Lactobacillus salivarius AACE1 and application thereof in preparation of fermented gastrodia elata

Abstract

The invention provides lactobacillus salivarius AACE1, and belongs to the technical field of probiotic fermentation and application, the lactobacillus salivarius AACE1 is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.20700. The lactobacillus salivarius AACE1 is a lactobacillus salivarius strain. When the gastrodia elata extract is applied to preparation of fermented gastrodia elata, a fermentation product has good antioxidant activity, can effectively inhibit skin from secreting proinflammatory response cytokines and relieve or treat damage of ultraviolet (UVB) to the skin, and is remarkable in effect. The invention also provides an application of the lactobacillus salivarius AACE1 in preparation of fermented gastrodia elata.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotic fermentation and application, and particularly relates to Lactobacillus salivarius AACE1 and application thereof in the preparation of fermented Gastrodia elata. Background Art

[0002] As the largest organ in the human body, the skin has biological functions such as preserving electrolytes and regulating body temperature. It also contains a large number of blood vessels and nerves for sensing external environmental stimuli, absorption and excretion. However, the skin is directly exposed to the external environment and is very susceptible to external damage, including photoaging caused by ultraviolet (UV) radiation.

[0003] In my country's high-altitude areas, as well as some areas close to tropical and temperate zones, it is common for the skin to be overexposed to ultraviolet (UV) radiation, so photoaging of the skin is more likely to occur in such areas. The UV region covers a wavelength range of 100-400nm and is divided into three bands, namely UVA (315-400nm), UVB (280-315nm), and UVC (100-280nm). When sunlight passes through the atmosphere, all UVC and about 90% of UVB are absorbed by the atmosphere, so the ultraviolet (UV) reaching the surface is mainly composed of UVA and a small amount of UVB. UVA is a long-wave ultraviolet ray with strong penetrating ability. It can penetrate the stratum corneum, epidermis, dermis and even subcutaneous tissue into the skin. UVB has a weak ability to penetrate the skin, mainly causing damage to the epidermis and superficial dermis. It is also widely concerned because it can be directly absorbed by DNA and form specific light damage, and has strong carcinogenicity and mutagenicity. Summary of the invention

[0004] In order to solve the problem of skin damage caused by ultraviolet UVB, the present invention provides a Lactobacillus salivarius AACE1, which is used in the preparation of fermented Gastrodia elata. The fermentation product has good antioxidant activity, can effectively inhibit the skin from secreting pro-inflammatory cytokines, and alleviate or treat skin damage caused by ultraviolet UVB, with significant effects.

[0005] The present invention also provides an application of Lactobacillus salivarius AACE1 in the preparation of fermented Gastrodia elata.

[0006] The present invention is achieved through the following technical solutions:

[0007] The invention provides a Lactobacillus salivarius AACE1 (Lactobacillus salivarius). The Lactobacillus salivarius AACE1 is preserved in the General Microbiological Center of China Microbiological Culture Collection Administration, and the preservation number is CGMCC No.20700.

[0008] Based on the same inventive concept, the present invention provides an application of Lactobacillus salivarius AACE1 in the preparation of fermented Gastrodia elata.

[0009] Based on the same inventive concept, the present invention provides a fermented Gastrodia elata, which is prepared by fermenting Gastrodia elata cell disrupted liquid with Lactobacillus salivarius.

[0010] Furthermore, the Lactobacillus salivarius includes the above-mentioned Lactobacillus salivarius AACE1.

[0011] Based on the same inventive concept, the present invention provides a method for preparing fermented Gastrodia elata, the preparation method comprising:

[0012] Inoculating the above-mentioned Lactobacillus salivarius AACE1 into the Gastrodia elata cell disrupted liquid and fermenting for a period of time to obtain a fermentation liquid;

[0013] The fermentation liquid is sterilized to obtain fermented Gastrodia elata.

[0014] Furthermore, the Gastrodia elata cell disruption solution is prepared by the following method:

[0015] Mixing fresh Gastrodia elata with water and breaking the wall to obtain Gastrodia elata homogenate;

[0016] The Gastrodia elata homogenate is sterilized to obtain a Gastrodia elata cell disruption solution.

