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

Abstract

The application provides a lactobacillus salivarius AACE1, belongs to the technical field of probiotic fermentation and application, and is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.20700.The lactobacillus salivarius AACE1 is applied to the preparation of fermented Gastrodia elata, and the fermentation product has good antioxidant activity, can effectively inhibit the secretion of pro-inflammatory cytokines of skin, and can relieve or treat the damage of ultraviolet UVB to skin with remarkable effect.The application also provides application of the lactobacillus salivarius AACE1 in the preparation of fermented Gastrodia elata.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of probiotic fermentation and application, and particularly relates to a lactobacillus salivarius AACE1 and application thereof in preparation of fermented gastrodia elata. BACKGROUND

[0002] Skin, as the largest organ of human body, has biological functions such as electrolyte preservation and body temperature regulation, and contains a large number of blood vessels and nerves for sensing external environmental stimuli, absorption and excretion. However, skin is directly exposed to the external environment and is easily damaged by the external environment, and such damage includes photoaging caused by ultraviolet (UV) radiation.

[0003] In high-altitude areas of China and some areas close to the tropics and temperate zones, it is common for skin to be excessively exposed to ultraviolet (UV) radiation, so photoaging of skin is more likely to occur in such areas. The UV region covers a wavelength range of 100-400 nm, and is divided into three bands, namely UVA (315-400 nm), UVB (280-315 nm) and UVC (100-280 nm). When sunlight passes through the atmosphere, all UVC and about 90% of UVB are absorbed by the atmosphere, so the ultraviolet (UV) that reaches the ground is mainly composed of UVA and a small amount of UVB. UVA is long-wave ultraviolet light, which has strong penetrating power and can penetrate the stratum corneum, epidermis, dermis and even subcutaneous tissue into the skin. UVB has weak penetrating power on the skin and mainly causes damage to the epidermis and superficial dermis, and is widely concerned because it can be directly absorbed by DNA and form specific photo-damage, which has strong carcinogenicity and mutagenicity. SUMMARY

[0004] In order to solve the problem of skin damage caused by ultraviolet UVB, the present application provides a lactobacillus salivarius AACE1, which is applied to the preparation of fermented gastrodia elata. The fermentation product has good antioxidant activity, can effectively inhibit the secretion of pro-inflammatory cytokines in the skin, and can relieve or treat the damage of ultraviolet UVB to the skin, and the effect is remarkable.

[0005] The application also provides application of the lactobacillus salivarius AACE1 in preparation of fermented gastrodia elata.

[0006] The application is achieved by the following technical solutions:

[0007] The application provides a lactobacillus salivarius AACE1, which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 20700.

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

[0009] Based on the same inventive concept, the application provides fermented Gastrodia elata, which is prepared by fermenting cell crushing liquid of Gastrodia elata by Lactobacillus salivarius.

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

[0011] Based on the same inventive concept, the application provides a preparation method of fermented Gastrodia elata, which includes the following steps:

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

[0013] Sterilizing the fermentation liquid to obtain fermented Gastrodia elata.

[0014] Further, the cell crushing liquid of Gastrodia elata is prepared by the following method:

[0015] Mixing fresh Gastrodia elata with water and then performing wall breaking treatment to obtain Gastrodia elata homogenate liquid;

[0016] Sterilizing the Gastrodia elata homogenate liquid to obtain the cell crushing liquid of Gastrodia elata.

[0017] Preferably, the step of mixing fresh Gastrodia elata with water and then performing wall breaking treatment to obtain Gastrodia elata homogenate liquid specifically includes:

[0018] After the fresh Gastrodia elata is cut and mixed with water, the mixture is placed in a wall breaking device, and wall breaking is performed at 20000±2000 rpm for 8.5±1.5 min to obtain the Gastrodia elata homogenate liquid;

[0019] The mass ratio of the fresh Gastrodia elata to water is 1:4.

