Phytobacterium plantarum YN-03, product prepared from same and application of product

Abstract

The invention provides a plant lactobacillus YN-03, a product prepared from the plant lactobacillus YN-03 and application, and belongs to the technical field of biological medicine, the plant lactobacillus YN-03 is preserved in China Center for Type Culture Collection on October 22, 2024, and the preservation number is CCTCC NO: M 20242301; the name of Latin is Lactiplantibacillus plantarum YN-03, and the name of Latin is Lactiplantibacillus plantarum YN-03; the gene sequence of the 16srRNA of the phytobacterium plantarum YN-03 is as shown in SEQ ID NO. 1, and the phytobacterium plantarum YN-03 has good gastrointestinal fluid and cholate tolerance. According to the plant lactobacillus YN-03, the product prepared from the plant lactobacillus YN-03 and the application of the plant lactobacillus YN-03, the plant lactobacillus YN-03 has the characteristic of high yield of GABA (gamma-aminobutyric acid) and shows potential application value in the aspect of relieving anxiety and depression.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to Lactobacillus plantarum YN-03 and products prepared therefrom and applications thereof. Background Art

[0002] Probiotics play a vital role in the body. They have multiple physiological functions, such as adjusting the balance of intestinal flora, lowering cholesterol, anti-oxidation, anti-infection and enhancing human immunity. These characteristics make probiotics have broad application prospects in the fields of food and medicine.

[0003] Among them, Lactobacillus plantarum, as an important microorganism in the gastrointestinal tract, is particularly critical for intestinal health. Numerous clinical, in vivo and in vitro studies have confirmed that Lactobacillus plantarum has shown significant effects in the prevention and treatment of inflammatory bowel disease, irritable bowel syndrome, cancer, diarrhea, lactose intolerance and other diseases. It can produce organic acids such as lactic acid, lower the pH value of the intestine, thereby inhibiting the growth and reproduction of harmful bacteria and creating a good environment for the survival of beneficial bacteria. At the same time, Lactobacillus plantarum can also enhance the immunity of the intestinal mucosa, promote the repair and regeneration of intestinal epithelial cells, and maintain the normal physiological function of the intestine.

[0004] However, in addition to its significant contribution to intestinal health, recent studies have also found that probiotics (including Lactobacillus plantarum) may also have some association with mental health. Depression, as an important mental health issue, is closely related to many physical and psychological diseases, such as cardiovascular disease, sleep disorders and cognitive impairment. A large number of people around the world are suffering from depression.

[0005] Diet is considered one of the most effective ways to prevent depression, and the intake of probiotics may have a positive effect on mood regulation. For example, gamma-aminobutyric acid (GABA) is an important neurotransmitter that plays an important role in regulating mood, relieving anxiety and improving sleep. Although there are currently few studies on the direct connection between Lactobacillus plantarum and GABA, future exploration of this area may provide new perspectives and methods for the prevention and treatment of mental health problems such as depression.

[0006] In the extensive research on probiotics, although we have gained a deep understanding of the physiological functions of many strains (including Lactobacillus plantarum), the research on some specific strains is still in its infancy. For example, no specific research on Lactobacillus plantarum YN-03 has been reported. Summary of the invention

[0007] The purpose of the present invention is to provide a Lactobacillus plantarum YN-03 and products and applications prepared therefrom. The Lactobacillus plantarum YN-03 has good gastrointestinal fluid and bile salt tolerance and has the characteristics of high GABA production, and shows potential application value in alleviating anxiety and depression.

[0008] To achieve the above object, the present invention provides a plant lactobacillus YN-03, which was deposited in the China Center for Type Culture Collection on October 22, 2024, with a deposit number of CCTCC NO: M 20242301; its Latin name is Lactiplantibacillus plantarum YN-03;

[0009] The gene sequence of 16srRNA of the Lactobacillus plantarum YN-03 is shown in SEQ ID NO.1.

[0010] The present invention also provides a use of the above-mentioned Lactobacillus plantarum YN-03 in preparing a product for alleviating anxiety and depression.