[0017] Preferably, the mixing of fresh Gastrodia elata with water and then performing wall-breaking treatment to obtain Gastrodia elata homogenate comprises:

[0018] Chop fresh Gastrodia elata into small pieces and mix with water, then place in a wall breaking device, break the wall at 20000±2000rpm for 8.5±1.5min, and obtain Gastrodia elata homogenate;

[0019] Wherein, the mass ratio of the fresh Gastrodia elata to water is 1:4;

[0020] The step of sterilizing the Gastrodia elata homogenate to obtain a Gastrodia elata cell disruption solution specifically comprises:

[0021] The Gastrodia elata homogenate is heated to 75±5° C., kept warm for 30±5 min, and then cooled to obtain a Gastrodia elata cell disruption solution.

[0022] Furthermore, the method of inoculating the above-mentioned Lactobacillus salivarius AACE1 into the Gastrodia elata cell disruption solution and fermenting for a period of time to obtain the fermentation solution specifically comprises:

[0023] The bacterial liquid of the above-mentioned Lactobacillus salivarius AACE1 is inoculated into the Gastrodia elata cell disrupted liquid, and the fermentation is carried out in a closed manner at 37±1°C for 5 to 7 days, and the fermentation liquid is obtained after the pH is stabilized at 3.4 to 3.8 and a specific sour taste of lactic acid is produced;

[0024] The mass ratio of the bacterial solution to the Gastrodia elata cell disruption solution is 1:(19-20), and the number of viable bacteria in the bacterial solution is 7.9×10 8 CFU / mL~9.0×10 8 CFU / mL.

[0025] Optionally, sterilizing the fermentation liquid to obtain fermented Gastrodia elata specifically comprises:

[0026] The fermentation liquid is heated to 75±5° C. and kept warm for 30±5 min to kill bacteria, thereby obtaining fermented Gastrodia elata.

[0027] Based on the same inventive concept, the present invention provides the use of the above-mentioned fermented Gastrodia elata as or in the preparation of a therapeutic drug for UVB photoaging or UVB photodamage.

[0028] Furthermore, the symptoms of UVB photoaging or UVB photodamage include at least one of skin pigmentation, scaling, erythema, increased wrinkles, skin thickening, decreased elasticity, skin ulceration, edema and reduction of skin appendages caused by UVB irradiation.

[0029] Based on the same inventive concept, the present invention provides the use of the above-mentioned fermented Gastrodia elata as or in the preparation of skin care products against UVB photoaging or UVB photodamage.

[0030] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0031] 1. The present invention discloses a Lactobacillus salivarius AACE1, which has been deposited in the General Microbiology Center of China Microbiological Culture Collection Administration Committee with a deposit number of CGMCC No.20700. The Lactobacillus salivarius AACE1 can be applied to ferment Gastrodia elata cell disrupted liquid, and the fermentation product has good antioxidant activity, can effectively inhibit the secretion of pro-inflammatory cytokines by the skin, and can alleviate or treat the damage of ultraviolet UVB to the skin, with remarkable effect.

[0032] 2. The present invention discloses an application of Lactobacillus salivarius AACE1 in the preparation of fermented Gastrodia elata. Lactobacillus salivarius AACE1 is inoculated into Gastrodia elata cell disruption liquid, and the fermented product is named fermented Gastrodia elata. Applying the fermented Gastrodia elata can alleviate or even treat symptoms such as skin pigmentation, scaling, erythema, increased wrinkles, skin thickening, reduced elasticity, skin ulceration and edema caused by acute UVB light damage, as well as some conditions such as acanthosis, lymphocyte and granulocyte infiltration, fibrous connective tissue hyperplasia, and reduction of skin appendages. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0034] Figure 1 The changes over time in the appearance score of the skin of mice with acute UVB photoaging treated with AACE1 fermented Gastrodia elata.

[0035] Figure 2 This is a comparison chart of the effect of AACE1 fermented Gastrodia elata in alleviating UVB damage to the skin.

[0036] Figure 3 H&E staining results of AACE1 fermented Gastrodia elata in treating acute UVB photodamage in mice skin tissue.

