Vegetable protein meat capable of improving glycolipid metabolism status

Abstract

The invention belongs to the field of biological manufacturing, and particularly relates to vegetable protein meat capable of improving glycolipid metabolism conditions. Chlorogenic acid; laccase; wherein the vegetable protein product is prepared from the raw materials through a cross-linking reaction. The vegetable protein product disclosed by the invention is obtained by specifically modifying the vegetable protein by combining chlorogenic acid and laccase, and the product not only can effectively control the weight gain, but also can systematically improve the glucose and lipid metabolism disorder caused by high-fat diet; the method mainly comprises the aspects of abnormal blood sugar level, abnormal blood fat level, liver fatty degeneration, intestinal flora imbalance and the like, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of biomanufacturing, and specifically relates to a plant-based meat protein that can improve glucose and lipid metabolism. Background Technology

[0002] As the proportion of high-fat and high-sugar foods in modern diets continues to increase, the proportion of obese people is also rising. Specifically, long-term intake of high-fat and high-sugar foods will disrupt the body's energy balance, promote excessive proliferation and hypertrophy of adipose tissue, and cause dysfunction of fat cells, leading to a series of metabolic complications such as insulin resistance, dyslipidemia, and non-alcoholic fatty liver disease.

[0003] Currently, interventions and treatments for obesity and its metabolic complications mainly include lifestyle management, drug therapy, and surgery. However, adherence to lifestyle management is generally low, drug therapy carries certain risks of side effects and weight rebound is likely after discontinuation, and surgery is highly invasive.

[0004] Therefore, there is an urgent need to develop a treatment strategy that has high patient compliance and can effectively intervene in obesity and its metabolic complications. Summary of the Invention

[0005] This invention aims to at least partially address one of the technical problems existing in the prior art. To this end, this invention provides a plant-based meat that can improve glucose and lipid metabolism. The plant-based protein product of this invention is obtained by specifically modifying plant proteins using a combination of chlorogenic acid and laccase. This product not only effectively controls weight gain but also systematically improves glucose and lipid metabolism disorders induced by a high-fat diet, mainly including abnormal blood glucose levels, abnormal blood lipid levels, hepatic steatosis, and intestinal flora imbalance, showing broad application prospects.

[0006] In a first aspect, the present invention provides a plant protein product. According to an embodiment of the present invention, the plant protein product comprises the following raw materials: plant protein; chlorogenic acid; laccase; wherein the plant protein product is obtained by cross-linking the raw materials. The plant protein product according to an embodiment of the present invention is obtained by specifically modifying the plant protein with a combination of chlorogenic acid and laccase. This product effectively intervenes in weight gain caused by obesity, and also intervenes in glucose and lipid metabolism disorders usually linked to obesity, mainly including abnormal blood glucose levels, abnormal blood lipid levels, hepatic steatosis, and intestinal flora imbalance.

[0007] According to embodiments of the present invention, the above-mentioned plant protein products may also have at least one of the following additional technical features: According to embodiments of the present invention, the plant protein includes one or more of soy protein isolate, pea protein, and wheat protein.

[0008] According to an embodiment of the present invention, the mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2).

[0009] In a second aspect, the present invention provides a food, medicine, or health product. According to embodiments of the present invention, the food, medicine, or health product includes: the plant protein product described in the first aspect.

[0010] Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this food, medicine, or health product, and will not be repeated here.

[0011] In a third aspect, the present invention provides a method for preparing the plant protein product described in the first aspect. According to an embodiment of the present invention, the method includes the following steps: contacting a modified system with the plant protein to perform a cross-linking reaction to obtain the plant protein product; wherein the modified system includes laccase and chlorogenic acid. The method according to the embodiments of the present invention has advantages such as being mild, green and safe, and producing plant protein products with excellent performance.

[0012] According to embodiments of the present invention, the above method may further include at least one of the following additional technical features: According to an embodiment of the present invention, the mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2).

[0013] According to an embodiment of the present invention, the contact method is gradient heating treatment, the initial temperature of the gradient heating treatment is 20~40℃, the final temperature of the gradient heating treatment is 135~150℃, and each temperature gradient is 25~35℃.

[0014] In a fourth aspect, the invention proposes the use of the plant protein products described in the first aspect in the preparation of a medicament. According to embodiments of the invention, the medicament is used for the prevention, relief, adjunctive treatment, or treatment of obesity and related metabolic syndromes.

[0015] Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this application, and will not be repeated here.

