Application of Pantoea ananatis

Abstract

The invention discloses an application of Pantoea ananatis, and relates to the technical field of microorganisms. Aiming at the defects of the existing obesity intervention means, the invention provides a brand-new solution: Pantoea anatitis adapted to a high-altitude environment is used as a core functional strain, through an intestinal flora driving mechanism, the expression of UCP1 protein in brown adipose tissue (BAT) is activated, the heat production function of the BAT is enhanced, and meanwhile, the storage of white adipose tissue (WAT) and the level of liver triglyceride (TG) are reduced; the weight-losing effect is realized. The strain can play a role through an'intestinal flora-BAT 'axis without cold exposure, has no obvious influence on liver functions, and is excellent in safety. The invention discloses a flora-mediated non-cold-dependent BAT activation way for the first time, can be used for preparing products such as weight-reducing medicines and health-care foods, has the advantages of mechanism innovation, exact effect and wide application, and provides a new technical choice for obesity intervention.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more particularly to a pineapple pancreatic bacteria. Pantoea ananatis Applications. Background Technology

[0002] Obesity has become a serious public health challenge worldwide. Statistics from the World Health Organization (WHO) show that there are over 1.9 billion overweight people globally, including 650 million obese individuals, and the incidence rate continues to rise annually. Obesity not only affects appearance and mental health but also serves as a core risk factor, significantly increasing the risk of chronic diseases such as diabetes, cardiovascular disease, and cancer, creating a vicious cycle of "obesity-chronic disease" that poses a serious threat to human health. Current clinical interventions for obesity mainly consist of traditional treatments, including diet control and exercise intervention, drug intervention, and metabolic surgery. However, diet control and exercise intervention rely heavily on the patient's long-term self-discipline, and most patients experience a 60%-80% weight rebound rate due to difficulty in maintaining the regimen. Furthermore, these methods cannot address the root cause of intestinal metabolic disorders. While weight-loss drugs, such as GLP-1 receptor agonists, can achieve a 10%-20% weight loss, they suffer from significant individual differences, muscle loss, and gastrointestinal adverse reactions. Moreover, their long-term safety is not fully understood, failing to meet the demand for broad-spectrum treatment with low side effects. Metabolic surgery is only suitable for severely obese patients and carries risks such as surgical trauma, postoperative complications, and impaired energy absorption. It also requires long-term lifestyle interventions after surgery, limiting its applicability and incurring high medical costs.

[0003] Therefore, developing other methods to control obesity is of great significance. Summary of the Invention

[0004] In view of this, the main objective of the present invention is to provide a pancreatic fungus for pineapple. Pantoea ananatis The application of this technology aims to solve the problems existing in current technologies.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows: In a first aspect, the present invention provides pancreatic fungus for pineapples. Pantoea ananatis Its fermentation broth or microbial agents containing it are used in the preparation of weight loss-related products.

[0006] In this invention, Pantothecin pineapple Pantoea ananatis This is a potential probiotic adapted to high-altitude environments, discovered to activate brown adipose tissue and promote systemic fat reduction through a gut microbiota-driven mechanism. This breakthrough provides a safe and effective alternative to cold-induced thermogenesis, with profound implications for obesity intervention. Through multi-omics and gavage experiments, this invention demonstrates... Pantoea ananatis canIt can activate brown adipose tissue (BAT), reduce the storage of white adipose tissue (WAT), and enhance triglyceride metabolism. It is worth noting that... Pantoea ananatis These effects are achieved without cold exposure, thus establishing the "gut microbiota-BAT" axis. This invention overcomes a key bottleneck in microbiota-based therapy, demonstrating a strain with excellent host adaptability and consistent metabolic benefits. This invention is the first to reveal a cold-exposure-independent, microbiota-mediated BAT activation pathway, bringing transformative progress to metabolic therapy.

[0007] Furthermore, the weight loss-related products are used to prevent or improve obesity caused by a high-fat diet.

[0008] In a second aspect, the present invention provides the aforementioned pancreatic mycorrhizal fungus of pineapple. Pantoea ananatis Its fermentation broth or its microbial agents in the preparation of lipid-lowering products.

[0009] Furthermore, the blood lipids include triglycerides.

