Separation reaction apparatus for enzymatic proteolysis

Abstract

The utility model relates to the technical field of separation reactor provides a kind of separation reaction device for enzyme protein hydrolysis, it include: reaction tank, temporary storage tank, filter membrane, first collecting jar and second collecting jar;Filter membrane is installed in temporary storage tank, to divide the inside of temporary storage tank into first chamber and second chamber, reaction tank and first collecting jar are selectively fluid communication with first chamber, second collecting jar and second chamber are selectively fluid communication. In this way, by integrating filter membrane in temporary storage tank, reaction product can be separated in real time, to avoid the accumulation of small molecule product, so as to reduce the inhibition to enzyme activity;Realize that reaction and separation are carried out simultaneously, reduce subsequent purification step, improve overall process efficiency;By separating product and enzyme in real time, reduce the probability of enzyme deactivation, significantly improve the reuse rate of enzyme, reduce production cost;Optimize overall structure, reduce subsequent step, reduce the dependence on additional equipment, realize more efficient production process.

Description

Technical Field

[0001] This utility model relates to the field of separation reactor technology, and in particular to a separation reaction device for enzyme protein hydrolysis. Background Technology

[0002] Enzymatic protein hydrolysis technology utilizes enzymes to catalyze the breakdown of proteins into small peptides and amino acids. It is widely used in food, pharmaceuticals, and animal feed to improve the functional properties, nutritional value, and absorption efficiency of products. Existing enzymatic protein hydrolysis technologies typically employ traditional reactors, such as stirred tank reactors and continuous tubular reactors. These traditional reactors have relatively simple designs, relying on the natural mixing and stirring of the solution to promote the reaction.

[0003] However, these traditional reactors have significant drawbacks: First, reaction inhibition: As the enzymatic reaction proceeds, small peptides and amino acids gradually accumulate in the reaction products. These products bind to the enzyme, reducing its activity and slowing the reaction rate, thus affecting hydrolysis efficiency. This "product inhibition effect" is a common problem in existing technologies, making it impossible to increase yield quickly. Second, separation and purification are difficult: In traditional reactors, small molecule products coexist with the enzyme in the reaction system, making efficient separation difficult. Due to the lack of immediate separation methods, complex separation and purification processes, such as centrifugation and ultrafiltration, are usually required after the reaction. This not only increases the number of process steps but also leads to low enzyme reuse rates and high production costs. Third, low equipment efficiency: Traditional reactors mostly rely on simple stirring or flow designs, making it impossible to achieve simultaneous reaction and separation. Low solution mixing efficiency results in long reaction times, insufficient overall capacity, and limits the scale and efficiency of industrial production. Utility Model Content

[0004] This invention provides a separation reaction device for enzyme protein hydrolysis, which solves the problem that traditional reactors in the prior art cannot simultaneously achieve reaction and separation.

[0005] This invention provides a separation reaction device for enzyme protein hydrolysis, comprising: a reaction vessel, a temporary storage vessel, a filter membrane, a first collection vessel, and a second collection vessel; The filter membrane is installed on the temporary storage tank to divide the interior of the temporary storage tank into a first chamber and a second chamber. The reaction tank and the first collection tank are selectively fluidly connected to the first chamber, and the second collection tank is selectively fluidly connected to the second chamber. The material in the first chamber enters the second chamber after being filtered by the filter membrane.

[0006] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the separation reaction device for enzyme protein hydrolysis further includes a material circulation pipeline, one end of which is connected to the reaction tank and the other end of which is connected to the first chamber.

[0007] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the material circulation pipeline includes a first pipeline and a pump body, one end of the first pipeline is connected to the reaction tank, the other end of the first pipeline is connected to the first chamber, and the pump body is installed on the first pipeline.

[0008] According to the separation reaction device for enzyme protein hydrolysis provided by this utility model, the material circulation pipeline further includes a first valve body and a second valve body, both of which are installed on the first pipeline and are located on both sides of the pump body.