[0016] According to embodiments of the present invention, the above-described uses may further include at least one of the following additional technical features: According to embodiments of the present invention, the associated metabolic syndrome includes one or more of hyperlipidemia, non-alcoholic fatty liver disease, and insulin resistance.

[0017] According to embodiments of the present invention, the drug has at least one of the following uses: controlling body weight; regulating blood lipid levels; regulating blood glucose levels; improving hepatic lipid deposition; improving glucose tolerance; improving insulin sensitivity; improving organ coefficients; improving adipose tissue deformation; and improving intestinal flora imbalance; wherein the organ coefficients include one or more of the following: heart coefficient, liver coefficient, spleen coefficient, and kidney coefficient; and the blood lipid level indicators are selected from one or more of the following: total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

[0018] In a fifth aspect, the present invention proposes the use of the plant protein products described in the first aspect in the preparation of health supplements. According to embodiments of the present invention, the health supplement has at least one of the following uses: regulating blood lipids and / or assisting in lowering blood lipids; regulating blood sugar and / or assisting in lowering blood sugar.

[0019] Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this application, and will not be repeated here.

[0020] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0021] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1 This is a statistical chart showing the weight gain of mice in each group in Example 1 of the present invention. In the chart, A shows the weight gain of mice in each group from week 1 to week 8, and B shows the weight gain of mice in each group. "a", "b", "d", "bc" and "cd" represent significant differences between the groups. Figure 2 The diagram shows the statistical results of organ coefficients of mice in each group in Example 1 of the present invention. A is the statistical results of liver coefficient of mice in each group, B is the statistical results of inguinal fat index of mice in each group, and C is the statistical results of epididymal fat index of mice in each group. "a", "b", "c", "d", "ab" and "bc" represent significant differences between groups. Figure 3 The figures shown are statistical results of serum lipid metabolism indicators of mice in each group in Example 1 of the present invention. Among them, A is the statistical result of triglyceride (TG) content in the serum of mice in each group, B is the statistical result of total cholesterol (TC) content in the serum of mice in each group, C is the statistical result of high-density lipoprotein cholesterol (HDL-C) content in the serum of mice in each group, and D is the statistical result of low-density lipoprotein cholesterol (LDL-C) content in the serum of mice in each group. Figure 4 The images show the pathological morphology of liver samples from mice in each group in Example 1 of this invention. From left to right, the images show the pathological morphology of liver samples from mice in the normal control group (CON), high-fat group (HFD), unmodified group (SHME), enzyme-modified group (LHME), and positive control group (MET). The scale bar is 100 μm. Figure 5 The images show the pathological morphological observation results of epididymal adipose tissue samples from mice in each group in Example 1 of the present invention. From left to right, the images show the pathological morphological observation results of epididymal adipose tissue samples from mice in the normal control group (CON), high-fat group (HFD), unmodified group (SHME), enzyme-modified group (LHME), and positive control group (MET). The scale bar is 100 μm. Figure 6 The graph shows the statistical results of insulin levels in each group of mice in Example 1 of the present invention. In the graph, A is the test result of serum insulin content level in each group of mice, B is the statistical result of insulin resistance index in each group of mice, and "a", "b", "c" and "d" represent significant differences between the groups. Figure 7 The graph shows the glucose tolerance test results of each group of mice in Example 1 of the present invention. In the graph, A is the blood glucose level test result of each group of mice in the OGTT, and B is the statistical result of the area under the curve (AUC) of each group of mice in the OGTT. "a", "b", "c", "d" and "bc" represent significant differences between the groups. Figure 8 The graphs show the statistical results of the gut microbiota of mice in each group in Example 1 of the present invention. In the graphs, A represents the statistical results of the Chao1 index of each group of mice, B represents the statistical results of the ACE index of each group of mice, C represents the statistical results of the Shannon index of each group of mice, D represents the statistical results of the Simpson index of each group of mice, E represents the statistical results of the β diversity PCA of each group of mice, and F represents the statistical results of the NMDS of each group of mice. "a", "b" and "ab" represent significant differences between the groups. Detailed Implementation

[0022] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0023] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0024] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0025] In this document, the terms “comprising” or “including” are open-ended expressions, meaning that they include the contents specified in this invention, but do not exclude other aspects.

[0026] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0027] Terms and Definitions In this article, the term "plant protein product" refers to a product obtained by using plant-derived protein as the main raw material and then performing a cross-linking reaction with chlorogenic acid and laccase.

