Tuna meat-sourced imidazole dipeptide with lactic acid degradation effect and preparation method of tuna meat-sourced imidazole dipeptide
By preparing imidazole dipeptide derived from tuna meat and bayberry extract, the activity of lactate dehydrogenase was inhibited, solving the problems of muscle acidification and fatigue caused by lactate accumulation, and achieving rapid metabolism of lactate and relief of fatigue.
Patent Information
- Application Number
- CN202511258780.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-04
- Publication Date
- 2025-12-12
AI Technical Summary
During strenuous exercise, lactic acid buildup leads to muscle acidification and fatigue, and current technologies struggle to effectively inhibit lactate dehydrogenase activity and accelerate lactate metabolism.
Using tuna meat as raw material, imidazole dipeptide was prepared through enzymatic hydrolysis, fermentation and ultrafiltration membrane filtration. Combined with bayberry extract and citric acid, it inhibited lactate dehydrogenase activity and accelerated lactate metabolism.
The synergistic effect of imidazole dipeptide and bayberry extract effectively reduces lactic acid buildup after exercise and promotes fatigue recovery through immediate acid-base buffering and long-term metabolic functions.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of peptide preparation, and in particular to an imidazole dipeptide from tuna meat with lactic acid degradation efficacy and a preparation method thereof. BACKGROUND
[0002] During intense exercise, humans and animals will produce a large amount of acidic substances (such as lactic acid), and lactic acid is the end product of glycolysis, which is reduced from pyruvic acid under the catalysis of lactate dehydrogenase (LDH). When the exercise intensity exceeds the anaerobic threshold, the oxygen demand of muscle exceeds the oxygen supply capacity, and the mitochondria cannot oxidize pyruvic acid in time, resulting in its conversion into lactic acid. The accumulation of lactic acid will cause the pH to drop, leading to muscle acidification, inhibition of enzyme activity, and fatigue, and also causing short-term soreness. However, biologists have found that pelagic fish such as tuna and bonito migrate at high speed for tens of thousands of kilometers without causing the accumulation of acidic substances in the body. Scientific research has found that the imidazole dipeptide composition in tuna and bonito can rapidly reduce the acidic substances in the fish body.
[0003] Therefore, the present application provides a preparation method of an imidazole dipeptide from tuna meat, and the prepared imidazole dipeptide can inhibit lactate dehydrogenase activity, reduce post-exercise lactic acid accumulation, and be used for preparing products for degrading lactic acid and relieving fatigue. SUMMARY
[0004] The purpose of the present application is to provide an imidazole dipeptide from tuna meat with lactic acid degradation efficacy and a preparation method thereof, which can inhibit lactate dehydrogenase activity, reduce post-exercise lactic acid accumulation, and be used for preparing products for degrading lactic acid and relieving fatigue.
[0005] To achieve the above purpose, the present application provides a preparation method of an imidazole dipeptide from tuna meat with lactic acid degradation efficacy, comprising the following steps:
[0006] Step 1, raw material pretreatment, selecting the dorsal and ventral meat of fresh albacore tuna, adding ice cubes to stir into meat paste, then adding distilled water, heating and filtering to obtain filtrate I;
[0007] Step 2, adding alkaline protease to the filtrate I for enzymolysis, after the enzymolysis is completed, inactivating and centrifuging to obtain an enzymolysis liquid;
[0008] Step 3, mixing the enzymolysis liquid with waxberry extract, and adding a carbon source, a nitrogen source and a stabilizer, inoculating Lactobacillus plantarum for fermentation, and after the fermentation is completed, filtering and sterilizing to obtain filtrate II;
[0009] Step 4, filtering the filtrate II using two ultrafiltration membranes with different sizes to obtain filtrate III;
[0010] Step 5, the filtrate III is spray-dried to obtain the imidazole dipeptide powder.
[0011] Further, in step 1, the ratio of the meat paste and distilled water is meat paste: distilled water = 1g: 8-12mL, and the water bath heating condition is 30-40℃ for 30min.
[0012] Further, in step 2, the enzymolysis condition is that the alkaline protease is added in an amount of 0.3% under the condition of 40-45℃ and pH 7.5-8.5 for 1-2h.
