Method for producing colla corii asini peptide through double-enzymolysis extrusion and application

Abstract

The invention relates to the technical field of colla corii asini peptides, and particularly discloses a method for producing colla corii asini peptides through double-enzymolysis extrusion and application. A method for producing colla corii asini peptide through double-enzymolysis extrusion comprises the following steps: (1) washing a colla corii asini raw material, crushing, mixing with water, and pulping to obtain premixed slurry; (2) adding an enzyme preparation into the premixed slurry for enzymolysis to obtain enzymolysis slurry; (3) performing twin-screw extrusion treatment on the enzymolysis slurry, controlling the liquid-material ratio to be 1: (2-5), and controlling the temperature of an extrusion screw to be 60-100 DEG C; (4) carrying out heat-preservation hydrolysis treatment on the enzymatic hydrolysate obtained by extrusion, and then carrying out enzyme deactivation treatment to obtain enzyme-deactivated slurry; and (5) treating the enzyme-deactivated slurry by a colloid mill and a homogenizer, and then filtering and sterilizing at high temperature to obtain the donkey-hide gelatin peptide product. According to the method for producing the donkey-hide gelatin peptide through double-enzymolysis extrusion, the molecular mass of the donkey-hide gelatin after enzymolysis can be remarkably reduced, and the total peptide content of the donkey-hide gelatin is excellently increased.

Description

Technical Field

[0001] The present application relates to the technical field of donkey-hide gelatin peptides, and more specifically, to a method for producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion and its application. Background Art

[0002] Donkey-hide gelatin is a gelatin block made from the skin of the donkey, an equine animal, which is bleached, soaked, dehaired, and then boiled. Modern research shows that donkey-hide gelatin contains collagen, various amino acids, polysaccharides, volatile substances and inorganic substances, various trace elements, hyaluronic acid, dermatan sulfate, etc. It can improve sleep quality, enhance memory, increase calcium intake in the body, and has anti-fatigue, blood-tonifying and blood-clotting, and cancer prevention and treatment effects.

[0003] Studies have shown that the protein content in donkey-hide gelatin is about 60-80%, mainly collagen and its degraded polypeptides. Donkey-hide gelatin is beneficial to human health, but the relative molecular mass of proteins and polypeptides in donkey-hide gelatin is relatively large, mainly concentrated in 6×10 3 - 2×10 5 Large proteins are not easily absorbed and utilized by the human body. Because the human body cannot directly absorb large proteins, they must be hydrolyzed into small peptides by proteases within the body before they can be absorbed. For people with weak digestive systems, taking donkey-hide gelatin requires a large amount of proteases to hydrolyze it, which places a heavy burden on the digestive system and severely impairs the absorption of the gelatin's active ingredients, resulting in a lack of therapeutic efficacy and even worsening the condition. Therefore, breaking down the large protein molecules in donkey-hide gelatin into small peptides facilitates rapid absorption by the body, maximizing its nutritional and tonic effects.

[0004] Methods for breaking down the protein macromolecules in donkey-hide gelatin into small peptides primarily include enzymatic hydrolysis, acid hydrolysis, and fermentation. Enzymatic hydrolysis not only preserves the rich nutritional value of donkey-hide gelatin but also is widely used because the enzyme preparation is biodegradable, reducing waste and aligning with green production practices. Enzymatic hydrolysis is used to break down the macromolecule proteins in donkey-hide gelatin into small peptides. The specific steps include raw material pretreatment, enzymatic hydrolysis, filtration, decolorization, and drying. Multiple enzymatic hydrolysis steps, such as alkaline protease, neutral protease, and pepsin, are often used to produce small peptides that are easily digested and absorbed by the human body.

[0005] Regarding the above-mentioned related technologies, the inventors believe that some proteins are difficult to completely decompose into small molecular peptides by relying solely on the action of enzymes. Therefore, after enzymatic hydrolysis, the molecular weight of donkey-hide gelatin still needs to be reduced, and the total peptide content of donkey-hide gelatin still needs to be improved. Summary of the Invention

[0006] In order to significantly reduce the molecular weight of donkey-hide gelatin after enzymatic hydrolysis and bring about an excellent increase in the total peptide content of donkey-hide gelatin, the present application provides a method and application for producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion.