[0028] In this paper, the term "chlorogenic acid" refers to a naturally occurring phenolic compound with the chemical name 3-(3,4-dihydroxycinnamoyl)quinic acid, which belongs to the hydroxycinnamic acid class of substances. In this invention, it serves as a key component, working in conjunction with laccase to specifically modify plant-derived proteins.

[0029] In this document, the term "laccase" refers to a copper-containing polyphenol oxidase that can catalyze the oxidation of various phenolic substances. In this invention, it acts as a biocatalyst, specifically catalyzing the oxidation of chlorogenic acid phenolic hydroxyl groups, initiating a subsequent cross-linking reaction with plant proteins, thereby obtaining the plant protein products described in this invention.

[0030] In this article, the term "crosslinking reaction" refers to the chemical reaction process in which chlorogenic acid molecules and plant protein molecules form covalent bonds under the biocatalysis of laccase, constructing a protein-chlorogenic acid-protein three-dimensional network structure. This reaction process is mild and does not require the addition of toxic chemical crosslinking agents.

[0031] In this article, the term "obesity" refers to a disease state caused by long-term intake of high-fat and high-sugar foods. It is mainly manifested in multiple aspects such as weight gain, excessive accumulation and abnormal distribution of adipose tissue. Furthermore, these core pathological changes can extend and aggravate a series of interrelated metabolic abnormalities, namely "obesity-related metabolic syndrome", which mainly includes various types such as glucose metabolism disorders (such as elevated fasting blood glucose, impaired glucose tolerance, and insulin resistance), lipid metabolism disorders (such as hypertriglyceridemia and low high-density lipoprotein cholesterol), hypertension, gut microbiota imbalance, and non-alcoholic fatty liver disease.

[0032] In this article, the term "metformin" refers to a biguanide insulin sensitizer (chemical name: 1,1-dimethylbiguanide) that is widely used in clinical practice. In the embodiments of this invention, it is used as a positive control drug to compare the relevant performance of the plant protein products of this invention, such as the effect on improving weight gain and the effect on improving abnormal blood lipid levels.

[0033] Plant protein products This invention proposes a plant protein product. According to an embodiment of the invention, the plant protein product comprises the following raw materials: plant protein; chlorogenic acid; laccase; wherein the plant protein product is obtained by cross-linking the raw materials. The plant protein product according to an embodiment of the invention is obtained by specifically modifying the plant protein with a combination of chlorogenic acid and laccase. This product effectively intervenes in weight gain caused by obesity, and also intervenes in glucose and lipid metabolism disorders that are usually linked to obesity, mainly including abnormal blood glucose levels, abnormal blood lipid levels, hepatic steatosis, and intestinal flora imbalance.

[0034] According to embodiments of the present invention, the plant protein includes one or more of soy protein isolate, pea protein, and wheat protein. Therefore, the plant protein product is suitable for a variety of plant protein raw materials.

[0035] It should be noted that the types of plant proteins applicable to the plant protein products of the present invention are not limited to the specific types listed above. Other plant proteins that can be specifically modified by the combination of chlorogenic acid and laccase, such as chickpea protein, rice protein, and corn protein, are all within the scope of protection of the present invention.

[0036] According to an embodiment of the present invention, the mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2). Therefore, by optimizing and controlling the addition ratio of each component in the plant protein product, while adjusting the solubility and other properties of the plant protein, its related biological activities (such as the effect of regulating blood lipid levels) are further optimized. For example, the mass ratio of the plant protein, the chlorogenic acid, and the laccase is 800:1:0.5, 900:1:0.5, 1000:1:0.5, 1100:1:0.5, 1200:1:0.5, 800:2:0.5, 800:3:0.5, 800:4:0.5, 800:5:0.5, 800... The ratios are 6:0.5, 800:1:0.6, 800:1:0.7, 800:1:0.8, 800:1:0.9, 800:1:1, 800:1:1.1, 800:1:1.2, 800:1:1.3, 800:1:1.4, 800:1:1.5, 800:1:1.6, 800:1:1.7, 800:1:1.8, 800:1:1.9, 800:1:2, preferably (900~1000):(1~4):(1~2), and more preferably 10000:4:1.

[0037] It should be noted that the present invention does not limit the specific form of the plant protein product, which can be any physical form such as powder, granules, paste, textured protein, high moisture extrudate, etc., as long as it contains the plant protein product obtained by modifying the plant protein with chlorogenic acid and laccase, it falls within the protection scope of the present invention.