[0009] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the separation reaction device for enzyme protein hydrolysis further includes a stirring assembly, which is installed in the reaction tank and configured to stir the material located in the reaction tank.

[0010] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the separation reaction device for enzyme protein hydrolysis further includes a third valve body, the reaction vessel is in fluid communication with the first chamber through a second pipe, and the third valve body is installed on the second pipe.

[0011] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the separation reaction device for enzyme protein hydrolysis further includes a fourth valve body, the first collection tank is in fluid communication with the first chamber through a third pipe, and the fourth valve body is installed on the third pipe.

[0012] According to the present invention, a separation reaction device for enzyme protein hydrolysis is provided, wherein the separation reaction device for enzyme protein hydrolysis further includes a fifth valve body, the second collection tank is in fluid communication with the second chamber through a fourth pipe, and the fifth valve body is installed on the fourth pipe.

[0013] According to the separation reaction apparatus for enzyme protein hydrolysis provided by this utility model, the reaction vessel is provided with a feed inlet; and / or, The reaction vessel is equipped with an air inlet and an air outlet.

[0014] According to the separation reaction apparatus for enzyme protein hydrolysis provided by this utility model, the reaction vessel is equipped with a flow rate sensor; and / or, The reaction vessel is equipped with a temperature sensor; and / or, The reaction vessel is equipped with a pH sensor.

[0015] This invention provides a separation reaction device for enzyme-protein hydrolysis. Milk-derived proteins and proteases are added to a reaction vessel, where an enzymatic hydrolysis reaction takes place. After a period of time, the reaction vessel connects to a first chamber, and the second chamber connects to a second collection vessel. At this point, small peptides in the reaction product pass through a filter membrane and enter the second collection vessel. After another period of time, the first collection vessel connects to the first chamber, allowing large amino acids to enter. Thus, by integrating a filter membrane into the storage vessel, reaction products can be separated in real time, avoiding the accumulation of small molecule products and reducing inhibition of enzyme activity. Simultaneous reaction and separation reduce subsequent purification steps and improve overall process efficiency. Real-time separation of products and enzymes reduces the probability of enzyme inactivation, significantly increasing enzyme reusability and reducing production costs. The optimized overall structure reduces subsequent steps and dependence on additional equipment, achieving a more efficient production process. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0017] Figure 1 This is a schematic diagram of the separation reaction device for enzyme protein hydrolysis provided by this utility model.

[0018] Figure label: 1. Reaction vessel; 11. pH sensor; 12. Temperature sensor; 13. Flow rate sensor; 14. Stirring blades; 15. Inspection port; 16. Feed inlet; 17. Stirring motor; 18. Air inlet; 19. Air outlet; 2. Temporary storage tank; 21. First chamber; 22. Second chamber; 3. First collection tank; 4. Second collection tank; 5. Material circulation pipeline; 51. First pipeline; 52. Pump body; 53. First valve body; 54. Second valve body; 6. Filter membrane; 7. Third valve body; 8. Fourth valve body; 9. Fifth valve body. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of this utility model clearer, the technical solutions of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model.

[0020] The following is combined Figure 1 This invention describes a separation reaction apparatus for the hydrolysis of enzyme proteins.

[0021] like Figure 1 As shown, the separation reaction device for enzyme protein hydrolysis according to an embodiment of the present invention includes: a reaction tank 1, a temporary storage tank 2, a filter membrane 6, a first collection tank 3, and a second collection tank 4.

[0022] The filter membrane 6 is installed on the temporary storage tank 2 to divide the internal space of the temporary storage tank 2 into a first chamber 21 and a second chamber 22. The reaction tank 1 and the first collection tank 3 are selectively fluidly connected to the first chamber 21, and the second collection tank 4 is fluidly selectively connected to the second chamber 22. The material in the first chamber 21 enters the second chamber 22 after being filtered by the filter membrane 6.

