Agoose carnosine extraction preparation process and application thereof

Abstract

The invention discloses an goose carnosine extraction preparation process and application thereof, and belongs to the technical field of goose carnosine extraction.The goose carnosine extraction preparation process comprises the following steps that firstly, tuna bones and bone marrow are separated, and pretreatment is conducted; 2, ultrasonic high-speed stirring treatment of tuna bone meal and bone marrow; 3, heat extraction; 4, membrane separation; and 5, mixing the target peptide with the liquid in the step 1, and then drying to obtain the goose carnosine with high stability. According to the method for extracting the tuna goose carnosine, the extraction rate is higher than 78%, the purine removal rate is higher than 55%, and the DPPH free radical scavenging rate is higher than 75%. The stabilizing agent composed of the malic acid, the betaine and the rhamnose not only can perform acidification and fermentation pretreatment on tuna bones, but also can play a role in stabilizing an goose carnosine sample, meanwhile, the extraction rate of tuna goose carnosine is finally improved, and the purine content can be effectively reduced by combining membrane filtration. High-voltage current treatment is performed before membrane separation, so that the DPPH free radical scavenging rate of the tuna goose carnosine can be increased.

Description

Technical Field

[0001] The invention relates to the technical field of anserine extraction, and in particular to an anserine extraction preparation process and application thereof. Background Art

[0002] Anserine (β-alanyl-1-methyl L-histidine) is a highly stable water-soluble dipeptide that mainly exists in the skeletal muscle tissue and brain tissue of vertebrates.

[0003] At present, the research on anserine mainly focuses on the extraction of fish anserine (such as salmon, tuna) and poultry muscles. For example, CN 112592315 B discloses a method for preparing high-content tuna anserine by membrane separation, wherein a crude tuna dry powder is mixed with a choline chloride-based low eutectic solvent to obtain a first precipitate and collect the supernatant; the first precipitate is mixed with a detergent and washed to obtain a supernatant; the supernatant is mixed and reacted to obtain a second precipitate, the second precipitate is mixed with a stabilizer solution, homogenized, and dried into powder to obtain a tuna anserine product.

[0004] However, the method uses tuna pulping to mix the fish meat and fish bones for extraction, and the extraction rate still needs to be further improved, and it is difficult to extract the fish bones in a targeted manner. At the same time, in order to maintain the stability and solubility of the obtained product anserine, a stabilizer is added, but the stabilizer only works in the product stage and has no effect in the early stage.

[0005] Based on this, the present invention designs an extraction and preparation process of anserine and its application to solve the above problems. Summary of the invention

[0006] In view of the above-mentioned shortcomings of the prior art, the present invention provides a process for extracting and preparing anserin and its application.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] A process for extracting and preparing anserine comprises the following steps:

[0009] Step 1: Using tuna bones as raw materials, separating tuna bones and bone marrow, and then pre-treating them respectively;

[0010] Pretreatment of tuna bones: crushing tuna bones into tuna bone powder, placing them in a 1.5-2.6% malic acid aqueous solution, acidifying them at 55-60° C. for 10-20 minutes, adding betaine and rhamnose and mixing them evenly; then fermenting them, inoculating yeast, fermenting them at 34-36° C. for 8-10 hours, and then inactivating them at high temperature, filtering them to obtain liquid and fermentation products, and the liquid is used in step five;

[0011] Pretreatment of tuna bone marrow: steam the tuna bone marrow in 125-130° C. steam for 1.4-2.2 hours to release the soluble peptides in the tuna bone marrow, then add 1.0-1.5w% alkaline protease (Alcalase), and inactivate it after enzymolysis at 45-50° C. for 2-7 hours to obtain an enzymolysis product;

[0012] Step 2, ultrasonic high-speed stirring treatment of tuna bone powder and bone marrow: mixing the fermentation product of tuna bone powder with the enzymatic hydrolysis product of tuna bone marrow, and ultrasonically stirring the mixture at room temperature of 20 to 25° C., with the stirring speed controlled at 2500 to 3000 r / min and the ultrasonic power controlled at 150 to 185 W to obtain a mixture;

[0013] Step 3, heat extraction: add hot water containing fructose to the mixture, stirring while adding, and add it within 10 minutes, the volume ratio of hot water to the mixture is 2.2-4.5:1; after the hot water is completely added, keep it above 75℃ and continue to stir and extract for 0.6-1.2 hours, centrifuge at 2500-3000rmp for 5-10 minutes, and take the liquid phase;

[0014] Step 4: membrane separation: firstly, a high voltage current with a frequency of 4200-4800 Hz and a voltage of 800-1000 V is passed through the liquid phase for 10-20 seconds, and then the power is turned off. Then, the liquid phase is subjected to membrane separation twice in sequence using an ultrafiltration membrane and an organic membrane to remove macromolecular proteins and polypeptides, etc., to obtain the target peptide;

[0015] The ultrafiltration membrane uses 3000-5000a ultrafiltration membrane;

[0016] The organic membrane uses 180-230Da organic membrane;

[0017] Step 5: Mix the target peptide with the liquid in step 1, use the liquid in step 1 as a fermentation stabilizer, mix well and freeze-dry to obtain anserine with high stability.

