ and 150-200 rpm in a shaker for 22-26 hours, more preferably at 36-38℃ and 160-190 rpm in a shaker for 23-25 ​​hours, and even more preferably at 37℃ and 180 rpm in a shaker for 24 hours. When fermenting the traditional Chinese medicine compound liquid, the fermented *Lactobacillus mucinus* A27-1 is a bacterial suspension of *Lactobacillus mucinus* A27-1, and the colony concentration of the bacterial suspension is 1×10⁻⁶. 8 The volume ratio of the *Lactobacillus fermentatus* A27-1 bacterial suspension to the traditional Chinese medicine compound solution is 3-5:100, more preferably 3.5-4.5:100, and even more preferably 4:100. The preparation method of the *Lactobacillus fermentatus* A27-1 bacterial suspension includes culturing thawed *Lactobacillus fermentatus* A27-1 on MRS agar plates at 35-40℃ for 12-24 hours to obtain cultured *Lactobacillus fermentatus*. Colonies of the cultured *Lactobacillus fermentatus* are inoculated into MRS broth medium and cultured at 35-40℃ and 150-200 rpm for 22-26 hours. The resulting culture is then inoculated into MRS broth medium at a volume percentage of 3%-5% and cultured and activated at 35-40℃ and 150-200 rpm to obtain the *Lactobacillus fermentatus* A27-1 bacterial suspension.

[0033] In this invention, after fermenting the traditional Chinese medicine compound liquid using *Lactobacillus myxitis* A27-1, the culture broth obtained after fermentation further includes centrifugation, collection of the supernatant, and filtration of the supernatant to obtain a filtrate. The centrifugation can be performed at 4°C and 5000 rpm for 10 minutes. The filtration is done using a 0.22 µm filter. The obtained filtrate is the traditional Chinese medicine fermentation broth.

[0034] In this invention, the herbal compound liquid comprises the following raw materials in parts by weight: 80-150 parts of processed Polygonum multiflorum, 80-150 parts of Sapindus mukorossi, 60-120 parts of Artemisia argyi, 60-120 parts of Angelica sinensis, 60-120 parts of Ganoderma lucidum, 60-120 parts of Gleditsia sinensis, 60-120 parts of Mentha haplocalyx, 60-120 parts of Sophora flavescens, 60-120 parts of Smilax glabra, 60-120 parts of Astragalus membranaceus, 60-120 parts of Rehmannia glutinosa, 60-120 parts of Ligustrum lucidum, 60-120 parts of Clematis chinensis, 60-120 parts of Carthamus tinctorius, 60-120 parts of Platycladus orientalis, 40-100 parts of Psoralea corylifolia, 150-200 parts of Eclipta prostrata, and 60-120 parts of tea seed cake.

[0035] In this invention, as a preferred embodiment, the herbal compound liquid is made from the following raw materials in parts by weight: 80-150 parts of processed Polygonum multiflorum, 80-150 parts of Sapindus mukorossi, 60-120 parts of Artemisia argyi, 60-120 parts of Angelica sinensis, 60-120 parts of Ganoderma lucidum, 60-120 parts of Gleditsia sinensis, 60-120 parts of Mentha haplocalyx, 60-120 parts of Sophora flavescens, 60-120 parts of Smilax glabra, 60-120 parts of Astragalus membranaceus, 60-120 parts of Rehmannia glutinosa, 60-120 parts of Ligustrum lucidum, 60-120 parts of Clematis chinensis, 60-120 parts of Carthamus tinctorius, 60-120 parts of Platycladus orientalis, 40-100 parts of Psoralea corylifolia, 150-200 parts of Eclipta prostrata, and 60-120 parts of tea seed cake. Further preferred ingredients include the following parts by weight: 100-130 parts of processed Polygonum multiflorum, 100-130 parts of Sapindus mukorossi, 70-100 parts of Artemisia argyi, 70-100 parts of Angelica sinensis, 70-100 parts of Ganoderma lucidum, 70-100 parts of Gleditsia sinensis, 70-100 parts of Mentha haplocalyx, 70-100 parts of Sophora flavescens, 70-100 parts of Smilax glabra, 70-100 parts of Astragalus membranaceus, 70-100 parts of Rehmannia glutinosa, 70-100 parts of Ligustrum lucidum, 70-100 parts of Clematis chinensis, 70-100 parts of Carthamus tinctorius, and 70-100 parts of Platycladus orientalis. 100 parts, 50-80 parts of Psoralea corylifolia, 160-180 parts of Eclipta prostrata and 70-100 parts of tea seed cake, more preferably, including the following raw materials in parts by weight: 115 parts of processed Polygonum multiflorum, 115 parts of Sapindus mukorossi, 90 parts of Artemisia argyi, 90 parts of Angelica sinensis, 90 parts of Ganoderma lucidum, 90 parts of Gleditsia sinensis, 90 parts of Mentha haplocalyx, 90 parts of Sophora flavescens, 90 parts of Smilax glabra, 90 parts of Astragalus membranaceus, 90 parts of Rehmannia glutinosa, 90 parts of Ligustrum lucidum, 90 parts of Clematis chinensis, 90 parts of Carthamus tinctorius, 90 parts of Platycladus orientalis, 70 parts of Psoralea corylifolia, 175 parts of Eclipta prostrata and 90 parts of tea seed cake.

[0036] The prepared Polygonum multiflorum is made using a black bean juice method. After processing with black bean juice, its properties become slightly warm, and its effects of tonifying the liver and kidneys, nourishing essence and blood, darkening hair, and strengthening muscles and bones are enhanced.