[0011] Therefore, the present invention adopts the above-mentioned Lactobacillus plantarum YN-03 and the products and applications prepared therefrom, and the beneficial technical effects are as follows:

[0012] (1) Lactobacillus plantarum YN-03 has good gastrointestinal fluid tolerance, bile salt tolerance, and good GABA production;

[0013] (2) As an important microorganism in the gastrointestinal tract, Lactobacillus plantarum YN-03 plays a vital role in protecting intestinal health. It can produce organic acids such as lactic acid, lower the pH value of the intestine, inhibit the growth and reproduction of harmful bacteria, and create a good environment for the survival of beneficial bacteria. At the same time, it can also enhance the immunity of the intestinal mucosa, promote the repair and regeneration of intestinal epithelial cells, and maintain the normal physiological function of the intestine. These characteristics make Lactobacillus plantarum YN-03 have broad application prospects in the fields of food and medical health.

[0014] (3) Lactobacillus plantarum YN-03 has a significant effect in relieving anxiety and depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is the phylogenetic tree of Lactobacillus plantarum YN-03;

[0016] Figure 2 To group and design the experimental plan;

[0017] Figure 3 To establish the CUMS mouse model process;

[0018] Figure 4The weight changes of mice in different treatment groups; Figure 4 In the table, CON is the blank control group, CUMS+NS is the model group, CUMS+F is the drug group, P represents the probiotic group, and PY represents the probiotic fermented milk group;

[0019] Figure 5 The effects of different interventions on TNF-α in the serum of CUMS mice;

[0020] Figure 6 The changes in sucrose preference of mice in different treatment groups;

[0021] Figure 7 The changes in immobility time of mice in different treatment groups. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0023] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0024] Embodiment 1

[0025] Isolation of Lactobacillus plantarum YN-03.

[0026] The yogurt sample was diluted in multiples and 10 -5 and 10 -6 200 μL of the dilution gradient sample is evenly spread on the MRS broth medium (solid) plate, and then taken back to the laboratory and cultured at 42°C for 48 hours. Pick single colonies with different morphology, size, and color and inoculate them in MRS broth medium (liquid), and culture them in a 42°C constant temperature incubator for 24-36 hours. After the strain grows well, perform Gram staining and microscopic examination. Save the isolated strains and extract the strain genomic DNA for subsequent determination and analysis.

[0027] The formula of MRS broth medium (solid) is: 10g soy peptone, 5g beef extract, 4g yeast powder, 20g glucose, 1mL Tween-80, 2g disodium hydrogen phosphate, 5g anhydrous sodium acetate, 2g triammine citrate, 0.02g manganese sulfate, 0.1g magnesium sulfate, 15g agar powder, and distilled water to make up to 1L. Adjust the initial pH to 6.20 and sterilize at 121℃ for 15min.

[0028] The formula of MRS broth medium (liquid) is: soy peptone 10g, beef extract 5g, yeast powder 4g, glucose 20g, Tween-80 1mL, disodium hydrogen phosphate 2g, anhydrous sodium acetate 5g, triammine citrate 2g, manganese sulfate 0.02g, magnesium sulfate 0.1g, distilled water to make up to 1L, adjust the initial pH to 6.20, and sterilize at 121℃ for 15min.

[0029] The present invention has no particular limitation on the sources of soy peptone, beef extract, yeast powder, glucose, Tween-80, disodium hydrogen phosphate, anhydrous sodium acetate, triammonium citrate, manganese sulfate, magnesium sulfate, and agar powder, and conventional commercial products well known to those skilled in the art can be used.

[0030] Strain identification was carried out by physiological and biochemical identification, 16srRNA molecular identification and whole genome sequencing.

[0031] The frozen test strain was inoculated into the preculture medium, cultured at 42°C for 24 to 36 hours, and subcultured for 2 to 3 generations. Then, 30 mL of the bacterial culture at the end of logarithmic growth was placed in a sterile tube and centrifuged. After centrifugation at 8000×g for 5 minutes (4°C), the bacteria were collected, the supernatant was discarded, and the genomic DNA of the strain was extracted using the CTAB freeze-thaw method specifically for lactic acid bacteria.

[0032] The composition of the preculture medium: peptone 10g, yeast extract powder 4g, beef extract powder 8g, glucose 20g, anhydrous sodium acetate 5g, triammine citrate 2g, dimethyl phosphate 2g, Tween-801mL, magnesium sulfate heptahydrate 0.2g, manganese sulfate tetrahydrate 0.05g, distilled water 1L, cysteine ​​salt 0.5g, pH adjusted to about 6.2, 21°C, 15min sterilization.

[0033] The isolated strain was subjected to physiological and biochemical identification and 16srRNA molecular identification according to the Bergey Manual of Bacterial Identification, using bacterial API50CH (BioMérieux). DNA samples of the strain were extracted and 16srRNA molecular identification was performed by Shanghai Meiji Biopharmaceutical Technology Co., Ltd. The molecular sequence was compared with NCBI database blastn, confirming that YN-03 was plantarum Lactobacillus. The phylogenetic tree is shown in Figure 1 .