[0018] Furthermore, in step 1, the weight ratio of the tuna bone powder to the 1.5-2.6% malic acid aqueous solution is 1:5.6-10.

[0019] Furthermore, in step 1, the molar weight ratio of malic acid, betaine and rhamnose is 2.5-4.0:3.2-4.3:2.0-2.3.

[0020] Furthermore, in step 1, the tuna bones are crushed into tuna bone powder of 100-200 mesh.

[0021] Furthermore, in step 1, 6.6×10 5 ~8.4×10 5CFU / mL of yeast.

[0022] Furthermore, in step 2, the weight ratio of tuna bones to tuna marrow is 1:0.2-0.4kg.

[0023] Furthermore, in step 3, the hot water containing fructose is prepared by dissolving fructose in 100° C. water with an amount of 50 to 100 times its weight.

[0024] In order to better achieve the purpose of the present invention, the present invention also provides an application of the anserine extraction and preparation process in the preparation of a low-purine tuna anserine product.

[0025] In order to better achieve the purpose of the present invention, the present invention also provides an anserine prepared by the extraction and preparation process.

[0026] In order to better achieve the purpose of the present invention, the present invention also provides a use of the anserine in the preparation of antioxidant products.

[0027] Compared with the prior art, the present invention has the following beneficial effects: the present invention uses tuna bones as raw materials to extract tuna anserine, and the extraction rate is higher than 78%, the purine removal rate is higher than 55%, and the DPPH free radical scavenging rate is higher than 75%. The stabilizer composed of malic acid, betaine and rhamnose can be used for acidification and pre-fermentation pretreatment of tuna bones in the early stage to improve the fermentation activity of yeast, and can be used to stabilize the anserine sample in the later stage, so that the tuna anserine has high solubility and no residue after dissolution, and is ultimately beneficial to improving the extraction rate of tuna anserine, and can effectively reduce the purine content by combining with membrane filtration. Before performing two membrane separations, high voltage current treatment is performed in advance, which is beneficial to further decomposition of the liquid phase and improves the DPPH free radical scavenging rate of tuna anserine. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention is clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Embodiment 1: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 100 mesh tuna bone powder, putting into 10 kg of 1.5% malic acid aqueous solution, acidifying at 60° C. for 10 min, then adding betaine and rhamnose to mix, wherein the molar weight ratio of malic acid, betaine and rhamnose is 4.0:3.2:2.3; then fermenting, inoculating 6.6×10 5 CFU / mL yeast, fermented at 36°C for 8h and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.4kg tuna bone marrow was put into 125°C steam and boiled for 2.2h to release the soluble peptides in the tuna bone marrow, then 1.0w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 50°C for 2h and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 25°C, the stirring speed was controlled at 2500r / min, and the ultrasonic power was controlled at 185W to obtain a mixture; hot water containing fructose (fructose dissolved in The method comprises the following steps: adding 50 times its weight of 100°C water to the mixture, stirring and adding, and the addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 4.5:1; after the hot water is completely added, the mixture is kept at 80°C for further stirring and extraction for 0.6 hours, centrifuged at 3000 rpm for 5 minutes, and the liquid phase is obtained; firstly, a high-voltage current with a frequency of 4800 Hz and a voltage of 800 V is passed through the liquid phase for 20 seconds, and then the power is turned off, and then a 3000a ultrafiltration membrane and a 230Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after uniform mixing, freeze-drying is performed to obtain anserine with high stability.