[0037] This invention does not have any particular limitation on the source of Polygonum multiflorum, Artemisia argyi, Angelica sinensis, Ganoderma lucidum, Gleditsia sinensis, Mentha haplocalyx, Sophora flavescens, Smilax glabra, Astragalus membranaceus, Rehmannia glutinosa, Ligustrum lucidum, Clematis chinensis, Carthamus tinctorius, Platycladus orientalis, Psoralea corylifolia, Eclipta prostrata and tea seed cake, and they are prepared using commercially available products in the field or conventional preparation methods.

[0038] In this invention, the preparation method of the traditional Chinese medicine compound liquid includes: The prepared Polygonum multiflorum, Sapindus mukorossi, Artemisia argyi, Angelica sinensis, Ganoderma lucidum, Gleditsia sinensis, Mentha haplocalyx, Sophora flavescens, Smilax glabra, Astragalus membranaceus, Rehmannia glutinosa, Ligustrum lucidum, Clematis chinensis, Carthamus tinctorius, Platycladus orientalis, Psoralea corylifolia, Eclipta prostrata, and tea seed cake are pulverized, sieved, and mixed to obtain a mixture. The mixture is then mixed with water, soaked, and boiled over high heat while stirring. It is then simmered over low heat, filtered, and the filtrate is collected to obtain a traditional Chinese medicine compound liquid.

[0039] In this invention, the mass ratio of the mixture to the volume of water is preferably 1g:8~15mL, more preferably 1g:10~12mL, and even more preferably 1g:10mL; the soaking method is preferably soaking at 20~25℃ for 1~3h, more preferably soaking at 20~25℃ for 1.5~2.5h, such as soaking at 22℃ for 2h; the stirring rate is preferably 80~200rpm / min, more preferably 100~150rpm / min, and even more preferably 120rpm / min; the boiling time is preferably 1.5~2.5h, more preferably 1.7~2.2h, such as 2h. The number of decoctions is 1~3 times, such as 2 times, and the decoction time for each time is preferably 2~4h, more preferably 2.5~3.5h, and even more preferably 3h. The mesh size of the sieve is 10~30 mesh, more preferably 15~25 mesh, and even more preferably 20 mesh. When the decoction is performed twice, the mixture is mixed with water at a mass-to-volume ratio of 1g:10mL, soaked at 22℃ for 2 hours, stirred at 120rpm / min, boiled over high heat for 2 hours, and then simmered over low heat for 3 hours to obtain the first decoction. The first filtrate and the first residue are collected. The first residue is mixed with water at a mass-to-volume ratio of 1g:9mL, boiled over high heat for 2 hours, and then simmered over low heat for 3 hours to obtain the second decoction. The second filtrate is collected, and the first and second filtrates are combined to obtain the total filtrate. This invention does not specifically limit the method of vacuum concentration; conventional vacuum concentration methods in the art can be used.

[0040] The fermented *Lactobacillus mucinus* A27-1 of this invention can survive in a traditional Chinese medicine (TCM) compound solution, and the resulting TCM fermentation broth can significantly inhibit *Candida albicans*, *Staphylococcus aureus*, and *Propionibacterium acnes*, especially enhancing the inhibitory effect of the TCM compound solution on these bacteria. The TCM fermentation broth of this invention also has antioxidant properties, further enhancing the antioxidant capacity of the TCM compound solution.

[0041] This invention provides an application of the above-mentioned traditional Chinese medicine fermentation liquid in at least one of the following (1) to (7): (1) Inhibits skin pathogens; (2) Preparation of drugs to inhibit skin pathogens; (3) Preparation of antioxidants; (4) Preparation of drugs to inhibit skin inflammation; (5) Prepare skin care products; (6) Enhance the ability of the traditional Chinese medicine compound solution to inhibit skin pathogens; (7) Prepare drugs that enhance the ability of traditional Chinese medicine compound liquid to inhibit skin pathogens.

[0042] In this invention, the pathogenic bacteria include one or more of Candida albicans, Staphylococcus aureus, and Propionibacterium acnes. The Candida albicans is a clinical strain of Candida albicans (…). Candida albicans )115 and standard strain of Candida albicans ( Candida albicans One or two of ATCC 10231. The *Staphylococcus aureus* is *Staphylococcus aureus* (…). Staphylococcus aureus ATCC25923. The Propionibacterium acnes mentioned is Propionibacterium acnes (ATCC25923). Propionibacterium acnes ATCC6919.

[0043] This invention provides a method for increasing the content of metabolites in a traditional Chinese medicine compound solution, wherein the traditional Chinese medicine compound solution is the aforementioned traditional Chinese medicine compound solution; The method includes the step of fermenting a traditional Chinese medicine compound solution using the aforementioned fermented Lactobacillus mucinus A27-1.

[0044] In this invention, the fermentation of the traditional Chinese medicine compound liquid can be carried out using the above-mentioned method for preparing traditional Chinese medicine fermentation liquid, and the specific methods will not be described in detail here.

[0045] In this invention, the metabolites include one or more of hexadecylsphingosine, oxymatrine, shikonin, gallic acid, (R)-red acid, D-(+)-phenyllactic acid, and azelaic acid.

[0046] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0047] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0048] In the following embodiments, the preparation method of the processed Polygonum multiflorum is as follows: Slice the raw Polygonum multiflorum and set aside. Take 10 kg of black beans and set aside.

