Preparation method and application of tomato seed small molecular protein with muscle attenuation resisting activity

Abstract

The invention relates to a preparation method and application of tomato seed small molecular protein with muscle attenuation resisting activity, and belongs to the field of biotechnology active ingredient development. The preparation method comprises the following steps: cleaning and drying tomato seeds, carrying out superfine grinding treatment, and preparing the small molecular protein with muscle attenuation resisting activity from the tomato seeds through alkali treatment and low-temperature enzymolysis of biological enzyme. Experimental results show that the tomato seed micromolecular protein obtained by the invention has the efficacy of improving muscle attenuation, has the advantages of safety, long-term use, no toxic or side effect and the like, and can be applied to the fields of functional foods, health care products and medicines for improving muscle attenuation and related symptoms thereof. The invention provides theoretical basis and technical support for developing novel muscle attenuation resistant products, and has wide application prospect and market value.

Description

Technical Field

[0001] The present invention belongs to the field of development and application of new resource proteins of biologically active ingredients, and specifically relates to a preparation method and application of a small molecule protein with anti-muscle attenuation activity. Background Art

[0002] With the development of society and the improvement of people's living standards, health issues have received more and more attention. In my country, sarcopenia has become an important issue affecting the quality of life of middle-aged and elderly people. According to statistics, about 30% of the middle-aged and elderly people in my country have varying degrees of sarcopenia. Sarcopenia not only leads to a decline in physical function, but also increases the risk of falls, fractures, etc., seriously affecting the quality of life of patients. Therefore, finding an effective method to fight sarcopenia has become a hot topic in current research.

[0003] Among the numerous studies on anti-muscle loss, plant-derived active ingredients have received extensive attention due to their natural, safe, and low-side effects. As a common vegetable, tomatoes are rich in nutrients, such as lycopene, vitamin C, vitamin E, etc., and have good health benefits. In recent years, studies have found that tomato seeds contain a class of small molecule proteins with anti-muscle loss activity, which provides new ideas for the development of new anti-muscle loss products.

[0004] However, the extraction, purification and activity research of tomato seed small molecule protein is still in its infancy. In the prior art, the extraction methods of tomato seed protein mainly include water extraction, acid-base method, ultrasonic-assisted method, etc., but these methods have problems such as low extraction rate, low purity, and serious loss of active ingredients. In addition, there are relatively few studies on the anti-muscle attenuation activity of tomato seed small molecule protein in the prior art, which limits its development in practical applications.

[0005] In view of the above problems, the present invention provides a method for preparing a tomato seed small molecule protein with anti-muscle attenuation activity and its application. The present invention aims to provide an efficient and simple method for preparing a tomato seed small molecule protein, and to study its anti-muscle attenuation activity, so as to provide a theoretical basis and technical support for the development of new anti-muscle attenuation products. The present invention has the following advantages: high extraction rate, good purity, and small loss of active ingredients; simple operation and easy industrialization; significant anti-muscle attenuation activity and broad application prospects. Summary of the invention

[0006] In order to overcome the problems existing in the prior art, the purpose of the present invention is to provide a method for preparing a tomato seed-derived small molecule protein with anti-muscle attenuation activity and its application.

[0007] The purpose of the present invention is achieved by at least one of the following technical solutions.

[0008] The present invention provides a preparation method of a small molecule protein with anti-muscle attenuation activity and application thereof.

[0009] A method for preparing a tomato seed small molecule protein having an anti-muscle attenuation effect comprises the following steps:

[0010] (1) Tomato seed defatting: The ultrafinely crushed tomato seed powder is defatted by steam explosion-wall breaking subcritical technology;

[0011] (2) Crude extraction of tomato seed protein: Mix the defatted tomato seed powder with distilled water, perform preliminary extraction in an alkaline environment, and collect the supernatant;

[0012] (3) Enzymatic hydrolysis of crude grain protein: adding the first protease and the second protease to the supernatant obtained in step (2) to perform segmented enzymatic hydrolysis, and collecting the supernatant;

[0013] (4) Purification and preparation of small molecule protein: The protein solution obtained in step (3) is vacuum concentrated at 40-60°C, desalted, and vacuum freeze-dried to obtain a small molecule protein with anti-muscle attenuation activity.

