A strain of Brevibacillus laterosporus with blood sugar-lowering and fat-reducing effects and its application

Abstract

The present invention discloses a strain of Brevibacillus laterosporus with blood sugar and fat reduction efficacy and its application. The strain is Brevibacillus laterosporus ASD0001, with a deposit number of GDMCC No: 63890. The present invention independently screened a Brevibacillus laterosporus strain from soil. The strain has the efficacy of reducing the weight of obese animals, inhibiting adipose tissue formation, and improving glucose tolerance and insulin resistance in obese animals. The strain can be used to prepare blood sugar and fat reduction products or products for preventing and treating diabetes, and has good market application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to a strain of Brevibacillus laterosporus with blood sugar-lowering and fat-reducing effects and applications thereof. Background Art

[0002] As people's living standards improve, the proportion of obese people is increasing, and obesity has become one of the most serious public health problems worldwide. The World Obesity Association's latest "Global Obesity Map 2023" shows that by 2035, more than 4 billion people in the world will be obese or overweight, accounting for 51% of the total population. The causes of obesity are diverse. In addition to the well-known problems with diet structure, there are also emotions, exercise frequency, lifestyle habits, etc. Relying solely on diet to regulate obesity is very difficult and unsustainable. However, obesity is not only related to appearance issues, but is also crucial to human health. For example, obesity is often accompanied by the occurrence of cardiovascular, liver, kidney and other diseases. Therefore, how to control the occurrence of obesity is becoming increasingly important for human health.

[0003] Besides weight loss measures like diet, exercise, and a regular lifestyle, medication is a relatively easy way to manage obesity. Common medications include digestive enzyme inhibitors, such as orlistat, which bind to pancreatic lipase in the intestines, preventing the breakdown and absorption of glycerol and fat in food. Appetite suppressants, such as dexfenfluramine, sibutramine, and fenfluramine, suppress appetite by stimulating the production and release of neurotransmitters and inhibiting their reuptake, essentially reducing food intake. However, these chemically synthesized weight loss drugs can have serious side effects. For example, sibutramine can cause dry mouth, constipation, dizziness, and insomnia. Since 2010, the State Food and Drug Administration has banned the production, sale, and use of sibutramine preparations and APIs in my country. Orlistat can also cause adverse gastrointestinal reactions, including flatulence, urgency during bowel movements, increased bowel movement frequency, and fatty stools. Recently, semaglutide has attracted much attention in the weight loss market. It can achieve the dual effects of lowering blood sugar and reducing weight. Its main mechanism is that it can activate the GLP-1 (glucagon-like peptide-1) receptor, promote insulin secretion in a glucose concentration-dependent manner, inhibit glucagon secretion, delay gastric emptying and increase satiety, and suppress appetite by inhibiting the action of the hypothalamic feeding center, thereby achieving the effect of lowering blood sugar and reducing weight. However, it also has certain side effects.

[0004] Therefore, developing a weight loss product with few side effects and good effects is a category that is currently in short supply in the market. Summary of the Invention

[0005] The first object of the present invention is to provide a strain of Brevibacillus laterosporus ASD0001 with the efficacy of lowering blood sugar and reducing fat. The strain has been deposited in the Guangdong Provincial Microbiological Culture Collection Center (GDMCC), address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province, Postal Code: 510070, with a deposit number of GDMCC No: 63890 and a deposit date of October 19, 2023.

[0006] The second object of the present invention is to provide the use of the above-mentioned Brevibacillus laterosporus ASD0001 in the preparation of products with blood sugar and fat reduction functions.

[0007] Preferably, the active ingredient of the blood sugar and fat reducing functional product comprises the bacteria of Brevibacillus laterosporus ASD0001 or its fermentation product.

[0008] Preferably, the blood sugar and fat reducing functional product includes a medicine.

[0009] The third object of the present invention is to provide the use of the above-mentioned Brevibacillus laterosporus ASD0001 in the preparation of products for preventing and treating diabetes.

[0010] Preferably, the product for preventing and treating diabetes comprises a medicine.

[0011] The fourth object of the present invention is to provide a blood sugar and fat reducing functional product, which contains the bacteria of Brevibacillus laterosporus ASD0001 or its fermentation product as an active ingredient.