[0037] Figure 4 Masson staining results of AACE1 fermented Gastrodia elata in treating acute UVB photodamage in mouse skin tissue. DETAILED DESCRIPTION

[0038] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0039] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0040] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0041] The overall idea of ​​the present invention is as follows:

[0042] Lactobacillus salivarius belongs to the family Lactobacillaceae and the genus Lactobacillus. It is widely found in the intestines of humans and animals and has strong probiotic potential. Moreover, Lactobacillus salivarius has the Qualified Presumption of Safety (QPS) status of the European Food Safety Authority and is also one of the strains approved for use in food production and processing by the Ministry of Health of my country in Announcement No. 65 of 2010, with high biosafety.

[0043] Gastrodia elata Bl. is a perennial herbaceous plant of the genus Gastrodia in the orchid family. It is also a traditional precious Chinese medicine. It was officially included in the catalog of "medicine and food homology" in November 2023. It is widely distributed in Southwest my country, East China, Central China, Northeast China and other places. It has a long history of medicinal use and significant clinical efficacy, attracting a large number of scholars to explore. At present, the pharmacological effects of Gastrodia elata are constantly being excavated, mainly in analgesia and sedation, anti-epileptic, neuroprotection, antioxidant and other aspects.

[0044] The present invention proposes a Lactobacillus salivarius AACE1 and an application thereof in the preparation of fermented gastrodia elata. The fermented gastrodia elata with Lactobacillus salivarius is a product obtained by fermenting gastrodia elata with Lactobacillus salivarius under suitable conditions. In short, gastrodia elata is co-cultured with lactic acid bacteria as a nutrient source to obtain a fermentation product. The final product collection is referred to as fermented gastrodia elata. At present, few people have published similar articles and reports in domestic and foreign literature reports, and there is no standardized operation for the preparation of fermented gastrodia elata. The present invention is formed by attempting to explore and using under this background.

[0045] The following will describe in detail the Lactobacillus salivarius AACE1 of the present invention and its application in the preparation of fermented Gastrodia elata in combination with the embodiments and experimental data.

[0046] Example 1

[0047] This embodiment provides a method for preparing AACE1 fermented Gastrodia elata, which is specifically as follows:

[0048] 1) After activating the preserved Lactobacillus salivarius AACE1, inoculate it in MRS medium and culture it at 37°C for 48 h. Observe the turbidity and pH changes of the bacterial solution, then perform microscopic morphological observation and Gram staining, and then dilute and spread the sample to detect the biomass of the viable bacteria. The number of viable bacteria was 7.9×10 8 CFU / mL~9.0×10 8 CFU / mL is appropriate.

[0049] 2) Chop the fresh Gastrodia elata into small pieces, add them into a wall-breaking machine at a water-to-material ratio of 1 (fresh Gastrodia elata): 4 (water) and mix well. After putting them into the wall-breaking machine, set the machine to 20,000 rpm for 7 minutes until the Gastrodia elata completely forms a paste-like homogenate, heat it to 75°C for 30 minutes, and after the temperature drops to room temperature, inoculate it in an ultra-clean workbench at a ratio of 1 (AACE1 bacterial solution): 20 (Gastrodia elata homogenate). After fully mixing, dispense them into small bottles and culture them in a closed manner at 37°C for 5-7 days. During this period, samples are taken every 24 hours and placed in a sterilized 5 mL EP tube to observe the fermentation situation. At the same time, the pH value is tested. When the pH of the sample is stable at 3.4-3.8 and a specific lactic acid taste is produced, the sample preparation is considered to be complete.

[0050] 3) Heat the sample at 75±5℃ for 30±5min to kill bacteria. After cooling to room temperature, use a pipette to dispense the sample into a sterilized 2ml EP tube and quench with liquid nitrogen (the purpose is to keep it in the state of the finished product and avoid inconsistency caused by time factors). After completion, immediately store it in a -80℃ freezer for later use.

[0051] Example 2

[0052] Modeling of acute UVB photoaging and treatment with Lactobacillus salivarius AACE1-fermented Gastrodia elata.

[0053] 1. Grouping and administration of experimental animals

[0054] Experimental animals: SPF-grade C57BL / 6J male mice, 5 weeks old, weighing 18-22 g, were provided by Spefoc (Beijing) Biotechnology Co., Ltd.

[0055] Experimental groups: The animals were randomly divided into 6 groups, including blank group, model group, positive control group (carbomer moist gel), Lactobacillus salivarius AACE1 group, Lactobacillus salivarius AACE1 fermented Gastrodia elata group, and Gastrodia elata group.