[0013] Further, in step 3, the preparation method of the waxberry extract is as follows:
[0014] The waxberry is cored, added with distilled water to be made into a slurry, and then extracted by heating in a water bath at 60℃ for 30-45min, followed by filtration and pH adjustment to 5.5-6.5.
[0015] Further, in step 3, the volume ratio of the enzymolysis solution and the waxberry extract is enzymolysis solution:waxberry extract = 1:2-4, the carbon source is 5% glucose, the nitrogen source is 0.5% yeast extract, and the stabilizer is 0.5% trehalose.
[0016] Further, in step 3, during the fermentation, the inoculation amount of the lactobacillus plantarum is 3%, the fermentation temperature is 35-37℃, and the fermentation time is 24-48h.
[0017] Further, in step 4, firstly, the filtrate II is filtered by using a 1500-5000Da ultrafiltration membrane to obtain a permeate; and then the filtered filtrate is filtered by using a 150-300Da ultrafiltration membrane to obtain a retentate.
[0018] Further, the application further provides the imidazole dipeptide powder prepared by the preparation method of the imidazole dipeptide with the lactic acid degradation effect from tuna meat.
[0019] Further, the effective components of the imidazole dipeptide powder include imidazole dipeptide, citric acid and anthocyanin.
[0020] Further, the application further provides the application of the imidazole dipeptide powder in the preparation of a lactic acid degradation product.
[0021] The imidazole dipeptide with the lactic acid degradation effect from tuna meat and the preparation method thereof have the following advantages and positive effects:
[0022] 1. The imidazole dipeptide in the application can inhibit the activity of lactate dehydrogenase, reduce the accumulation of lactic acid after exercise, and citric acid directly participates in the tricarboxylic acid cycle to accelerate the metabolism of lactic acid into CO2 and water.
[0023] 2. This invention utilizes microbial fermentation to prepare imidazole dipeptides. During fermentation, microorganisms decompose the cell walls of bayberries, releasing more active ingredients. For example, myricetin increases the number and function of mitochondria through the AMPK and PGC-1α pathways. Imidazole dipeptides immediately neutralize the acidic environment, preventing low pH from inhibiting mitochondrial enzyme activity. The imidazole dipeptides and bayberry extract work synergistically, effectively promoting lactic acid clearance and conversion through a dual strategy of "immediate acid-base buffering and protection" and "long-term metabolic enhancement," thus accelerating the recovery from exercise-induced fatigue. Therefore, the imidazole dipeptides prepared in this invention can be used to prepare products that degrade lactic acid and relieve fatigue.
[0024] The technical solution of the present invention will be further described in detail below through embodiments. Detailed Implementation
[0025] The technical solution of the present invention will be further described below through embodiments.
[0026] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.
[0027] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention. Experimental methods in the following embodiments that do not specify specific conditions are generally determined according to national standards. Experimental instruments, equipment, and reagents in the following embodiments that do not specify their sources are all commercially available materials.
[0028] Unless otherwise defined or stated, all technical and scientific terms used in this invention have the same meaning as those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein can be applied to the methods of this invention. It should be noted that, without conflict, the embodiments and features described in this invention can be combined with each other. *Lactobacillus plantarum* was obtained commercially.
[0029] Example 1
[0030] A method for preparing an imidazole dipeptide derived from tuna meat that has the function of degrading lactic acid includes the following steps:
[0031] Step 1, raw material pretreatment: Select the back and belly meat of fresh bigeye tuna, add ice cubes and grind into minced meat. Take 10g of minced meat, add 100mL of distilled water, heat in a water bath at 30-40℃ for 30min, and filter to obtain filtrate I.
[0032] Step 2: Add 0.3% alkaline protease to filtrate I and hydrolyze for 1 hour at 45°C and pH 7.5-8.5. After hydrolysis, inactivate and centrifuge to obtain the hydrolysate.
