Use for sweet-tasting protein

Abstract

Provided is a use for a sweet-tasting protein in reducing body weight, the sweet-tasting protein being brazzein or thaumatin. The provided sweet-tasting protein significantly inhibits high-fat diet-induced weight gain in subjects, and alleviates to a certain extent symptoms of insulin resistance caused by obesity, allowing for resumption of normal insulin secretion in subjects, lowering blood lipids, and ameliorating liver damage caused by high-fat diets, indicating that the sweet-tasting protein is effective in reducing body fat and decreasing body weight.

Description

Uses of sweet protein

[0001] This application claims priority to an earlier application filed on October 28, 2024, with patent application number 2024115080676, entitled "Use of Sweet Proteins". The entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of biomedicine, and more specifically to the use of a sweet protein. Background Technology

[0003] Excessive sugar intake is one of the greatest challenges to human health in the world today, leading to increasingly prominent problems such as obesity, overweight, and diabetes. With rising health awareness, low-sugar and sugar-reduced diets have become a shared pursuit for consumers and businesses. Plant-based sweetened proteins are high-sweetness, low-calorie non-sugar protein sweeteners. As a sucrose substitute, they do not cause diseases such as high blood pressure, high cholesterol, or diabetes, and have enormous market potential in the food and pharmaceutical industries.

[0004] Brazzein is a sweet protein extracted from the fruit of the West African climbing plant *Pentadiplandra brazzeana* (Baillon). Composed of 54 amino acids, it is characterized as a monomeric protein with a molecular weight of 6.5 kDa. Its molecule contains four pairs of disulfide bonds (Cys4-Cys52, Cys16-Cys37, Cys22-Cys47, and Cys26-Cys49), which form the basis of its excellent stability. By weight, brazzein is 500 to 2000 times sweeter than sucrose, with a clean and lingering sweetness similar to sucrose. Compared to other sweet proteins, brazzein has the smallest molecular weight, the best water solubility, and retains its sweetness even after heat treatment at 80°C for 4 hours in aqueous solution. It also exhibits good thermal and pH stability, making it suitable for many industrial food manufacturing processes.

[0005] Thaumatin is an extremely sweet substance extracted from the nut shell of the plant *Thaurnatocuccus danielli*, and is a natural protein. There are two main types of thaumatin: Thaumatin I, a compound composed of 207 amino acids linked in a straight chain, and Thaumatin II, composed of 198 amino acids. Thaumatin is approximately 2000 times sweeter than sucrose. Although very sweet, its sweetness is completely different from sucrose, and its sweetness develops very slowly. The sweetness lasts for a long time, leaving a licorice-like aftertaste. Thaumatin is highly soluble in water, heat-stable, and stable under acidic conditions, and is widely used in food manufacturing processes. However, current research on sweet proteins mostly focuses on their use as sugar substitutes to improve the flavor of food or beverages, lacking research on the functional aspects of the sweet protein molecules themselves. Summary of the Invention

[0006] To address the aforementioned problems, the present invention provides an application of a sweet protein in weight loss, wherein the sweet protein is Brazil sweetener or sema sweetener.

[0007] On the other hand, the present invention provides the use of sweet proteins in reducing, alleviating or reversing obesity-related problems, wherein the sweet proteins are Brazil sweet or sema sweet.

[0008] In one implementation, the application is for weight reduction.

[0009] In one implementation, the application is for alleviating glucose tolerance;

[0010] Preferably, the application is to alleviate glucose tolerance caused by obesity.

[0011] In one implementation, the application is to lower serum blood glucose;

[0012] Preferably, the application is to reduce the increase in serum blood glucose caused by obesity.

[0013] In one implementation, the application is to restore insulin secretion;

[0014] Preferably, the application is to restore insulin secretion caused by obesity.

[0015] In one embodiment, the application is to lower blood lipids; preferably, the blood lipids are plasma free fatty acids, cholesterol, or triglycerides.

[0016] In one implementation, the application is for the recovery of liver damage.

[0017] On the other hand, the present invention provides the application of a sweet protein in alleviating glucose tolerance, wherein the sweet protein is caramel or sematriol. In some embodiments of the present invention, glucose tolerance caused by obesity is alleviated.