[0038] Food, medicine or health products This invention provides a food, medicine, or health product. According to embodiments of the invention, the food, medicine, or health product includes the aforementioned plant protein product.

[0039] Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this food, medicine, or health product, and will not be repeated here.

[0040] For example, when the aforementioned plant protein products are added to food, their forms include, but are not limited to, imitation meat products, protein powder, meal replacement shakes, energy bars, noodles, bread, biscuits, etc.; when the aforementioned plant protein products are added to pharmaceuticals, they can be used as the main active ingredient, combined with a pharmaceutically acceptable carrier, to prepare pharmaceutical preparations for the prevention or treatment of diseases such as obesity and non-alcoholic fatty liver disease, such as oral preparations (capsules, tablets), etc.; when the aforementioned plant protein products are added to health supplements, the soluble plant protein products can be compounded with other beneficial ingredients (such as prebiotics) to prepare oral liquids. It should be noted that the food, pharmaceutical, and health supplement forms specifically listed above are merely illustrative examples. Those skilled in the art will understand that any form made from the plant protein products described in this invention through processing falls within the scope of protection of this invention.

[0041] method This invention provides a method for preparing the aforementioned plant protein product. According to an embodiment of the invention, the method includes the following steps: contacting a modified system with the plant protein to perform a cross-linking reaction to obtain the plant protein product; wherein the modified system includes laccase and chlorogenic acid. The method according to the embodiments of the invention has advantages such as being mild, green and safe, and producing plant protein products with excellent performance.

[0042] According to embodiments of the present invention, the mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2). Exemplarily, the mass ratios of the plant protein, the chlorogenic acid, and the laccase are 800:1:0.5, 900:1:0.5, 1000:1:0.5, 1100:1:0.5, 1200:1:0.5, 800:2:0.5, 800:3:0.5, 800:4:0.5, 800:5:0.5, 800:6:0.5, 800:1:0.6, 800:1:0.7, and 800:1:0. 8, 800:1:0.9, 800:1:1, 800:1:1.1, 800:1:1.2, 800:1:1.3, 800:1:1.4, 800:1:1.5, 800:1:1.6, 800:1:1.7, 800:1:1.8, 800:1:1.9, 800:1:2, preferably (900~1000):(1~4):(1~2), more preferably 10000:4:1.

[0043] According to an embodiment of the present invention, the contact method is a gradient heating treatment, wherein the initial temperature of the gradient heating treatment is 20~40℃, the final temperature of the gradient heating treatment is 135~150℃, and each temperature gradient is 25~35℃. Thus, by optimizing the contact conditions, the degree of cross-linking and the degree of protein tissue formation are synergistically optimized, resulting in a more uniform, dense, and elastic fibrous structure in the final plant protein product, and a more superior therapeutic effect (such as regulating blood lipid levels). For example, the initial temperature of the gradient heating treatment is 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, 32℃, 34℃, 36℃, 38℃, or 40℃, preferably 26~36℃, and more preferably 28~32℃; the gradient heating treatment... The endpoint temperatures are 135℃, 136℃, 137℃, 138℃, 139℃, 140℃, 141℃, 142℃, 143℃, 144℃, 145℃, 146℃, 147℃, 148℃, 149℃, and 150℃, preferably 140~150℃, and more preferably 146~150℃; each temperature gradient is 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, 31℃, 32℃, 33℃, 34℃, and 35℃, preferably 28~32℃, and more preferably 29~31℃.

[0044] It should be noted that in order to obtain the final plant protein product, after the gradient heating treatment, it is also necessary to cool it to fix its shape.

[0045] Uses in drug preparation This invention proposes the use of the aforementioned plant protein products in the preparation of pharmaceuticals. According to embodiments of the invention, the pharmaceuticals are used for the prevention, relief, adjunctive treatment, or treatment of obesity and related metabolic syndromes.

[0046] Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this application, and will not be repeated here.

[0047] According to embodiments of the present invention, the associated metabolic syndrome includes one or more of hyperlipidemia, non-alcoholic fatty liver disease, and insulin resistance.

[0048] It should be noted that the obesity-related metabolic syndromes specifically listed above are merely illustrative examples. All types of metabolic complications (i.e., related metabolic syndromes) caused by obesity or high-fat / high-sugar diets, such as glucose and lipid metabolism disorders, fall within the scope of protection of this invention.