[0049] (1) Making black bean juice: Take 10 kg of black beans, add water and boil twice, combine the first and second filtrates. First boiling: Boil for about 4 hours, add 60 kg of water, filter to obtain the first filtrate and bean residue, boil down to about 15 kg of juice (first filtrate). Second boiling: Boil the bean residue for about 3 hours, add 40 kg of water, filter to obtain the second filtrate, boil down to about 10 kg of juice (second filtrate). Total: about 25 kg of black bean juice is obtained.

[0050] (2) Soaking or mixing: Mix 100kg of raw Polygonum multiflorum slices with 25kg of black bean juice and let it soak until the juice is absorbed.

[0051] (3) Steaming / stewing: Steamed Polygonum multiflorum was obtained by high-pressure steaming at 121℃ for 3 hours.

[0052] (4) Sun drying: Spread out the steamed Polygonum multiflorum to dry completely, avoiding exposure to the sun to prevent cracking.

[0053] (5) Cyclic operation: Repeat steps (3) and (4) to steam and dry 5 times until the inside and outside are black and the texture is soft, thus obtaining processed Polygonum multiflorum.

[0054] Example 1 Isolation, screening and identification of fermenting Lactobacillus mucilaginosus (1) The fermented mucinous lactobacillus was isolated from traditional fermented pickled vegetables in Guizhou Province.

[0055] (2) Screening for fermenting *Lactobacillus mucinus*: Screening for bacteria with anti-*Helicobacter pylori* (… Helicobacter pylori Hp suspensions were prepared from strains with Hp activity and spread onto culture plates. The plates were perforated with a sterile punch and divided into experimental and control groups. The experimental group was added with the supernatant of the fermentation broth of the candidate strain, while the control group was added with MRS liquid culture medium. The plates were incubated at 37°C in a microaerophilic environment for 72 hours. The diameter of the inhibition zone was measured. The diameter of the inhibition zone of strain A27-1 was 14 mm, and strain A27-1 was obtained through screening.

[0056] (3) Morphological identification of the strain: After treatment, the supernatant of the strain was inoculated into MRS solid medium and cultured in a constant temperature incubator at 37℃ for 24h. After the culture was completed, single colonies of different sizes and shapes were picked and the strain was isolated and purified multiple times by streak plate method and Gram staining was performed. The morphology of the bacteria was observed under a microscope.

[0057] Figure 1 The results showed that strain A27-1 was a Gram-positive bacillus.

[0058] (4) Molecular identification of the strain: Genomic DNA of the target strain was extracted as a template, and PCR amplification was performed using 27F (5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID NO.1) and 1492R (5'-GGTTACCTTGTTACGACTT-3', SEQ ID NO.2) as primers. PCR amplification system: 10µL of 2×Taq PCR MasterMixⅡ, 0.5µL each of primers 27F and 1492R, 1µL of DNA template, and 8µL of double-distilled water (ddH2O). PCR amplification program: 94℃ for 3min; 94℃ for 30s, 58℃ for 30s, 72℃ for 90s, for a total of 30 cycles, with a final induction at 72℃ for 5min, and storage at 4℃. The amplified products were tested and qualified by 0.8% agarose gel electrophoresis and sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing to obtain 16S rDNA. The sequencing results were submitted to the NCBI GenBank database for homology comparison using BLAST, and a phylogenetic tree was constructed using Mega11.0 software.

[0059] BLAST sequence alignment of the 16S rDNA on the NCBI website revealed that strain A27-1 is related to *Lactobacillus fermentans* (…). Lactobacillus fermentum The 16S rDNA sequence of the sample showed over 97.57% homology, through... Figure 2 The phylogenetic tree results revealed that *Lactobacillus fermentatus* A27-1 and *Lactobacillus fermentatus* (… Lactobacillus fermentum The reference strains of strain A27-1 clustered closely, indicating that strain A27-1 belongs to... Lactobacillus fermentum This refers to Lactobacillus fermentans A27-1.

[0060] The *Lactobacillus fermentatus* identified in this embodiment was named A27-1 (denoted as *Lactobacillus fermentatus* A27-1 or strain A27-1), and classified as *Lactobacillus fermentatus*. Lactobacillus fermentum It was deposited on January 9, 2026 at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 37341.

[0061] Example 2 Preparation of Traditional Chinese Medicine Fermentation Broth Using *Lactobacillus mucinus* A27-1 (Example 1) and Verification of Its Effects 1.1 A traditional Chinese medicine composition, made from the following raw materials in the following proportions: Prepared Polygonum multiflorum 115g, Sapindus mukorossi 115g, Artemisia argyi 90g, Angelica sinensis 90g, Ganoderma lucidum 90g, Gleditsia sinensis 90g, Mentha haplocalyx 90g, Sophora flavescens 90g, Smilax glabra 90g, Astragalus membranaceus 90g, Rehmannia glutinosa 90g, Ligustrum lucidum 90g, Clematis chinensis 90g, Carthamus tinctorius 90g, Platycladus orientalis 90g, Psoralea corylifolia 70g, Eclipta prostrata 175g, and tea seed cake 90g.

[0062] A method for preparing a traditional Chinese medicine compound liquid, comprising the following steps: (1) Grind 115g of prepared Polygonum multiflorum, 115g of Sapindus mukorossi, 90g of Artemisia argyi, 90g of Angelica sinensis, 90g of Ganoderma lucidum, 90g of Gleditsia sinensis, 90g of Mentha haplocalyx, 90g of Sophora flavescens, 90g of Smilax glabra, 90g of Astragalus membranaceus, 90g of Rehmannia glutinosa, 90g of Ligustrum lucidum, 90g of Clematis chinensis, 90g of Carthamus tinctorius, 90g of Platycladus orientalis, 70g of Psoralea corylifolia, 175g of Eclipta prostrata and 90g of tea bran into powder, pass through a 20-mesh sieve, mix them together to obtain a mixture.