[0020] The step of sterilizing the Gastrodia elata homogenate liquid to obtain the cell crushing liquid of Gastrodia elata specifically includes:

[0021] The Gastrodia elata homogenate liquid is heated to 75±5℃, and after incubation for 30±5 min, the Gastrodia elata homogenate liquid is cooled to obtain the cell crushing liquid of Gastrodia elata.

[0022] Further, the step of inoculating the above-mentioned Lactobacillus salivarius AACE1 into the cell crushing liquid of Gastrodia elata for fermentation for a period of time to obtain fermentation liquid specifically includes:

[0023] The bacterial liquid of the above-mentioned Lactobacillus salivarius AACE1 is inoculated into the cell crushing liquid of Gastrodia elata, and closed fermentation is performed at 37±1℃ for 5-7 days; after the pH is stabilized at 3.4-3.8 and specific sour taste of lactic acid is generated, the fermentation liquid is obtained;

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

[0025] Optionally, the fermentation liquid is subjected to sterilization treatment to obtain fermented Gastrodia elata, and the method specifically comprises the following steps:

[0026] The fermentation liquid is heated to 75±5 DEG C and kept for 30±5 min to kill bacteria, so as to obtain fermented Gastrodia elata.

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

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

[0029] Based on the same inventive concept, the application provides application of the above-mentioned fermented Gastrodia elata as or in preparation of an anti-UVB photoaging or UVB photodamage skin care product.

[0030] The one or more technical solutions in the embodiments of the application have at least the following technical effects or advantages:

[0031] 1. The Lactobacillus salivarius AACE1 has been preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 20700, which can be applied to a cell broken liquid of fermented Gastrodia elata, and the fermentation product has good antioxidant activity and can effectively inhibit the secretion of pro-inflammatory cytokines of the skin, relieve or treat the damage of ultraviolet UVB to the skin, and the effect is remarkable.

[0032] 2. The application of the Lactobacillus salivarius AACE1 in preparation of fermented Gastrodia elata, the Lactobacillus salivarius AACE1 is inoculated into a cell broken liquid of Gastrodia elata, and the product obtained by fermentation is named fermented Gastrodia elata, and smearing the fermented Gastrodia elata can relieve or even treat the symptoms of skin pigmentation, scaling, erythema, increased wrinkles, thickened skin, decreased elasticity, skin ulceration and edema caused by UVB acute photodamage, as well as some conditions such as thickening of the stratum spinosum, lymphocyte and granulocyte infiltration, fibrous connective tissue proliferation and reduced skin appendages. DETAILED DESCRIPTION

[0033] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following will briefly introduce the drawings needed in the embodiments description. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can be obtained by those skilled in the art without any creative effort on the basis of these drawings.

[0034] Figure 1 Appearance score of AACE1 fermented Gastrodiae Rhizoma treated UVB acute photoaging mouse skin over time.

[0035] Figure 2 Comparison chart of AACE1 fermented Gastrodiae Rhizoma alleviating UVB damage to skin.

[0036] Figure 3 H&E staining results of AACE1 fermented Gastrodiae Rhizoma treated UVB acute photoaging mouse skin tissue.

[0037] Figure 4 Masson staining results of AACE1 fermented Gastrodiae Rhizoma treated UVB acute photoaging mouse skin tissue. DETAILED DESCRIPTION

[0038] The advantages and various effects of the present application will be more clearly presented by the following specific embodiments and examples. Those skilled in the art should understand that these specific embodiments and examples are used to illustrate the present application, not to limit the present application.

[0039] Throughout the specification, unless otherwise specifically indicated, the terms used herein are understood as having the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art to which the present application belongs. If there is a conflict, the present specification takes precedence.

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

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

[0042] Lactobacillus salivarius belongs to the family of Lactobacillaceae and the genus of Lactobacillus, and widely exists in the intestinal tract of humans and animals, and has strong probiotic potential. Moreover, Lactobacillus salivarius has the status of qualified safety presumption (QPS) of the European Food Safety Authority, and is one of the food production and processing strains approved by the Ministry of Health of the People's Republic of China in the No. 65 Announcement in 2010, and has high biological safety.