[0030] Embodiment 2: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 200 mesh tuna bone powder, putting into 5.6 kg of 2.6% malic acid aqueous solution, acidifying at 55° C. for 20 min, then adding betaine and rhamnose to mix, the molar weight ratio of malic acid, betaine and rhamnose is 2.5:4.3:2.0; then fermenting, inoculating 8.4×10 5CFU / mL yeast, fermented at 34°C for 10 hours and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.2 kg tuna bone marrow was put into 130°C steam and boiled for 1.4 hours to release the soluble peptides in the tuna bone marrow, and then 1.5w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 45°C for 7 hours and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 20°C, the stirring speed was controlled at 3000r / min, and the ultrasonic power was controlled at 150W to obtain a mixture; hot water containing fructose (fructose dissolved in it) was added to the mixture, and the mixture was stirred ... The mixture is prepared by adding 100 times its weight of 100°C water to the mixture, stirring and adding within 10 minutes, and the volume ratio of hot water to the mixture is 2.2:1; after the hot water is completely added, it is maintained at 76°C and stirred for extraction for 1.2 hours, centrifuged at 2500rmp for 10 minutes, and the liquid phase is taken; first, a high-voltage current with a frequency of 4200Hz and a voltage of 1000V is passed through the liquid phase for 10s, and then the power is turned off, and then a 5000a ultrafiltration membrane and a 180Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0031] Embodiment 3: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 180 mesh tuna bone powder, putting into 7.0 kg of 2.2% malic acid aqueous solution, acidifying at 57° C. for 15 min, then adding betaine and rhamnose to mix, the molar weight ratio of malic acid, betaine and rhamnose is 3.0:3.6:2.1; then fermenting, inoculating 7.1×10 5CFU / mL yeast, fermented at 35°C for 9h and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.3kg tuna bone marrow was put into 128°C steam and boiled for 1.8h to release the soluble peptides in the tuna bone marrow, then 1.4w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 47°C for 5h and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 22°C, the stirring speed was controlled at 2800r / min, and the ultrasonic power was controlled at 175W to obtain a mixture; hot water containing fructose (fructose soluble The mixture is prepared by adding 60 times its weight in 100°C water, stirring it, and the addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 3.5:1; after the hot water is completely added, it is maintained at 82°C for 1 hour of stirring and extraction, and centrifuged at 2800rmp for 7 minutes to obtain a liquid phase; firstly, a high-voltage current with a frequency of 4500Hz and a voltage of 900V is passed through the liquid phase for 15s, and then the power is turned off, and then a 3500a ultrafiltration membrane and a 210Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0032] Embodiment 4: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 120 mesh tuna bone powder, putting into 6.3 kg of 1.8% malic acid aqueous solution, acidifying at 56° C. for 12 min, then adding betaine and rhamnose to mix, wherein the molar weight ratio of malic acid, betaine and rhamnose is 2.7:3.5:2.0; then fermenting, inoculating 6.9×10 55CFU / mL yeast, fermented at 34.5℃ for 8.6h and then inactivated at high temperature, filtered to obtain liquid and fermentation product, the liquid being used as a fermentation stabilizer; 0.25kg tuna bone marrow was put into 127℃ steam and boiled for 1.5h to release soluble peptides in the tuna bone marrow, then 1.1w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 46℃ for 3.5h and then inactivated to obtain enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 20.5℃, the stirring speed was controlled at 2600r / min, and the ultrasonic power was controlled at 155W to obtain a mixture; hot water containing fructose ( fructose dissolved in 70 times its weight of 100°C water) is added to the mixture while stirring, and the addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 2.5:1; after the hot water is completely added, it is maintained at 78°C and stirred for extraction for 0.8h, centrifuged at 2700rmp for 6min, and the liquid phase is taken; first, a high-voltage current with a frequency of 4500Hz and a voltage of 800V is passed through the liquid phase for 13s, and then the power is turned off, and then a 3000a ultrafiltration membrane and a 200Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0033] Embodiment 5: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 170 mesh tuna bone powder, putting into 9 kg of 2.5% malic acid aqueous solution, acidifying at 59° C. for 18 min, then adding betaine and rhamnose to mix, wherein the molar weight ratio of malic acid, betaine and rhamnose is 3.8:3.3:2.2; then