[0049] According to embodiments of the present invention, the drug has at least one of the following uses: controlling body weight; regulating blood lipid levels; regulating blood glucose levels; improving hepatic lipid deposition; improving glucose tolerance; improving insulin sensitivity; improving organ coefficients; improving adipose tissue deformation; and improving intestinal flora imbalance; wherein the organ coefficients include one or more of the following: heart coefficient, liver coefficient, spleen coefficient, and kidney coefficient; and the blood lipid level indicators are selected from one or more of the following: total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

[0050] Uses in the preparation of health products Those skilled in the art will understand that the features and advantages described above for plant protein products also apply to this application, and will not be repeated here.

[0051] This invention proposes the use of the aforementioned plant protein products in the preparation of health supplements. According to embodiments of the invention, the health supplement has at least one of the following uses: regulating blood lipids and / or assisting in lowering blood lipids; regulating blood sugar and / or assisting in lowering blood sugar.

[0052] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0053] Example 1: Experiment on the effects of plant-based meat products on glucose and lipid metabolism disorders I. Preparation of Plant-Based Meat Products 1. Unmodified plant-based meat products Plant protein (purchased from Shandong Yuwang Ecological Food Co., Ltd., product number 901W) was subjected to high-moisture extrusion. The extrusion conditions were: moisture content of 40%~60%, screw speed of 220 r / min, gradient heating temperature of 30℃, 60℃, 90℃, 120℃ and 150℃ respectively, and a flat and long die. After cooling and molding, the product was then sheared. Thus, unmodified plant-based meat products were prepared.

[0054] 2. Chlorogenic acid + laccase modification - plant-based meat products Take plant protein, chlorogenic acid (purchased from Shaanxi Ronglin Biotechnology Co., Ltd., product number RLSW20240115), and laccase (purchased from Shanghai Macklin Biochemical Co., Ltd., product number L871938), and mix them in an addition ratio of 10000:4:1 by mass to obtain a mixture. Subject the mixture to high-moisture extrusion. Among them, the extrusion conditions are a moisture content of 40% - 60%, a screw speed of 220 r / min, and the heating temperatures of gradient heating are set at 30°C, 60°C, 90°C, 120°C, and 150°C respectively, and the die head is a flat long die; then it is formed through a cooling die and then subjected to shearing treatment; thus, chlorogenic acid + laccase-modified plant meat products are prepared.

[0055] II. Experiment on the effects of different plant meat products on glycolipid metabolism disorders 1. Experimental animals Experimental animals: 40 SPF-grade C57BL / 6J male mice (6 weeks old, weighing 18 - 22 g, all purchased from Henan Sk贝斯 Biotechnology Co., Ltd., and the experimental animal use license is SYXK (Yu) 2021 - 0003); Feeding conditions: The experimental animals are housed in the Key Laboratory of Animal Immunology of Henan Academy of Agricultural Sciences, with a temperature of 24 ± 2°C, a relative humidity of 60 ± 10%, and a 12 h / 12h light-dark alternating cycle. All mice are first fed with ordinary feed for 1 week for adaptation and then used for subsequent experiments.

[0056] 2. Experimental grouping The whole experiment strictly complies with animal welfare and ethical requirements, and the animal welfare and ethics approval number is LLSC4102502011.

[0057] Randomly divide the 40 experimental mice into 5 groups (n = 8), namely the normal control group (CON), the high-fat group (HFD), the unmodified group (SHME), the enzyme-modified group (LHME), and the positive control group (MET). The feed, water, and cages are changed every other day, and the body weight and food intake of each group of mice are measured and recorded weekly. Among them, the specific treatment operations of each group of mice are as follows: Normal control group (CON): Feed 25 - 30 g of ordinary feed (purchased from Changzhou Mouse One and Mouse Two Biotechnology Co., Ltd.) daily and drink normal water for 8 consecutive weeks; High-fat group (HFD): Feed 25 - 30 g of high-fat feed (purchased from Changzhou Mouse One and Mouse Two Biotechnology Co., Ltd.) daily and drink normal water for 8 consecutive weeks; Unmodified group (SHME): Feed 25 - 3 g of high-fat feed daily (where 20 wt% of the high-fat feed is replaced by the unmodified plant meat products prepared in step one), drink normal water, and feed continuously for 8 weeks; Enzyme-modified group (LHME): fed 25-30 g of high-fat diet daily (of which 20 wt% of the high-fat diet was replaced with chlorogenic acid + laccase-modified plant meat product prepared in step one), with normal drinking water, for 8 consecutive weeks; Positive control group (MET): fed 25-30 g of high-fat diet daily, and gavaged with 2 mL of 2 g / L physiological saline containing metformin daily, with normal drinking water, for 8 consecutive weeks.