[0007] In the first aspect, the present application provides a method for producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion, which adopts the following technical solution: A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion, comprising the following steps: (1) Washing and crushing the raw material of donkey-hide gelatin, and mixing and beating with water to obtain a premixed slurry; (2) adding an enzyme preparation to the premixed slurry obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis slurry; (3) The enzymatic hydrolysis slurry of step (2) is subjected to twin-screw extrusion, the liquid-to-solid ratio is controlled to be 1:(2-5), and the extrusion screw temperature is controlled at 60-100°C; (4) subjecting the enzymatic hydrolyzate obtained by extrusion in step (3) to a heat preservation and hydrolysis treatment, and then to an enzyme inactivation treatment to obtain an enzyme inactivation slurry; (5) The enzyme-inactivated slurry obtained in step (4) is filtered through a sieve, processed through a colloid mill and a homogenizer, and then filtered and sterilized at high temperature to obtain the donkey-hide gelatin peptide product.

[0008] By adopting the above technical solution, the donkey-hide gelatin raw material is processed to obtain a premixed slurry, and the collagen peptide bonds can be specifically cut by enzyme preparations to decompose the large-molecule collagen into small-molecule peptide segments; and in this process, the high temperature (60-100°C) and high shear force of twin-screw extrusion fully denature and stretch the donkey-hide gelatin protein, exposing more enzyme action sites, and the enzymatic reaction rate is increased by more than 40%. In addition, the extrusion process realizes the simultaneous enzymatic reaction and mechanical energy input, which can reduce the residual large molecules, thereby significantly reducing the molecular weight of the donkey-hide gelatin after enzymatic hydrolysis, and bringing about an excellent increase in the total peptide content of donkey-hide gelatin; subsequent enzyme inactivation treatment is performed to terminate the reaction in time to prevent excessive hydrolysis from causing peptide inactivation, and filtration can accurately intercept the target molecular weight peptide segment, and the colloid mill and homogenizer are used in combination to reduce the peptide segment particle size to the micron level, and finally a donkey-hide gelatin peptide product with better application quality is obtained. Therefore, in the method of producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion, the synergistic effect of enzymatic hydrolysis and extrusion can make the protein more completely decomposed into small molecular peptides, and the protein that is more difficult to completely decompose into small molecular peptides can also be deeply and effectively acted upon, so that the obtained donkey-hide gelatin peptide product not only has a smaller molecular weight of donkey-hide gelatin, but also has a significantly higher total peptide content of donkey-hide gelatin, breaking through the technical bottleneck of traditional donkey-hide gelatin enzymatic hydrolysis, and at the same time achieving a double leap in production efficiency and product quality, providing core technical support for the industrial production of highly bioactive donkey-hide gelatin peptides.

[0009] Preferably, in step (2), the enzyme preparation is a combination of one or more of mesophilic α-amylase, saccharifying enzyme and protease.

[0010] Preferably, in step (2), the enzyme preparation is composed of the following enzyme raw materials in corresponding weight ratios in the premixed slurry: Medium-temperature α-amylase 0.3-0.5%; Glucoamylase 0.3-0.5%; Protease 0.1-0.3%.

[0011] By employing this technical solution, proteases directly target and cleave the peptide bonds of donkey-hide gelatin collagen, degrading the macromolecules into small peptide segments. Combined with the high extrusion temperature (60-100°C), the protein is fully denatured, increasing enzymatic hydrolysis efficiency by over 40%. Intermediate-temperature α-amylases and saccharifying enzymes efficiently hydrolyze starchy impurities at extrusion temperatures of 60-100°C, perfectly matching the extrusion screw temperature (60-100°C), enabling simultaneous enzymatic hydrolysis and mechanical extrusion. The synergistic combination of these different enzymes achieves temperature adaptability, substrate specificity, and stage-by-stage synergy, significantly reducing the molecular weight of the hydrolyzed donkey-hide gelatin and significantly increasing its total peptide content, ultimately resulting in a high-quality donkey-hide gelatin peptide product.

[0012] Preferably, step (4) is specifically configured as follows: the enzymatic hydrolyzate obtained by extrusion in step (3) is subjected to a heat-insulating hydrolysis treatment at a temperature of 60-70° C. for a time of 35-60 min, and then an enzyme-inactivating treatment is performed to obtain an enzyme-inactivating slurry.