[0034] The whole genome of Lactobacillus plantarum YN-03 was sequenced using the Illumina HiSeq sequencing platform. After assembly and alignment, its genome length was 38971bp, the G+C content was 45.61%, it contained 8 plasmids, and the chromosome genome contained 3229 coding genes.

[0035] Isolation and screening of high-yielding GABA lactic acid bacteria.

[0036] Preparation of γ-aminobutyric acid standard curve: standard preparation of 0, 0.2, 0.4, 0.6, 0.8, 1 mg / mL γ-aminobutyric acid standard solutions, then take out 1.0 mL from the standard solutions of different concentrations, add 1.0 mL of 0.01 mol / L sodium tetraborate buffer (pH 9), 1.0 mL of 6% redistilled phenol solution, 2 mL of 7.5% sodium hypochlorite solution, mix well, heat in a boiling water bath for 10 min, take out and immediately put into ice water and let stand for 5 min, after the solution turns blue-green, add 1.5 mL of 60% ethanol solution, shake and mix continuously, and then measure its absorbance at 645 nm. Draw a standard curve with absorbance as the ordinate and the concentration of each standard solution (mg / mL) as the abscissa, and establish a regression equation.

[0037] Activation and initial screening of lactic acid strains: The lactic acid bacteria strains stored at -80°C were transferred to MRS medium and cultured at 37°C for 24 hours. After the strains were activated for three generations, they were inoculated into GYP seed medium and cultured for 24 hours, and then inoculated into GYP fermentation medium and cultured for 36 hours to obtain fermentation liquid, which was centrifuged at 12000r / min for 10 minutes to obtain supernatant. 1mL of supernatant was taken and the ability of 80 lactic acid bacteria to produce GABA was determined according to the standard curve determination method, and lactic acid bacteria strains with high GABA production were selected.

[0038] GABA production capacity: The concentration of GABA produced by Lactobacillus plantarum YN-03 was determined according to the standard curve determination method. The results are shown in Table 1 below:

[0039] Table 1 GABA concentration produced by Lactobacillus plantarum YN-03

[0040] Strain number GABA concentration (mg / mL) YN-03 1.900±0.02

[0041] As shown in Table 1, Lactobacillus plantarum YN-03 has a good ability to produce GABA, and the concentration of GABA produced is 1.900 mg / mL as determined by the standard curve determination method.

[0042] According to the separation and screening of high GABA production capacity in this example, it can be seen that Lactobacillus plantarum YN-03 has good GABA production capacity.

[0043] Colony morphology characteristics.

[0044] The plant lactobacillus YN-03 of the present invention forms milky white opaque colonies with a diameter of 0.5 to 1 mm, round and flat or micro-papillary, flat edges, on an MRS solid culture medium.

[0045] In the clean bench, use an inoculation loop to pick the purified plant lactobacillus plantarum YN-03 sludge for smearing, and then perform Gram staining after it dries. The morphology and staining characteristics of the strain were observed and recorded under a microscope. The morphology of this strain under a microscope is Gram-positive bacteria, with round or oval bodies, mostly single, paired, chained or agglomerated; on MRS broth medium (solid), the colony diameter and colony edges are neat, creamy white and opaque.

[0046] The plant lactobacillus YN-03 of the present invention has the following biological characteristics: the bacteria are rod-shaped under a microscope, and the arrangement state is also single, paired, chain or clustered, but not in an obvious chain structure, without flagella or cilia, and will not move.

[0047] Gastrointestinal fluid tolerance of Lactobacillus plantarum YN-03.

[0048] Acid resistance test: The fermentation liquid of the activated two-generation high-yield GABA lactic acid bacteria strain was centrifuged at 4°C, 4000r / min for 10 minutes, the bacteria were collected, and the bacteria were washed once with sterilized PBS buffer. Then the bacterial concentration was adjusted to OD 600 =1, PBS was used as blank control. The bacterial suspension was inoculated into MRS broth medium (liquid) with a pH of 2.0 at 3%, mixed, and cultured at 37°C. The bacterial suspension at 0h and 3h was taken to determine the number of viable bacteria and to determine the bile salt tolerance of the strain. Each group was repeated 3 times.