fermenting, inoculating 8.1×10 5CFU / mL yeast, fermented at 35.5°C for 9h and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.32kg tuna bone marrow was put into 129°C steam and boiled for 1.9h to release the soluble peptides in the tuna bone marrow, then 1.4w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 48°C for 6h and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 23°C, the stirring speed was controlled at 2900r / min, and the ultrasonic power was controlled at 180W to obtain a mixture; hot water containing fructose (fructose soluble The mixture is prepared by adding 1% ethanol (prepared in 80 times its weight of 100°C water) to the mixture, stirring and adding, and the addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 4.2:1; after the hot water is completely added, it is maintained at 77°C and stirred for extraction for 0.7h, centrifuged at 2600rmp for 9min, and the liquid phase is taken; first, a high-voltage current with a frequency of 4600Hz and a voltage of 950V is passed through the liquid phase for 17s, and then the power is turned off, and then a 5000a ultrafiltration membrane and a 190Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0034] Embodiment 6: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 170 mesh tuna bone powder, putting into 7.6 kg of 2.1% malic acid aqueous solution, acidifying at 58.8° C. for 14 min, then adding betaine and rhamnose to mix, the molar weight ratio of malic acid, betaine and rhamnose is 2.6:4.2:2.1; then fermenting, inoculating 7.4×10 5CFU / mL yeast, fermented at 34.3°C for 8.1h and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.22kg tuna bone marrow was put into 126.5°C steam and boiled for 1.8h to release the soluble peptides in the tuna bone marrow, then 1.25w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 46.5°C for 3.5h and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 21.5°C, the stirring speed was controlled at 2600r / min, and the ultrasonic power was controlled at 170W to obtain a mixture; Hot water (fructose dissolved in 90 times its weight of 100°C water) is added to the mixture while stirring. The addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 3.6:1; after the hot water is completely added, it is maintained at 76°C and stirred for extraction for 1 hour, centrifuged at 2850rmp for 6.5 minutes, and the liquid phase is taken; first, a high-voltage current with a frequency of 4650Hz and a voltage of 850V is passed through the liquid phase for 18s, and then the power is turned off, and then a 3000a ultrafiltration membrane and a 185Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0035] Embodiment 7: In some embodiments, an extraction and preparation process of anserine is provided, using tuna bones as raw materials, separating tuna bones and bone marrow; crushing 1 kg of tuna bones into 140 mesh tuna bone powder, putting into 8.5 kg of 2.4% malic acid aqueous solution, acidifying at 57° C. for 12 min, then adding betaine and rhamnose to mix, the molar weight ratio of malic acid, betaine and rhamnose is 3.7:4.1:2.1; then fermenting, inoculating 6.9×10 5CFU / mL yeast, fermented at 34.8°C for 9.5h and then inactivated at high temperature, filtered to obtain a liquid and a fermentation product, the liquid being used as a fermentation stabilizer; 0.35kg tuna bone marrow was steamed in 129.4°C steam for 2h to release the soluble peptides in the tuna bone marrow, then 1.3w% alkaline protease (Alcalase) was added, and enzymolysis was performed at 47.5°C for 3.6h and then inactivated to obtain an enzymolysis product; the fermentation product of tuna bone meal was mixed with the enzymolysis product of tuna bone marrow, and ultrasonic stirring was performed at room temperature of 23.5°C, the stirring speed was controlled at 2650r / min, and the ultrasonic power was controlled at 155W to obtain a mixture; hot water containing fructose was added to the mixture, and the mixture was stirred at 2650r / min and 155W to obtain a mixture; (fructose dissolved in 85 times its weight of 100°C water) is added to the mixture while stirring, and the addition is completed within 10 minutes, and the volume ratio of hot water to the mixture is 2.8:1; after the hot water is completely added, it is maintained at 79°C and stirred for extraction for 0.7h, centrifuged at 2950rmp for 8min, and the liquid phase is taken; first, a high-voltage current with a frequency of 4210Hz and a voltage of 820V is passed through the liquid phase for 20s, and then the power is turned off, and then a 5000a ultrafiltration membrane and a 220Da organic membrane are used to perform membrane separation on the liquid phase twice in sequence to remove large molecular proteins and polypeptides, etc., to obtain the target peptide; the target peptide is mixed with the above-mentioned fermentation stabilizer, and after mixing evenly, freeze-dried to obtain anserine with high stability.