[0023] Among them, filter membrane 6 can be a nanofiltration membrane, with a pore size of 1 nm or larger, generally between 1 and 2 nm, allowing solvent molecules or certain low molecular weight solutes or low-valence ions to pass through. Its molecular weight cutoff is between that of reverse osmosis membranes and ultrafiltration membranes, and its operating pressure is usually low, so it is also called "low-pressure reverse osmosis" or "loose reverse osmosis".

[0024] Nanofiltration membranes are mostly composite membranes with an asymmetric structure. They consist of a porous support layer and a dense separation layer. The porous support layer provides support and its mechanism is sieving. The dense separation layer plays the main separation role. The surface separation layer is composed of polyelectrolytes, thus exhibiting a certain retention rate for inorganic salts. The separation mechanism can be explained using the dissolution-diffusion theory.

[0025] Nanofiltration membranes are made from a variety of materials, including organic and inorganic nanofiltration membranes. Organic nanofiltration membranes are prepared using organic polymer materials as the substrate, such as cellulose acetate, polyamides, sulfonated polysulfones and sulfonated polyethersulfones, polycarbonates, and polyesters. These materials exhibit good film-forming properties, chemical stability, and mechanical strength. Inorganic nanofiltration membranes, such as alumina and silica, have significant advantages in heat resistance, chemical resistance, and service life. In addition, there are organic-inorganic composite nanofiltration membranes and multilayer composite nanofiltration membranes, which combine the advantages of both organic and inorganic materials, exhibiting excellent separation performance and service life.

[0026] It should be noted that the large amino acids produced by the enzyme-catalyzed protein decomposition can enter the first collection tank 3, while the small peptides can pass through the filter membrane 6 and enter the second collection tank 4.

[0027] In practical applications, milk proteins and proteases are added to reaction vessel 1 and undergo enzymatic hydrolysis. After a period of time, reaction vessel 1 is connected to the first chamber 21, and the second chamber 22 is connected to the second collection vessel 4. At this time, small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection vessel 4. After another period of time, the first collection vessel 3 is connected to the first chamber 21, and large molecule amino acids can enter the first collection vessel 3. Thus, by integrating the filter membrane 6 into the temporary storage vessel 2, the reaction products can be separated in real time, avoiding the accumulation of small molecule products and thus reducing the inhibition of enzyme activity; the reaction and separation are carried out simultaneously, reducing subsequent purification steps and improving the overall process efficiency; by separating the product and enzyme in real time, the probability of enzyme inactivation is reduced, significantly improving the enzyme reuse rate and reducing production costs; the overall structure is optimized, subsequent steps are reduced, dependence on additional equipment is reduced, and a more efficient production process is achieved.

[0028] To ensure uniform mixing of materials and further improve reaction efficiency, such as Figure 1 As shown, the separation reaction apparatus for enzyme protein hydrolysis also includes a material circulation pipeline 5, one end of which is connected to the reaction tank 1, and the other end of which is connected to the first chamber 21. The material circulation pipeline 5 allows the material to circulate between the reaction tank 1 and the first chamber 21.

[0029] Specifically, milk protein and protease are added to reaction vessel 1 and enzymatic hydrolysis is carried out in reaction vessel 1. After a period of time, reaction vessel 1 is connected to the first chamber 21 and the second chamber 22 is connected to the second collection vessel 4. At this time, small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection vessel 4. After that, the connection between reaction vessel 1 and the first chamber 21 is disconnected, and the connection between the second chamber 22 and the second collection vessel 4 is disconnected. The material circulation pipeline 5 is opened, and the material in the first chamber 21 returns to reaction vessel 1 to continue the reaction. After a period of time, reaction vessel 1 is connected to the first chamber 21 and the second chamber 22 is connected to the second collection vessel 4. Small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection vessel 4. After another period of time, the first collection vessel 3 is connected to the first chamber 21, and large molecule amino acids can enter the first collection vessel 3.