[0018] On the other hand, the present invention provides the application of sweet protein in lowering serum blood glucose or restoring insulin secretion, wherein the sweet protein is carbamate or semamate.

[0019] In some embodiments of the present invention, the application is to reduce the elevation of serum blood glucose caused by obesity or to restore the decline in insulin secretion caused by obesity.

[0020] On the other hand, the present invention provides the application of a sweet protein in lowering blood lipids, wherein the sweet protein is Brazil brittle or sematrandezine. In some embodiments of the present invention, it reduces elevated blood lipids caused by obesity.

[0021] In one embodiment, the blood lipids are plasma free fatty acids (FFA), cholesterol, or triglycerides (TG).

[0022] On the other hand, the present invention provides the application of a sweet protein in the recovery of liver damage, wherein the sweet protein is carrageenan or sematrandez. In some embodiments of the invention, liver damage caused by obesity is recovered.

[0023] In one embodiment of the invention, the obesity is caused by a high-fat and / or high-sugar diet.

[0024] In one implementation, the Brazilian sweetener or soma sweetener is obtained through recombinant expression.

[0025] In one embodiment, the amino acid sequence of the Brazilian sweet has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 1.

[0026] In one embodiment, the amino acid sequence of the Brazilian sweet is any one of SEQ ID NO: 1-8, SEQ ID NO: 10-13;

[0027] Preferably, the amino acid sequence of the Brazilian sweet is SEQ ID NO: 7.

[0028] In one embodiment, the amino acid sequence of the semathy has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 9.

[0029] In one embodiment, the amino acid sequence of the semathy is SEQ ID NO: 9.

[0030] In one embodiment, the sweet protein is added to an edible food product.

[0031] In one implementation, the edible food is a food, beverage, health product, dietary supplement, or medicine.

[0032] In one embodiment, the beverage is fermented milk, carbonated beverage, or special purpose beverage.

[0033] On the other hand, the present invention provides a sweet protein for weight loss, said sweet protein being Brazil sweet or Soma sweet.

[0034] On the other hand, the present invention provides a method for reducing weight, which includes administering a sweet protein, said sweet protein being Brazil sweet or sema sweet, to a subject in need.

[0035] On the other hand, the present invention provides a sweet protein for alleviating glucose tolerance, said sweet protein being Brazil sweetener or soma sweetener.

[0036] In some embodiments of the present invention, the sweet protein is a sweet protein that alleviates glucose tolerance caused by obesity.

[0037] On the other hand, the present invention provides a method for alleviating glucose tolerance, which includes administering a sweet protein, said sweet protein being carbomer or semamer, to a subject in need.

[0038] In some embodiments of the present invention, the method is a method for alleviating glucose tolerance caused by obesity.

[0039] On the other hand, the present invention provides a sweet protein for lowering serum blood glucose or restoring insulin secretion, said sweet protein being caramel or sematriol.

[0040] In some embodiments of the present invention, the sweet protein is a sweet protein that reduces the increase in serum blood glucose caused by obesity or restores the decrease in insulin secretion caused by obesity.

[0041] On the other hand, the present invention provides a method for lowering serum blood glucose or restoring insulin secretion, comprising administering a sweet protein, said sweet protein being carbomer or sematriol, to a subject in need.

[0042] In some embodiments of the present invention, the method is a method for reducing the increase in serum blood glucose caused by obesity or restoring the decrease in insulin secretion caused by obesity.

[0043] On the other hand, the present invention provides a sweet protein for lowering blood lipids, said sweet protein being Brazil sweetener or sema sweetener.

[0044] In some embodiments of the present invention, the sweet protein is a sweet protein that reduces the increase in blood lipids caused by obesity.

[0045] In one embodiment, the blood lipids are plasma free fatty acids (FFA), cholesterol, or triglycerides (TG).

[0046] On the other hand, the present invention provides a method for lowering blood lipids, which includes administering a sweet protein, said sweet protein being Brazil sweet or sema sweet, to a subject in need.

[0047] In some embodiments of the present invention, the method is a method for reducing the increase in blood lipids caused by obesity.

[0048] On the other hand, the present invention provides a sweet protein for restoring liver damage, said sweet protein being Brazil sweetener or sema sweetener.