[0012] A fifth object of the present invention is to provide a product for preventing and treating diabetes, comprising the bacteria of Brevibacillus laterosporus ASD0001 or a fermentation product thereof as an active ingredient.

[0013] The present invention has the following beneficial effects:

[0014] The present invention independently screened out a Brevibacillus laterosporus from the soil, which has the effects of reducing the weight of obese animals, inhibiting the formation of adipose tissue, and improving the glucose tolerance and insulin resistance of obese animals. It can be used to prepare products with blood sugar and fat reduction functions or products for preventing and treating diabetes, and has good market application prospects.

[0015] Brevibacillus laterosporus ASD0001 was deposited on October 19, 2023 in the Guangdong Provincial Microbiological Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, Postal Code: 510070, with the deposit number: GDMCC No: 63890. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Figure 3. Body weight differences between BL1 and ASD0001 mice after four weeks of high-fat diet feeding and six weeks of gavage feeding. A: Comparison of body size between the four groups of mice at week 6. From left to right: control group (C), model group (H), BL1 group (BL1), ASD0001 group (ASD0001); B: Body weights of the four groups of mice at week 6. * Represents the significant difference in body weight between the ASD0001 group, the model group, and the BL1 group. * represents p<0.05, ** Represents p < 0.01.

[0017] Figure 2 Figure 2. Differences in inguinal white adipose tissue (iWAT) weights between BL1 and ASD0001 mice after four weeks of high-fat diet feeding and six weeks of gavage feeding. A: Images of iWAT from four groups of mice. From left to right: control group (C), model group (H), BL1 group (BL1), and ASD0001 group (ASD0001); B: iWAT weights of the four groups of mice. ## The difference between group H and group C was extremely significant (p < 0.01). ** This indicates that the difference between ASD0001 and group H and group BL1 is extremely significant, p < 0.01.

[0018] Figure 3 Figure 2. The difference in epididymal white adipose tissue (eWAT) weight between BL1 and ASD0001 mice after 4 weeks of high-fat diet feeding and 6 weeks of gavage feeding. A: Photographs of eWAT from four groups of mice. From left to right: control group (C), model group (H), BL1 group (BL1), and ASD0001 group (ASD0001); B: eWAT weight measurements from four groups of mice. ## The difference between group H and group C was extremely significant (p < 0.01). ** Indicates that the difference between ASD0001 and H group is extremely significant, p < 0.01, * Indicates significant difference between ASD0001 and BL1 groups, p < 0.05.

[0019] Figure 4 Oil Red O staining of liver tissue from BL1 and ASD0001 mice fed a high-fat diet for 4 weeks and then gavage-fed for 6 weeks. In the figure, C: control group, H: model group, BL1: BL1-treated group, ASD0001: ASD0001-treated group.

[0020] Figure 5 The difference in glucose tolerance between BL1 and ASD0001 mice after being fed a high-fat diet for 4 weeks and then gavage-fed for 6 weeks. In the figure, C: control group, H: model group, BL1: BL1-treated group, ASD0001: ASD0001-treated group, ##The difference between group H and group C was extremely significant (p < 0.01). ** Indicates that the difference between ASD0001 and H groups is extremely significant, p < 0.01.

[0021] Figure 6 The difference in insulin resistance between BL1 and ASD0001 mice after being fed a high-fat diet for 4 weeks and then gavage-fed for 6 weeks. In the figure, C: control group, H: model group, BL1: BL1-treated group, ASD0001: ASD0001-treated group, ## The difference between group H and group C was extremely significant (p < 0.01). * Indicates that there is a significant difference between ASD0001 and H groups, p < 0.05. DETAILED DESCRIPTION

[0022] The present invention will be further described below with reference to the accompanying drawings and specific examples, but the examples do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0023] The strain Brevibacillus laterosporus BL1 involved in the following examples has a deposit number of GDMCC No: 62699 and has been disclosed in the document Weng GY, Huang J, et al. Brevibacillus laterosporus BL1, apromoting probiotic, prevents obesity and modulates gut microbiota in mice fed a high-fat diet. Frontiers in Nutrition. 2022 Nov 24: 9: 1050025.