[0030] In practical applications, such as Figure 1As shown, the material circulation pipeline 5 includes a first pipe 51 and a pump body 52. ​​One end of the first pipe 51 is connected to the reaction tank 1, and the other end of the first pipe 51 is connected to the first chamber 21. The pump body 52 is installed on the first pipe 51. The material circulation pipeline 5 also includes a first valve body 53 and a second valve body 54. Both the first valve body 53 and the second valve body 54 are installed on the first pipe 51 and are located on both sides of the pump body 52, respectively.

[0031] For example, one end of the first pipe 51 is connected to the reaction vessel 1 near the top, and the other end of the first pipe 51 is connected to the temporary storage tank 2 near the filter membrane 6, corresponding to the first chamber 21.

[0032] The first valve body 53 and the second valve body 54 can be solenoid valves.

[0033] To ensure uniform mixing of materials and further improve reaction efficiency, such as Figure 1 As shown, the separation reaction apparatus for enzyme protein hydrolysis also includes a stirring assembly, which is installed in the reaction vessel 1 and configured to stir the materials located in the reaction vessel 1.

[0034] The working principle of the mixing assembly is that the mixing motor 17 provides power to drive the drive wheel to rotate, and then the power is transmitted to the driven wheel and the mixing shaft through the transmission device. The mixing shaft drives the mixing blades 14 to rotate at high speed or reciprocate, so as to fully mix the materials.

[0035] Specifically, milk protein and protease are added to reaction vessel 1 and enzymatic hydrolysis is carried out in reaction vessel 1. After a period of time, reaction vessel 1 is connected to the first chamber 21 and the second chamber 22 is connected to the second collection vessel 4. At this time, small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection vessel 4. After that, the connection between reaction vessel 1 and the first chamber 21 is disconnected, and the connection between the second chamber 22 and the second collection vessel 4 is disconnected. The material circulation pipeline 5 is turned on, and the material in the first chamber 21 is returned to reaction vessel 1 to continue the reaction. The stirring component is turned on. After a period of time, reaction vessel 1 is connected to the first chamber 21 and the second chamber 22 is connected to the second collection vessel 4. Small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection vessel 4. After another period of time, the first collection vessel 3 is connected to the first chamber 21, and large molecule amino acids can enter the first collection vessel 3.

[0036] In practical applications, such as Figure 1 As shown, the separation reaction apparatus for enzyme protein hydrolysis also includes a third valve body 7. The reaction vessel 1 is in fluid communication with the first chamber 21 through a second pipe, and the third valve body 7 is installed in the second pipe. The third valve body 7 can be a solenoid valve.

[0037] It should be noted that the third valve body 7 can control the opening and closing of the second pipeline, thus enabling selective fluid communication between the reaction vessel 1 and the first chamber 21.

[0038] In practical applications, such as Figure 1 As shown, the separation reaction device for enzyme protein hydrolysis also includes a fourth valve body 8. The first collection tank 3 is in fluid communication with the first chamber 21 through a third pipe, and the fourth valve body 8 is installed in the third pipe. The fourth valve body 8 can be a solenoid valve.

[0039] It should be noted that the fourth valve body 8 can control the opening and closing of the third pipeline, thus enabling selective fluid communication between the first collection tank 3 and the first chamber 21.

[0040] In practical applications, such as Figure 1 As shown, the separation reaction device for enzyme protein hydrolysis also includes a fifth valve body 9. The second collection tank 4 is in fluid communication with the second chamber 22 through a fourth pipe, and the fifth valve body 9 is installed in the fourth pipe. The fifth valve body 9 can be a solenoid valve.

[0041] It should be noted that the fifth valve body 9 can control the opening and closing of the fourth pipeline, thus enabling selective fluid communication between the second collection tank 4 and the second chamber 22.