[0049] In some embodiments of the present invention, the sweet protein is a sweet protein that restores liver damage caused by obesity.

[0050] On the other hand, the present invention provides a method for restoring liver injury, which includes administering a sweet protein, said sweet protein being carbomer or semamer, to a subject in need.

[0051] In some embodiments of the present invention, the method is a method for restoring liver damage caused by obesity.

[0052] On the other hand, the present invention provides a sweet protein for reducing, alleviating or reversing obesity-related problems, said sweet protein being Brazil sweetener or sema sweetener.

[0053] In one embodiment, the sweet protein is a weight-reducing sweet protein.

[0054] In one embodiment, the sweet protein is a sweet protein that alleviates glucose tolerance;

[0055] Preferably, the sweet protein is a sweet protein that alleviates glucose tolerance caused by obesity.

[0056] In one embodiment, the sweet protein is a sweet protein that lowers serum blood glucose or restores insulin secretion;

[0057] Preferably, the sweet protein is a sweet protein that reduces the increase in serum blood glucose caused by obesity.

[0058] In one embodiment, the sweet protein is a sweet protein that restores insulin secretion;

[0059] Preferably, the sweet protein is a sweet protein that restores insulin secretion lost due to obesity.

[0060] In one embodiment, the sweet protein is a lipid-lowering sweet protein; preferably, the blood lipids are plasma free fatty acids, cholesterol, or triglycerides.

[0061] In one embodiment, the sweet protein is a sweet protein that restores liver damage.

[0062] On the other hand, the present invention provides a method for reducing, alleviating or reversing obesity-related problems, comprising administering a sweet protein, said sweet protein being Brazil sweet or sema sweet, to a subject in need.

[0063] In one implementation, the method is a method for reducing body weight.

[0064] In one implementation, the method is a method for alleviating glucose tolerance;

[0065] Preferably, the method is a method for alleviating glucose tolerance caused by obesity.

[0066] In one implementation, the method is a method for lowering serum blood glucose or restoring insulin secretion;

[0067] Preferably, the method is a method for reducing the increase in serum blood glucose caused by obesity.

[0068] In one implementation, the method is a method for restoring insulin secretion;

[0069] Preferably, the method is a method for restoring insulin secretion caused by obesity.

[0070] In one embodiment, the method is a method for lowering blood lipids; preferably, the blood lipids are plasma free fatty acids, cholesterol, or triglycerides.

[0071] In one implementation, the method is a method for restoring liver damage. Beneficial effects

[0072] The saccharin provided by this invention significantly inhibited weight gain in subjects caused by high-fat and / or high-sugar diets, could alleviate insulin resistance symptoms caused by obesity to a certain extent, restored normal insulin secretion in subjects, reduced blood lipids, and restored liver damage caused by high-fat and / or high-sugar diets, indicating that saccharin has the effect of reducing body fat and weight loss. Attached Figure Description

[0073] Figure 1 shows the results of the oral glucose tolerance test (OGTT) at 16 weeks in mice of the LF-1, LF-2, HF-1, HF-2 and HF1-HF4 groups.

[0074] Figure 2 shows the weight of mesenteric and epididymal fat in mice of the LF-1, LF-2, HF-1, HF-2 and HF1-HF4 groups at 19 weeks.

[0075] Figure 3 shows the glucose content (A) and insulin expression (B) in the serum of mice in the LF-1, LF-2, HF-1, HF-2 and HF1-HF4 groups after sacrifice at 19 weeks.

[0076] Figure 4 shows the effects of Brazil sweetener and samaram intake on blood lipids in mice. (A) shows the effect on plasma free fatty acids (FFA); (B) shows the effect on cholesterol; and (C) shows the effect on triglycerides (TG).

[0077] Figure 5 shows the alanine aminotransferase (ALT) levels of mice in the LF-1, LF-2, HF-1, HF-2, and HF1-HF4 groups at 19 weeks. Detailed Implementation

[0078] Unless otherwise stated, all percentages, parts and ratios used herein are by weight of the total composition. Unless otherwise stated, all such weights relating to the listed ingredients are based on the level of active substance and therefore do not include solvents or byproducts that may be present in commercially available materials.