[0024] Example 1: Lipid-lowering effect of Brevibacillus laterosporus ASD0001

[0025] Forty 5-week-old male C57BL / 6 mice (purchased from Zhuhai Best Biotechnology Co., Ltd.) were individually housed in a controlled environment (24±2°C, 45-60% relative humidity, 12h / 12h light-dark cycle) with free access to food throughout the experiment. After acclimation for one week, the mice were randomly divided into four groups (n=10 per group), namely the control group (C), the model group (H), the BL1 group (BL1), and the ASD0001 group (ASD0001). The mice in the control group were fed with a normal diet (Article No. XTCON50J, purchased from Xiehe Biological Co., Ltd.), while the mice in the model group, BL1 group, and ASD0001 group were all fed with a high-fat diet (Article No. XTHF60, purchased from Xiehe Biological Co., Ltd.). After 4 weeks of high-fat diet feeding, the mice in the BL1 group and the ASD0001 group were gavaged with the strain BL1 (1×10 9 CFU / d / mouse) and ASD0001 (1×10 9 CFU / d / mouse) for 6 consecutive weeks, while the mice in the control and model groups were gavaged with an equal volume of normal saline. The body weight (BW) of the mice was recorded weekly. At the end of the 6-week treatment, the body weight of the mice was measured. After the mice were sacrificed, the liver, inguinal white adipose tissue (iWAT) and epididymal white adipose tissue (eWAT) were obtained, and the weights of iWAT and eWAT were measured. The liver was fixed and stained with Oil Red O to observe the level of fat formation in the liver tissue. The results are shown in Figure 1-4 .

[0026] Figure 1 The results showed that after 4 weeks of high-fat diet feeding (the 0th week for weight calculation), the body weight of mice was significantly higher than that of mice in the control group C. After 6 weeks of gavage feeding with BL1 and ASD0001, the body weight of mice in the model group H was significantly fatter than that of mice in the control group C. The body weight of mice in the BL1 and ASD0001 groups was significantly thinner than that of mice in the model group H ( Figure 1 A). Weekly weight measurements of mice in each group revealed that, from 1 to 6 weeks of gavage, the BL1 and ASD0001 groups maintained lower weights than the model H group. After 5 and 6 weeks of gavage, the ASD0001 group exhibited significantly lower weights than the model H group and the BL1 group. This suggests that, compared with the BL1 strain, ASD0001 is more effective in controlling the weight of obese mice.

[0027] Figure 2The study showed that mice were fed a high-fat diet for 4 weeks and then gavage-fed with BL1 and ASD0001 for 6 weeks. The mice were then dissected and their iWAT weights measured. The results showed that the iWAT weight of the model group H was significantly higher than that of the control group C. The iWAT weights of the mice gavage-fed with ASD0001 and BL1 were significantly lower than those of the model group H, and the iWAT weight of the ASD0001 group was significantly lower than that of the BL1 group. This suggests that ASD0001 is more effective than the BL1 strain in reducing iWAT formation in mice.

[0028] Figure 3 The study showed that mice were fed a high-fat diet for 4 weeks and then gavage-fed with BL1 and ASD0001 for 6 weeks. The mice in each group were dissected and their eWAT weights were measured. The results showed that the eWAT weight of the model group H was significantly higher than that of the control group C. The eWAT weights of the mice gavage-fed with ASD0001 and BL1 were significantly lower than those of the model group H, and the eWAT weight of the ASD0001 group was significantly lower than that of the BL1 group. This suggests that ASD0001 is more effective than the BL1 strain in reducing eWAT formation in mice.

[0029] Figure 4 The study shows that mice were fed a high-fat diet for 4 weeks and then gavage-fed with BL1 and ASD0001 for 6 weeks. Liver tissue was dissected from each group and the levels of fat formation in the liver tissues of the four groups were compared. As shown in the figure, the level of fat formation in the model group H was significantly higher than that in the control group C. Both the gavage-fed ASD0001 and BL1 groups effectively reduced fat formation in the mice, indicating that ASD0001 is more effective than the BL1 strain in reducing liver fat formation.