[0010] In a third aspect, the present invention provides pancreatic fungus for pineapples. Pantoea ananatis Its fermentation broth or microbial agents containing it are used in the preparation of products that regulate intestinal flora.

[0011] In a fourth aspect, the present invention provides the above-described pancreatic mycorrhizal fungus of pineapple. Pantoea ananatis Its use, or its fermentation broth or microbial agents containing it, in the preparation of products that activate brown adipose tissue, reduce white adipose tissue storage, and / or enhance triglyceride metabolism.

[0012] Furthermore, the product may include food, pharmaceuticals, or additives.

[0013] Furthermore, the dosage form of the medicine includes any one of the following: powder, granules, tablets, pills, capsules, suspension, emulsion, syrup, aerosol, or suppository; The food products include any one of the following: dietary supplements, prebiotics, solid beverages, and dairy products.

[0014] Furthermore, the product also contains pharmaceutically acceptable carriers or food-acceptable excipients.

[0015] Pineapple Pantothecin Pantoea ananatis Commonly reported as a plant pathogen, it is also widely found in the gut microbiota of animals. This invention is the first to discover the activating effect of this bacterium on BAT, and it may be a more promising probiotic specifically for weight loss.

[0016] The beneficial effects of this invention include at least the following: (1) This invention is the first to reveal a "gut microbiota-brown adipose tissue (BAT)" activation pathway independent of cold exposure, utilizing the adaptation to high-altitude environments. Pantoea ananatis By activating the thermogenesis function of BAT through the gut microbiota-driven mechanism, fat metabolism regulation can be achieved without relying on cold stimulation, which solves the core problem of the limited application scenarios of traditional cold-induced thermogenesis and provides a brand-new technical direction for obesity intervention.

[0017] (2) Experimental data of this invention confirm that, Pantoea ananatis It can significantly reduce the body weight of animals in a high-fat diet model. p <0.05), while reducing the storage of white adipose tissue (WAT) (including subcutaneous fat, paratesticular fat, and abdominal fat), p <0.01), and effectively reduces liver triglyceride (TG) levels. It can also enhance the thermogenesis capacity of BAT by activating UCP1 protein expression, achieving a synergistic effect of "weight loss + lipid reduction + improvement of fat distribution". The weight loss effect is stable and statistically significant.

[0018] (3) The present invention Pantoea ananatis As a potential probiotic, oral administration of this product has no significant effect on the core liver function indicators (alanine aminotransferase ALT and aspartate aminotransferase AST) in model animals. It avoids the gastrointestinal adverse reactions and muscle loss that may be caused by traditional weight loss drugs, and does not require the trauma and complications of metabolic surgery. It meets the needs of broad-spectrum obesity treatment with low side effects, is suitable for long-term use, has excellent overall safety, and has no obvious toxic side effects.

[0019] (4) It does not rely on the patient's long-term self-discipline and can be taken orally. It is suitable for the prevention and improvement of obesity caused by high-fat diet. It avoids the problem of high rebound rate of diet and exercise intervention and breaks through the limitation of the scope of application of metabolic surgery. It can be developed into various product forms such as weight loss drugs and health food to meet the obesity intervention needs of different groups. It has broad application prospects. Attached Figure Description

[0020] Figure 1 for Pantoea ananatis Results of reducing triglyceride levels and activating brown adipose tissue. (a) Oral gavage P. ananatis Schematic diagram of experimental design affecting lipid metabolism in C57BL / 6J mice (n=8). (bg) Oral gavage P. ananatisSubsequent observed changes: (b) body weight over time, (c) body weight, (d) brown adipose tissue weight, (e) subcutaneous fat, (f) paratesticular fat, (g) abdominal fat. Levels of alanine aminotransferase (ALT) (h), aspartate aminotransferase (AST) (i), total bile acids (CHOL) (j), and triglycerides (TG) (k) in the liver. (l) Paraffin section of brown adipose tissue. (m) P. ananatis Expression of ucp1 in brown adipose tissue of mice fed a high-fat diet by gavage (HFD). Detailed Implementation

[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0022] Furthermore, the technical solutions of the various embodiments of the present invention can be combined with each other, but only if they are feasible for those skilled in the art. If the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