[0014] Furthermore, as a preferred embodiment of the small molecule protein of the effective ingredient of the present invention, the small molecule protein is prepared by an alkali method coupled with a low-temperature biological enzymatic hydrolysis method.

[0015] Furthermore, the tomato seed powder in step (1) refers to a component dried at 50-60° C. to a constant weight and having a particle size of ≤355 μm after crushing.

[0016] Furthermore, the protein content of the tomato seed raw material in step (1) is ≥ 25%.

[0017] Furthermore, the defatted tomato seed powder in step (1) has an oil content of ≤5%.

[0018] Furthermore, the alkaline environment in step (2) is pH 9-12, the extraction time is 0.5-2h, and the solid-liquid ratio is 1:10-30 (g / mL).

[0019] Furthermore, the supernatant in step (2) is obtained by centrifugation at 4000-6000×g for 10-30 min.

[0020] (3) Enzymatic hydrolysis of crude grain protein: Add the first protease and the second protease to the supernatant obtained in step (2) to perform segmented and directional enzymatic hydrolysis, and collect the supernatant.

[0021] Furthermore, in step (3), the first protease is Bacillus licheniformis protease, and the second protease is composite flavor protease.

[0022] Furthermore, the segmented enzymatic hydrolysis conditions in step (3) are: enzymatic hydrolysis time 2 to 4 hours, enzymatic hydrolysis temperature 45 to 65° C., and enzyme addition amount 5000 to 10000 U / g.

[0023] Furthermore, the supernatant in step (3) is obtained by centrifugation at 4000-6000×g for 10-30 min.

[0024] Furthermore, in step (3), the desalination refers to the removal of small molecules by membrane permeation or ultrafiltration, and the upper limit of the removal molecular weight is 1000-3000Da.

[0025] Furthermore, the mechanism of action of the above raw material component protease segmented directional enzymolysis is as follows:

[0026] Some common sequence features of small molecule proteins play an important role in inflammatory responses, such as the presence of hydrophobic amino acids (Val, Ile, Pro) and positively charged amino acids (His, Arg, Lys). Previous studies have shown that small molecule proteins with a high content of hydrophobic amino acids, mainly located at the N-terminus of the small molecule proteins, and positively charged amino acids at the C-terminus, exhibit higher anti-inflammatory activity.

[0027] Bacillus licheniformis protease: It is a kind of alkaline protease. Because its active center contains serine, it is also called serine protease. It can not only hydrolyze peptide bonds, but also hydrolyze amide bonds, ester bonds, and transesterify and transpeptide. The first protease is classified as an endonuclease, especially for peptide bonds containing hydrophobic amino acid residues. It has a specific cutting effect, and can obtain a large number of small molecule proteins containing hydrophobic amino acids in the C segment or N terminal.

[0028] Composite flavor protease: Contains two active enzymes, polypeptide exo- and endo-enzymes, and is mainly used to reduce or avoid the production of bitter peptides during hydrolysis, or degrade bitter peptides into amino acids, enhance or improve the taste and flavor of the hydrolyzate, and can supplement the first protease, effectively breaking the long-chain structure of the protein and generating more small molecule proteins.

[0029] Furthermore, the small molecule protein has been shown through experiments to have the effect of improving muscle attenuation.

[0030] Furthermore, the obtained tomato seed small molecule protein has a good improvement effect on the decreased swallowing frequency and motor ability of nematodes during aging. The effect was most significant on the 9th day of intervention, which was increased by 30% compared with the control group; at the same time, it can significantly increase the body swinging frequency of nematodes, which was increased by 25% compared with the control group.

[0031] Furthermore, the obtained tomato seed small molecule protein has a positive effect on weight recovery of sarcopenic mice, and can significantly improve the decline in muscle strength of sarcopenic mice, and has the effect of improving muscle attenuation.

[0032] Furthermore, the present invention provides the use of the above-mentioned small molecule protein in the preparation of products for delaying muscle attenuation.