[0058] 3. Sample collection and processing After the experiment, the mice in each group were fasted for 12 hours but allowed to drink water. Their final weight was recorded, and they were then anesthetized and euthanized. Then, the liver, inguinal fat, and epididymal fat tissue were dissected and separated, weighed, and the organ coefficient and fat index were calculated. The formula for calculating the organ coefficient is shown in Equation 1, and the formula for calculating the fat index is shown in Equation 2.

[0059] Organ Index (%) = (Organ Mass (g) / Mouse Body Weight (g)) × 100 Equation 1 Fat index (%) = (Fat mass (g) / Body weight (g)) × 100 (Formula 2) Serum, liver tissue, and epididymal adipose tissue were collected from each group of mice for subsequent testing; Fecal samples were collected from each group of mice and stored at -80°C for gut microbiota analysis.

[0060] 4. Detection indicators Serum lipid metabolism indicators: The levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in the serum of mice in each group (collected at the end of week 8 of the experiment) were measured using a BS-240VET fully automated biochemical analyzer. The reagent kits used for each test were purchased from Nanjing Jiancheng Biotechnology Co., Ltd.

[0061] Blood glucose and insulin levels: Samples were taken at week 8 of the experiment to test blood glucose and insulin levels. The test indicators included fasting blood glucose (FBG) level and serum insulin (INS) level to calculate the insulin resistance index (HOMA-IR). The formula for calculating the insulin resistance index is shown in Equation 3.

[0062] Insulin resistance index = (FBG * INS) / 22.5 (Equation 3) In addition, an oral glucose tolerance test (OGTT) was performed on mice in each group at week 8 of the experiment. The specific experimental procedure is as follows: After each group of mice was administered glucose by gavage, blood glucose levels were measured at 0 min, 30 min, 60 min, and 120 min, and the area under the curve (AUC) was calculated.

[0063] Histological observation: The liver and epididymal adipose tissue obtained in step 3 were dehydrated, embedded, and prepared into paraffin sections. After HE staining, the tissue morphology and changes in fat vacuoles were observed under a microscope.

[0064] Gut microbiota analysis: The fecal samples obtained in step 3 were sequenced using the Illumina MiSeq PE300 platform to analyze the richness (Chao1 index, ACE index), diversity (Shannon index, Simpson index), and β diversity (PCA, NMDS).

[0065] 5. Statistical Analysis The data obtained are expressed as mean ± standard deviation and statistical analysis was performed using SPSS 24.0 software.

[0066] Data that follow a normal distribution and have homogeneity of variance are analyzed using one-way ANOVA; otherwise, nonparametric tests are used. P <0.05 indicates that the difference is statistically significant.

[0067] 6. Experimental Results (1) Weight gain The statistics on the weight gain of mice in each group are shown below. Figure 1 .

[0068] The results showed that compared with the normal control group, the weight gain of the high-fat group mice was significantly increased. Compared with the high-fat group mice, the weight gain was significantly reduced after feeding them a high-fat diet containing unmodified plant-based meat products (unmodified group) or a high-fat diet containing chlorogenic acid and laccase-modified plant-based meat products (enzyme-modified group). The control of weight gain in the enzyme-modified group mice was close to that in the metformin drug intervention group (positive control group).

[0069] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can effectively inhibit weight gain caused by a high-fat diet. Among them, chlorogenic acid + laccase-modified plant-based meat products have a better effect on controlling weight gain.

[0070] (2) Organ coefficient The statistical results of organ coefficients for each group of mice are shown in the figure. Figure 2 .

[0071] The results showed that compared with the normal control group, the liver weight, inguinal fat index and epididymal fat index of the high-fat group mice were significantly increased. After feeding the high-fat diet containing unmodified plant-based meat products (unmodified group), the inguinal fat index and epididymal fat index were significantly reduced. After feeding the high-fat diet containing chlorogenic acid and laccase-modified plant-based meat products (enzyme-modified group), the liver weight, inguinal fat index and epididymal fat index were significantly reduced, which were close to the relevant parameter levels of the metformin drug intervention group (positive control group).

[0072] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can effectively reduce fat accumulation in organs, with chlorogenic acid + laccase-modified plant-based meat products showing better results.