[0056] The treatment methods of experimental animals in each group are as follows:

[0057] (1) Blank group: The mice were depilated on their backs (area 2 cm × 2 cm), and no other experimental treatment was performed;

[0058] (2) Model group: The back of mice was depilated (area 2 cm × 2 cm), and the back of mice was irradiated with UVB according to the dose;

[0059] (3) Positive control group: The back of the mice was depilated (area 2 cm × 2 cm), and the back of the mice was irradiated with UVB according to the dose. Then, 0.2 mL of carbomer wet gel was evenly applied to the back of the mice at regular intervals every day. The drug was only administered once a day.

[0060] (4) Lactobacillus salivarius AACE1 group: The backs of mice were depilated (area 2 cm×2 cm), and the backs of mice were irradiated with UVB according to the dosage for modeling. Then, 0.2 mL of Lactobacillus salivarius AACE1 bacterial solution (preparation method see step 1 of Example 1)) was evenly applied to the backs of mice once a day;

[0061] (5) Gastrodia elata group: The backs of mice were depilated (area 2 cm×2 cm), and the backs of mice were irradiated with UVB according to the dosage for modeling. Then, 0.2 mL of Gastrodia elata homogenate was evenly applied to the backs of mice once a day (the preparation method is shown in step 2 of Example 1);

[0062] (6) Lactobacillus salivarius AACE1 fermented Gastrodia elata group: The backs of mice were depilated (area 2 cm×2 cm), and the backs of mice were irradiated with UVB according to the dosage for modeling. Then, 0.2 mL of Lactobacillus salivarius AACE1 fermented Gastrodia elata prepared in Example 1 was evenly applied to the backs of mice once a day.

[0063] 2. Construction of UV-damaged mouse model

[0064] Mice were anesthetized with Avertin (tribromoethanol) (20 μL / g). After the mice were anesthetized, the hair on the back of the mice was shaved with a shaver, and the excess short hair was removed with a depilatory cream. The residual depilatory cream was then scrubbed with sterile saline, and then disinfected with 75% alcohol after being wiped clean. The skin of the mice was observed to ensure that the skin of the mice to be modeled had no wounds, damage, or scratches.

[0065] Blindfold the mouse with gauze and fix the mouse on the cardboard with a tie. The UV aging box ZW-40 (Shaoxing Zhicheng Instrument Co., Ltd.) was used. The lamp was preheated for 10 minutes before modeling. Two 15W UVB lamps were used to irradiate the mouse skin at a distance of 15 cm from the back for 2 days, 30 minutes each time, and the skin damage of the mouse was observed. When the mouse skin showed symptoms such as rashes, edema, and ulcers, the modeling was successful. The modeling and drug administration of each group of mice are shown in Table 1.

[0066] Table 1 Mouse modeling and drug administration

[0067] Group drug Dosage Irradiation conditions Blank group PBS Phosphate Buffered Saline 0.2mL / piece No irradiation Model Group PBS Phosphate Buffered Saline 0.2mL / piece UVB irradiation Positive drug group Carbomer Moist Gel 0.2mL / piece UVB irradiation Gastrodia Group Gastrodia elata homogenate 0.2mL / piece UVB irradiation AACE1 group AACE1 bacterial solution 0.2mL / piece UVB irradiation AACE1 fermented Gastrodia elata group AACE1 fermented Gastrodia elata 0.2mL / piece UVB irradiation

[0068] Example 3

[0069] Therapeutic effect and evaluation of Gastrodia elata fermented with Lactobacillus salivarius AACE1 on acute UVB sunburn.

[0070] 1. Effect of Lactobacillus salivarius AACE1 fermented Gastrodia elata on acute UVB sunburn

[0071] During the experiment, the drug administration was strictly carried out and the changes in the back skin of the mice were closely observed (see Figure 2 ), take photos and record them every day.

[0072] (1) Day 5 of modeling and drug administration:

[0073] from Figure 2 It can be seen that on the 5th day of drug administration, mice in all groups except the blank group had obvious erythema, accompanied by increased skin wrinkles, significant decrease in elasticity, and skin thickening, indicating that all groups were successfully modeled.