[0033] Furthermore, the present invention provides the use of the above-mentioned small molecule protein in the preparation of products for improving muscle motility.

[0034] As a preferred embodiment of the application of the present invention, the products include medicines, foods, health products, cosmetics and biological products.

[0035] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0036] 1. The preparation method of the small molecule protein provided by the present invention is simple, the chemical properties are stable, and it has the effect of significantly improving muscle attenuation. It has the advantages of being safe, long-term use, and having no side effects.

[0037] 2. The small molecule bioactive protein provided by the present invention can be applied to products for improving muscle attenuation and its related symptoms, providing a new technical means for research and application in related fields. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on the swallowing frequency of nematodes;

[0039] Figure 2 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on the body swing frequency of nematodes;

[0040] Figure 3 Effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on the body weight of mice;

[0041] Figure 4 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on the pulling force of the hind limbs of mice;

[0042] Figure 5 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on the muscle mass of mice. DETAILED DESCRIPTION

[0043] The specific implementation of the present invention is further described below in conjunction with the accompanying drawings and examples, but the implementation and protection of the present invention are not limited thereto. It should be noted that if there are processes that are not particularly described in detail below, they can be implemented or understood by those skilled in the art with reference to the prior art. The reagents or instruments used without indicating the manufacturer can all be obtained from commercial channels.

[0044] Example 1

[0045] A method for preparing a tomato seed-derived small molecule protein: tomato seed powder is mixed with distilled water at a material-water ratio of 1:10, 1M NaOH is used to adjust the pH to 12.0, oscillate at 45°C for 2 hours, and collect the supernatant after centrifugation at 5500×g for 10 minutes. Repeat the above steps for re-extraction once. Combine the supernatants, add 5000U of the first protease, oscillate in a water bath at 50°C for 2 hours, inactivate the enzyme in a boiling water bath for 10 minutes, add 5000U of the second protease to repeat the enzymolysis once, inactivate the enzyme, collect the supernatant after centrifugation at 5500×g for 10 minutes, concentrate in a vacuum at 55°C, dialyze for 48 hours, and freeze-dry for 48 hours.

[0046] Example 2

[0047] A method for preparing a tomato seed-derived small molecule protein: tomato seed powder is mixed with distilled water at a material-water ratio of 1:20, 1M NaOH is used to adjust the pH to 9.0, oscillate at 55°C for 2 hours, and collect the supernatant after centrifugation at 4000×g for 30 minutes. Repeat the above steps for re-extraction once. Combine the supernatants, add 3000U of the first protease, oscillate in a water bath at 55°C for 3 hours, inactivate the enzyme in a boiling water bath for 10 minutes, add 3000U of the second protease to repeat the enzymolysis once, inactivate the enzyme, collect the supernatant after centrifugation at 4000×g for 20 minutes, concentrate in a vacuum at 40°C, dialyze for 24 hours, and freeze-dry for 48 hours.

[0048] Example 3

[0049] A method for preparing a tomato seed-derived small molecule protein: tomato seed powder is mixed with distilled water at a material-water ratio of 1:30, 1M NaOH is used to adjust the pH to 10.0, oscillate at 60°C for 2 hours, and collect the supernatant after centrifugation at 6000×g for 15 minutes. Repeat the above steps for re-extraction once. Combine the supernatants, add 10,000U of the first protease, oscillate in a water bath at 60°C for 4 hours, inactivate the enzyme in a boiling water bath for 10 minutes, add 10,000U of the second protease to repeat the enzymolysis once, inactivate the enzyme, collect the supernatant after centrifugation at 6000×g for 15 minutes, and freeze-dry for 48 hours.

[0050] Comparative Example 1

[0051] Preparation of tomato seed source small molecule protein: Mix tomato seed powder with distilled water at a material-water ratio of 1:10, adjust the pH to 9.0 with 1M NaOH, oscillate at 45°C for 2h, centrifuge at 8000rpm for 10min, and collect the supernatant. Repeat the above steps for re-extraction once. Combine the supernatants, adjust the pH to 4.0 with 1M HCl. Centrifuge at 8000rpm for 10min and collect the precipitate. After the precipitate is redissolved, add 1M NaOH to the solution, adjust the pH to 7.0, concentrate under vacuum at 55°C, desalt for 48h, and freeze-dry for 48h.