[0033] Step 3, mix the enzyme solution with the bayberry extract solution 1:2, add 5% glucose, 0.5% yeast extract and trehalose, inoculate 3% Lactobacillus plantarum, and ferment, the fermentation temperature is 35-37℃, the fermentation time is 24h, after the fermentation is completed, filter sterilization to obtain filtrate II;
[0034] The preparation method of the bayberry extract solution is as follows:
[0035] The bayberry is cored, and distilled water is added to make a slurry, which is heated in a water bath at 60℃ for 30-45min, then filtered, and the pH value is adjusted to 5.5-6.5;
[0036] Step 4, filter the filtrate II with two different size ultrafiltration membranes to obtain filtrate III;
[0037] The specific operation is as follows:
[0038] First, filter the filtrate II with a 1500-5000Da ultrafiltration membrane to obtain a permeate;
[0039] Then filter the filtrate with a 150-300Da ultrafiltration membrane to obtain a retentate;
[0040] Step 5, spray dry the filtrate III to obtain imidazole dipeptide powder.
[0041] Example 2
[0042] A preparation method of an imidazole dipeptide with lactic acid degradation effect from tuna meat, comprising the following steps:
[0043] Step 1, raw material pretreatment, select the dorsal and ventral meat of fresh albacore tuna, add ice cubes to stir into a meat paste, take 10g of meat paste, add 100mL of distilled water, heat in a water bath at 30-40℃ for 30min, filter to obtain filtrate I;
[0044] Step 2, add 0.3% alkaline protease to the filtrate I, and enzymatically digest at 45℃, pH 7.5-8.5 for 1h, after the enzymatic digestion is completed, inactivate and centrifuge to obtain an enzyme solution;
[0045] Step 3, mix the enzyme solution with the bayberry extract solution 1:3, add 5% glucose, 0.5% yeast extract and trehalose, inoculate 3% Lactobacillus plantarum, and ferment, the fermentation temperature is 35-37℃, the fermentation time is 24h, after the fermentation is completed, filter sterilization to obtain filtrate II;
[0046] The preparation method of the bayberry extract solution is as follows:
[0047] The red bayberry is cored, and is beaten into a slurry with distilled water. The slurry is heated in a water bath at 60°C for 30-45 min, and then filtered. The pH value is adjusted to 5.5-6.5.
[0048] Step 4, the filtrate II is filtered by using two ultrafiltration membranes with different sizes to obtain a filtrate III.
[0049] The specific operation is as follows:
[0050] First, the filtrate II is filtered by using a 1500-5000 Da ultrafiltration membrane to obtain a permeate.
[0051] Then, the filtrate is filtered by using a 150-300 Da ultrafiltration membrane to obtain a retentate.
[0052] Step 5, the filtrate III is spray dried to obtain an imidazole dipeptide powder.
[0053] Example 3
[0054] A preparation method of an imidazole dipeptide with lactic acid degradation effect from tuna meat, comprising the following steps:
[0055] Step 1, raw material pretreatment, select the dorsal and ventral meat of fresh albacore tuna, add ice cubes to stir into a meat paste, take 10 g of the meat paste, add 100 mL of distilled water, heat in a water bath at 30-40°C for 30 min, and filter to obtain a filtrate I.
[0056] Step 2, add 0.3% alkaline protease to the filtrate I, and perform enzymolysis at 45°C and pH 7.5-8.5 for 1 h. After the enzymolysis is completed, inactivate and centrifuge to obtain an enzymolysis liquid.
[0057] Step 3, mix the enzymolysis liquid with a red bayberry extract at a ratio of 1:4, and add 5% glucose, 0.5% yeast extract and trehalose. Inoculate 3% Lactobacillus plantarum, and perform fermentation at a temperature of 35-37°C for 24 h. After the fermentation is completed, filter and sterilize to obtain a filtrate II.
[0058] The preparation method of the red bayberry extract is as follows:
[0059] The red bayberry is cored, and is beaten into a slurry with distilled water. The slurry is heated in a water bath at 60°C for 30-45 min, and then filtered. The pH value is adjusted to 5.5-6.5.
[0060] Step 4, the filtrate II is filtered by using two ultrafiltration membranes with different sizes to obtain a filtrate III.
[0061] The specific operation is as follows:
[0062] First, the filtrate II is filtered by using a 1500-5000 Da ultrafiltration membrane to obtain a permeate.
[0063] The filtrate is then filtered using a 150-300 Da ultrafiltration membrane to obtain a retentate;
[0064] Step 5, the filtrate III is spray dried to obtain imidazole dipeptide powder.