[0074] (2) Day 10 of modeling and drug administration:

[0075] Model group: accompanied by large areas of erythema, medium brown precipitation, obvious skin wrinkles, accompanied by edema, crusts and necrosis, and obvious skin thickening.

[0076] Positive drug group: accompanied by moderate erythema, scattered light brown pigmentation, obvious skin wrinkles, poor elasticity, and slight thickening of the skin.

[0077] AACE1 bacterial fluid group: The skin was accompanied by moderate erythema, medium brown precipitation, edema, crust and necrosis, obvious skin wrinkles, poor elasticity and obvious thickening of the skin.

[0078] AACE1 bacterial liquid fermented Gastrodia elata group: It can be seen that on the tenth day, there was no obvious erythema and pigmentation on the skin of mice in the AACE1 bacterial liquid fermented Gastrodia elata group, only mild skin wrinkles and mild skin thickening.

[0079] Gastrodia elata group: The skin was accompanied by crusts and necrosis, medium brown precipitation, moderate skin wrinkles and poor skin elasticity, and the skin was significantly thickened.

[0080] (3) Modeling and drug administration day 18:

[0081] Model group: The wound was still unhealed with necrosis, the skin at the wound was thickened, and light brown pigment was deposited.

[0082] Positive drug group: There were no obvious wounds on the skin tissue, the skin had slight wrinkles, no obvious pigmented erythema, the skin elasticity was good, and the thickness was normal.

[0083] AACE1 bacterial liquid group: The skin tissue was still accompanied by wounds and necrotic areas, the skin elasticity at the wound was poor, and the skin was slightly thickened.

[0084] AACE1 bacterial liquid fermented Gastrodia elata group: The skin morphology and structure were intact, there was no pigmentation, no erythema, the skin was smooth, the elasticity was normal, the skin thickness was thin, and the recovery was good.

[0085] Gastrodia elata group: There were unhealed wounds with necrosis, thickening of the skin at the wound site with slight wrinkles, and light brown pigmentation.

[0086] A comprehensive comparison of the macroscopic mouse skin data shows that the wounds of the AACE1 bacterial solution fermented Gastrodia elata group and the positive drug group had healed on the 18th day, while the model group, Gastrodia elata group and AACE1 bacterial solution still had wounds. The recovery of the AACE1 bacterial solution fermented Gastrodia elata group was better than that of the positive group.

[0087] 2. Mouse skin appearance scoring criteria and results

[0088] After the irradiation, seven people who did not participate in the experiment used a double-blind method to score the degree of skin damage in the irradiated area on the back of the mice in accordance with the "Technical Specifications for Safety of Cosmetics". The scoring criteria are shown in Table 2:

[0089] Table 2 Mouse skin appearance scoring criteria

[0090]

[0091] The scores are obtained from Table 2 and the results are as follows: Figure 1 As shown, Figure 1 M is the model group; P is the positive drug group (Carbomer wet gel); G is the Gastrodia group; LS is the Lactobacillus salivarius AACE1 bacterial solution group; GL is the Lactobacillus salivarius AACE1 fermented Gastrodia group; CK is the blank group. As can be seen from the figure, the effect of the Lactobacillus salivarius AACE1 fermented Gastrodia group is almost the same as that of the positive drug group, which is better than the bacterial solution group, model group and Gastrodia group.

[0092] Example 4

[0093] Effects of Gastrodia elata fermented with Lactobacillus salivarius AACE1 on pathological changes of skin cells and collagen fibers in mice.

[0094] In order to study the effect of Lactobacillus salivarius AACE1 fermented Gastrodia elata on the skin histopathology of UVB-irradiated mice, the back skin of the mice in the above groups was collected to make tissue sections and performed H&E staining and Masson staining.