[0052] Comparative Example 2

[0053] Preparation of tomato seed source small molecule protein: Mix tomato seed powder with distilled water at a material-water ratio of 1:10, adjust the pH to 12.0 with 1M NaOH, shake at 45°C for 2h, centrifuge at 8000rpm for 10min, and collect the supernatant. Repeat the above steps for re-extraction once. Combine the supernatants, add 5000U neutral protease, shake in a water bath at 50°C for 2h, inactivate the enzyme in a boiling water bath for 10min, add 5000U bromelain and repeat the enzymolysis once, inactivate the enzyme, centrifuge at 8000rpm for 10min, collect the supernatant, and freeze-dry for 48h.

[0054] Comparative Example 3

[0055] Preparation of tomato seed source small molecule protein: Mix tomato seed powder with distilled water at a material-water ratio of 1:10, adjust the pH to 10.0 with 1M NaOH, oscillate at 55°C for 2h, centrifuge at 8000rpm for 10min, and collect the supernatant. Repeat the above steps for re-extraction once. Combine the supernatants, add 5000U neutral protease, oscillate in a water bath at 50°C for 2h, inactivate the enzyme in a boiling water bath for 10min, add 5000U papain and repeat the enzymolysis once, inactivate the enzyme, centrifuge at 8000rpm for 10min, collect the supernatant, and freeze-dry for 48h.

[0056] 1. Evaluation of the anti-muscle decay efficacy of tomato seed small molecule protein

[0057] 1.1 Based on the nematode model

[0058] Detection of nematode body swing frequency:

[0059] The body swing frequency of Caenorhabditis elegans was used to analyze the anti-muscle attenuation effect of the tomato seed protein prepared by the methods described in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3. The tomato seed protein prepared in Comparative Example 2 was added to the food of the synchronized nematodes, and 20 nematodes were randomly selected on the 8th day to measure their swing rate using a stereo microscope. The test nematodes were placed in M9 buffer and swam freely for 30 seconds to adapt to it before the test began. One swing of the head or tail of the nematode was counted as one time, and the record was recorded for 30 seconds.

[0060] Nematode swallowing frequency detection:

[0061] The swallowing frequency of Caenorhabditis elegans was used to analyze the anti-muscle attenuation effect of the tomato seed protein prepared by the methods described in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3. The tomato seed protein prepared in Comparative Example 2 was added to the food of the synchronized nematodes, and 20 nematodes were randomly selected on the 1st, 5th and 9th days to measure their pharyngeal swallowing frequency using a stereo microscope. The pharyngeal pump beats up and down a cycle as a complete beat, and the number of pharyngeal pump beats was recorded every 30 seconds.

[0062] 1.2 Based on mouse model

[0063] Mouse body weight determination:

[0064] By comparing the changes in the weight of mice, the effect of tomato seed protein prepared by the methods described in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 on improving muscle attenuation in mice was analyzed. The tomato seed protein prepared in Comparative Example 2 was gavaged into sarcopenic mice for 3 weeks. During the experiment, the food intake and water intake of the mice were weighed and the weight of the mice was recorded every day, and the data difference of each group of mice was calculated. The terminal weight of the mice was recorded at the end of the experiment.

[0065] Mouse muscle mass determination:

[0066] By comparing the changes in mouse muscle mass, the effects of tomato seed protein prepared by the methods described in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 on improving mouse muscle attenuation were analyzed. The tomato seed protein prepared in Comparative Example 2 was gavaged into sarcopenic mice for 3 weeks. At the end of the experiment, the mice were anesthetized, and the gastrocnemius, soleus, tibialis anterior and extensor digitorum muscles of the legs were dissected and taken. The blood stains were washed off in physiological saline, and the muscles were temporarily placed on ice covered with filter paper to absorb the water, and the weight was quickly weighed and recorded (accurate to 0.0001 g).