[0042] Specifically, the process is as follows: First step: Close all valves and pump 52. Add milk protein, protease, and gas to reaction tank 1 for enzymatic hydrolysis. Second step: After a period of time, open the third valve 7 and the fifth valve 9, connecting reaction tank 1 to the first chamber 21 and the second collection tank 4 to the second chamber 22. At this time, small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection tank 4. Third step: Close the fifth valve 9 and the third valve 7, and turn on the stirring motor 17, the first valve 53, the second valve 54, and pump 52. The material continues to react in reaction tank 1. Fourth step: After another period of time, open the third valve 7 and the fifth valve 9, connecting reaction tank 1 to the first chamber 21 and the second collection tank 4 to the second chamber 22. Small molecule peptides in the reaction product pass through the filter membrane 6 and enter the second collection tank 4. Fifth step: Open the fourth valve 8, connecting the first collection tank 3 to the first chamber 21, allowing large molecule amino acids to enter the first collection tank 3. In this case, steps three and four can be executed again sequentially between steps four and five.

[0043] In optional embodiments, such as Figure 1 As shown, the top of the reaction vessel 1 is provided with a feed inlet 16 and an inspection port 15. In this way, both milk proteins and proteases can be added to the reaction vessel 1 through the feed inlet.

[0044] In addition, the top of the reaction vessel 1 is provided with an air inlet 18 and an air outlet 19.

[0045] The inlet 18 is used to supply oxygen or reaction gases, mix and stir to enhance mass transfer, and regulate reaction conditions. The outlet 19 is used to discharge excess gas, remove reaction byproducts, control the gas phase composition, and maintain stable reactor pressure.

[0046] Factors affecting protease hydrolysis mainly include pH, temperature, enzyme dosage, and substrate concentration. Enzymes exhibit maximum activity within a specific pH range. During protease hydrolysis, the pH changes as the hydrolysis progresses. Therefore, pH needs to be controlled during hydrolysis to maintain it within the enzyme's optimal range of action, thereby improving hydrolysis efficiency. Generally, higher temperatures lead to faster chemical reactions. However, enzymes are proteins, and excessively high temperatures can cause protein denaturation, gradually reducing activity. Therefore, protease hydrolysis must be performed at an appropriate temperature. Under certain conditions, higher enzyme concentrations result in faster reaction rates. Theoretically, enzyme dosage is unrelated to the degree of hydrolysis, but actual hydrolysis conditions are not always the most stable conditions for the protease. Therefore, increasing the enzyme dosage often leads to an increase in the degree of hydrolysis, and there is a certain enzyme dosage threshold. After reaching this threshold, the degree of hydrolysis stabilizes and no longer increases with further increases in enzyme dosage. Higher substrate concentrations increase the chance of collision between the enzyme and substrate, thus increasing the enzymatic reaction rate. However, when the substrate concentration increases to a certain level, the reaction rate will no longer increase with the increase of substrate concentration due to the limited number of enzymes; at this point, the reaction rate reaches its maximum value.

[0047] Therefore, such as Figure 1 As shown, reaction vessel 1 is equipped with a flow rate sensor 13, a temperature sensor 12, and a pH sensor 11. pH monitoring serves the following purposes: maintaining the optimal pH environment for the enzyme, controlling reaction equilibrium, extending enzyme lifespan, and controlling product quality. The pH value is generally between 6 and 7. pH adjustment measures include adding a pH adjuster to adjust the pH of the reaction solution. Commonly used pH adjusters in this field include sodium hydroxide, potassium hydroxide, and hydrochloric acid. Temperature monitoring serves the following purposes: maintaining enzyme activity and stability, optimizing reaction rate, preventing side reactions, and ensuring equipment safety. For example, reaction vessel 1 is surrounded by a jacket. Cooling or heating media are added to the jacket to maintain the temperature inside reaction vessel 1 at the target temperature. Flow rate detection serves the following purposes: optimizing reaction efficiency, ensuring system stability, controlling residence time, regulating mass transfer rate, energy saving and process optimization, real-time feedback, and safety assurance. Flow rate adjustment can be achieved by controlling the stirring motor 17 via the control board.

[0048] In practical applications, the separation reaction device for enzyme protein hydrolysis also includes a control main board, and the pump body 52, the first valve body 53, the second valve body 54, the third valve body 7, the fourth valve body 8, the fifth valve body 9, the stirring motor 17, the flow rate sensor 13, the temperature sensor 12 and the pH sensor 11 are all electrically connected to the control main board.