[0043] Gastrodia elata Bl. is a perennial herb of Orchidaceae Gastrodia genus, also a traditional precious Chinese medicine, has been officially listed in the "food and medicine homologous" material directory in November 2023, widely distributed in China's southwest, east China, central China, northeast and other places, 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 continuously explored, which mainly show good performance in analgesia, antiepileptic, neuroprotection, antioxidant and other aspects.

[0044] The application provides saliva lactobacillus AACE1 and application of the saliva lactobacillus AACE1 in preparation of fermented Gastrodia elata. The fermented Gastrodia elata is obtained by fermenting Gastrodia elata under suitable conditions by using saliva lactobacillus. In brief, Gastrodia elata is used as a nutrient source to co-culture with lactic acid bacteria to obtain a fermentation product, and the final product is collectively referred to as fermented Gastrodia elata. At present, few similar articles are reported in the literature at home and abroad, and no standardized operation for preparing fermented Gastrodia elata is found. The application is formed under the background of trial and exploration.

[0045] The application of saliva lactobacillus AACE1 in preparation of fermented Gastrodia elata will be described in detail in combination with examples and experimental data.

[0046] Example 1

[0047] The application provides a preparation method of AACE1 fermented Gastrodia elata, and the preparation method specifically comprises the following steps:

[0048] 1) after activation of the preserved saliva lactobacillus AACE1, the saliva lactobacillus AACE1 is inoculated into MRS culture medium and cultured at 37 DEG C for 48 hours, the turbidity and pH change of the bacterial liquid are observed, microscopic morphological observation and gram staining are performed, and then the viable biomass is detected by dilution and coating, so that the viable count is 7.9x10 8 CFU / mL to 9.0x10 8 CFU / mL.

[0049] 2) fresh Gastrodia elata is cut into pieces, mixed in a cell wall breaking machine at a ratio of 1 (fresh Gastrodia elata) to 4 (water), and then put into the cell wall breaking machine, and set to 20000 rpm mode for 7 minutes until the Gastrodia elata is completely formed into a paste homogenate, and heated to 75 DEG C for 30 minutes; after the temperature is reduced to room temperature, inoculation is carried out in a clean bench at a ratio of 1 (AACE1 bacterial liquid) to 20 (Gastrodia elata homogenate), and then mixed thoroughly and divided into small bottles, and then cultured at 37 DEG C in a sealed state for 5-7 days; during the culture, the sample is taken out every 24 hours, placed in a sterilized 5mL EP tube, so that the fermentation condition is observed, and the pH value is detected at the same time; when the sample pH is stable at 3.4-3.8, and a specific lactic acid smell is generated, the sample preparation is considered to be completed.

[0050] 3) The sample was heated at 75±5℃ for 30±5min to kill the bacteria, and after cooling to room temperature, the sample was aliquoted into 2ml EP tubes using a pipette, and then quenched with liquid nitrogen (the purpose is to keep the product in a just-made state, to avoid differences caused by time factors), and immediately placed in a -80℃ freezer for preservation.

[0051] Example 2

[0052] Acute UVB photoaging modeling and treatment with Gastrodon AACE1 fermented Gastrodia elata.

[0053] 1. Grouping and administration of experimental animals

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

[0055] Experimental grouping: randomly divided into 6 groups, including a blank group, a model group, a positive control group (carbomer wet gel), a Gastrodon AACE1 group, a Gastrodon AACE1 fermented Gastrodia elata group, and a Gastrodia elata group.

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

[0057] (1) Blank group: the mouse back was shaved (area of 2cm x 2cm), and no other experimental treatment was performed;

[0058] (2) Model group: the mouse back was shaved (area of 2cm x 2cm), and the mouse back was treated with UVB irradiation modeling according to the dosage;

[0059] (3) Positive control group: the mouse back was shaved (area of 2cm x 2cm), and after the mouse back was treated with UVB irradiation modeling according to the dosage, the mouse back was evenly coated with carbomer wet gel 0.2mL every day, and the administration was performed only once a day;