[0013] By adopting the above technical solution, some incompletely hydrolyzed intermediates still exist in the enzymatic hydrolysate after extrusion. Keeping it warm at 60-70°C can maintain the activity of the enzyme preparation and further degrade the large molecular peptides. At the same time, this temperature range is higher than the critical inactivation point of most proteases (55-60°C), which can gradually reduce the enzyme activity, reduce the load for subsequent enzyme inactivation treatment, and avoid the destruction of the peptide structure due to sudden heating, so as to ensure the excellent and stable quality of the final donkey-hide gelatin peptide product.

[0014] Preferably, in step (4), during the enzyme inactivation treatment, the enzyme inactivation temperature is 90-95° C., and the enzyme inactivation time is 15-20 min.

[0015] By adopting the above technical solution, the above enzyme inactivation temperature can directly destroy the three-dimensional structure of the enzyme protein, so that the enzyme preparation is completely inactivated within the above enzyme inactivation time, the enzymatic reaction process is terminated, and the residual enzyme activity is prevented from interfering with the subsequent process; at the same time, the above temperature is lower than the aggregation temperature of donkey-hide gelatin peptide, which can avoid peptide chain cross-linking or denaturation precipitation, thereby ensuring the stability of the final donkey-hide gelatin peptide product.

[0016] Preferably, in step (5), the rotation speed of the colloid mill is 1000-2000 r / min and the temperature is 55-65°C.

[0017] By adopting the above technical solution, the rotation speed of the above colloid mill can avoid the serious frictional heating phenomenon, thereby avoiding local high temperature causing peptide chain aggregation; at the same time, the above operation can also reduce the particle size of the peptide segment to the micron level, and the distribution is more uniform, and can avoid excessive mechanical force to destroy the active structure of small molecule peptides, ensuring that the final quality of the donkey-hide gelatin peptide product is excellent and stable.

[0018] Preferably, in step (5), during the homogenizer treatment, the homogenization temperature is 55-65°C, the first-stage homogenization pressure is 60-100 MPa, and the second-stage homogenization pressure is 40-60 MPa.

[0019] By adopting the above technical solution, the homogenization temperature is 55-65°C, which can not only avoid collagen peptide precipitation caused by low temperature, but also avoid peptide chain cross-linking or denaturation caused by high temperature; in the first-level homogenization, high-pressure shear force above 60MPa can completely disintegrate collagen aggregates and improve the dissolution rate of peptide segments; in the second-level homogenization, medium pressure of 40-60MPa promotes uniform dispersion of broken particles and prevents re-aggregation; and step-by-step pressure reduction can also reduce the damage of mechanical shear to functional peptide segments; therefore, the above operation is conducive to obtaining high-quality and stable donkey-hide gelatin peptide products.

[0020] Preferably, in step (5), the filtration operation is performed using a 100-300 mesh sieve.

[0021] By adopting the above technical solution, the sieve of the above specifications can quickly remove incompletely enzymatically hydrolyzed collagen clumps, colloidal aggregates and mechanical impurities, ensuring that the final quality of the donkey-hide gelatin peptide product is excellent and stable.

[0022] In a second aspect, the present application provides an application of double enzymatic hydrolysis and extrusion to produce donkey-hide gelatin peptides, using the following technical solution: The invention discloses an application of double enzymatic hydrolysis and extrusion in producing donkey-hide gelatin peptides. Donkey-hide gelatin and glutinous rice are added to donkey-hide gelatin raw materials, and a double enzymatic hydrolysis and extrusion method is used to produce donkey-hide gelatin peptides to obtain donkey-hide gelatin and glutinous rice plant-based products.

[0023] By employing this technical solution, blood glutinous rice and donkey-hide gelatin peptides form a "plant-animal" dual-protein system, improving amino acid balance and iron bioavailability. The active ingredients of the traditional Chinese medicine components are released through enzymatic hydrolysis, and together with the donkey-hide gelatin peptides, they form a "blood-tonifying + immune-modulating" complex functional network. Therefore, by adding blood glutinous rice and traditional Chinese medicine to the donkey-hide gelatin raw material to produce a plant-based donkey-hide gelatin and blood glutinous rice product, the advantages of donkey-hide gelatin, blood glutinous rice, and traditional Chinese medicine are combined to achieve functional diversification. Furthermore, the enzyme preparations used include cellulase and pectinase, which can degrade the plant cell wall polysaccharides in the donkey-hide gelatin raw material, reduce colloid blockage, release more collagen-exposed sites, and ensure efficient protease contact.