[0063] (2) Mix 1735g of the mixture with 17.35L of water and soak at 22℃ for 2h. Stir at 120rpm / min, boil over high heat for 2h, and then simmer over low heat for 3h to obtain the first decoction. Filter and collect the first filtrate and the first filter residue. Add water to the first filter residue at a mass-volume ratio of 1g:9mL and mix. Boil over high heat for 2h, and then simmer over low heat for 3h to obtain the second decoction. Filter and collect the second filtrate. Combine the first and second filtrates to obtain the total filtrate. Concentrate under reduced pressure at 60℃ to 1 / 4 of the total filtrate volume to obtain the traditional Chinese medicine compound liquid.

[0064] 1.2 Preliminary screening of probiotic strains and preparation of traditional Chinese medicine fermentation broth Studies have found that not all lactic acid bacteria strains can survive in traditional Chinese medicine compound liquids. Therefore, preliminary screening of probiotic strains is conducted in order to obtain lactic acid bacteria strains that can survive in traditional Chinese medicine fermentation liquids.

[0065] 1.2.1 Revival and Subculturing of Lactic Acid Bacteria Lactic acid bacteria strains stored at -20℃ were thawed in a 37℃ water bath. After thorough mixing, the resulting lactic acid bacteria culture was evenly spread onto MRS agar plates and incubated overnight at 37℃. A loopful of colony was scraped and transferred to MRS broth medium, and then incubated at 37℃ with shaking at 180 rpm for 24 h. After incubation, the mixed culture was streaked in three zones on MRS agar plates and incubated overnight at 37℃. Single colonies were picked and incubated in MRS liquid medium at 37℃ and 180 rpm for 24 h to obtain the culture. The culture was then inoculated into fresh MRS broth medium at a volume percentage of 4% and activated at 37℃ and 180 rpm for 24 h to obtain the lactic acid bacteria culture.

[0066] The clean bench was irradiated with ultraviolet light for 30 minutes before and after the experiment to ensure sterility.

[0067] 1.2.2 Preliminary Screening of Probiotic Fermented Traditional Chinese Medicine Compound Liquid In fact, research has found that not all lactic acid bacteria can survive in the traditional Chinese medicine compound liquid described in this invention. Therefore, preliminary screening of the probiotic fermented traditional Chinese medicine compound liquid is conducted to screen for lactic acid bacteria that can survive in the traditional Chinese medicine compound liquid.

[0068] First, add 500 µL of the traditional Chinese medicine compound solution prepared in section 1.1 above to a 5 mL centrifuge tube. Then, add 100 µL of the lactic acid bacteria solution prepared in section 1.2.1 above to the centrifuge tube. Label the centrifuge tube cap with the bacterial strain name and date. Incubate overnight at 37℃ and 180 rpm for 24 h. After 24 h, observe whether the liquid in the centrifuge tube is turbid. If it is turbid, dilute and spot the sample onto a plate. Spot the lactic acid bacteria onto MRS solid medium, i.e., use sterile physiological saline to perform serial dilutions of the sample to 10⁻⁶. -7 Four dilutions were selected for each sample, with three replicates for each gradient. 5 µL of the dilution was pipetted onto the MRS solid medium in the separated areas and incubated overnight for 24 h. If colonies grew, the dilutions with colony counts of 30 to 300 were selected for counting.

[0069] According to the initial screening results, it was found that the fermented Lactobacillus mucinus A27-1 from Example 1 could survive in the traditional Chinese medicine compound solution.

[0070] 1.2.3 Preparation of Traditional Chinese Medicine Fermentation Broth The fermented Lactobacillus mucinus A27-1 from Example 1 was activated according to section 1.2.1 to obtain fermented Lactobacillus mucinus A27-1 bacterial culture.

[0071] The concentration of Lactobacillus fermentation culture A27-1 was adjusted to 1×10⁻⁶. 8 CFU / mL was inoculated into 5 mL of the traditional Chinese medicine compound solution prepared in section 1.1 above at a volume percentage of 4%, and cultured at 37℃ and 180 rpm for 24 h to obtain the culture broth. Then, the culture broth was centrifuged at 5000 rpm for 10 min at 4℃. The collected supernatant was filtered through a 0.22 µm filter. After filtration, the filtrate was dispensed into 1.5 mL EP tubes, with 1 mL of filtrate in each tube, to obtain the fermentation broth A27-1 for fermentation of traditional Chinese medicine.

[0072] 1.3 Determination of antioxidant capacity of traditional Chinese medicine fermentation broth 1.3.1 Determination of DPPH free radical scavenging ability Sample group: 2 mL of fermentation broth A27-1 (denoted as A27-1) and 2 mL of the herbal compound solution (denoted as TCM) prepared in part 1.1 were mixed with 1 mL of DPPH ethanol solution (0.2 mmol / L) in 5 mL centrifuge tubes, reacted at 37℃ in the dark for 30 min, centrifuged at 5000 rpm for 5 min, and the supernatant was collected in a new 5 mL centrifuge tube. OD of the sample group was measured. 517 Let it be denoted as A1.

[0073] The blank group used an equal volume of anhydrous ethanol to replace the 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) ethanol solution, and the absorbance value at the same wavelength was recorded as A0; the control group used an equal volume of ultrapure water to replace the traditional Chinese medicine fermentation broth, and the absorbance value at the same wavelength was recorded as A2.