[0073] (3) Serum lipid metabolism indicators The statistical results of serum lipid metabolism indicators of mice in each group are shown in the figure. Figure 3 .

[0074] The results showed that compared with the normal control group, the high-fat group mice had significantly higher levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol, and lower levels of high-density lipoprotein cholesterol. This suggests that a high-fat diet can cause lipid metabolism disorders and increase the risk of cardiovascular disease. Compared with the high-fat group mice, it was found that feeding mice with a high-fat diet containing unmodified plant-based meat products (unmodified group) or a high-fat diet containing chlorogenic acid and laccase-modified plant-based meat products (enzyme-modified group) significantly reduced the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol, while increasing the level of high-density lipoprotein cholesterol. The levels of each parameter were close to those of the metformin-treated group mice (positive control group).

[0075] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can effectively regulate serum lipid levels and improve lipid metabolism. Among them, chlorogenic acid + laccase-modified plant-based meat products have a better effect.

[0076] (4) Pathological morphology of liver and epididymal adipose tissue The pathological morphological observation results of liver samples from each group of mice are shown in the figure. Figure 4 The pathological morphological observation results of epididymal adipose tissue samples from each group of mice are shown in the figure. Figure 5 .

[0077] The results showed that: (1) Liver: The liver cells of the normal control group mice were uniform in morphology, tightly connected, with intact nuclei, blue-purple color, and no obvious lipid droplets (lipid vacuoles); while the liver cells of the high-fat group mice were damaged, with uneven cell arrangement, reduced number of nuclei, and larger and more numerous fat vacuoles in the liver cells, indicating that lipid accumulation in the liver of the high-fat group mice was serious; while the liver cells of the unmodified group mice were more intact in morphology, with a relatively increased number of nuclei, and some fat vacuoles were still present in the liver tissue, with a reduced area of ​​lipid droplets; at the same time, the liver cells of the enzyme-modified group mice were more intact in morphology, with clear and increased number of nuclei, fewer and smaller fat vacuoles, and the liver cell state was similar to that of the metformin drug intervention treatment group (positive control group). The cells of both groups were close to the liver cells of the normal control group (see details). Figure 4 (2) Epididymal adipose tissue: In the normal control group, the adipocytes in the epididymal adipose tissue of mice were smaller in outline, more compact in arrangement, and more numerous; while in the high-fat group, lipid accumulation was severe, the adipocytes were disordered and loosely arranged, and the diameter of the adipocytes was significantly increased; compared with the high-fat group, the fat accumulation in the unmodified group and the enzyme-modified group was inhibited, and their adipocyte status was similar to that of the metformin drug intervention treatment group (positive control group). The cells in both groups were close to the adipocytes in the epididymal adipose tissue of the normal control group (see details). Figure 5 ).

[0078] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can inhibit weight gain and visceral fat accumulation caused by a high-fat diet, regulate lipid metabolism disorders, and improve pathological damage to liver and epididymal adipose tissue. Among them, chlorogenic acid + laccase-modified plant-based meat products have better effects and are close to the intervention effect of metformin.

[0079] (5) Blood glucose and insulin levels The statistical results of insulin levels in each group of mice are shown below. Figure 6 The results of glucose tolerance tests in each group of mice are shown in the figure. Figure 7 .

[0080] The results showed that: (1) Insulin levels: Compared with the normal control group, the serum insulin content and insulin resistance index (HOMA-IR) of the high-fat group mice were significantly increased, indicating that the high-fat diet successfully induced hyperinsulinemia and insulin resistance in mice; at the same time, it was found that after feeding the high-fat diet containing unmodified plant-based meat products (unmodified group) or the high-fat diet containing chlorogenic acid + laccase modified plant-based meat products (enzyme modified group), the serum insulin content and insulin resistance index of both groups decreased significantly in a dose-dependent manner. Among them, the improvement effect of chlorogenic acid + laccase modified plant-based meat products was more significant (see details). Figure 6(2) Glucose tolerance: After 30 min of glucose gavage, the blood glucose values ​​of mice in each group reached the highest value. Among them, the blood glucose of mice in the high-fat group rose rapidly but fell slowly after gavage. At the same time, compared with the high-fat group, the AUC values ​​of the unmodified group and the enzyme-modified group were significantly reduced. Among them, the AUC value of the enzyme-modified group was reduced more significantly and was similar to that of the metformin drug intervention treatment group (positive control group).