[0060] (4) Gastrodon AACE1 group: the mouse back was shaved (area of 2cm x 2cm), and after the mouse back was treated with UVB irradiation modeling according to the dosage, the mouse back was evenly coated with Gastrodon AACE1 liquid 0.2mL once a day (preparation method see Example 1 step 1));

[0061] (5) Gastrodia elata group: the mouse back was shaved (area of 2cm x 2cm), and after the mouse back was treated with UVB irradiation modeling according to the dosage, the mouse back was evenly coated with Gastrodia elata homogenate liquid 0.2mL once a day (preparation method see Example 1 step 2));

[0062] (6) Lactobacillus salivarius AACE1 fermented Gastrodiae Rhizoma group: After the mice were modeled by UVB irradiation on the back (area of 2 cm x 2 cm) according to the dose, the mice were uniformly smeared with 0.2 mL of Lactobacillus salivarius AACE1 fermented Gastrodiae Rhizoma prepared in Example 1 on the back once a day.

[0063] 2. Construction of UV damage mouse model

[0064] The mice were anesthetized using Avertin (tribromoethanol) (20 μL / g), and after the mice were anesthetized, the hair on the back of the mice was removed using a hair clipper, and the remaining fine and short hair was removed using depilatory cream. The residual depilatory cream was then removed by washing with sterile normal saline, and the skin was cleaned. After cleaning, the skin was disinfected with 75% alcohol. The skin of the mice to be modeled was observed to ensure that there were no wounds, damage, or scratches.

[0065] The eyes of the mice were covered with gauze, and the mice were fixed on a paperboard with a string. A UV aging box ZW-40 (Shaoxing Chisheng Instrument Co., Ltd.) was used, and the lamp was preheated for 10 min before modeling. A UVB lamp with a power of 2 x 15 W was used to irradiate the skin of the mice at a distance of 15 cm, for 30 min each time for 2 days. The skin damage of the mice was observed. When the skin of the mice showed redness, edema, and ulceration, the modeling was considered successful. The modeling and administration of the mice in each group are shown in Table 1.

[0066] Table 1. Modeling and administration of mice

[0067] Group Drug Dosage Irradiation condition Blank group PBS phosphate buffer solution 0.2 mL / each No irradiation Model group PBS phosphate buffer solution 0.2 mL / each UVB irradiation Positive drug group Carbomer wet gel 0.2 mL / each UVB irradiation Gastrodia elata group Gastrodia elata homogenate 0.2 mL / each UVB irradiation AACE1 group AACE1 bacteria solution 0.2 mL / each UVB irradiation AACE1 fermented Gastrodia elata group AACE1 fermented Gastrodia elata 0.2 mL / each UVB irradiation

[0068] Example 3

[0069] Therapeutic effect and evaluation of Lactobacillus salivarius AACE1 fermented Gastrodiae Rhizoma on UVB acute sunburn.

[0070] 1. Effect of Lactobacillus salivarius AACE1 fermented Gastrodiae Rhizoma on UVB acute sunburn

[0071] The administration was strictly performed during the experiment, and the changes in the skin of the back of the mice were closely observed (see Figure 2 for details). The mice were photographed and recorded every day.

[0072] (1) 5th day after modeling and administration:

[0073] From Figure 2 it can be seen that on the 5th day after modeling and administration, the mice in all groups except the blank group showed obvious erythema, and at the same time, the number of skin wrinkles increased, the elasticity decreased significantly, and the skin thickened, indicating that the modeling of all groups was successful.

[0074] (2) 10th day after modeling and administration:

[0075] Model group: with large area of erythema, medium brown deposit, skin fold obvious, with edema, scab and necrosis, skin obviously thickened.

[0076] Positive drug group: with moderate erythema, scattered distribution of light brown pigment deposit, skin fold obvious, poor elasticity, skin slightly thickened.

[0077] AACE1 bacteria liquid group: skin with moderate erythema, medium brown deposit, with edema, scab and necrosis, skin fold obvious, poor elasticity, skin obviously thickened.