[0079] definition

[0080] The terminology set forth herein is used only to describe embodiments and should not be construed as limiting the scope of this disclosure as a whole. Unless otherwise stated, “a,” “the,” and “at least one” are used interchangeably. Furthermore, unless the context clearly indicates otherwise, the singular forms “a” and “the” include their plural forms as used in the specification and appended claims.

[0081] Throughout this specification, when defining numerical ranges for specific features of the invention, the invention relates to and explicitly incorporates each specific subrange therein. Furthermore, throughout this specification, when defining a group of substances for specific features of the invention, the invention relates to and explicitly incorporates each specific subgroup therein. Any specified range or group should be understood as a simplified representation of each member of the range or group, and of each possible subrange or subgroup covered therein.

[0082] As used herein, "sweet protein" refers to a protein with a strong sweet taste, suitable for sweetening food, beverages, and / or pharmaceutical products intended for human consumption. By weight, the sweet protein of the present invention, when compared to a 1% sucrose solution, can be at least 1000 times sweeter than sucrose, preferably at least 2000 times, more preferably at least 5000 times, and even more preferably 10000 times sweeter.

[0083] As used herein, "brassine," also known as brassine protein, brassine sweet protein, or brassine, refers to a sweet protein extracted from the fruit of the West African climbing plant *Pentadiplandra brazzeana* (Baillon) and described in WO9531547, or its recombinant form. In nature, brassine occurs in three different forms, with or without a Gln residue or pyroglutamic acid at its N-terminus. In the context of this invention, the wild-type brassine sequence comprises the amino acid sequence of SEQ ID NO: 1, without a Gln residue or pyroglutamic acid at its N-terminus. The source of the brassine described in this invention is not limited, including wild-type brassine or its naturally occurring mutants or recombinant analogs; the production method is not limited, including recombinant expression, extraction, chemical synthesis, in vivo or in vitro acquisition, etc. In one embodiment, the Brazilian sweetener of the present invention is preferably recombinant Brazilian sweetener, which refers to Brazilian sweetener produced by recombinant host cells, wherein the amino acid sequence of the Brazilian sweetener is any one of SEQ ID NO: 1 to 8, or has a sequence identity with SEQ ID NO: 1 of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or more, preferably SEQ ID NO: 7.

[0084] As used herein, "somatian" refers to a super-sweet substance that can be extracted from the nut shell of the natural plant Thaurnatocuccus danielli. There are two main types of somatian: somatian I is a compound composed of 207 amino acids linked in a straight chain, and somatian II is composed of 198 amino acids. The somatian of this invention is preferably somatian II, and its source is not limited, including wild-type somatian or its naturally occurring mutants or recombinant analogs of somatian. The production method is not limited, including recombinant expression, extraction, chemical synthesis, in vivo or in vitro acquisition, etc. In one embodiment, the somatian of this invention is preferably recombinant somatian, referring to somatian produced by recombinant host cells, wherein the amino acid sequence of said somatian is SEQ ID NO: 9, or has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 9.

[0085] As used herein, a “mutant” of Brazil sweet refers to a sweet protein with a modified amino acid sequence compared to the amino acid sequence of Brazil sweet (SEQ ID NO: 1), i.e., an amino acid sequence in which one or more amino acids are substituted, deleted, and / or inserted. A “mutant” of sema sweet refers to a sweet protein with a modified amino acid sequence compared to the amino acid sequence of sema sweet (SEQ ID NO: 9), i.e., an amino acid sequence in which one or more amino acids are substituted, deleted, and / or inserted.

[0086] As used herein, “expression” means, in the context of this invention, any step involving the production of the sweet protein of this invention, including but not limited to transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0087] As used herein, “sequence identity” refers to the correlation between two amino acid sequences or two nucleotide sequences, described by the parameter “sequence identity.” When aligned using the CLUSTALW algorithm with preset parameters, a specific sequence has at least a certain percentage of amino acid residues that are identical to a specified reference sequence. The preset parameters for the CLUSTALW algorithm are: deletion counts are the residues that are not identical to the reference sequence, including deletions occurring at any end. For example, a 500-amino acid peptide variant lacking five amino acid residues at the C-terminus has a 99% (495 / 500 identical residues × 100) percentage of sequence identity relative to the parent peptide. Such variants are covered by the phrase “variants with at least 99% sequence identity to the parent.”