[0049] Bile salt tolerance test: The lactic acid bacteria strain with high GABA production was activated for two generations, and the fermentation liquid of the second generation strain was centrifuged at 4°C and 4000r / min for 10 minutes to collect the bacteria, and then washed once with sterilized PBS buffer. The concentration of the bacteria was adjusted to OD 600 =1, PBS was used as blank control. The prepared bacterial suspension was inoculated into MRS broth medium (liquid) with a bile salt concentration of 0.3% at 3%, mixed evenly, and the live bacterial counts were measured at 0, 3, 6, and 9 hours at 37°C to determine the bile salt tolerance of the strains, with 3 replicates for each group.

[0050] Gastric juice resistance test: Take 2mL of 1mol / L hydrochloric acid and adjust the pH to 2.0 with distilled water. Add pepsin at a ratio of 1g / 100mL, fully dissolve, and sterilize with a 0.20μm microporous filter membrane. Inoculate the activated bacterial solution into the treated artificial gastric juice at a volume fraction of 10%, mix well, and culture in a constant temperature shaker at 37℃ and 100r / min. Use the spreading dilution method to determine the survival rate of high-yield GABA lactic acid bacteria after 3h of culture in artificial gastric juice, and repeat the determination three times.

[0051] Intestinal fluid resistance test: Take 0.85g KH2PO4 and add it to 62.5L distilled water, adjust the pH to 7.6 with 1mol / LNaOH solution, sterilize the prepared solution by high pressure, add trypsin at a ratio of 1g / 100mL on a clean bench, fully dissolve, and sterilize with a 0.20μm microporous filter membrane. Inoculate the activated bacterial solution into the treated artificial intestinal fluid at an inoculum volume of 10%, mix well, and culture in a constant temperature shaker at 37℃ and 100r / min, and determine the number of viable bacteria at 0 and 3h by the pouring method, and repeat the determination three times.

[0052] Tolerance effect: After the bile salt concentration of Lactobacillus plantarum YN-03 was 0.3%, pH 2.0 and treated with simulated gastric fluid and simulated intestinal fluid, the results are shown in Table 2 below:

[0053] Table 2 Survival of Lactobacillus plantarum YN-03 in simulated gastrointestinal digestive fluid

[0054]

[0055]

[0056] As shown in Table 2, Lactobacillus plantarum YN-03 has good tolerance characteristics, with a survival rate of 99.5% in 0.3% bile salt, 98% at pH 2.0, 81% after digestion in artificial gastric juice for 3 hours, and 98% after treatment in simulated intestinal juice for 3 hours.

[0057] According to the gastrointestinal fluid tolerance effect of this example, it can be seen that Lactobacillus plantarum YN-03 has a good tolerance to gastrointestinal fluid in the intestine and has probiotic properties.

[0058] Embodiment 2

[0059] Functional gene characteristics of Lactobacillus plantarum YN-03.

[0060] The KEGG database was used to perform functional annotation on the whole genome of Lactobacillus plantarum YN-03. KEGG is a professional database that compares and analyzes bacterial genes and metabolic pathways at the molecular level. It can explore the molecular level of bacteria and study the relationship between functional genes and biological molecules in various biological pathways. The KEGG database divides the biological pathways of microorganisms into six categories: Metabolism, Genetic Information Processing, Environmental Information Processing, Cellular Processes, Organismal Systems, and Human Diseases.

[0061] The tolerance-related functional genes of Lactobacillus plantarum YN-03 were annotated, and 10 acid tolerance-related genes (lysC, dapA, argC, argH, metE, argR, argJ, argS, murE, lysS); 8 heat tolerance-related genes (dnaK, hrcA, dnaJ, dnaA, dnaG, dnaB, dnaE, dnaN); 2 cold tolerance-related genes (cspA, clpC); and 2 bile salt tolerance-related genes (purD, gcvH) were annotated.

[0062] Genes related to probiotic functions were detected in the genome of Lactobacillus plantarum YN-03 and annotated to obtain functional genes related to carbohydrate active enzymes, glutamate dehydrogenase, lactic acid, and acid and bile resistance. The specific genes are shown in Table 3.

[0063] Table 3 Probiotic-related genes

[0064]

[0065] Embodiment 3

[0066] Improvement effect of Lactobacillus plantarum YN-03 on depressive behavior in CUMS mice.

[0067] Experimental design.