[0078] AACE1 bacteria liquid fermented Gastrodia group: on the tenth day, the skin of mice in the AACE1 bacteria liquid fermented Gastrodia group showed no obvious erythema and pigment deposit, only with mild skin fold and slight skin thickening.

[0079] Gastrodia group: skin with scab and necrosis, medium brown deposit, moderate skin fold and poor skin elasticity, skin obviously thickened.

[0080] (3) On the 18th day of modeling and administration:

[0081] Model group: still with necrotic non-healing wounds, skin thickening at the wound site, light brown pigment deposition.

[0082] Positive drug group: no obvious wound in skin tissue, skin with mild fold, no obvious pigment erythema, skin elasticity acceptable, normal thickness.

[0083] AACE1 bacteria liquid group: skin tissue still with wound and necrotic site, poor skin elasticity at the wound site, slight skin thickening.

[0084] AACE1 bacteria liquid fermented Gastrodia group: skin morphology structure is complete, no pigment deposition, no erythema, smooth skin, normal elasticity, thin skin thickness, good recovery.

[0085] Gastrodia group: still with necrotic non-healing wounds, skin thickening at the wound site with mild fold, light brown pigment deposition.

[0086] By comparing the macroscopic mouse skin data, it can be seen that the wound of AACE1 bacteria liquid fermented Gastrodia group and positive drug group has healed on the 18th day, and the model group and Gastrodia group and AACE1 bacteria liquid still have wounds. The recovery of AACE1 bacteria liquid fermented Gastrodia group is better than that of the positive group.

[0087] 2. Mouse skin appearance score standard and results

[0088] After the end of irradiation, according to the "Cosmetic Safety Technical Specifications", seven people who did not participate in this test were taken by double-blind method to score the skin damage degree of the mouse back irradiation area, the scoring standard is shown in Table 2:

[0089] Table 2 Mouse skin apparent scoring standard

[0090]

[0091] The score value obtained from Table 2 is plotted, and the results are shown in Figure 1 , 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 liquid group; GL is the Lactobacillus salivarius AACE1 fermented Gastrodia group; CK is the blank group. As can be seen from the figure, the Lactobacillus salivarius AACE1 fermented Gastrodia group and the positive drug group have almost the same effect, which is better than the liquid group, the model group and the Gastrodia group.

[0092] Example 4

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

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

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

[0096] On the 5th day, the skin epidermis of each group was incomplete, the structure was disordered, the boundary was not clear, there was parakeratosis and hyperkeratosis, and the dermis was necrotic; the nucleus was fragmented and showed a structureless eosinophilic substance; a small amount of fibrous connective tissue proliferation was visible in the dermis and subcutis; a small amount of granulocytes and lymphocytes infiltrated. On the 18th day, except for the skin of the positive drug group, the blank group and the Gastrodia elata fermented by Lactobacillus salivarius AACE1 group, the skin of the other groups was not completely recovered. In the Gastrodia elata group (G), the skin was thin and uneven, the spinous layer was moderately thick, a large amount of new granulation tissue proliferation was visible for repair, a large amount of fibrous connective tissue proliferation was visible, and the skin appendages at the damage site were reduced; in the positive drug group (P), the epidermis was complete and relatively thin; a small amount of fibrous connective tissue proliferation was visible in the superficial layer of the dermis, and skin appendages such as hair follicles and sebaceous glands were distributed sporadically; in the model group (M), a small amount of fibrous connective tissue proliferation was visible in the superficial layer of the dermis, and skin appendages such as hair follicles and sebaceous glands were distributed sporadically; in the Lactobacillus salivarius AACE1 liquid group (LS), the keratin layer of the skin tissue was proliferated, the skin layer was significantly thickened, a large amount of connective tissue proliferation was visible in the dermis, and there was a small amount of lymphocyte and granulocyte infiltration; in the Gastrodia elata fermented by Lactobacillus salivarius AACE1 group (GL), the epidermis was complete, the squamous epithelial cells were normal in shape and structure and arranged closely, and the subcutaneous muscle layer was complete and uniform in thickness. Therefore, the skin of the mice treated with Gastrodia elata fermented by Lactobacillus salivarius AACE1 recovered the best, and was almost the same as the positive drug (for details, see Figure 3 ).