[0081] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can improve insulin resistance induced by a high-fat diet by regulating serum insulin levels. In addition, both can improve glucose intolerance induced by a high-fat diet, with chlorogenic acid + laccase-modified plant-based meat products showing better effects.

[0082] (6) Intestinal flora The results of gut microbiota statistics for each group of mice are shown below. Figure 8 .

[0083] The results showed that: (1) α-diversity: Compared with the normal control group (CON), the richness (Chao1 index, ACE index) and diversity (Shannon index) of the gut microbiota in the high-fat diet group (HFD) mice were significantly reduced, and the species distribution evenness decreased (Simpson index increased), indicating that the high-fat diet led to the simplification and imbalance of the gut microbiota structure. After intervention with unmodified plant-based meat products or chlorogenic acid + laccase modified plant-based meat products, the above indicators were significantly improved. Among them, the intervention with chlorogenic acid + laccase modified plant-based meat products had a better effect, and its effect was closer to that of the positive drug treatment group (see details). Figure 8 (A~D); (2) β diversity (PCA, NMDS analysis): The intestinal flora structure of the HFD group mice was significantly separated from that of the CON group, confirming that a high-fat diet would cause an overall shift in the composition of the flora. After intervention with unmodified plant-based meat products or chlorogenic acid + laccase-modified plant-based meat products, the distribution of the sample points was closer to that of the CON group mice. This indicates that intervention with unmodified plant-based meat products or chlorogenic acid + laccase-modified plant-based meat products can, to some extent, reverse the disorder of the intestinal flora structure induced by a high-fat diet and promote its return to a normal state. Among them, the intervention effect of chlorogenic acid + laccase-modified plant-based meat products was better, and its effect was closer to that of the positive drug treatment group (see details). Figure 8 (E~F in the text).

[0084] The above results indicate that consuming unmodified plant-based meat products and chlorogenic acid + laccase-modified plant-based meat products can effectively improve the intestinal flora imbalance induced by a high-fat diet, with chlorogenic acid + laccase-modified plant-based meat products showing better results.

[0085] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0086] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A plant protein product, characterized in that, The plant protein product comprises the following raw materials: Plant protein; Chlorogenic acid; Laccase; The plant protein product is obtained from the raw material through a cross-linking reaction.

2. The plant protein product according to claim 1, characterized in that, The plant protein includes one or more of soy protein isolate, pea protein, and wheat protein.

3. The plant protein product according to claim 1, characterized in that, The mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2).

4. A food, medicine, or health product, characterized in that, The food, medicine, or health product mentioned includes: The plant protein product according to any one of claims 1 to 3.

5. A method for preparing the plant protein product according to any one of claims 1 to 3, characterized in that, The method includes the following steps: The modified system is brought into contact with the plant protein to carry out a cross-linking reaction, thereby obtaining the plant protein product; The modification system includes laccase and chlorogenic acid.

6. The method according to claim 5, characterized in that, The mass ratio of the plant protein, the chlorogenic acid, and the laccase is (800~1200):(1~6):(0.5~2). Optionally, the contact method is gradient heating treatment, wherein the initial temperature of the gradient heating treatment is 20~40℃, the final temperature of the gradient heating treatment is 135~150℃, and each temperature gradient is 25~35℃.

7. The use of the plant protein product according to any one of claims 1 to 3 in the preparation of a medicine, characterized in that, The drug is used to prevent, alleviate, assist in the treatment of or treat obesity and related metabolic syndrome.

8. The use according to claim 7, characterized in that, The associated metabolic syndromes include one or more of hyperlipidemia, non-alcoholic fatty liver disease, and insulin resistance.

9. The use according to claim 7, characterized in that, The drug has at least one of the following uses: Control your weight; Regulate blood lipid levels; Regulate blood sugar levels; Improves liver lipid deposition; Improve the body's glucose tolerance; Improves the body's insulin sensitivity; Improve organ coefficient; Improve adipose tissue deformity; Improve gut microbiota imbalance; The organ coefficients include one or more of the following: heart coefficient, liver coefficient, spleen coefficient, and kidney coefficient; The blood lipid level indicators are selected from one or more of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

10. The use of the plant protein product according to any one of claims 1 to 3 in the preparation of health products, characterized in that, The health product has at least one of the following uses: Regulates blood lipids and / or assists in lowering blood lipids; Regulates blood sugar and / or helps lower blood sugar.

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