[0074] The formula for DPPH free radical scavenging rate is as follows: Clearance rate (%) = [1 - (A1 - A2) ÷ A0] × 100%.

[0075] 1.3.2 Determination of superoxide anion radical scavenging ability Sample group: Take 0.2 mL of the fermentation broth A27-1 (denoted as A27-1) and 0.2 mL of the herbal compound solution (denoted as TCM) prepared in part 1.1, and mix them with 1 mL of Tris-HCl buffer (50 mmol / L, pH 8.0), respectively, and incubate at 37℃ for 10 min. Add 0.1 mL of pyrogallol solution (25 mmol / L) and incubate at 37℃ for 4 min. Finally, add 0.2 mL of 1 mol / L HCl to terminate the reaction, and measure the absorbance at 320 nm using an ELISA reader.

[0076] The traditional Chinese medicine fermentation broth A27-1 in the sample group was replaced with 1 mg / mL VC (denoted as VC) as a positive control group.

[0077] Replace the traditional Chinese medicine fermentation broth A27-1 in the sample group with the same volume of distilled water as a blank control group.

[0078] In addition, an equal volume of distilled water was used to replace the pyrogallol solution in the sample group or the positive control group as the sample background group.

[0079] The formula for superoxide anion radical scavenging rate is as follows: Clearance rate (%) = [A 空白 -(A 样品 -A 背景 )]÷A 空白 ×100%.

[0080] A 空白 : refers to the absorbance of the blank control group (water + pyrogallol); A 样品 : refers to the absorbance of the sample group or positive control group (each sample or VC + pyrogallol); A 背景 This refers to the absorbance of the sample background group (each sample or VC+water).

[0081] Figure 3 The results from the A27-1 strain indicate that, compared to TCM, the fermentation broth from strain A27-1 has a stronger scavenging ability.

[0082] Figure 3 The results from group B showed that the superoxide anion radical scavenging capacity of the TCM group was approximately 45%. Compared with TCM, the fermentation broth of traditional Chinese medicine obtained from strain A27-1 fermentation showed the highest superoxide anion radical scavenging rate.

[0083] In conclusion, the fermentation broth of traditional Chinese medicine obtained by fermentation with strain A27-1 can significantly improve the antioxidant capacity of the traditional Chinese medicine compound solution.

[0084] 1.4 Determination of the antibacterial activity of traditional Chinese medicine fermentation broth This embodiment uses Staphylococcus aureus (Staphylococcus aureus) Staphylococcus aureus ATCC25923, clinical strain of Candida albicans ( Candida albicans )115 (abbreviated as clinical strain Candida albicans 115), standard strain Candida albicans ( Candida albicans ATCC10231 (abbreviated as standard strain Candida albicans 10231), Propionibacterium acnes ( Propionibacterium acnes The antibacterial activity of the fermentation broth A27-1 prepared by ATCC6919 in part 1.2.3 was determined.

[0085] Among them, the clinical strain Candida albicans 115 was isolated from vaginal secretions that were positive for Helicobacter pylori serum antibodies at Guiyang Jinyang Hospital.

[0086] 1.4.1 Determination of antibacterial activity using the micro-broth dilution method (1) Resuscitation and passage of Staphylococcus aureus Staphylococcus aureus strain ATCC25923, stored at -20℃, was rapidly thawed in a 37℃ water bath. After thorough mixing, the Staphylococcus aureus culture was evenly spread onto LB agar plates and incubated overnight at 37℃. A loopful of colony was scraped and transferred to LB broth medium, and then incubated at 37℃ with shaking at 180 rpm for 24 h. After incubation, the cultures were mixed, streaked in three zones on an agar plate, and incubated overnight at 37℃. Single colonies were picked and incubated in LB liquid medium (also known as LB broth medium) at 37℃ with shaking for 24 h. The culture was then inoculated at a volume percentage of 4% into fresh LB broth medium and activated at 37℃ with shaking at 180 rpm for 24 h to obtain the Staphylococcus aureus culture.

[0087] The biosafety cabinet was irradiated with ultraviolet light for 30 minutes before and after the experiment to ensure sterility.

[0088] (2) Resuscitation and subculturing of Candida albicans Take clinical strain Candida albicans 115 or standard strain Candida albicans 10231 stored at -20℃ and thaw them rapidly in a 37℃ water bath. After mixing, spread the inoculum evenly onto Sabouraud dextrose agar and incubate overnight at 37℃. Scrape a loopful and inoculate it into Sabouraud dextrose liquid agar, incubate for 24 h at 37℃ and 180 rpm on a shaker. Remove the inoculum, mix well, streak in three zones on agar plates, and incubate overnight at 37℃. Pick a single colony and incubate it in Sabouraud dextrose liquid agar at 37℃ and 180 rpm for 24 h. Inoculate 4% (v / v) of the colony into Sabouraud dextrose liquid agar and activate for 24 h at 37℃ and 180 rpm on a shaker to obtain Candida albicans 115 or Candida albicans 10231 inoculum.

[0089] The clean bench was irradiated with ultraviolet light for 30 minutes before and after the experiment to ensure sterility.