[0095] The results of H&E staining showed that:

[0096] On the 5th day, the epidermis of the skin in each group was incomplete, with disordered structure, unclear boundaries, incomplete keratinization and hyperkeratinization, dermal necrosis, fragmented nuclei, and unstructured eosinophilic substances; a small amount of fibrous connective tissue hyperplasia was seen in the dermis and subcutaneous tissue; a small amount of granulocytes and lymphocytes infiltrated. On the 18th day, except for the positive drug group, the blank group and the Lactobacillus salivarius AACE1 fermented Gastrodia group, the skin of the other groups had not recovered completely. In the Gastrodia elata group (G), the skin thickness was uneven, with moderate acanthosis, more new granulation tissue proliferation and repair, more fibrous connective tissue proliferation, and fewer skin appendages at the injured site; in the positive drug group (P), the woven epidermis was intact and thin; very little fibrous connective tissue proliferation was seen in the superficial dermis, and the hair follicles and sebaceous glands of the skin appendages were scattered; in the model group (M), very little fibrous connective tissue proliferation was seen in the superficial dermis, and the hair follicles and sebaceous glands of the skin appendages were scattered; in the Lactobacillus salivarius AACE1 bacterial fluid group (LS), the stratum corneum proliferation was seen in the skin tissue, the cortex was significantly thickened, and more connective tissue proliferation was seen in the dermis, accompanied by dotted lymphocytes and granulocytes infiltration; in the Lactobacillus salivarius AACE1 fermented Gastrodia elata group (GL), the epidermis structure was intact, the squamous epithelial cells had normal morphology and structure, and were tightly arranged, and the subcutaneous muscle layer structure was intact with uniform thickness. It can be seen that the skin recovery of mice treated with Lactobacillus salivarius AACE1 fermented Gastrodia elata was the best, almost on par with the positive drug (see Figure 3 ).

[0097] The results of Masson staining showed that:

[0098] The collagen fibers in the skin tissue were sky blue to bright dark blue, and the others were stained red. On the 5th day, it was found that the collagen fibers in the granulation tissue of each group were loosely arranged and irregular, and the color was light. On the 18th day, it was found that in the Gastrodia elata group (G), more fine collagen fibers were observed in the injured part of the skin tissue; in the positive drug group (P), a very small amount of fine collagen fibers were observed in the superficial dermis of the skin tissue; in the model group (M), a small amount of fine collagen fibers were observed in the superficial dermis, staggered, and stained blue; in the Lactobacillus salivarius AACE1 bacterial liquid group (LS), a large number of collagen fibers stained blue were observed in the dermis of the skin tissue, and more collagen fibers were observed; in the Lactobacillus salivarius AACE1 fermented Gastrodia elata group (GL), a large number of collagen fibers stained blue were observed in the dermis of the skin tissue, and the collagen fibers were arranged regularly, and there was no obvious abnormality in the content. The results show that the Lactobacillus salivarius AACE1 fermented Gastrodia elata group was far superior to the other groups in terms of skin collagen fiber recovery, and was even slightly better than the positive drug group (see Figure 4 ).

[0099] Example 5

[0100] Isolation, identification and conservation of Lactobacillus salivarius AACE1.

[0101] 1. Isolation of Lactobacillus salivarius AACE1

[0102] A healthy Yunnan black-bone chicken was obtained from the Yunnan Provincial College Feed Antibiotic Substitution Research Engineering Technology Center Experimental Base. After dissection, a 2-5 cm section of the small intestine was cut off and placed in a 5.0 mL sterile EP tube. It was rinsed three times with sterile water, fully ground, and filled to 3 mL with sterile water. Then, the grinding solution was diluted to 10 with sterile water. -5 , respectively draw 100μL 10 -3 , 10 -4 and 10 -5 The ground and diluted bacterial suspension was spread on MRS solid medium containing calcium carbonate, and after constant temperature culture at 37°C for 24 hours, the suspected lactic acid bacteria strains were obtained and numbered. Finally, the numbered suspected lactic acid bacteria strains were purified and cultured multiple times, 50% glycerol was added, and stored in a -80°C refrigerator for future use.

[0103] 2. Identification of Lactobacillus salivarius AACE1

[0104] According to TS I NGKE TSP701-50 Bacter ia Genomic DNA Kit (Beijing Qingke Biotechnology Co., Ltd.) instructions for extracting bacterial DNA. Use universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGYTACCTTGTTA CGACTT-3') to amplify the 16S rRNA gene of the isolated strain. After 1.0% agarose gel electrophoresis, the PCR amplification product was sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing within the same day. The sequencing results were compared with BLAST in NCBI GenBank to determine the strain. The specific sequencing results are as follows. The research results show that the 16S rRNA sequence of the strain is highly consistent with the sequence similarity of Lactobacillus salivarius 3158, with a similarity of 99%, and it is identified as Lactobacillus salivarius.