[0097] The Masson staining results showed that:

[0098] The collagen fibers of the skin tissue were sky blue to bright dark blue, and the others were dyed red; on the 5th day, it was found that the collagen fibers in the granulation tissue of each group were arranged irregularly and loosely, and the color was light. On the 18th day, it was found that in the Gastrodia elata group (G), a large amount of fine collagen fibers proliferated in the damaged site of the skin tissue; in the positive drug group (P), a small amount of fine collagen fibers proliferated in the superficial layer of the dermis of the skin tissue; in the model group (M), a small amount of fine collagen fibers proliferated in the superficial layer of the dermis, and the collagen fibers were arranged in a staggered manner and dyed blue; in the Lactobacillus salivarius AACE1 liquid group (LS), a large amount of collagen fibers dyed blue were visible in the dermis of the skin tissue, and a large amount of collagen fiber proliferation was visible; in the Gastrodia elata fermented by Lactobacillus salivarius AACE1 group (GL), a large amount of collagen fibers dyed blue were visible in the dermis of the skin tissue, the collagen fibers were arranged regularly, and there was no obvious abnormality in the content. From the results, it can be seen that the Gastrodia elata fermented by Lactobacillus salivarius AACE1 group is far superior to other groups in the recovery of skin collagen fibers, and is even slightly better than the positive drug group (for details, see Figure 4 ).

[0099] Example 5

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

[0101] 1. Isolation of AACE1 from Lactobacillus salivarius

[0102] A healthy Yunnan Silkie chicken was obtained from the experimental base of the Yunnan Provincial Higher Education Institutions' Engineering Technology Center for Research on Antibiotic Substitution in Feed. After dissection, a 2-5 cm section of the small intestine was removed and placed in a 5.0 mL sterile EP tube. The tube was repeatedly rinsed three times with sterile water, thoroughly ground, and then brought to a final volume of 3 mL with sterile water. The grinding solution was then serially diluted with sterile water to a final volume of 10. -5 Take 100 μL of each of the 10 -3 10 -4 and 10 -5 The diluted bacterial suspension was spread onto MRS solid medium containing calcium carbonate and incubated at 37°C for 24 hours to obtain suspected lactic acid bacteria strains, which were then numbered. Finally, the numbered suspected lactic acid bacteria strains were purified and cultured multiple times, then 50% glycerol was added, and the samples were stored at -80°C for later use.

[0103] 2. Identification of Lactobacillus salivarius AACE1

[0104] According to TS I NGKE TSP701-50 Bacterial DNA was extracted using the Bacteria Genomic DNA Kit (Beijing Qingke Biotechnology Co., Ltd.). The 16S rRNA gene of the isolated strain was amplified using universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGYTACCTTGTTA CGACTT-3'). The PCR amplification products were detected by 1.0% agarose gel electrophoresis and then sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing on the same day. The sequencing results were aligned using BLAST from NCBI GenBank to confirm the strain. The specific sequencing results are as follows: the 16S rRNA sequence of the strain showed a high degree of similarity (99%) to *Lactobacillus salivarius* 3158, confirming its identification as *Lactobacillus salivarius*.

[0105]

[0106] 3. Lactobacillus salivarius AACE1

[0107] Lactobacillus salivarius AACE1, Accession No. CGMCC No. 20700, Accession Unit: China General Microbiological Culture Collection Center, Accession Address: No. 1, Beichen West Road, Chaoyang District, Beijing.

[0108] Finally, it is to be understood that the term "including", "comprising", and variations thereof, are intended to be broad and encompass the terms "consisting of" and "consisting essentially of" unless otherwise noted. Stated another way, nothing in the specification is to be construed as requiring the inclusion of any element in the process, method, article, or apparatus unless explicitly stated.