[0090] (3) Resuscitation and passage of Propionibacterium acnes Propionibacterium acnes strain ATCC6919, stored at -20℃, was rapidly thawed in a 37℃ water bath. After mixing, the bacterial culture was evenly spread onto bovine brain heart extract (BHI) solid medium and incubated overnight in a 37℃ anaerobic incubator. A loopful was then inoculated into BHI liquid medium and incubated for 24 h in an anaerobic incubator. The culture was then removed, mixed, streaked into three zones on agar plates, and incubated overnight in an anaerobic incubator. Single colonies were picked and incubated in BHI liquid medium for 24 h in an anaerobic incubator. After inoculation, a 4% (v / v) inoculum was added to BHI liquid medium and incubated for 24 h in an anaerobic incubator to activate the propionibacterium acnes culture.

[0091] The clean bench was irradiated with ultraviolet light for 30 minutes before and after the experiment to ensure sterility.

[0092] (4) Determination of antibacterial ability by micro-broth dilution method Activated Staphylococcus aureus, Candida albicans 115, or Candida albicans 10231, and Propionibacterium acnes bacterial suspensions were sequentially inoculated into LB liquid medium, Sabouraud dextrose agar, Sabouraud dextrose agar, and BHI medium, respectively. The cultures were incubated overnight at 37°C. After centrifugation to remove the medium, the bacterial cells were resuspended in physiological saline, and the concentration of each bacterial suspension was adjusted to 1×10⁻⁶ using a turbidimetric tube. 8 CFU / mL is reserved. Add 100 µL of traditional Chinese medicine fermentation broth A27-1 to each well of a 96-well plate, then add the appropriate amount of culture medium to each well to a total of 190 µL. Mix well, and then take 10 µL of 1×10⁻⁶ CFU / mL for later use. 8 CFU / mL bacterial suspensions were added to each well. The mixture was incubated at 37°C and 120 rpm / min for 12 h in a shaker. The absorbance (OD) at 600 nm was measured using a multi-mode microplate reader. 600 The antibacterial rate is calculated using the following formula to evaluate the antibacterial effect.

[0093] During the antibacterial activity assay, three groups were set up: a blank control, a negative control, and a positive control. The blank control consisted of no traditional Chinese medicine fermentation broth A27-1 added, with the volume supplemented to 190 µL using various culture media (referred to as blank bacterial solution). The negative control group consisted of 100 µL of traditional Chinese medicine fermentation broth A27-1 replaced with 100 µL of traditional Chinese medicine compound solution.

[0094] Positive control group: Amphotericin was used as a positive control for the inhibition of Candida albicans, amoxicillin was used as a positive control for the inhibition of Staphylococcus aureus, and erythromycin was used as a positive control for the inhibition of Propionibacterium acnes. The above 100µL of traditional Chinese medicine fermentation broth A27-1 was replaced with 100µL of 4µg / mL amphotericin, amoxicillin or erythromycin.

[0095] Antibacterial rate (%) = (Absorbance value of blank bacterial solution - Absorbance value of sample bacterial solution) ÷ Absorbance value of blank bacterial solution × 100%.

[0096] Figure 4 The results of the study showed that the inhibition rate of the traditional Chinese medicine compound solution (TCM) against the clinical strain of Candida albicans 115 was approximately 15.68%, while the inhibition rate of the antibiotic amphotericin B against the clinical strain of Candida albicans 115 was over 95%. However, the traditional Chinese medicine fermentation broth A27-1 showed a significantly better inhibitory effect on the clinical strain of Candida albicans 115 than the traditional Chinese medicine compound solution (TCM).

[0097] Figure 4 The results of B showed that the Chinese herbal compound solution (TCM) inhibited the standard strain of Candida albicans 10231 by approximately 24.90%, and the antibiotic amphotericin B also inhibited the standard strain of Candida albicans 10231 by over 95%. The Chinese herbal fermentation broth A27-1 was significantly more effective than the Chinese herbal compound solution (TCM) in inhibiting the standard strain of Candida albicans 10231.

[0098] In summary, the fermentation broth of traditional Chinese medicine obtained by strain A27-1 has the ability to promote the inhibition of Candida albicans growth by the traditional Chinese medicine compound solution, and has a good antibacterial effect against Candida albicans.

[0099] Figure 5 The results showed that the traditional Chinese medicine compound solution (TCM) had a certain inhibitory effect on Staphylococcus aureus, with an inhibition rate of 65%, while the traditional Chinese medicine fermentation broth A27-1 had an inhibition rate of 83% against Staphylococcus aureus. Compared with the traditional Chinese medicine compound solution, the traditional Chinese medicine fermentation broth A27-1 had a significantly higher inhibitory effect on Staphylococcus aureus.

[0100] Figure 6 The results showed that, compared with the traditional Chinese medicine compound solution, the antibacterial rate of the fermented traditional Chinese medicine broth A27-1 against Propionibacterium acnes was significantly higher than that of the traditional Chinese medicine compound solution, indicating that the fermented traditional Chinese medicine broth obtained by fermentation of strain A27-1 has the ability to promote the inhibition of Propionibacterium acnes by the traditional Chinese medicine compound solution.