[0030] Example 2: Hypoglycemic Effect of Brevibacillus laterosporus ASD0001

[0031] Forty 5-week-old male C57BL / 6 mice were individually housed in a controlled environment (24±2°C, 45-60% relative humidity, 12h / 12h light-dark cycle) with free access to food throughout the experiment. After acclimation for one week, the mice were randomly divided into four groups (n=10 per group), namely the control group (C), the model group (H), the BL1 group (BL1), and the ASD0001 group (ASD0001). The mice in the control group were fed with a normal diet (Article No. XTCON50J, purchased from Kyowa Biotechnology Co., Ltd.), while the mice in the model group, BL1 group, and ASD0001 group were all fed with a high-fat diet (Article No. XTHF60, purchased from Kyowa Biotechnology Co., Ltd.). After four weeks of high-fat diet feeding, the mice in the BL1 group and the ASD0001 group were gavage-treated with the strain BL1 (1×10 9 CFU / d / mouse) and ASD0001 (1×10 9CFU / d / mouse), and were gavaged for 6 consecutive weeks, while the mice in the control group and model group were gavaged with an equal volume of normal saline. Intraperitoneal glucose tolerance test (IGTT) and insulin tolerance test (ITT) were performed at week 7. Blood glucose levels in the tail vein were detected using a blood glucose meter. In IGTT analysis, mice were intraperitoneally injected with glucose (2g / kg body weight) after fasting overnight, and then the blood glucose levels were measured at 0, 15, 30, 60, 90 and 120 minutes after injection. In ITT analysis, mice were intraperitoneally injected with insulin (0.75U / kg body weight) after fasting for 6 hours, and then the blood glucose levels were measured at 0, 15, 30, 60, 90 and 120 minutes after injection. The results are shown in Figure 5-6 .

[0032] Figure 5 The results showed that after mice were fed a high-fat diet for 4 weeks, they were gavage-fed with ASD0001 and BL1 for 6 weeks. Overnight fasted mice were intraperitoneally injected with glucose (2g / kg body weight), and blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes. The glucose levels in the model group H were significantly higher than those in the control group C. At 30, 60, 90, and 120 minutes after glucose injection, the blood glucose levels in the ASD0001 and BL1 groups were significantly lower than those in the model group H. For high-fat-induced obese mice, ASD0001 was more effective in improving glucose tolerance than the BL1 strain.

[0033] Figure 6 The study showed that mice were fed a high-fat diet for 4 weeks and then gavage-fed with ASD0001 and BL1 for 6 weeks. After fasting for 6 hours, the mice were intraperitoneally injected with insulin (0.75 U / kg body weight). Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes. Blood glucose levels in the model group H were higher than those in the control group C at all times. At 15 and 120 minutes after insulin injection, blood glucose levels in the ASD0001 group were lower than those in the model group H and BL1 groups. In high-fat-induced obese mice, ASD0001 was more effective than the BL1 strain in improving insulin resistance.

[0034] The above are merely preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be construed as limiting the present invention, and the scope of protection of the present invention should be determined by the scope defined in the claims. Persons skilled in the art will appreciate that improvements and modifications may be made without departing from the spirit and scope of the present invention, and such improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Brevibacillus laterosporus ( Brevibacillus laterosporus ) ASD0001, deposit number: GDMCC No: 63890.

2. Use of the Brevibacillus laterosporus ASD0001 according to claim 1 in the preparation of blood sugar and fat reducing functional medicines.

3. The use according to claim 2, characterized in that The active ingredient of the blood sugar and fat reducing functional medicine comprises the bacteria of Brevibacillus laterosporus ASD0001.

4. Use of the Brevibacillus laterosporus ASD0001 according to claim 1 in the preparation of medicines for preventing and treating diabetes.

5. A drug for reducing blood sugar and reducing fat, characterized in that: The invention comprises the bacterial cell of Brevibacillus laterosporus ASD0001 according to claim 1 as an active ingredient.

6. A medicine for preventing and treating diabetes, characterized in that: The invention comprises the bacterial cell of Brevibacillus laterosporus ASD0001 according to claim 1 as an active ingredient.

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