[0023] The following specific embodiments illustrate the solution proposed in this invention: laboratory animals Two-month-old specific pathogen-free (SPF) grade male C57BL / 6J mice (purchased from SPF Biotechnology Co., Ltd. and Beijing Vital River Laboratory Animal Technology Co., Ltd.) were housed at the Animal Experiment Center of the Institute of Zoology, Chinese Academy of Sciences (CNASLA0014). All animals underwent a 7-day acclimatization period before any treatment. The mouse housing temperature was maintained at 23 ± 0.5 ℃, with a light cycle of 14 hours of vertical light and 10 hours of darkness (lights turned on at 06:00), and humidity ranging from 30% to 70%. During the experiment, mice had free access to autoclaved water and food. Two diets were used: a standard diet (containing 18% protein, 3% fat, 12% fiber, and 47% carbohydrates, Beijing Keao Xieli Feed Co., Ltd.) and a high-fat diet (60% energy from fat, Research Diets, New Brunswick, NJ; D12492). At the end of the experiment, the animals were euthanized, and blood and organs were collected for further testing and analysis. All animal experiments were approved by the Animal Experiment Welfare and Ethics Committee of the Institute of Zoology, Chinese Academy of Sciences (Approval No.: IOZ-IACUC-2024-137).

[0024] Experimental Design Experiment 1: 7 macaques distributed at altitudes between 50 and 4317 meters ( M. mulatta Fecal samples were collected from 82 individuals in the population. New bacterial species were identified using metagenomic shotgun sequencing binning technology, the genomes of major species were assembled, and a gut microbiota gene catalog was constructed.

[0025] Experiment 2: To investigate the effects of gut microbiota and its metabolites at high altitudes, fecal microbiota from high- and low-altitude rhesus monkeys were transplanted into mice. A total of 26 male mice were randomly divided into three groups: high-altitude rhesus monkey fecal microbiota transplantation group (high-FMT, n = 9), low-altitude rhesus monkey fecal microbiota transplantation group (low-FMT, n = 9), and control group (control, transplanted with the same group's fecal microbiota, n = 8). All mice were administered 200 µL of a fresh compound antibiotic by gavage daily for 4 days. The antibiotic consisted of 100 µg / mL neomycin, 50 µg / mL streptomycin, and 100 U / mL penicillin (Sigma, Germany). During the experiment, all mice received 200 µL of fecal suspension daily for four weeks. Throughout the experiment, all mice were fed a standard diet.

[0026] Experiment 3: To investigate the significant enrichment of Pantotheca pineapple in high-altitude macaque populations (… P. ananatis To investigate the role of *U. pineapple* in lipid metabolism, a single-bacterial gavage experiment was conducted based on a high-fat diet (HFD). *U. pineapple* was aerobically cultured on BL agar plates at 30 °C. Bacterial cells were washed off the plates with sterile PBS, and the bacterial culture was then diluted to 10^9 colony-forming units (CFU). Twenty-four male C57BL / 6N mice were randomly assigned to three groups: a normal diet control group (ND-Con, n = 8), a high-fat diet control group (HF-Con, n = 8), and a high-fat diet plus *U. pineapple* group (HF-B, n = 8, 10^9 CFU of *U. pineapple*). *U. pineapple* was freshly prepared daily and administered by gavage once daily. Body weight and food intake were measured weekly during the 9-week study.

[0027] Experiment 4: To evaluate whether oral administration of *Panthera pineapple* effectively triggers thermogenesis in brown adipose tissue (BAT), a gavage test was conducted under a normal diet (ND). Twenty male C57BL / 6N mice were randomly assigned to two experimental groups (n = 10 per group): a control group receiving PBS (ND-PBS) and a group receiving 10^9 colony-forming units (CFU) of *Panthera pineapple* daily (ND-P. ananatis). After a one-week stabilization period, mice were administered gavage daily for five consecutive days. Core body temperature was monitored at 0.5 hours and 1 hour after gavage. Subsequently, mice underwent a 4 °C cold exposure, and core body temperature was recorded at 2 and 4 hours post-exposure. At 4 hours post-exposure, the temperature difference between groups was captured using an infrared thermal imager. Finally, BAT was collected for quantitative PCR (qPCR) analysis to assess key thermogenic genes (including...). Ucp1 , Pgc1α , Prdm16 and Cidea The expression levels of mRNA in BAT were measured to determine the regulatory patterns of upstream and downstream pathways involved in short-term BAT activation.