[0101] 1.5 Determination of the inhibitory effect of traditional Chinese medicine fermentation liquid on skin inflammation Human epidermal immortalized keratinocytes (HaCaT) are derived from the skin epidermis and are closely related to proliferation, differentiation, formation of the stratum corneum, and the skin barrier. This embodiment uses HaCaT to further evaluate the alleviating effect of a traditional Chinese medicine compound on skin inflammation, and the promoting effect of a fermented broth obtained from the A27-1 strain on skin inflammation. The specific experimental methods are as follows: (1) HaCaT cell resuscitation and passage culture Remove the HaCaT cell cryovials from the -80℃ freezer and immediately place them in a 37℃ water bath with rapid shaking to thaw completely within 1-2 minutes. Slowly add the cell suspension to a 15mL centrifuge tube containing 1mL of complete culture medium, mix gently, and centrifuge at 900rpm for 3 minutes. Discard the supernatant, add 1mL of complete culture medium to the centrifuge tube, and gently pipette to resuspend the cell pellet. Take 500μL of the resuspended cell suspension and seed it into two T25 culture flasks containing 4mL of complete culture medium. Gently shake to distribute the cells evenly and incubate at 37℃ with 5% CO2. After 24 hours, discard the old culture medium, wash once with PBS, and replace with fresh complete culture medium to continue culturing. When the cell confluence reaches 80%-90%, passage or 6-well plate plating experiments can be performed.

[0102] (2) Perforated plate laying and starvation treatment HaCaT cells in the logarithmic growth phase were digested with trypsin to prepare a single-cell suspension, and then 2 × 10⁶ cells were distributed per well. 5 Cells were seeded at a density of 1000 cells / well in 6-well plates and incubated at 37°C with 5% CO2 for 24 hours. When the cell confluence reached approximately 70%, the old culture medium was discarded and replaced with serum-free culture medium for 12 hours of starvation treatment to synchronize the cells.

[0103] (3) Construction of Pam3CSK4 inflammation model and sample intervention After the starvation treatment, cells were treated with Pam3CSK4 at a final concentration of 500 ng / mL for 4 h to establish an in vitro inflammation model.

[0104] Cell experiments were divided into control group, model group, traditional Chinese medicine compound solution group, and traditional Chinese medicine fermentation broth A27-1 group. The control group consisted of culture medium and cells not treated with Pam3CSK4; the model group consisted of cells treated with 500 ng / mL Pam3CSK4 for 4 h. The treatment methods for the traditional Chinese medicine compound solution group and the traditional Chinese medicine fermentation broth A27-1 group were as follows: After modeling, neutralized traditional Chinese medicine compound solution (the pH of the traditional Chinese medicine compound solution was adjusted to neutral, denoted as neutral (pH=7)-10 μL) and neutralized traditional Chinese medicine fermentation broth A27-1 (the pH of the traditional Chinese medicine fermentation broth A27-1 was adjusted to neutral, denoted as A27-1 (pH=7)-10 μL) were added to the in vitro inflammatory model cells, respectively, with a dosage of 10 μL and an intervention period of 1 h. Cells from each group were collected, total RNA was extracted using the Trizol method, cDNA was obtained by reverse transcription, and the mRNA expression level of the inflammatory factor IL-6 was detected by qPCR.

[0105] Figure 7 The results showed that, compared with the control group, the expression level of IL-6 mRNA in the Pam3CSK4-induced model group was significantly upregulated (p<0.001), indicating that the HaCaT cell inflammation model was successfully constructed and can be used for subsequent treatment efficacy evaluation.

[0106] Figure 7 The results showed that, 1 hour after treatment, the IL-6 mRNA expression levels in both the herbal compound liquid group and the herbal fermentation liquid A27-1 group were significantly lower than those in the model group, demonstrating an anti-inflammatory effect on the skin. The herbal fermentation liquid A27-1 group showed a better anti-inflammatory effect, with the largest decrease in IL-6 mRNA expression level, approaching the level of the blank control group, demonstrating stronger anti-inflammatory potential.

[0107] 1.6 Non-targeted metabolomics analysis of traditional Chinese medicine fermentation broth 1.6.1 Functional analysis of major metabolites in the metabolomics of traditional Chinese medicine fermentation broth under positive ion mode The fermented Chinese medicine broth A27-1 prepared in part 1.2.3 of this embodiment and the Chinese medicine compound liquid prepared in part 1.1 were subjected to non-targeted metabolomics analysis by chromatography-mass spectrometry to further study the advantageous metabolites that promote the anti-inflammatory and antibacterial properties of the Chinese medicine compound liquid.

[0108] The most enriched metabolites in different groups under positive ion mode are summarized, and the top 20 most abundant metabolites are visualized and presented in bar chart format, such as... Figure 8 As shown.

[0109] Figure 8 The results showed that, compared with the traditional Chinese medicine compound solution, the fermentation broth of A27-1 traditional Chinese medicine could increase the abundance of the dominant metabolites hexadecasphinganine and oxymatrine, especially in the fermentation group in which strain A27-1 participated, which was the most dominant. Figures 3-5 The results show that the dominant metabolites have great potential in promoting anti-inflammatory and antibacterial effects.

[0110] 1.6.2 Analysis of metabolites with scalp care potential under positive ion mode Figure 9 The results showed that the dominant metabolites with scalp care potential in different groups were morin, oleamide, parthenolide, and andrographolide. After fermentation in A27-1, the parthenolide content in the A27-1 fermented herbal broth remained relatively high. The content of andrographolide, which has anti-inflammatory and antiviral activities, decreased after fermentation in A27-1, but the proportion of metabolites with potent anti-inflammatory, antioxidant, anti-allergic, and skin-repairing effects, such as (+)-shikonin, increased. This provides a basis for further research into its potential role in inhibiting Staphylococcus aureus, Candida albicans, and Propionibacterium acnes, as well as in assisting with the care of skin infections, folliculitis, and seborrheic dermatitis.

[0111] In summary, the beneficial metabolites that deliver the potential benefits of scalp care are mainly concentrated in group A27-1.