[0024] Preferably, the Chinese medicinal materials are composed of Codonopsis pilosula, Rehmannia root, Astragalus membranaceus and Angelica sinensis.

[0025] By adopting the above technical solutions, Codonopsis pilosula replenishes Qi and produces blood, Rehmannia root nourishes Yin and replenishes blood, Astragalus replenishes Qi and strengthens the exterior, and Angelica promotes blood circulation and regulates menstruation. Together with donkey-hide gelatin collagen peptides, they form an optimized combination of "double replenishment of Qi and blood + functional synergy", producing a more comprehensive donkey-hide gelatin and glutinous rice plant-based product.

[0026] In summary, this application has the following beneficial effects: 1. In the method for producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion, the synergistic effect of enzymatic hydrolysis and extrusion enables proteins to be relatively completely decomposed into small-molecule peptide segments. Proteins that are relatively difficult to completely decompose into small-molecule peptide segments can also be deeply and effectively affected, thereby significantly reducing the molecular weight of donkey-hide gelatin after enzymatic hydrolysis and achieving an excellent increase in the total peptide content of donkey-hide gelatin, thereby obtaining a high-quality donkey-hide gelatin peptide product. 2. This application achieves temperature adaptability, substrate specificity and stage synergy through the synergistic combination of medium-temperature α-amylase, saccharifying enzyme and protease, and realizes a good combination of enzymatic hydrolysis reaction and twin-screw extrusion treatment, thereby significantly reducing the molecular weight of donkey-hide gelatin after enzymatic hydrolysis and bringing about an excellent improvement in the total peptide content of donkey-hide gelatin, ultimately obtaining a donkey-hide gelatin peptide product with better application quality. DETAILED DESCRIPTION

[0027] The present application is further described in detail below with reference to the following examples and comparative examples.

[0028] Example Example 1 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion, comprising the following steps: (1) Washing and crushing the donkey-hide gelatin raw material, and mixing and beating with water in a weight ratio of 1:4 to obtain a premixed slurry; (2) adding an enzyme preparation to the premixed slurry obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis slurry; (3) The enzymatic hydrolysis slurry of step (2) is subjected to twin-screw extrusion treatment, and the liquid-to-material ratio is controlled to be 1:3.5. The extrusion screw temperature zones 1 to 7 are: 50°C, 55°C, 60°C, 70°C, 75°C, 80°C, and 85°C, respectively; (4) subjecting the enzymatic hydrolyzate obtained by extrusion in step (3) to a heat preservation and hydrolysis treatment, and then to an enzyme inactivation treatment to obtain an enzyme inactivation slurry; (5) The enzyme-inactivated slurry obtained in step (4) is processed by a colloid mill and a homogenizer, and then filtered and sterilized at 120° C. to obtain the donkey-hide gelatin peptide product.

[0029] Note: In the above operation, in step (2), the enzyme preparation is composed of the following enzyme raw materials in the premixed slurry in the corresponding weight ratio: medium-temperature α-amylase 0.4%, saccharifying enzyme 0.4%, and protease 0.2%. In step (4), the holding temperature of the hydrolysis treatment is 65°C and the holding time is 47.5 minutes; the enzyme inactivation temperature is 92.5°C and the enzyme inactivation time is 17.5 minutes. In step (5), the speed of the colloid mill is 1500r / min and the temperature is 60°C; during the homogenizer treatment, the homogenization temperature is 60°C, the first homogenization pressure is 80MPa, and the second homogenization pressure is 50MPa; the filtration operation is carried out using a 200-mesh screen.

[0030] Example 2 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (2), the enzyme preparation is composed of the following enzyme raw materials in corresponding weight proportions in the premixed slurry: 0.3% medium-temperature α-amylase, 0.3% saccharifying enzyme, and 0.1% protease.

[0031] Example 3 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (2), the enzyme preparation is composed of the following enzyme raw materials in corresponding weight proportions in the premixed slurry: 0.5% medium-temperature α-amylase, 0.5% saccharifying enzyme, and 0.3% protease.

[0032] Example 4 A method for producing donkey-hide gelatin peptide by double enzymatic extrusion is different from Example 1 in that, in step (3), the liquid-to-material ratio is controlled to be 1:2, and the extrusion screw temperature zones 1 to 7 are, in sequence: 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, and 85°C.