[0067] Mouse pulling force measurement:

[0068] The grip tester was used to measure the tensile force of mice to analyze the effect of tomato seed protein prepared by the methods described in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 on the muscle strength of mice. Before the formal experiment, the grip tester was fixed to a horizontal plane, and a piece of wire mesh was connected to the grip tester with a rubber band. At the beginning of the experiment, the mouse was placed on the platform so that it could grasp the middle of the wire mesh. After it grasped it firmly, the tail of the mouse was grasped and pulled horizontally backward. When the animal moves backward unintentionally, it will instinctively grasp the wire mesh to prevent retreat until the pulling force exceeds its maximum grip. Therefore, at this time, the grip tester automatically records the maximum value of the pulling force, displays it on the screen, and records it. Each mouse was tested 4 times, with an interval of 5 minutes for each mouse, and the test was carried out after the mouse recovered its strength. The entire experiment was completed by the same operator.

[0069] Experimental results analysis:

[0070] Figure 1 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2, and 3 on the swallowing frequency of nematodes. In comparison, the tomato seed small molecule protein obtained by the method of Example 1 significantly increased the swallowing frequency of nematodes on the 1st, 5th, and 9th days, with the most significant effect on the 9th day, which was 30% higher than the control group. As nematodes age, their swallowing frequency tends to decrease, and the tomato seed small molecule protein obtained by Example 1 can significantly slow down this trend, indicating that the small molecule protein obtained by the present invention has the effect of delaying muscle attenuation during aging.

[0071] Figure 2 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2, and 3 on the body swing frequency of nematodes. In comparison, the tomato seed small molecule protein obtained by the method of Example 1 has a more significant effect on improving the body swing frequency of nematodes, which is 25% higher than that of the control group. The muscle mass of nematodes gradually decreases with the occurrence of aging, which also leads to a decrease in their athletic ability. By observing and comparing the body swing frequencies of nematodes in different intervention groups, it is found that the small molecule protein obtained by Example 1 is more effective in improving the athletic ability of nematodes. The results show that the tomato seed small molecule protein prepared by the method of the present invention has the greatest effect on the body swing frequency of nematodes and has the potential to improve muscle attenuation.

[0072] Figure 3 The effect of the protein prepared in Example 1 and Comparative Example 1, Comparative Example 2, and Comparative Example 3 of the present invention on the body weight of mice. In the experiment, we constructed a mouse muscle attenuation model by physical disuse to simulate the process of sarcopenia. One week after modeling, we found that the body weight and hind limb tension of each group of mice decreased significantly, indicating that the muscle attenuation model was successfully constructed. In order to further analyze the improvement effect of the protein prepared in Example 1 and Comparative Example 1, Comparative Example 2, and Comparative Example 3 on muscle attenuation, we gavaged mice with the proteins prepared in Example 1 and Comparative Example 1, Comparative Example 2, and Comparative Example 3 for 3 weeks. The results showed that compared with the model group, after gavage with the tomato seed small molecule protein obtained by the method of Example 1, the weight of the mice no longer decreased and gradually recovered, indicating that the small molecule protein has a positive effect on the weight recovery of sarcopenic mice. The results show that the tomato seed small molecule protein prepared by the method of the present invention has the greatest effect on body weight and has the potential to improve muscle attenuation.

[0073] Figure 4The effects of the proteins prepared in Example 1 and Comparative Examples 1, 2 and 3 of the present invention on the pulling force of mice. The effects of the proteins prepared in Example 1 and Comparative Examples 1, 2 and 3 on the muscle strength of mice were further analyzed. The changes in the pulling force of mice were measured weekly by a tensile force meter. It was observed that after gavage with the small molecule protein obtained by the method of Example 1, the pulling force of the hind limbs of mice gradually recovered, which shows that the positive drug has a significant effect on the recovery of muscle attenuation. At the same time, the proteins of Comparative Examples 2 and 3 also have a certain improvement effect on the recovery of mouse muscle strength. The results show that the tomato seed small molecule protein prepared by the method of the present invention has the greatest effect on the pulling force of mice, can significantly improve the decline in muscle strength of sarcopenic mice, and has the effect of improving muscle attenuation.