[0068] Constructing the CUMS mouse model: Seven different stimulation methods were applied, including: fasting from food and water, tail clamping, tail suspension, day and night reversal, strobe, restraint and electric shock. One stimulation was randomly given every day. The same stimulation could not be applied for three consecutive days to ensure that the mice could not expect it. For the specific CUMS protocol, see Figure 3 .

[0069] Food and water fasting: Remove the mice's chow and water bottles for 24 h.

[0070] Tail clamp: Use a wooden clamp to clamp the mouse's tail 1 cm away from the tip for 60 seconds.

[0071] Reversal of day and night: adjust the day and night cycle, turn off the lights from 8:00 to 20:00 to create a dark environment during the day, and the mice are in a dark environment; turn on the lights from 20:00 to 8:00 (the next day) to create a bright environment at night, and the mice are in a bright environment.

[0072] Strobe: A strobe light of 400 times / min was placed in the mouse living environment for 12 hours.

[0073] Restraint: Mice were restrained in a 50 ml disposable syringe for 1 h.

[0074] Tail suspension: The mice were suspended upside down for 5 min.

[0075] Electric shock: Use a 36V piezoelectric device to shock the soles of the mice six times.

[0076] Animal grouping and basic process: 60 healthy male C57BL / 6J mice of SPF grade of 6-8 weeks old were selected and purchased from Sibeifu (Beijing) Biotechnology Co., Ltd. All the mice were purchased and raised under standard experimental conditions. Three mice were raised in a plastic cage (290×178×160mm, M1), the laboratory temperature was 21±1°C, the relative humidity was 55±2%, 12h bright, 12h dark cycle (lights on at 8:00 am and lights off at 20:00 pm), and the mice were free to eat and drink. All mice were fed adaptively for 1 week and then the behavioral experiment was carried out for 4 consecutive days. This example was carried out in accordance with the "Guide to the Care and Use of Laboratory Animals" published by the National Institutes of Health of the United States.

[0077] The mice were divided into 5 groups, each with 12 mice, namely: control + placebo group (CON+NS) fed with 0.9% NaCl 200μl / d, chronic stress + placebo group (CUMS+NS) fed with 0.9% NaCl 200μl / d, chronic stress + fluoxetine group (CUMS+F) fed with fluoxetine 200μl / d, chronic stress + Lactobacillus plantarum YN-03 group (CUMS+P) fed with YN-03 200μl / d and chronic stress + Lactobacillus plantarum YN-03 yogurt group (CUMS+PY) fed with Lactobacillus plantarum YN-03 yogurt 200μl / d. All chronic stress groups were fed with Lactobacillus plantarum YN-03 yogurt 200μl / d according to the daily diet. Figure 2 The stress procedure was applied and the corresponding intervention drugs were given by gavage. The control group did not receive any stress stimulation except adding feed and water, cleaning the mouse cage, and gavage with the corresponding drugs. After 28 days of CUMS modeling, all mice underwent behavioral tests for 4 consecutive days, and then were killed and specimens were collected.

[0078] Experimental results.

[0079] Effect of Lactobacillus plantarum YN-03 intervention on body weight of CUMS mice.

[0080] Effects of Lactobacillus plantarum YN-03 on body weight of CUMS mice Figure 4As shown in the figure, there was no significant difference in body weight among the groups at the beginning of the experiment (P<0.05), but during the 4-week stress period, the body weight of the model group mice was significantly lower than that of the control group mice of the same age (P<0.05). Especially in the first week of stress, due to the sudden unpredictable stimulation, the body weight of the model group mice decreased significantly; while the drug intervention group and the Lactobacillus plantarum YN-03 and Lactobacillus plantarum YN-03 yogurt intervention group did not show a significant downward trend, indicating that drug and Lactobacillus plantarum YN-03 and Lactobacillus plantarum YN-03 yogurt intervention had a certain alleviating effect on chronic unpredictable stimulation. Since the 2nd week, the body weight of mice in all groups has increased slightly, including the model group, indicating that the mice have adapted to the unpredictable stimulation to a certain extent. Since then, the body weight of all CUMS mice has stabilized, while the normal mouse group still maintains a slight increase, which suggests that the model group is already in an anxious state.

[0081] Effect of Lactobacillus plantarum YN-03 intervention on TNF-α in the serum of CUMS mice.