[0109] While the preferred embodiments of the application have been described, additional modifications and changes can occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, the particular embodiments are to be regarded as illustrative and not restrictive, and it is intended to include all such modifications and changes as falling within the scope of the application. Accordingly, the scope of the application should be determined by the following claims and their appropriate legal equivalents.

[0110] Obviously, numerous modifications and variations of the present application are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims and their legal equivalents, the application can be practiced otherwise than as specifically described.

Claims

1. A Lactobacillus salivarius AACE1, characterized in that, The Lactobacillus salivarius AACE1 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 20700.

2. Use of the Lactobacillus salivarius AACE1 in claim 1 in the preparation of fermented Gastrodia elata.

3. A fermented Gastrodia elata, characterized in that, The fermented Gastrodia elata is prepared by fermenting Gastrodia elata cell crushing liquid with Lactobacillus salivarius. The Lactobacillus salivarius is the Lactobacillus salivarius AACE1 in claim 1.

4. A method for preparing fermented Gastrodia elata, characterized by, The preparation method comprises: Inoculating the Lactobacillus salivarius AACE1 in claim 1 into Gastrodia elata cell crushing liquid for fermentation for a period of time to obtain fermentation liquid; Sterilizing the fermentation liquid to obtain fermented Gastrodia elata.

5. The method for preparing fermented Gastrodia elata according to claim 4, characterized in that, The Gastrodia elata cell crushing liquid is prepared by the following method: Mixing fresh Gastrodia elata with water and performing wall breaking treatment to obtain Gastrodia elata homogenate liquid; Sterilizing the Gastrodia elata homogenate liquid to obtain Gastrodia elata cell crushing liquid.

6. The method of claim 5, wherein the fermented Gastrodia elata is prepared by the steps of: The mixing of fresh Gastrodia elata with water and the wall breaking treatment to obtain Gastrodia elata homogenate liquid specifically comprises: After the fresh Gastrodia elata is crushed and mixed with water, the mixture is placed in a wall breaking device and wall breaking is performed at 20000±2000 rpm for 8.5±1.5 min to obtain Gastrodia elata homogenate liquid; The mass ratio of the fresh Gastrodia elata to water is 1:4; The sterilizing of the Gastrodia elata homogenate liquid to obtain Gastrodia elata cell crushing liquid specifically comprises: The Gastrodia elata homogenate liquid is heated to 75±5℃, kept for 30±5 min, and then cooled to obtain Gastrodia elata cell crushing liquid.

7. The method for preparing fermented Gastrodia elata according to claim 4, characterized in that, The inoculation of the Lactobacillus salivarius AACE1 in claim 1 into Gastrodia elata cell crushing liquid for fermentation for a period of time to obtain fermentation liquid specifically comprises: The bacterial liquid of the Lactobacillus salivarius AACE1 in claim 1 is inoculated into Gastrodia elata cell crushing liquid, and closed fermentation is performed at 37±1℃ for 5-7 days until the pH is stabilized at 3.4-3.8 and specific lactic acid taste is generated, thereby obtaining fermentation liquid; The mass ratio of the bacterial liquid to the Gastrodia cell broken liquid is 1:(19-20), and the number of viable bacteria in the bacterial liquid is 7.9x10 8 CFU / mL-9.0x10 8 CFU / mL.

8. The method for preparing fermented Gastrodia elata according to claim 4, characterized in that, The sterilizing of the fermentation liquid to obtain fermented Gastrodia elata specifically comprises: The fermentation liquid is heated to 75±5℃, kept for 30±5 min to kill bacteria, and then fermented Gastrodia elata is obtained.

9. Use of the fermented Gastrodia elata in claim 3 in the preparation of a therapeutic drug for UVB photoaging or UVB photodamage.

10. Use according to claim 9, characterized in that, The symptoms of the UVB photoaging or the UVB photodamage include at least one of skin pigmentation, scaling, erythema, increased wrinkles, thickened skin, decreased elasticity, skin ulceration, edema, and reduced skin appendages caused by UVB irradiation.

11. Use of the fermented Gastrodia elata in claim 3 in the preparation of an anti-UVB photoaging or UVB photodamage skin care product.

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