[0028] Macaque fecal sample collection A total of 82 fecal samples were collected from seven macaque populations at different altitudes: 7 samples from Mangkang County, Tibet (4317 m), 12 samples from Yajiang County, Ganzi Prefecture, Sichuan (4120 m), 13 samples from Nyingchi City, Tibet (3100 m), 10 samples from Shennongjia National Park, Hubei (1600 m), 9 samples from Jiyuan City, Henan (1058 m), 10 samples from Beijing Zoo (575 m), and 21 samples from Dangan Island, Guangdong (50 m). Immediately after collection, samples were placed in sterile 15 mL centrifuge tubes and stored on dry ice. Subsequently, the samples were transported under dry ice conditions (transportation time up to one week), and finally stored at -80 ℃ in a freezer at the Institute of Zoology, Chinese Academy of Sciences, for further analysis.

[0029] DNA extraction, sequencing, and data quality control Microbial DNA was extracted from fecal samples using the QIAamp DNA Soil Mini Kit (Qiagen, Valencia, CA, USA) according to standard protocol. The quality and quantity of extracted DNA were assessed using a Nanodrop spectrophotometer (ND-1000, Nanodrop Technologies, Wilmington, DE, United States) and agarose gel electrophoresis. DNA samples were stored at -20 °C for later use. Shotgun sequencing was performed using the Illumina NovaSeq 6000 platform, generating at least 10 Gb of sequencing data per sample with a read length of 150 bp.

[0030] Metagenome assembly Raw sequencing reads were quality-trimmed using Trimmomatic version 0.36. Sequences with an average quality below 20 within a 4-base sliding window and reads shorter than 70 bp were removed. Subsequently, to eliminate host contamination, the data were aligned to the rhesus monkey genome (assembly number GCF_003339765.1) using Bowtie2 (v2.3.5). This process ensured the acquisition of clean, uncontaminated sequencing data. Metagenomic assembly was performed using MEGAHIT (v1.1.3) with default parameters, and sequence data for each sample were assembled separately. Gene prediction was then performed using MetaGeneMark in contigs longer than 300 bp.

[0031] Genome Reconstruction Gut microbial genome reconstruction was performed using the functional modules of metaWRAP (v1.1.1), which provides various modules for analyzing metagenomic bins. The default minimum contig length for bin construction using metaBAT2, Maxbin2, and CONCOCT was set to 1000 bp. Subsequently, the bin_refinement module of metaWRAP was used to optimize metagenomic assembly genomes (MAGs). The best MAGs were selected based on the highest scoring function, resulting in the construction of 31,096 MAGs.

[0032] MAGs redundancy removal and species-level clustering Redundancy was removed from all 31,096 MAGs using dRep (v2.2.3) with the parameters set to "-comp80 -con 10 -str 100 -strW 0". Of these, 2,922 non-redundant MAGs met the CheckM (v1.0.12) quality assessment criteria (integrity greater than 80% and contamination less than 10%). dRep was able to identify and select the best representative genomes from a set of highly similar genomes. Furthermore, using dRep's 'cluster' tool, MAGs were classified into species-level genome bins (SGBs) based on a 95% average nucleotide identity (ANI) threshold. SGBs with at least one reference genome in the Genome Taxonomy Database (GTDB) were classified as known SGBs, while those without a reference genome were classified as unknown SGBs. A total of 1,157 representative MAGs were identified, including 261 known SGBs and 896 unknown SGBs. Each MAG conformed to the Metagenomic Assembly Genome Core Information Standard (MIMAG), which requires each MAG to contain 23S, 16S, and 5S rRNA genes, at least 18 tRNA genes, with an integrity of over 90% and a contamination of less than 5%. These genetic elements were identified using Prokka.

[0033] MAGs and taxonomic unit abundance estimation Clean reads for each sample were aligned to representative MAGs, and the abundance of each MAG in each sample was quantified using the QUANT_bin module of metaWRAP. Based on the abundance and phylogenetic relationships of MAGs, they were classified into different taxonomic levels such as phylum, class, order, family, and genus. The abundance of MAGs was then summarized to generate abundance tables for each taxonomic level. Notably, in the bacterial abundance calculation of the metagenomic composition of colon contents in FMT mice, a macaque MAG library and the same quantitative method were used to compare the bacterial populations before and after transplantation.

