Composite biological enzyme preparation device and preparation method thereof

Abstract

The invention discloses a composite biological enzyme preparation device and a preparation method thereof. The preparation method comprises the following steps: S1, preparing raw materials; s2, mixing and processing; s3, reaction treatment; s4, separating and collecting; s5, sampling and detecting; the S1 raw material preparation comprises the following steps: preparing amylase, protease and lipase, respectively weighing the raw materials with certain mass by using a high-precision electronic balance, dissolving the amylase in a proper amount of phosphate buffer, dissolving the protease in a proper amount of trihydroxymethyl aminomethane buffer, and uniformly stirring; then dissolving lipase in a proper amount of sodium acetate buffer agent to prepare a raw material solution with proper concentration; through the design, bio-enzyme raw materials are uniformly mixed, the component proportion of the composite bio-enzyme is improved, meanwhile, the reaction temperature and the pH value can be accurately monitored and controlled in real time, a proper environment is provided for the bio-enzyme reaction, and the activity and the yield of the bio-enzyme are effectively improved.

Description

Technical Field

[0001] The present invention relates to the technical field of compound bioenzyme production, and specifically to a compound bioenzyme preparation device and a preparation method thereof. Background Art

[0002] Compound bioenzyme is a biological preparation composed of various different types of bioenzymes combined in a specific ratio. It combines the functions of multiple bioenzymes, and each component works synergistically to achieve complex functions that are difficult to achieve by a single bioenzyme or to improve the efficiency of a specific reaction. Compound bioenzymes are widely used in many fields such as food, medicine, and environmental protection. Currently, there are some problems in the preparation process of compound bioenzymes. The control of the reaction conditions (such as temperature, pH value) of compound bioenzymes is not precise enough, resulting in a decrease in bioenzyme activity and also limiting the yield. Therefore, a compound bioenzyme preparation device and a preparation method thereof are proposed to solve the above problems. Summary of the Invention

[0003] In order to make up for the deficiencies of the prior art and solve the problem that the control of the reaction conditions (such as temperature, pH value) of compound bioenzymes is not precise enough, the present invention proposes a compound bioenzyme preparation device and a preparation method thereof.

[0004] A compound bioenzyme preparation device and a preparation method thereof include S1 raw material preparation; S2 mixing and processing; S3 reaction treatment; S4 separation and collection; S5 sampling and detection; the S1 raw material preparation includes: preparing amylase, protease, and lipase, respectively weighing a certain mass of each raw material using a high-precision electronic balance, dissolving the amylase in an appropriate amount of phosphate buffer, then dissolving the protease in an appropriate amount of tris(hydroxymethyl)aminomethane buffer, and then dissolving the lipase in an appropriate amount of sodium acetate buffer to prepare raw material solutions with appropriate concentrations.

[0005] Preferably, the S2 mixing and processing includes a mixing tank. The mixing tank is provided with a plurality of feed ports at the top for inputting different bioenzyme raw material solutions. A stirring paddle is provided inside the mixing tank, and the stirring paddle is connected to a motor; the amylase raw material solution, protease raw material solution, and lipase raw material solution are input into the mixing tank through the feed ports at the top of the mixing tank, the motor is started, and the stirring paddle stirs at a speed of 500 revolutions per minute for 30 minutes to fully mix the two raw materials.

[0006] Preferably, the S3 reaction treatment includes a reaction kettle, in which a temperature sensor and a pH sensor are installed for real-time monitoring of the reaction temperature and pH value respectively. The outside of the reaction kettle is wrapped with a heating jacket and a cooling pipe to control the reaction temperature through the heating jacket and the cooling pipe. The reaction kettle is also provided with an acid-base addition port, and the pH value of the reaction system can be automatically adjusted by adding acid-base solution according to the monitoring result of the pH sensor. The reaction kettle is connected to a mixing tank through a pipeline; the mixed solution is transported to the reaction kettle, the temperature sensor and the pH sensor are turned on, and the reaction temperature is controlled at 37°C - 40°C through the heating jacket or the cooling pipe. When the pH sensor monitors that the pH value of the reaction system deviates from the predetermined range, acid-base solution is added through the acid-base addition port to adjust the pH value to 7.0 - 7.5, and the raw material solution reacts under this condition for 5 hours.