[0112] 1.6.3 Basic Metabolite Analysis of Traditional Chinese Medicine Fermentation Broth under Negative Ion Mode Figure 10The results showed that the dominant metabolites were gallic acid, alpha-isopropylmalate, 9,10,13-trihydroxyoctadecenoic acid (9,10,13-TriHOME), (R)-leucic acid, and D-(+)-phenyllactic acid. Compared with the compound herbal solution (NC), the gallic acid content in the fermented herbal broth obtained from fermentation A27-1 was increased. Compared with the compound herbal solution (NC), the (R)-leucic acid and D-(+)-phenyllactic acid in the fermented herbal broth group obtained from fermentation A27-1 (denoted as A271) showed a significant increasing trend, indicating that the fermented herbal broth obtained from fermentation A27-1 has superior anti-inflammatory efficacy and antibacterial effect.

[0113] 1.6.4 Analysis of potential metabolites for anti-inflammatory and antibacterial scalp care under negative ion mode Figure 11 The results showed that, compared with the herbal compound solution (NC), the A27-1 herbal fermentation broth group exhibited the most significant increase in myricetin 3-O-rhamnoside content, indicating that the herbal fermentation broth obtained from strain A27-1 fermentation promoted the abundance of myricetin. Compared with the herbal compound solution (NC), the A27-1 herbal fermentation broth also promoted the content of azelaic acid, indicating that the A27-1 herbal fermentation broth has the effect of promoting the content of azelaic acid, an effective component in traditional Chinese medicine that contributes to scalp care.

[0114] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A fermenting Lactobacillus mucinus A27-1, characterized in that, The preservation number of the fermenting Lactobacillus mucinus A27-1 is CGMCC No. 37341.

2. A traditional Chinese medicine fermentation liquid, characterized in that, The fermented Chinese medicine broth is obtained by fermenting the Chinese medicine compound broth using Lactobacillus mucinus A27-1 as described in claim 1.

3. The traditional Chinese medicine fermentation liquid according to claim 2, characterized in that, The fermentation method is to culture the food at 35~40℃ and 150~200rpm in a shaker for 22~26h. The culture medium obtained after fermentation also includes centrifugation, collection of supernatant, filtration of supernatant, and obtaining filtrate.

4. The traditional Chinese medicine fermentation liquid according to claim 2, characterized in that, The herbal compound liquid comprises the following raw materials in parts by weight: Prepared Polygonum multiflorum 80-150 parts, Sapindus mukorossi 80-150 parts, Artemisia argyi 60-120 parts, Angelica sinensis 60-120 parts, Ganoderma lucidum 60-120 parts, Gleditsia sinensis 60-120 parts, Mentha haplocalyx 60-120 parts, Sophora flavescens 60-120 parts, Smilax glabra 60-120 parts, Astragalus membranaceus 60-120 parts, Rehmannia glutinosa 60-120 parts, Ligustrum lucidum 60-120 parts, Clematis chinensis 60-120 parts, Carthamus tinctorius 60-120 parts, Platycladus orientalis 60-120 parts, Psoralea corylifolia 40-100 parts, Eclipta prostrata 150-200 parts, and tea seed cake 60-120 parts.

5. The traditional Chinese medicine fermentation liquid according to claim 4, characterized in that, The preparation method of the traditional Chinese medicine compound liquid includes: The prepared Polygonum multiflorum, Sapindus mukorossi, Artemisia argyi, Angelica sinensis, Ganoderma lucidum, Gleditsia sinensis, Mentha haplocalyx, Sophora flavescens, Smilax glabra, Astragalus membranaceus, Rehmannia glutinosa, Ligustrum lucidum, Clematis chinensis, Carthamus tinctorius, Platycladus orientalis, Psoralea corylifolia, Eclipta prostrata, and tea seed cake are pulverized, sieved, and mixed to obtain a mixture. The mixture is then mixed with water, soaked, and boiled over high heat while stirring. It is then simmered over low heat, filtered, and the filtrate is collected to obtain a traditional Chinese medicine compound liquid.

6. The traditional Chinese medicine fermentation liquid according to claim 5, characterized in that, The mass ratio of the mixture to the volume of water is 1g:8~15mL; the soaking method is soaking at 20~25℃ for 1~3h; the stirring rate is 80~200rpm / min; the boiling time is 1.5~2.5h; the decocting is performed 1~3 times, and each decocting time is 2~4h.

7. The use of the traditional Chinese medicine fermentation liquid according to any one of claims 2 to 6 in at least one of the following (1) to (7): (1) Inhibits skin pathogens; (2) Preparation of drugs to inhibit skin pathogens; (3) Preparation of antioxidants; (4) Preparation of drugs to inhibit skin inflammation; (5) Prepare skin care products; (6) Enhance the ability of the traditional Chinese medicine compound solution to inhibit skin pathogens; (7) Prepare drugs that enhance the ability of traditional Chinese medicine compound liquid to inhibit skin pathogens.

8. The application according to claim 7, characterized in that, The pathogens include one or more of Candida albicans, Staphylococcus aureus, and Propionibacterium acnes.

9. A method for increasing the content of metabolites in a traditional Chinese medicine compound liquid, characterized in that, The herbal compound solution is the herbal compound solution described in any one of claims 2 to 6; The method includes the step of fermenting the traditional Chinese medicine compound liquid using the fermented Lactobacillus myxobolus A27-1 as described in claim 1.

10. The method according to claim 9, characterized in that, The metabolites include one or more of hexadecylsphingosine, oxymatrine, shikonin, gallic acid, (R)-red acid, D-(+)-phenyllactic acid, and azelaic acid.

Images