[0034] Constructing a gene catalog of macaque gut microbiota To generate a comprehensive functional gene library of rhesus monkey gut microbiota, the gut microbiota genes of each individual were merged. First, MEGAHIT (v1.1.3) was used to generate contigs from the filtered data of each individual. Subsequently, MetaGeneMark was used to predict genes in contigs longer than 300 bp. Redundancy was removed using CD-HIT software with the parameters set to "-c 0.95 -aS 0.9 -g 1.0". A gene catalog containing 22,803,526 non-redundant genes was finally generated.

[0035] FMT mouse body weight, food intake, and tissue weight measurements During the experiment, body weight and food intake were measured daily at 9:00 AM using an electronic balance. Food intake (g) was calculated by subtracting the weight of remaining feed from the initial feed weight. After the transplantation experiment, the mice were euthanized using carbon dioxide, and necessary samples were collected, including: cadaver, brown fat, white fat, heart, liver, spleen, lungs, kidneys, blood, and colon contents.

[0036] Determination of blood glucose, blood lipids, total bile acids and insulin in FMT mice Serum glucose, total bile acids, total cholesterol (CHOL), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) levels were measured using kits from Beijing Xinchuangyuan Biotechnology Co., Ltd. Insulin levels were quantified using a male rat radioimmunoassay (RIA) kit from the China Institute of Atomic Energy, with a detection range of 5-160 μIU / mL.

[0037] Hematoxylin-eosin staining of small intestine and BAT Small intestinal samples from FMT mice were fixed at room temperature with 4% buffered paraformaldehyde and then embedded in paraffin. Tissue sections were 5–6 µm thick and stained with hematoxylin and eosin (H&E). Villous length and crypt depth were analyzed using ImageJ software. BAT (bacterial lipids) samples from mice administered *Ureaplasma bromelain* via gavage were paraffin-embedded, sectioned, and subjected to H&E staining and scanning to investigate the effect of *Ureaplasma bromelain* on the lipid content of BAT.

[0038] Targeted metabolomics analysis of short-chain fatty acids (SCFAs) in the colonic contents of FMT mice To compare the metabolic capacity of SCFAs produced by the gut microbiota of rhesus monkeys at high and low altitudes, colonic contents of FMT mice were homogenized in water containing glass beads, centrifuged, and then analyzed by GC-MS. A Thermo Fisher Scientific gas chromatograph was used for analysis, and specific parameters for injection, column temperature, and mass spectrometry detection were set. The concentration of target compounds in the samples was determined using a standard curve. Metabolites with concentrations below 0 were reported as not detected (ND). To assess the technical accuracy of each experiment, the relative standard deviation of the peak areas of all compounds detected in the quality control (QC) samples was calculated.

[0039] LC-MS / MS-based lipidomics analysis of liver and colon contents To compare the lipid metabolism capacity of the gut microbiota of rhesus monkeys at different altitudes, colonic contents and liver samples were collected for lipidomics analysis. Lipid extraction and mass spectrometry analysis were performed by Applied Protein Technology. Individual samples from each group were pooled to generate QC samples to ensure system stability and data reliability. LC-MS / MS analysis was performed using a Q Exactive Plus mass spectrometer (Thermo Fisher Scientific) coupled with a UHPLC Nexera LC-30A (SHIMADZU) system. The chromatographic column was a Waters ACQUITY UPLC CSH C18 (2.1 mm × 10 cm, particle size 1.7 µm). The column temperature was maintained at 45 ℃, and the flow rate was 300 μL / min. Mobile phase A consisted of acetonitrile aqueous solution (acetonitrile:water = 6:4, v / v) + 0.1% formic acid + 0.1 mM ammonium formate, and mobile phase B consisted of acetonitrile-isopropanol solution (acetonitrile:isopropanol = 1:9, v / v) + 0.1% formic acid + 0.1 mM ammonium formate. Electrospray ionization (ESI) was used for detection in both positive and negative ion modes. Mass spectrometry analysis was performed using a Q Exactive series mass spectrometer with the following ESI source conditions: heater temperature 300 °C; jacket gas flow rate 45 arb; auxiliary gas flow rate 15 arb; tail gas flow rate 1 arb; spray voltage 3000 V; capillary temperature 350 °C; S-lens RF level 50%; MS1 scan range 200-1800. Lipid identification, peak extraction, alignment, and quantification were performed using LipidSearch software (v4.1, Thermo Fisher Scientific). The extracted ion characteristics only consider variables that have more than 50% non-zero measurements in at least one set.