[0105]

[0106] 3. Preservation of Lactobacillus salivarius AACE1

[0107] Lactobacillus salivarius AACE1, deposit number: CGMCC No.20700, deposit unit: General Microbiology Center of China Culture Collection Administration, deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0108] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0109] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0110] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A Lactobacillus salivarius AACE1, characterized in that: The Lactobacillus salivarius AACE1 is deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, and the deposit number is CGMCC No.20700.

2. Use of Lactobacillus salivarius AACE1 as claimed in claim 1 in preparing fermented Gastrodia elata.

3. A fermented Gastrodia elata, characterized in that: The fermented gastrodia elata is prepared by fermenting gastrodia elata cell disrupted liquid with Lactobacillus salivarius.

4. The fermented Gastrodia elata according to claim 3, characterized in that: The Lactobacillus salivarius includes the Lactobacillus salivarius AACE1 described in claim 1.

5. A method for preparing fermented Gastrodia elata, characterized in that: The preparation method comprises: Inoculating the Lactobacillus salivarius AACE1 described in claim 1 into the Gastrodia elata cell disrupted liquid and fermenting for a period of time to obtain a fermentation liquid; The fermentation liquid is sterilized to obtain fermented Gastrodia elata.

6. The method for preparing fermented Gastrodia elata according to claim 5, characterized in that: The Gastrodia elata cell disruption solution is prepared by the following method: The fresh Gastrodia elata is mixed with water and then subjected to cell wall breaking treatment to obtain a Gastrodia elata homogenate; The Gastrodia elata homogenate is sterilized to obtain a Gastrodia elata cell disruption solution.

7. The method for preparing fermented Gastrodia elata according to claim 6, characterized in that: The step of mixing fresh Gastrodia elata with water and then breaking the wall of the mixture to obtain a Gastrodia elata homogenate comprises: Chop fresh Gastrodia elata into small pieces and mix with water, then place in a wall breaking device, break the wall at 20000±2000rpm for 8.5±1.5min, and obtain Gastrodia elata homogenate; Wherein, the mass ratio of the fresh Gastrodia elata to water is 1:4; The step of sterilizing the Gastrodia elata homogenate to obtain a Gastrodia elata cell disruption solution specifically comprises: The Gastrodia elata homogenate is heated to 75±5° C., kept warm for 30±5 min, and then cooled to obtain a Gastrodia elata cell disruption solution.

8. The method for preparing fermented Gastrodia elata according to claim 5, characterized in that: The method of inoculating the Lactobacillus salivarius AACE1 described in claim 1 into the Gastrodia elata cell disrupted liquid and fermenting for a period of time to obtain the fermentation liquid specifically comprises: Inoculate the bacterial liquid of Lactobacillus salivarius AACE1 described in claim 1 into the Gastrodia elata cell disrupted liquid, and ferment in a closed manner at 37±1° C. for 5 to 7 days, until the pH is stabilized at 3.4 to 3.8 and a specific sour taste of lactic acid is produced, thereby obtaining a fermentation liquid; The mass ratio of the bacterial solution to the Gastrodia elata cell disruption solution is 1:(19-20), and the number of viable bacteria in the bacterial solution is 7.9×10 8 CFU / mL~9.0×10 8 CFU / mL.

9. The method for preparing fermented Gastrodia elata according to claim 5, characterized in that: The step of sterilizing the fermentation liquid to obtain fermented Gastrodia elata specifically comprises: The fermentation liquid is heated to 75±5° C. and kept warm for 30±5 min to kill bacteria, thereby obtaining fermented Gastrodia elata.

10. Use of the fermented Gastrodia elata according to claim 3 or 4 as or in the preparation of a therapeutic drug for UVB photoaging or UVB photodamage.

11. The use according to claim 10, characterized in that: The symptoms of UVB photoaging or UVB photodamage include at least one of skin pigmentation, scaling, erythema, increased wrinkles, skin thickening, decreased elasticity, skin ulceration, edema and reduction of skin appendages caused by UVB irradiation.

12. Use of the fermented Gastrodia elata according to claim 3 or 4 as or in the preparation of skin care products against UVB photoaging or UVB light damage.

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