[0033] Example 5 A method for producing donkey-hide gelatin peptide by double enzymatic extrusion is different from Example 1 in that, in step (3), the liquid-to-material ratio is controlled to be 1:5, and the extrusion screw temperature zones 1 to 7 are, in sequence: 65°C, 75°C, 80°C, 85°C, 90°C, 95°C, and 100°C.

[0034] Example 6 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (4), the holding temperature of the hydrolysis treatment is 60°C and the holding time is 60 minutes.

[0035] Example 7 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (4), the holding temperature of the hydrolysis treatment is 70°C and the holding time is 35 minutes.

[0036] Example 8 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (4), the enzyme inactivation temperature is 90°C and the enzyme inactivation time is 20 minutes.

[0037] Example 9 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (4), the enzyme inactivation temperature is 95°C and the enzyme inactivation time is 15 minutes.

[0038] Example 10 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), the rotation speed of the colloid mill is 1000 r / min and the temperature is 65°C.

[0039] Example 11 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), the rotation speed of the colloid mill is 2000 r / min and the temperature is 55°C.

[0040] Example 12 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), during the homogenizer treatment, the homogenization temperature is 55°C, the first-level homogenization pressure is 60 MPa, and the second-level homogenization pressure is 40 MPa.

[0041] Example 13 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), during the homogenizer treatment, the homogenization temperature is 65°C, the first-level homogenization pressure is 100 MPa, and the second-level homogenization pressure is 60 MPa.

[0042] Example 14 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), the filtration operation is performed using a 100-mesh screen.

[0043] Example 15 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that, in step (5), the filtration operation is performed using a 300-mesh screen.

[0044] Example 16 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion is different from Example 1 in that in step (5), the medium-temperature α-amylase and saccharifying enzyme in the enzyme preparation are replaced by proteases in the corresponding dosage ratio.

[0045] Comparative Example Comparative Example 1 A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion, which differs from Example 1 in that it comprises the following steps: (1) Washing and crushing the donkey-hide gelatin raw material, and mixing and beating with water in a weight ratio of 1:4 to obtain a premixed slurry; (2) adding an enzyme preparation to the premixed slurry obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis slurry; (3) subjecting the enzymatic hydrolysis slurry obtained in step (2) to a heat preservation hydrolysis treatment, and then to an enzyme inactivation treatment to obtain an enzyme inactivation slurry; (4) The enzyme-inactivated slurry obtained in step (3) was processed by a colloid mill and a homogenizer, and then filtered and sterilized at 120°C for 10 minutes to obtain the donkey-hide gelatin product.

[0046] Performance testing Test samples: Donkey-hide gelatin raw materials were purchased from Shandong Liaocheng Renkang Donkey-hide Gelatin Products Co., Ltd., with production license number SC13141092600022; medium-temperature α-amylase, saccharifying enzyme and protease were all purchased from Shanghai ELISA Biotechnology Co., Ltd.

[0047] Test method: Based on the above test samples, the method for producing donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion in Examples 1-16 and Comparative Example 1 was applied to obtain 17 donkey-hide gelatin peptide products.

[0048] Gel permeation chromatography was used to measure the molecular weight distribution of donkey-hide gelatin in 17 donkey-hide gelatin peptide products. The principle of gel permeation chromatography is to separate peptides of different molecular weights through a chromatographic column, and combine with the standard calibration curve to determine the molecular weight distribution range of collagen degradation products in donkey-hide gelatin. The proportion of donkey-hide gelatin with a molecular weight below 3,000 is recorded in Table 1.

[0049] The total donkey-hide gelatin peptide content of 17 donkey-hide gelatin peptide products was measured by high-performance liquid chromatography-mass spectrometry. The HPLC-MS method used a ChromCore 120 C18 5μm, 4.6×250mm column with octadecylsilane bonded silica gel as the filler, acetonitrile as the mobile phase A, and 0.1% formic acid solution as the mobile phase B. The gradient elution was performed at a flow rate of 0.3ml per minute. Scanning detection was performed in the electrospray ionization source positive ion mode and multiple reaction monitoring mode. The total donkey-hide gelatin peptide content is recorded in Table 1.

[0050] Table 1 Application test results of Examples 1-16 and Comparative Example 1 Combining Example 1 and Comparative Example 1 with Table 1, it can be seen that if twin-screw extrusion is not performed during the production of donkey-hide gelatin peptides by double enzymatic hydrolysis and extrusion, it is found that the proportion of donkey-hide gelatin molecular weight below 3,000 and the total peptide content of donkey-hide gelatin measured above are both poor; this shows that the synergistic effect of enzymatic hydrolysis and extrusion can significantly reduce the molecular weight of donkey-hide gelatin after enzymatic hydrolysis, and bring about an excellent improvement in the total peptide content of donkey-hide gelatin, thereby obtaining a donkey-hide gelatin peptide product of better quality.