[0040] LC-MS / MS-based untargeted metabolomics of colonic contents To identify differentially expressed metabolites associated with *Ureaplasma pineapple*, non-targeted metabolomics (LC-MS / MS) analysis was performed on mouse colonic contents following a single bacterial gavage. QC samples prepared from equal volumes of mixed experimental samples were used to calibrate the LC-MS system and monitor instrument performance, thus assessing system stability. LC-MS / MS analysis involved separation on a Hypersil Gold C18 column (2.1 mm × 10 cm, 1.9 µm particle size, Thermo Fisher Scientific) coupled with a Q Exactive™ HF / Q Exactive™ HF-X mass spectrometer and a Vanquish UHPLC system. Column temperature was maintained at 40 °C, and flow rate was 200 μL / min. Mobile phase A was water containing 0.1% formic acid, and mobile phase B was methanol. The mass spectrometer was operated in negative and positive scan modes (100–1,500 m / z) with the following ion source parameters: spray voltage 3,500 V; jacket gas flow rate 35 psi; auxiliary gas flow rate 10 L / min; capillary temperature 320 °C; S-lens RF level 60; and auxiliary gas heater temperature 350 °C. Data were imported into the CD3.3 database search software for processing and compared with the mzCloud, mzVault, and Masslist databases. Standardization was then performed to obtain relative peak areas. Compounds with a peak area coefficient of variation (CV) exceeding 30% in the QC samples were excluded, thus obtaining the final identification and relative quantification of metabolites.

[0041] Liver biochemical markers test To observe the effects of gavage administration of Pantothecin bromelain on liver health and lipid content in HFD mice, alanine aminotransferase (ALT), aspartate aminotransferase (AST), cholesterol (CHOL), and triglycerides (TG) were measured using the alanine substrate method, aspartate substrate method, CHOD-PAP method, and GPO-PAP method, respectively.

[0042] Western blotting of UCP1 in mouse BAT To investigate whether *Ureaplasma bromelain* can activate the energy metabolism function of BAT (biotinylated amino acids), UCP1 was extracted from the BAT of mice administered *Ureaplasma bromelain* via gavage. The differences in UCP1 content were then assessed by Western blotting (WB). 15 µg of protein was used for WB analysis. SDS-PAGE electrophoresis and membrane transfer were performed according to standard protocols. UCP1 antibody (Abcam Shanghai Trading Co., Ltd.) was diluted 1:2000 and incubated overnight at 4 °C. Subsequently, it was incubated for 1 hour at room temperature with a 1:5000 dilution of goat anti-rabbit HRP secondary antibody. The chemiluminescent reaction was initiated using ECL chemiluminescence buffer (Abclonal, China), and the results were visualized using the Bio-Rad ChemiDoc XRS+ system. For quantification of the Western blotting results, the cumulative optical density (IntDen) of the protein bands was analyzed using ImageJ software. The net cumulative optical density (NetIntDen) was obtained after background subtraction.

[0043] Example 1 Obtained through metagenomic molecular methods Pantoea ananatis Purchased from the company Pantoea ananatis This strain was purchased from Baosai Biotechnology Co., Ltd. The following is the instruction manual for this strain: Fungal strain name: Pantothecinus pineapple Pantoea ananatis Strain preservation information: Autonomous isolation Characteristics: Non-type strain, Class 1 safety microorganism Genotype: Non-whole genome sequenced strain Resistance information: None Culture conditions: LB medium, 30 degrees Celsius, aerobic medium Note: Bacteria isolated from plant source Number: U0090 Manufacturer: Bioscibio The above experiments revealed that: 1. Pantoea ananatis It can effectively prevent obesity To verify Pantoea ananatis ( P. ananatis The role of lipid metabolism in this invention was investigated by gavage administration to high-fat diet mice (HF-B). P. ananatis ( Figure 1 a) The results show that, P. ananatis It can significantly reduce the body weight of mice. Figure 1 b, c; p<0.05), brown fat (BAT) weight ( Figure 1 d; p<0.05) and white fat (WAT) weight (Figure 1 e–g; p<0.01). For further research P. ananatis The effects on lipid metabolism were investigated by measuring the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acids (CHOL), and triglycerides (TG) in mouse liver. The results showed that... P. ananatis It can significantly reduce TG levels in the liver, while having no significant effect on other indicators. Figure 1 h–k). This prompt P. ananatis It has a significant lipid-lowering effect.