[0065] Comparative Example 1
[0066] A preparation method of an imidazole dipeptide with lactic acid degradation efficacy from tuna meat, comprising the following steps:
[0067] Step 1, raw material pretreatment, fresh albacore tuna dorsal and ventral meat is selected, ice cubes are added and stirred into a meat paste, 10 g of the meat paste is taken, 100 mL of distilled water is added, and the water bath is heated at 30-40°C for 30 min, and then filtered to obtain filtrate I;
[0068] Step 2, 0.3% alkaline protease is added to the filtrate I, and the enzyme is hydrolyzed at 45°C and pH 7.5-8.5 for 1 h, after the enzyme hydrolysis is completed, the enzyme is inactivated and centrifuged to obtain an enzyme hydrolysate;
[0069] Step 3, the enzyme hydrolysate is mixed with lemon extract at a ratio of 1:2, and 5% glucose, 0.5% yeast extract and trehalose are added, 3% Lactobacillus plantarum is inoculated, and fermentation is carried out, the fermentation temperature is 35-37°C, the fermentation time is 24 h, after the fermentation is completed, the filtrate is sterilized to obtain filtrate II;
[0070] The preparation method of the lemon extract is as follows:
[0071] The lemon slices are cored and added with distilled water to make a slurry, which is heated in a water bath at 60°C for 30-45 min, and then filtered, and the pH value is adjusted to 5.5-6.5;
[0072] Step 4, the filtrate II is filtered using two ultrafiltration membranes of different sizes to obtain filtrate III;
[0073] The specific operation is as follows:
[0074] First, the filtrate II is filtered using a 1500-5000 Da ultrafiltration membrane to obtain a permeate;
[0075] Then the filtrate is filtered using a 150-300 Da ultrafiltration membrane to obtain a retentate;
[0076] Step 5, the filtrate III is spray dried to obtain imidazole dipeptide powder.
[0077] Comparative Example 2
[0078] A preparation method of an imidazole dipeptide with lactic acid degradation efficacy from tuna meat, comprising the following steps:
[0079] Step 1, raw material pretreatment, select the dorsal and ventral meat of fresh bigeye tuna, add ice cubes to stir into meat paste, take 10 g of meat paste, add 100 mL of distilled water, heat in a water bath at 30-40℃ for 30 min, filter to obtain filtrate I;
[0080] Step 2, add 0.3% alkaline protease to filtrate I, and hydrolyze at 45℃ and pH 7.5-8.5 for 1 h, after hydrolysis is completed, inactivate and centrifuge to obtain a hydrolysis liquid;
[0081] Step 3, filter the hydrolysis liquid using two different size ultrafiltration membranes to obtain filtrate II;
[0082] The specific operation is as follows:
[0083] First, filter the hydrolysis liquid using a 1500-5000 Da ultrafiltration membrane to obtain a permeate;
[0084] Then filter the filtrate using a 150-300 Da ultrafiltration membrane to obtain a retentate;
[0085] Step 5, spray dry filtrate III to obtain imidazole dipeptide powder.
[0086] Effect test
[0087] Select 60 SD rats of single sex and weighing 160-200 g, and randomly divide them into 6 groups, 10 rats in each group. Among them, 5 groups are fed with imidazole dipeptide powder prepared in Examples 1-3 and Comparative Examples 1-2 (2 g of imidazole dipeptide freeze-dried powder is dissolved in 10 mL of distilled water), and the other group is fed with an equal amount of distilled water as a control group.
[0088] Gavage for 15 days, 30 min after feeding on the 15th day, a lead wire with a mass of 4% of the body weight of the mouse is placed on the tail of the mouse to perform a weight-loaded swimming experiment. After the mouse swims with weight, it rests for 10 minutes, and the eyeball is removed to take blood, and the whole blood lactic acid is determined according to the kit instructions. The results are shown in Table 1.