[0088] As used in this article, "fermented milk" refers to a product with a lower pH value made from raw cow (sheep) milk or milk powder through sterilization and fermentation.

[0089] As used in this article, "carbonated beverage" refers to a beverage that is infused with carbon dioxide under certain conditions, excluding beverages that produce carbon dioxide through fermentation itself.

[0090] As used in this article, “special purpose beverage” refers to a liquid beverage containing specific ingredients that is suitable for the needs of all or certain populations.

[0091] As used herein, the sweet protein of the present invention can be administered by any method, including injection (e.g., subcutaneous, intraperitoneal, etc.), oral administration, or other methods of administration known in the art.

[0092] As used in this article, the term "subject" refers to a mammal. A mammal is a warm-blooded animal that typically has fur or hair and gives birth to live offspring and nurses them with milk. Mammals include humans, companion animals (e.g., dogs, cats), livestock (e.g., dairy cows, horses, sheep, pigs, goats), and wild animals.

[0093] Example

[0094] 1. Experimental Materials

[0095] Preparation of Brazil Sweet: The Brazil sweet used in this invention was obtained through recombinant expression using genetic engineering methods known in the art. The natural sequence of the Brazil sweet protein (UniProtKB / Swiss-Prot:P56552.1) from the West African climbing plant (Pentadiplandra brazzeana) is available from the NCBI website (https: / / www.ncbi.nlm.nih.gov / ). The amino acid sequence of the Brazil sweet recombinantly expressed in this application is SEQ ID NO: 7.

[0096] Thaumatine: The thaumatine of this invention was obtained through recombinant expression using genetic engineering methods known in the art. The natural sequence of thaumatine II from the African bamboo taro (Thaumatococcus daniellii) (UniProtKB / Swiss-Prot:P02884.1) is available from the NCBI website (https: / / www.ncbi.nlm.nih.gov / ). The amino acid sequence of the recombinantly expressed thaumatine II in this application is SEQ ID NO: 9.

[0097] Concentrated whey protein powder: purchased from HILMAR.

[0098] Citric acid: purchased from Shandong Yingxuan Industrial Co., Ltd.

[0099] C57BL / 6J male mice: 48 male mice aged 7-8 weeks were purchased from Beijing Vitali Laboratory Animal Technology Co., Ltd.

[0100] High-fat feed (HF) and low-fat feed (LF): Purchased from Xiaoshuyoutai (Beijing) Biotechnology Co., Ltd., with the following specific components (unit: g):

[0101] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0102] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0103] The required intake for mice was calculated based on the recommended adult daily intake (EDI) for both carrageenan and samaram. The EDI for samaram was 77 mg / 70 kg / day, and the EDI for carrageenan was 60 mg / 70 kg / day. The gavage volume for mice was calculated using the following formula: Gavage volume for mice = EDI / 70000 * mouse body weight * 11.5, where the mouse body weight was 20 g. The samples used in the mouse experiment are as follows:

[0104] Sample 1: whey protein powder 394.29mg, carbamate 0.79mg, citric acid 9.46mg.

[0105] Sample 2: whey protein powder 82.14mg, carbamate 0.16mg, citric acid 1.97mg.

[0106] Sample 3: whey protein powder 389.23mg, semathy 1.01mg, citric acid 8.95mg.

[0107] Sample 4: whey protein powder 97.31mg, semathy 0.25mg, citric acid 2.24mg.

[0108] Sample 5: whey protein powder 394.29 mg, citric acid 9.46 mg.

[0109] Sample 6: whey protein powder 389.23mg, citric acid 8.95mg.

[0110] 2 Animal experiments

[0111] 1) Mouse model construction

[0112] Mouse models were constructed according to the following protocol (Table 1):

[0113] Table 1: Mouse Model Construction Scheme

[0114] Mouse experimental grouping: Mice were randomly divided into 8 groups, with 6 mice in each group. LF-1, LF-2, HF-1 and HF-2 served as the control group, while HF1, HF2, HF3 and HF4 served as the experimental group.