[0044] Analysis of BAT slices showed that, compared with the untreated high-fat diet group (HF-Con), oral administration P. ananatis The BAT volume of the mice was closer to that of the normal diet group (ND-Con). Figure 1 l). Furthermore, Western blot analysis of UCP1 protein in BAT showed that oral administration... P. ananatis It can effectively activate the heat generation function of BAT (Baidu, Alibaba, Tencent). Figure 1 This effect (m) may be related to its anti-obesity effect.

[0045] Table 1. Changes in mice after oral administration of Pantoea ananatis

[0046] 2. Pantoea ananatis It can produce a variety of metabolites To investigate P. ananatis Potential metabolites that could activate BAT were identified, and the untargeted metabolome of colonic contents was compared between ND-Con and HF-Con, and between HF-B and HF-Con. Compared to HF-Con, ND-Con showed a significant enrichment of 132 metabolites (|fold change|>2; p<0.005). Furthermore, the HF-B group showed a significant enrichment of 72 metabolites compared to HF-Con. p <0.005). Notably, there are 12 overlapping metabolites between the two groups, including 1-methylguanosine, 2'-deoxyadenosine, coumarin, ferulic acid, adenine, D-(+)-maltose, and isoflavonic acid.

[0047] In summary, this invention utilizes macaques at different altitudes ( Macaca mulattaA temperature gradient was established, and fecal microbiota transplantation experiments were conducted to mitigate differences caused by dietary and environmental factors. This method enables the study of the gut microbiota's response to low temperatures, particularly whether it activates BAT in non-low-temperature environments, and to explore its potential mechanisms. Fecal samples were collected from wild rhesus macaque populations living at altitudes between 50 and 4000 meters, with annual average temperatures of 22.5°C and 5.8°C, respectively. This species represents an excellent model for studying the cold adaptation mechanisms of the gut microbiota. The results showed that at room temperature, the high-altitude gut microbiota significantly reduced white fat, serum total bile acids, and triglyceride levels. Metagenomic analysis identified the main lipid metabolic pathways associated with the high-altitude gut microbiota, highlighting... Pantoea ananatis It is a key bacterial species. Furthermore, a high-fat diet model was established using isogenetic male mice to demonstrate... Pantoea ananatis It can activate the UCP1 protein, thereby enhancing the thermogenesis capacity of BAT, reducing WAT content, and preventing obesity.

[0048] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0049] The sequence numbers of the above embodiments of the present invention are for descriptive purposes only and do not represent the superiority or inferiority of the embodiments.

[0050] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. Pantoea ananatis Pantoea ananatis or a fermentation broth thereof or a microbial agent containing the same in the preparation of a weight loss related product.

2. Use according to claim 1, characterized in that, The weight loss-related product is used to prevent or improve obesity caused by a high-fat diet.

3. Pantoea ananatis Pantoea ananatis or a fermentation broth thereof in the preparation of a hypolipidemic product.

4. Use according to claim 3, characterized in that, The blood lipid includes triglyceride.

5. Pantoea ananatis Pantoea ananatis or a fermentation broth thereof or a microbial agent containing the same for use in the preparation of a product for modulating the gut microbiota.

6. Pantoea ananatis Pantoea ananatis or a fermentation broth thereof or a bacterial agent containing the same in the manufacture of a product for activating brown adipose tissue, reducing white adipose tissue storage, and / or enhancing triglyceride metabolism.

7. Use according to any one of claims 1 to 6, characterized in that, The product includes food, medicine, or additive.

8. The use according to claim 7, wherein, The dosage form of the medicine includes any one of powder, granule, tablet, pill, capsule, suspension, emulsion, syrup, aerosol, or suppository. The food includes any one of dietary supplement, prebiotic, solid beverage, or dairy product.

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