[0051] Combining Example 1 and Example 2-3 with Table 1, it can be seen that the enzyme preparation is composed of the following enzyme raw materials in the premixed slurry in the corresponding weight ratio: medium-temperature α-amylase 0.3-0.5%, saccharifying enzyme 0.3-0.5%, and protease 0.1-0.3%, which can make the above-mentioned measured molecular weight of donkey-hide gelatin below 3,000 and the total peptide content of donkey-hide gelatin both perform better. Combining Example 16 with Table 1, it can be seen that if the enzyme preparation lacks the use of medium-temperature α-amylase and saccharifying enzyme, and only protease is used, it is found that the proportion of donkey-hide gelatin molecular weight below 3,000 and the total peptide content of donkey-hide gelatin will be significantly reduced. This shows that the synergistic cooperation between α-amylase, saccharifying enzyme and protease can achieve temperature adaptability, substrate specificity and stage synergy, thereby significantly reducing the molecular weight of donkey-hide gelatin after enzymatic hydrolysis and bringing about an excellent improvement in the total peptide content of donkey-hide gelatin.

[0052] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion, characterized in that: The following steps are involved: (1) Washing and crushing the raw material of donkey-hide gelatin, and mixing and beating with water to obtain a premixed slurry; (2) adding an enzyme preparation to the premixed slurry obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis slurry; (3) The enzymatic hydrolysis slurry of step (2) is subjected to twin-screw extrusion, the liquid-to-solid ratio is controlled to be 1:(2-5), and the extrusion screw temperature is controlled at 60-100°C; (4) subjecting the enzymatic hydrolyzate obtained by extrusion in step (3) to a heat preservation and hydrolysis treatment, and then to an enzyme inactivation treatment to obtain an enzyme inactivation slurry; (5) The enzyme-inactivated slurry obtained in step (4) is processed by a colloid mill and a homogenizer, and then filtered and sterilized at high temperature to obtain the donkey-hide gelatin peptide product.

2. The method for producing donkey-hide gelatin by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (2), the enzyme preparation is a combination of one or more of medium-temperature α-amylase, saccharifying enzyme and protease.

3. The method for producing donkey-hide gelatin by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (2), the enzyme preparation is composed of the following enzyme raw materials in corresponding weight ratios in the premixed slurry: Medium-temperature α-amylase 0.3-0.5%; Glucoamylase 0.3-0.5%; Protease 0.1-0.3%.

4. The method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: The specific configuration of step (4) is as follows: the enzymatic hydrolyzate obtained by extrusion in step (3) is subjected to a heat preservation hydrolysis treatment at a temperature of 60-70° C. for a heat preservation time of 35-60 min, and then an enzyme inactivation treatment is performed to obtain an enzyme inactivation slurry.

5. The method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (4), during the enzyme inactivation treatment, the enzyme inactivation temperature is 90-95°C and the enzyme inactivation time is 15-20 minutes.

6. The method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (5), the rotation speed of the colloid mill is 1000-2000 r / min and the temperature is 55-65°C.

7. The method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (5), during the homogenizer treatment, the homogenization temperature is 55-65°C, the first-stage homogenization pressure is 60-100 MPa, and the second-stage homogenization pressure is 40-60 MPa.

8. The method for producing donkey-hide gelatin peptide by double enzymatic hydrolysis and extrusion according to claim 1, characterized in that: In step (5), the filtration operation is performed using a 100-300 mesh sieve.

9. The use of double enzymatic hydrolysis and extrusion in producing donkey-hide gelatin peptide according to claim 1, characterized in that: Donkey-hide gelatin and glutinous rice plant-based products are prepared by adding glutinous rice and Chinese medicinal materials into the raw material of donkey-hide gelatin and applying the method of double enzymatic hydrolysis and extrusion to produce donkey-hide gelatin peptides.

10. The use of double enzymatic hydrolysis and extrusion in producing donkey-hide gelatin peptide according to claim 9, characterized in that: The Chinese medicinal materials consist of Codonopsis pilosula, Rehmannia root, Astragalus root and Angelica sinensis.

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