[0115] Mice were housed separately in an SPF-grade animal facility at a temperature maintained at 20-22°C and a relative humidity of 60%, with 12 hours of light per day. The living environment and bedding were kept clean, and the mice were fed a prescribed diet. Groups LF-1 and LF-2 were fed a low-fat diet, while the remaining groups were fed a high-fat diet. All mice underwent acclimatization for one week, followed by model establishment and drug treatment until week 19.

[0116] 2) Detection indicators

[0117] ① Record the average weekly body weight of mice in each group until week 19;

[0118] ② At 16 weeks, the glucose tolerance of mice in each group was detected, and an oral glucose tolerance test (OGTT) was performed. The oral glucose tolerance test was to fast the mice for 6 h and then weigh them. According to the body weight, a 20% glucose solution (2 g / kg body weight) was administered by gavage. At 30 min, 60 min, 90 min, and 120 min after gavage, the tail vein blood of the mice was taken, and the blood glucose level of the mice was detected using a blood glucose meter and blood glucose test strips. A GTT curve was made, and the area under the curve (AUC) from 0 min to 120 min was calculated.

[0119] ③ Blood collection and index analysis: After the experiment was completed, the mice were anesthetized with 2.5% paraformaldehyde, and blood was taken through the abdominal aorta. Serum was separated by centrifugation at 4°C and 2000 g for 15 min and stored at -80°C for further determination of the levels of free fatty acids (FFA), triglycerides (TG), cholesterol (TC), glucose, insulin, and alanine aminotransferase (ALT) in the serum.

[0120] ④ Tissue collection and index analysis: The liver, kidney, and white adipose tissue WAT (epididymal fat, mesenteric fat) were collected, weighed, and placed at -80°C for further analysis.

[0121] ⑤ Data processing and analysis: GraphPad Prism 10 was used for drawing, and SPSS19.0 software was used for variance analysis. The Duncan test method (P<0.05) was used for significant analysis of the results. The data representation method after processing was: mean ± standard deviation, and all experiments were repeated at least 3 times.

[0122] 3) Test results

[0123] ① Effect on mouse body weight

[0124] The changes in the average body weight of mice in each group over time are shown in Table 2. After 19 weeks of group feeding, there were significant differences in the body weights of mice in the high-fat diet groups (HF-1, HF-2) and the low-fat diet groups (LF-1, LF-2). Compared with the HF-1 group, both the licking and gavage groups of the thaumatin experimental groups HF1 and HF3 significantly inhibited the weight gain of mice caused by the high-fat diet. Compared with the HF-2 group, both the licking and gavage groups of the brazzein experimental groups HF2 and HF4 also significantly inhibited the weight gain of mice caused by the high-fat diet. At the 19th week, the body weights of the thaumatin experimental groups were lower than those of the brazzein experimental groups (HF1<HF2, HF3<HF4), indicating that the weight loss effect of the thaumatin experimental groups was better than that of the brazzein experimental groups.

[0125] Table 2: Changes in the average body weight of mice over time (unit: g)

[0126] ② Effects on glucose tolerance in mice

[0127] Figure 1 shows the results of the oral glucose tolerance test (OGTT) in each group of mice. Compared with the HF-1 and HF-2 groups, both the gavage and licking groups (HF1-HF4) of the Brazil nut and samaras nut experimental groups alleviated glucose tolerance in mice. Specifically, the Brazil nut experimental groups (HF2 and HF4) showed a stronger ability to alleviate glucose tolerance than the samaras nut experimental groups (HF1 and HF3), and the gavage groups (HF1 and HF2) showed a stronger ability to alleviate glucose tolerance than the corresponding licking groups (HF3 and HF4). These results indicate that glycoproteins can alleviate insulin resistance symptoms caused by obesity to some extent.

[0128] ③ Effects on mouse adipose tissue

[0129] Fat is mainly composed of white adipose tissue, with the epididymis and mesentery being the most abundant components. Figures 2(A) and (B) show the weight of mesenteric and epididymal fat in each group of mice, respectively. Compared with the HF-1 and HF-2 groups, the weight of fat in mice in the Brazil nut and sema nut experimental groups (HF1-HF4) via gavage and licking was reduced, showing an overall trend of reduced white adipose tissue weight. This is consistent with the pattern of body weight changes, indicating that the Brazil nut and sema nut experimental groups can reduce body weight to some extent by inhibiting lipogenesis.