[0089] Table 1 Lactic acid content
[0090]
[0091]
[0092] As shown in Table 1, compared with the control group, the lactic acid content in the blood of the rats in the example 1, 2 and 3 groups is obviously reduced, which indicates that the imidazole dipeptide powder prepared in example 1, 2 and 3 can obviously reduce the lactic acid content in the blood and relieve fatigue. Compared with the comparative example 1, the lactic acid content in the blood of the rats in the example 1 group is obviously reduced, which indicates that the imidazole dipeptide and the Myrica rubra extract have a synergistic effect, the imidazole dipeptide can reduce the accumulation of lactic acid after exercise, and at the same time, the citric acid directly participates in the tricarboxylic acid cycle to accelerate the metabolism of lactic acid into CO2 and water. Compared with the comparative example 2, the lactic acid content in the blood of the rats in the example 1 group is obviously reduced, which indicates that the microorganisms decompose the cell wall of Myrica rubra during the fermentation process to release more active ingredients, and at the same time, the short-chain fatty acids produced during the fermentation process improve the stability and absorption rate of the imidazole dipeptide, and the two have a synergistic effect to reduce the accumulation of lactic acid after exercise and relieve fatigue.
[0093] Therefore, the imidazole dipeptide from tuna meat and the preparation method thereof having the effect of degrading lactic acid can inhibit the activity of lactic acid dehydrogenase, reduce the accumulation of lactic acid after exercise, and at the same time, the citric acid accelerates the metabolism of lactic acid, which can be used for preparing products for degrading lactic acid.
[0094] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application but not to limit it, although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can still be modified or replaced by equivalents, and these modifications or equivalent replacements cannot make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present application.
Claims
1. A method for preparing an imidazolic dipeptide derived from tuna meat having a lactic acid degradation effect, characterized by, The method comprises the following steps: Step 1, raw material pretreatment, fresh big-eye tuna dorsal and ventral meat is selected, ice cubes are added to stir into a meat paste, then distilled water is added, heated and filtered to obtain filtrate I; Step 2, alkaline protease is added to the filtrate I for enzymolysis, after the enzymolysis is completed, inactivation and centrifugation are performed to obtain an enzymolysis liquid; Step 3, the enzymolysis liquid is mixed with a waxberry extract liquid, and a carbon source, a nitrogen source and a stabilizer are added, and then Lactobacillus plantarum is inoculated for fermentation, after the fermentation is completed, filtration and sterilization are performed to obtain filtrate II; Step 4, the filtrate II is filtered by using two ultrafiltration membranes with different sizes to obtain filtrate III; Step 5, the filtrate III is spray dried to obtain imidazole dipeptide powder.
2. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid degradation function according to claim 1, characterized in that: In step 1, the ratio of the meat paste to distilled water is 1g:8-12mL, and the water bath heating condition is 30-40℃ for 30min.
3. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid-degrading effects according to claim 1, characterized in that, In step 2, the enzymolysis condition is that the alkaline protease is added in an amount of 0.3% under the condition of 40-45℃ and pH 7.5-8.5 for 1-2h.
4. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid-degrading effects according to claim 1, characterized in that, In step 3, the preparation method of the waxberry extract liquid is as follows: The waxberry is cored, distilled water is added to beat into a slurry, and then the slurry is heated in a water bath at 60℃ for 30-45min, and then filtered and adjusted to pH 5.5-6.
5.
5. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid-degrading effects according to claim 1, characterized in that, In step 3, the volume ratio of the enzymolysis liquid to the waxberry extract liquid is 1:2-4, the carbon source is 5% glucose, the nitrogen source is 0.5% yeast extract, and the stabilizer is 0.5% trehalose.
6. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid-degrading effects according to claim 1, characterized in that, In step 3, during the fermentation, the inoculation amount of Lactobacillus plantarum is 3%, the fermentation temperature is 35-37℃, and the fermentation time is 24-48h.
7. The method for preparing an imidazole dipeptide derived from tuna meat with lactic acid degradation function according to claim 1, characterized in that: In step 4, firstly, the filtrate II is filtered by using a 1500-5000Da ultrafiltration membrane to obtain a permeate; and then the filtrate is filtered by using a 150-300Da ultrafiltration membrane to obtain a retentate.
8. Imidazole dipeptide powder prepared by the preparation method of imidazole dipeptide with lactic acid degradation function from tuna meat source according to any one of claims 1-7.
9. The imidazole dipeptide powder according to claim 8, characterized by: The effective components of the imidazole dipeptide powder include imidazole dipeptide, citric acid and anthocyanin.
10. Application of the imidazole dipeptide powder of claim 9 in the preparation of a lactic acid degradation product.