[0036] Comparative Example 1: The difference from Example 3 is that the pretreatment process of tuna bones is as follows: 1 kg of tuna bones are crushed into 180 mesh tuna bone powder and inoculated with 7.1×10 5 CFU / mL of yeast was fermented at 35°C for 9 hours and then inactivated by high temperature. Liquid and fermentation products were obtained after filtration, and the liquid was used as a fermentation stabilizer.

[0037] Comparative Example 2: The difference from Example 3 is that no high voltage current treatment is performed before the two membrane separations.

[0038] Test example: Liquid chromatography was used to detect the content of target tuna anserine in the product, and the extraction rate was calculated. The purine removal rate was detected. The antioxidant activity of the product anserine was also detected (DPPH free radical scavenging rate determination method). The results are shown in Table 1.

[0039] Anserine extraction rate = target tuna anserine content / total product weight * 100%

[0040] Table 1 Anserine sample content and purity test results

[0041]

[0042] Compared with comparative examples 1-2, the extraction rate of tuna anserine by the method of the present invention is higher than 78%, the purine removal rate is higher than 55%, and the DPPH free radical scavenging rate is higher than 75%. The stabilizer composed of malic acid, betaine and rhamnose can be used for acidification of tuna bones and pretreatment before fermentation to improve the fermentation activity of yeast in the early stage, and can be used to stabilize the anserine sample in the later stage, so that the tuna anserine has high solubility and no residue after dissolution, and is ultimately beneficial to improving the extraction rate of tuna anserine, and can effectively reduce the purine content by combining with membrane filtration. Before performing two membrane separations, high voltage current treatment is performed in advance, which is beneficial to further decomposition of the liquid phase and improves the DPPH free radical scavenging rate of tuna anserine.

[0043] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A process for extracting and preparing anserin, characterized in that: The following steps are involved: Step 1: Using tuna bones as raw materials, separating tuna bones and bone marrow, and then pre-treating them respectively; Pretreatment of tuna bones: crushing tuna bones into tuna bone powder, placing them in a 1.5-2.6% malic acid aqueous solution, acidifying them at 55-60° C. for 10-20 minutes, adding betaine and rhamnose and mixing them evenly; then fermenting them, inoculating yeast, fermenting them at 34-36° C. for 8-10 hours, and then inactivating them at high temperature, filtering them to obtain liquid and fermentation products, and the liquid is used in step five; Pretreatment of tuna bone marrow: steam the tuna bone marrow in 125-130° C. steam for 1.4-2.2 hours to release the soluble peptides in the tuna bone marrow, then add 1.0-1.5w% alkaline protease, and inactivate it after enzymolysis at 45-50° C. for 2-7 hours to obtain an enzymolysis product; Step 2, ultrasonic high-speed stirring treatment of tuna bone powder and bone marrow: mixing the fermentation product of tuna bone powder with the enzymatic hydrolysis product of tuna bone marrow, and ultrasonically stirring the mixture at room temperature of 20 to 25° C., with the stirring speed controlled at 2500 to 3000 r / min and the ultrasonic power controlled at 150 to 185 W to obtain a mixture; Step 3, heat extraction: add hot water containing fructose to the mixture, stirring while adding, and add it within 10 minutes, the volume ratio of hot water to the mixture is 2.2-4.5:1; after the hot water is completely added, keep it above 75℃ and continue to stir and extract for 0.6-1.2 hours, centrifuge at 2500-3000rmp for 5-10 minutes, and take the liquid phase; Step 4: membrane separation: firstly, a high voltage current with a frequency of 4200-4800 Hz and a voltage of 800-1000 V is passed through the liquid phase for 10-20 seconds, and then the power is turned off. Then, the liquid phase is subjected to membrane separation twice in sequence using an ultrafiltration membrane and an organic membrane to remove macromolecular proteins and polypeptides, etc., to obtain the target peptide; Step 5: Mix the target peptide with the liquid in step 1, use the liquid in step 1 as a fermentation stabilizer, mix well and freeze-dry to obtain anserine with high stability.

2. The process for extracting and preparing anserin according to claim 1, characterized in that: In step 1, the weight ratio of the tuna bone powder to the 1.5-2.6% malic acid aqueous solution is 1:5.6-10.

3. The process for extracting and preparing anserin according to claim 2, characterized in that: In step 1, the molar weight ratio of malic acid, betaine and rhamnose is 2.5-4.0:3.2-4.3:2.0-2.

3.

4. The process for extracting and preparing anserin according to claim 3, characterized in that: In step 1, the tuna bones are crushed into tuna bone powder of 100-200 mesh.

5. The process for extracting and preparing anserin according to claim 4, characterized in that: In step 1, 6.6×10 5 ~8.4×10 5 CFU / mL of yeast.

6. The process for extracting and preparing anserin according to claim 5, characterized in that: In step 2, the weight ratio of tuna bones to tuna marrow is 1:0.2-0.4kg.

7. The process for extracting and preparing anserin according to claim 6, characterized in that: In step 3, the hot water containing fructose is prepared by dissolving fructose in 100° C. water with an amount of 50 to 100 times its weight.

8. Use of the extraction and preparation process of anserine according to claim 7 in the preparation of a low-purine tuna anserine product.

9. An anserine prepared according to the extraction and preparation process of claim 7.

10. Use of the anserine according to claim 9 in the preparation of antioxidant products.

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