[0130] ④ Effects on serum blood glucose in mice

[0131] Figure 3(A) shows the glucose content measured in the serum of mice after sacrifice. Significant differences were observed between the LF-1, LF-2 and HF-1, HF-2 groups. Both the Brazil nut and samaras nut groups (HF1-HF4) significantly reduced serum blood glucose, with the gavage groups (HF1, HF2) showing a more significant reduction. Figure 3(B) shows the serum insulin expression in each group. The significant difference in insulin levels between the HF-1, HF-2 and LF-1, LF-2 groups indicates that the stimulation of insulin secretion by blood glucose decreased under a high-fat diet, leading to glucose tolerance. This corresponds to the higher blood glucose levels in the HF-1, HF-2 groups. However, both the Brazil nut and samaras nut groups (HF1-HF4) restored some normal insulin secretion, with the gavage groups (HF1, HF2) showing better recovery than the licking groups (HF3, HF4).

[0132] ⑤ Effects on blood lipids in mice

[0133] Figure 4 shows the effects of Brazil and samamel intake on blood lipids in mice, which are generally consistent with the effects on blood glucose. Compared with LF-1 and LF-2 groups, the HF-1 and HF-2 groups significantly increased the levels of plasma free fatty acids (FFA), cholesterol, and triglycerides (TG) in mice, exhibiting typical characteristics of obesity and metabolic disorders. The gavage and licking groups (HF1-HF4) of both the samamel and Brazil groups significantly reduced the above-mentioned blood lipid indicators, especially the gavage groups (HF1 and HF2), which showed more significant effects, approaching those of the low-fat diet group.

[0134] ⑥ Effects on mouse liver tissue

[0135] Alanine aminotransferase (ALT), also known as alanine aminotransferase, is mainly found in the liver and serum. Hepatocellular damage leads to elevated serum ALT levels, thus serum ALT levels can be used as a biomarker for evaluating liver damage. Figure 5 shows the ALT levels in each group of mice. It can be seen that the serum ALT levels in the HF-1 and HF-2 groups were higher than those in the LF-1 and LF-2 groups, indicating that a high-fat diet causes some degree of liver damage. The intake of Brazil nuts and samaras (HF1-HF2) both reduced serum ALT levels, suggesting that the intake of Brazil nuts and samaras can, to some extent, restore liver damage caused by a high-fat diet.

[0136] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0137] The amino acid sequence involved in this invention is as follows:

Claims

1. Application of sweet protein in weight loss, wherein the sweet protein is Brazil sweetener or sema sweetener.

2. The application of sweet protein in reducing, alleviating or reversing obesity-related problems, wherein the sweet protein is carrageenan or sematrandez; Preferably, the application is to alleviate glucose tolerance; Preferably, the application is to lower serum blood glucose; Preferably, the application is to restore insulin secretion; Preferably, the application is to lower blood lipids; more preferably, the blood lipids are plasma free fatty acids, cholesterol, or triglycerides. Preferably, the application is for the recovery of liver damage.

3. The application according to claim 1 or 2, wherein the Brazilian sweetener or soma sweetener is obtained by recombinant expression.

4. The application according to any one of claims 1-3, wherein the amino acid sequence of the Brazilian sweet has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:

1.

5. The application according to claim 4, wherein the amino acid sequence of the Brazilian sweet is any one of SEQ ID NO: 1-8, SEQ ID NO: 10-13.

6. The application according to claim 5, wherein the amino acid sequence of the Brazilian sweet is SEQ ID NO:

7.

7. The application according to any one of claims 1-6, wherein the amino acid sequence of the semathy has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:

9.

8. The application according to claim 7, wherein the amino acid sequence of the semathymidine is SEQ ID NO:

9.

9. The application according to any one of claims 1-8, wherein the sweet protein is added to an edible food.

10. The application according to claim 9, wherein the edible food is a food, beverage, health product, dietary supplement, or medicine; More preferably, the beverage is fermented milk, carbonated beverage, or beverage for special purposes.

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