[0074] Figure 5 The effects of the proteins prepared in Example 1 of the present invention and Comparative Examples 1, 2 and 3 on mouse muscle mass. In the experimental study, we conducted detailed sampling and weighing analysis on the leg muscles of mice to evaluate the effects of different treatment groups on the attenuation of gastrocnemius and soleus muscles. Through precise muscle weight measurement, it was found that compared with the model group, mice gavaged with the tomato seed small molecule protein obtained by the method of Example 1 showed significant improvement in the weight of gastrocnemius and soleus muscles.

[0075] Specifically, the weight of the gastrocnemius and soleus muscles of the mice in the model group was significantly reduced, which is consistent with the pathological characteristics of sarcopenia. However, the weight of the gastrocnemius and soleus muscles of the mice gavaged with the tomato seed small molecule protein obtained by the method of Example 1 showed a significant upward trend. The results show that the tomato seed small molecule protein prepared by the method of the present invention has the greatest effect on the muscle mass of mice and can significantly improve the muscle attenuation problem of sarcopenia mice.

[0076] In summary, the tomato seed small molecule protein prepared by the method of the present invention can not only improve muscle function, such as increasing the pulling force of the hind limbs, but also directly act on muscle tissue to promote the recovery of muscle weight and quality, thereby achieving an anti-muscle attenuation effect.

[0077] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for preparing a tomato seed small molecule protein having anti-muscle attenuation effect, characterized in that: The following steps are involved: (1) Tomato seed defatting: The ultrafinely crushed tomato seed powder is defatted by steam explosion-wall breaking subcritical technology; (2) Crude extraction of tomato seed protein: Mix the defatted tomato seed powder with distilled water, perform preliminary extraction in an alkaline environment, and collect the supernatant; (3) Enzymatic hydrolysis of crude grain protein: adding the first protease and the second protease to the supernatant obtained in step (2) to perform segmented enzymatic hydrolysis, and collecting the supernatant; (4) Purification and preparation of small molecule protein: The protein solution obtained in step (3) is vacuum concentrated at 40-60°C, desalted, and vacuum freeze-dried to obtain a small molecule protein with anti-muscle attenuation activity.

2. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: The tomato seed powder refers to a component that is dried at 50-60° C. to a constant weight and has a particle size of ≤355 μm after being crushed.

3. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: The alkaline environment has a pH of 9 to 12, the extraction time is 0.5 to 2 hours, and the solid-liquid ratio is 1:10 to 30 (g / mL).

4. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: The supernatant was obtained after centrifugation at 4000-6000×g for 10-30 min.

5. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: In step (3), the first protease is Bacillus licheniformis protease, and the second protease is composite flavor protease.

6. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: The conditions for the segmented enzymatic hydrolysis in step (3) are as follows: after the first protease hydrolyzes for 2 to 4 hours, the second protease is switched to continue hydrolyzing for 2 to 4 hours, the hydrolysis temperature is 45 to 65° C., and the enzyme addition amount is 5000 to 10000 U / g.

7. The method for preparing the tomato seed small molecule protein with anti-muscle attenuation effect according to claim 1, characterized in that: The desalination in step (4) refers to the removal of small molecules by membrane permeation or ultrafiltration, with the upper limit of the removal molecular weight being 1000 to 3000 Da.

8. The tomato seed small molecule protein with anti-muscle decay effect prepared by the preparation method according to claim 1 according to any one of claims 1 to 7, characterized in that: The obtained tomato seed small molecule protein has an improving effect on the decreased swallowing frequency and motor ability of nematodes during the aging process of nematodes. The effect was most significant on the 9th day of intervention, which was increased by 30% compared with the control group; at the same time, it significantly increased the body swing frequency of nematodes, which was increased by 25% compared with the control group; the tomato seed small molecule protein has a positive effect on the weight recovery of sarcopenic mice, and can significantly improve the decreased muscle strength of sarcopenic mice, and has the effect of improving muscle attenuation.

9. Use of the tomato seed small molecule protein as claimed in claim 8 in the preparation of products for delaying muscle attenuation or improving muscle mobility.

10. The use according to claim 9, characterized in that: The products include medicines, foods, health products, cosmetics and biological products.

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