[0049] The control board is configured to control the operation of the pump body 52, the first valve body 53, the second valve body 54, the third valve body 7, the fourth valve body 8, the fifth valve body 9 and the stirring motor 17 according to a preset program.

[0050] Thus, the separation reaction device for enzyme protein hydrolysis provided by this utility model has the following characteristics: A filter membrane 6 is integrated into the separation reaction device to separate reaction products (such as small molecule peptides) in real time, avoiding their accumulation in the system and reducing the inhibitory effect on enzyme activity. Through the selective filter membrane 6, small molecule products are removed from the reaction solution in a timely manner by utilizing their size characteristics, while retaining the enzyme and unreacted substrate in the reaction system, thereby ensuring the continuity and high efficiency of the reaction. The addition of a material circulation pipeline 5 enhances solution mixing efficiency, ensuring complete reaction of the reactants. The optimized mixing effect significantly shortens the reaction time and improves the overall hydrolysis efficiency. A continuous operation mode is achieved where reaction and separation are carried out simultaneously, eliminating the need for frequent interruptions of the reaction for separation or purification, greatly reducing manual intervention, optimizing production steps, and significantly improving the efficiency and economy of industrial production.

[0051] The following is an example illustrating the specific application of the enzyme protein hydrolysis separation reaction device provided by this invention. Milk-derived proteins, including casein and whey protein, are processed using proteases. The raw materials are milk-derived proteins with a protein content ranging from 20% to 100%, and commercially available milk-derived protein raw materials can be purchased.

[0052] Water is added to the milk protein raw material to completely dissolve it, and the dry matter content reaches 2%~10%, preferably 5~10%. Protease is added and enzymatic hydrolysis is carried out in the separation reaction device for enzyme protein hydrolysis provided by this utility model. The enzymatic hydrolysis conditions are 20~65℃, preferably 40~60℃, the stirring speed is 10~300 rpm, and the reaction time is 0.5~8 hours, preferably 1~4 hours.

[0053] Reaction solution 1: React in reaction vessel 1 for 1 hour.

[0054] Reaction liquid three: This is the liquid that flows into temporary storage tank 2 after reaction liquid one has reacted in reaction tank 1.

[0055] Reaction Solution 2: Reaction Solution 2 is a portion taken out after Reaction Solution 1 has reacted for 1 hour. It is kept at the same temperature as the membrane-passing liquid (Reaction Solution 3) during the membrane-passing process and continues to react until the membrane-passing is completed. After the membrane-passing is completed, Reaction Solution 2, along with the intercepted liquid and the filtrate, continues to react for another hour under the same conditions.

[0056] Intercepted liquid: Continue reaction in the first collection tank 3 for 1 hour.

[0057] Filtrate: Part of the liquid in the second collection tank 4.

[0058] Filtrate 1: The remaining liquid in the second collection tank 4 continues to react for 1 hour.

[0059] Example 1: A 5% casein content was used for enzymatic hydrolysis at 55°C for 2 hours, with a stirring speed of 200 rpm. Samples were taken from reaction vessel 1, temporary storage vessel 2, first collection vessel 3, and second collection vessel 4. Reaction vessel 1 was the upper layer, and temporary storage vessel 2, first collection vessel 3, and second collection vessel 4 were the lower layers. The test results for Example 1 are shown in the table below: Table 1

[0060] Comparative Example 1: Using a conventional reactor, the reaction conditions were identical to those in Example 1, except for the reactor itself. The test results for Comparative Example 1 are shown in the table below: Table 2

[0061] Example 2: The conditions were the same as in Example 1, except that the amount of enzyme added was halved. The test results for Example 2 are shown in the table below: Table 3

[0062] Comparative Example 2: Using a conventional reactor, the reaction conditions were identical to those in Example 2, except for the reactor itself. The test results for Comparative Example 2 are shown in the table below: Table 4