[0082] Increased levels of TNF-α (tumor necrosis factor) in serum are usually considered as one of the indicators for judging anxiety and depression. Figure 5 As shown in the results, after 4 weeks of CUMS modeling, there were significant differences in the levels of tumor necrosis factor-α (TNF-α) in the serum of mice in the blank control group, model group and intervention group (P < 0.05). The TNF-α content in the model group (CUMS+NS) was significantly higher than that in normal mice in the blank control group (P < 0.05), indicating that chronic unpredictable stress caused inflammation in the mice in the model group. Combined with the behavioral abnormalities of the mice in the model group, the anxiety and depression of the mice in the model group were verified. The TNF-α content in the drug group was close to that in the blank control group, indicating that fluoxetine can effectively prevent the occurrence of inflammation. At the same time, the TNF-α content in the serum of mice in the intervention group of Lactobacillus plantarum YN-03 (CUMS+P) and Lactobacillus plantarum YN-03 yogurt (CUMS+PY) was significantly lower than that in the model group, indicating that Lactobacillus plantarum YN-03 and Lactobacillus plantarum YN-03 yogurt can also appropriately alleviate the occurrence of inflammation, but the effect is not as good as the drug fluoxetine.

[0083] Effect of Lactobacillus plantarum YN-03 intervention on sucrose preference behavior of CUMS mice.

[0084] Effects of Lactobacillus plantarum YN-03 intervention on sucrose preference behavior of CUMS mice Figure 6As shown in the figure, after 4 weeks of CUMS modeling, the sucrose preference index of mice in the blank control group, model group and intervention group was significantly different (P < 0.05). The sucrose preference index of the model group (CUMS + NS) was significantly lower than that of normal mice in the blank control group (P < 0.05), indicating that chronic unpredictable stress caused anxiety in the model group mice and affected their preference for sucrose. The sucrose preference index of the drug group was close to that of the blank control group, indicating that fluoxetine did have a significant anti-anxiety effect and did not affect the sucrose preference of mice. At the same time, the sucrose preference index of the intervention group of Lactobacillus plantarum YN-03 (CUMS + P) and Lactobacillus plantarum YN-03 yogurt (CUMS + PY) was significantly higher than that of the model group, but also significantly lower than that of the blank control group and the drug group, indicating that Lactobacillus plantarum YN-03 and Lactobacillus plantarum YN-03 yogurt also have a certain anti-anxiety effect, but the effect is not as good as the drug fluoxetine, and can be used as a dietary therapy to assist in relieving anxiety behavior.

[0085] Effect of Lactobacillus plantarum YN-03 intervention on forced swimming behavior of CUMS mice.

[0086] The forced swimming test (FST) is based on the immobility time of mice in water to detect the effect of anti-anxiety drugs. The immobility time of anxious mice in water is significantly higher than that of normal mice. After drug intervention, the immobility time of anxious mice returns to the level of normal mice. Figure 7 As shown in the figure, after 4 weeks of CUMS modeling, there was a significant difference in the immobility time of mice in the blank control group, model group and intervention group (P < 0.05). The immobility time of mice in the model group (CUMS + NS) was significantly higher than that of normal mice in the blank control group (P < 0.05), indicating that chronic unpredictable mild stress caused anxiety in mice in the model group and affected their immobility time in the water. The immobility time of mice in the drug group was close to that of the blank control, indicating that fluoxetine did have a significant anti-anxiety effect and did not affect the immobility time of mice. At the same time, the immobility time of mice in the intervention group (Lactobacillus plantarum YN-03, Lactobacillus plantarum YN-03 yogurt) was significantly lower than that of the model group, and significantly higher than that of the blank control group and the drug group, indicating that Lactobacillus plantarum YN-03 and Lactobacillus plantarum YN-03 yogurt also have a certain anti-anxiety effect, but the effect is not as good as the drug fluoxetine.

[0087] It is worth noting that the contents not elaborated in detail in the present invention are all prior art and are well known to those skilled in the art.

[0088] Therefore, the present invention adopts the above-mentioned Lactobacillus plantarum YN-03 and the products and applications prepared therefrom. Lactobacillus plantarum YN-03 has the characteristic of high GABA production and shows potential application value in alleviating anxiety and depression.

[0089] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A Lactobacillus plantarum YN-03, characterized in that The plant lactobacillus YN-03 was deposited in the China Center for Type Culture Collection on October 22, 2024, with a deposit number of CCTCC NO: M 20242301; its Latin name is Lactiplantibacillus plantarum YN-03; The gene sequence of 16srRNA of the Lactobacillus plantarum YN-03 is shown in SEQ ID NO.

1.

2. A use of the plant lactobacillus YN-03 as claimed in claim 1 in the preparation of a product for relieving anxiety and depression.

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