[0063] As can be seen from Example 1 above, using the separation reaction device for enzymatic protein hydrolysis of this embodiment, the contents of various indicators in the obtained filtrate and filtrate one are basically the same, reflecting that the enzymatic hydrolysis reaction stopped after one hour of continued reaction in filtrate one. This indicates that the enzyme has been enriched in the retentate during the membrane process and has not been filtered out through the membrane. This reaction phenomenon proves that this device can enrich the enzyme in the retentate during the reaction process, preventing it from being filtered out through the membrane, and the resulting filtrate stops the enzymatic hydrolysis reaction. The content of peptides ≤2000 kDa in filtrate and filtrate one reaches 58.90 and 56.44 respectively, which is much higher than the content of peptides ≤2000 kDa in reaction solution one and reaction solution two (5.86 and 6.41 respectively). This result shows that the filtrate contains more short peptides and fewer large molecular proteins and large peptides. This indicates that the separation reaction device for enzymatic protein hydrolysis of this embodiment can achieve immediate separation of hydrolysis products, making the lower layer product of the reaction highly enriched with small molecular nitrogen sources that can be directly utilized by microorganisms. Since microorganisms can more easily absorb and utilize free amino acids and small peptides during the culture process, but have difficulty directly metabolizing large protein molecules and polypeptides, the lower layer product can serve as an ideal nitrogen source for microbial culture media.

[0064] Although the amount of enzyme used in Example 2 was reduced by 50%, the yield of short peptides still reached more than 80% of that in Example 1, and the indicators were not significantly different from those in Example 1. This demonstrates that the separation reaction device for enzyme protein hydrolysis of this invention has the effect of improving efficiency when the amount of enzyme used is reduced.

[0065] In contrast, the products of traditional enzymatic hydrolysis reactors (Comparative Example 1 and Comparative Example 2) still contain a large proportion of macromolecular proteins and peptides, which require subsequent separation and purification to obtain suitable small molecule products. This not only prolongs the reaction process and increases energy consumption and equipment burden, but also results in low protein utilization in the raw materials, thus limiting their practical value as a nitrogen source for culture media.

[0066] In this embodiment of the invention, through the optimized design of the separation reaction device for enzymatic protein hydrolysis and the filter membrane 6, products of different molecular weight ranges can be separated in real time during the enzymatic hydrolysis process, thereby producing products of different specifications in a single reaction. By adjusting the membrane pore size, the molecular weight range of the final product can also be flexibly controlled to meet the needs of different downstream applications.

[0067] It is particularly important to note that the efficiency of enzyme activity is not simply linearly related to the amount of enzyme added. Specifically, influencing factors include the amount of enzyme added, substrate concentration, and reaction time. Generally, under consistent substrate concentration and reaction conditions, when plotting two lines between increased enzyme addition and improved reaction efficiency, the slope of the increase in enzyme addition will be greater. This is because the reaction between enzyme and substrate molecules requires their binding within the reaction system. When the substrate concentration is high and there is a relative excess of substrate molecules, all added enzyme molecules can fully bind with the substrate molecules, and the degree of substrate hydrolysis is directly proportional to the amount of enzyme added. However, when the added enzyme molecules are in excess relative to the substrate molecules, the increase in enzyme addition and the increase in enzymatic hydrolysis are no longer directly proportional. In Example 2, compared to a conventional reactor, as the permeate flows out, the enzymes in the retrieval liquid, being large molecules that cannot pass through the membrane, accumulate in the retrieval liquid. Therefore, the final retrieval liquid, with indicators such as amino acid nitrogen and total amino acids, showing the degree of hydrolysis, will be much higher than in a conventional reactor. On the other hand, in both Example 2 and Example 1, the substrate protein in the effluent decreased as the permeate flowed out. In Example 1, due to the higher initial enzyme dosage, the reduction in substrate was greater, resulting in a relative enrichment of the protein substrate compared to Example 2, meaning the actual substrate concentration was increased. Therefore, with a relatively small amount of enzyme added, a greater degree of hydrolysis than a linear decrease in the added amount can be achieved.

[0068] Furthermore, the experimental data from Examples 1 and 2 show that the water content of the retrieval liquid is as high as 97.10% and 95.10%, respectively, indicating good product solubility. Comparison of the dry-based peptide and protein content data of the original solution and the retrieval liquid reveals a significant improvement in the yield of the enzymatic hydrolysis product from this reactor. Some insoluble substances may be generated during the enzymatic hydrolysis process; these insoluble substances, after separation, serve as enzymatic hydrolysis byproducts or waste, with low added value. Based on the ratio of the total soluble dry matter after hydrolysis to the original dry matter, the overall yield of the reactor can reach 97.20% and 95.90%, respectively, far exceeding the yields of 91.10% and 90.20% achieved with traditional equipment.

[0069] The high yield is attributed to the continuous flow of products through the membrane into the lower layer during the reaction, which promptly eliminates product inhibition effects and allows the hydrolysis reaction to continue, further improving the degree of hydrolysis and the final product conversion rate. Simultaneously, because the enzyme itself is a large protein molecule, it does not easily penetrate the membrane structure, thus allowing it to continuously accumulate in the upper layer of the reactor, maintaining a high local enzyme concentration, enhancing catalytic efficiency, and reducing enzyme waste. This design also facilitates continuous feeding and continuous reaction; raw materials can be continuously added from top to bottom, while products continuously flow out from the lower layer, achieving truly continuous production, effectively simplifying the production process, reducing operational complexity, and improving overall production efficiency and product stability.

[0070] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. A separation reaction apparatus for enzyme protein hydrolysis, characterized in that, include: The reaction vessel, temporary storage vessel, filter membrane, first collection vessel, and second collection vessel; The filter membrane is installed on the temporary storage tank to divide the interior of the temporary storage tank into a first chamber and a second chamber. The reaction tank and the first collection tank are selectively fluidly connected to the first chamber, and the second collection tank is selectively fluidly connected to the second chamber. The material in the first chamber enters the second chamber after being filtered by the filter membrane.

2. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The separation reaction device for enzyme protein hydrolysis further includes a material circulation pipeline, one end of which is connected to the reaction tank and the other end of which is connected to the first chamber.

3. The separation reaction apparatus for enzyme protein hydrolysis according to claim 2, characterized in that, The material circulation pipeline includes a first pipe and a pump body. One end of the first pipe is connected to the reaction vessel, and the other end of the first pipe is connected to the first chamber. The pump body is installed on the first pipe.

4. The separation reaction apparatus for enzyme protein hydrolysis according to claim 3, characterized in that, The material circulation pipeline also includes a first valve body and a second valve body, both of which are installed on the first pipeline and located on both sides of the pump body.

5. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The separation reaction apparatus for enzyme protein hydrolysis further includes a stirring assembly, which is installed in the reaction vessel and configured to stir the materials located in the reaction vessel.

6. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The separation reaction device for enzyme protein hydrolysis further includes a third valve body. The reaction vessel is in fluid communication with the first chamber through a second pipe, and the third valve body is installed on the second pipe.

7. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The separation reaction device for enzyme protein hydrolysis further includes a fourth valve body, and the first collection tank is in fluid communication with the first chamber through a third pipe, with the fourth valve body installed on the third pipe.

8. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The separation reaction device for enzyme protein hydrolysis further includes a fifth valve body, and the second collection tank is in fluid communication with the second chamber through a fourth pipe, with the fifth valve body installed on the fourth pipe.

9. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The reaction vessel is provided with a feed inlet; and / or, The reaction vessel is equipped with an air inlet and an air outlet.

10. The separation reaction apparatus for enzyme protein hydrolysis according to claim 1, characterized in that, The reaction vessel is equipped with a flow rate sensor; and / or, The reaction vessel is equipped with a temperature sensor; and / or, The reaction vessel is equipped with a pH sensor.

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