[0007] Preferably, the S4 separation and collection includes a centrifuge separator and a collection tank. The centrifuge separator is connected to the reaction kettle through a pipeline, and the centrifuge separator is connected to the collection tank through a pipeline. After the reaction is completed, the mixed liquid inside the reaction kettle is transported to the centrifuge separator and centrifuged at a speed of 3000 revolutions per minute for 20 minutes for solid-liquid separation. The separated liquid flows into the collection tank equipped with a filtering device, and after further removing impurities, a composite bio-enzyme product is obtained.

[0008] Preferably, for the S5 sampling and detection, the composite bio-enzyme product in the collection tank is quantitatively taken out multiple times, and each component in the composite bio-enzyme is separated and quantitatively analyzed by a high-performance liquid chromatograph; according to the chemical properties of different bio-enzymes, a suitable chromatographic column and mobile phase are selected, and by comparing with the retention time and peak area of the standard product, the types and contents of each component in the composite bio-enzyme are determined to ensure that it meets the predetermined component ratio. Through detection, it is determined that the component ratio of the composite bio-enzyme product meets the expectation and the bio-enzyme activity is relatively high.

[0009] The beneficial effects of the present invention are as follows:

[0010] Through the design of the present invention, the bio-enzyme raw materials are evenly mixed, the component ratio of the composite bio-enzyme is improved, and at the same time, the reaction temperature and pH value can be accurately monitored and controlled in real time, providing a suitable environment for the bio-enzyme reaction, effectively improving the activity and yield of the bio-enzyme. Description of the Drawings

[0011] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required for the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0012] Figure 1 It is a schematic diagram of the operation process of a composite bioenzyme preparation device of the present invention and its preparation. Specific embodiments

[0013] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0014] Please refer to Figure 1 As shown, a composite bioenzyme preparation device and its preparation method include S1 raw material preparation; S2 mixing and processing; S3 reaction treatment; S4 separation and collection; S5 sampling and detection; S1 raw material preparation includes: preparing amylase, protease, and lipase, respectively weighing a certain mass of each raw material using a high-precision electronic balance, dissolving the amylase in an appropriate amount of phosphate buffer, and the phosphate buffer is made by mixing sodium dihydrogen phosphate and disodium hydrogen phosphate in a certain proportion. The phosphate buffer can resist the influence of external acid-base changes on the pH value of the solution to a certain extent and maintain the pH value stable. Then dissolve the protease in an appropriate amount of tris(hydroxymethyl)aminomethane buffer. Tris(hydroxymethyl)aminomethane is a commonly used biological buffer. By adjusting the ratio of Tris to HCl, the pH value of the solution can be accurately controlled within a specific range to meet the activity requirements of the protease and avoid aggregation and precipitation. Then dissolve the lipase in an appropriate amount of sodium acetate buffer to make a raw material solution with an appropriate concentration. The sodium acetate buffer system can effectively maintain the stability of the solution within a specific pH range, so that the lipase remains active in a slightly acidic environment. By adjusting the ratio of acetic acid to sodium acetate, the pH value of the solution can be accurately controlled. To prevent microbial contamination.

[0015] The S2 hybrid processing includes a mixing tank. There are multiple feed ports at the top of the mixing tank. Different biological enzyme raw material solutions are sequentially injected into the interior of the mixing tank through the multiple feed ports. A stirring paddle is provided inside the mixing tank, and the stirring paddle is connected to a motor. The amylase raw material solution, protease raw material solution, and lipase raw material solution are input into the mixing tank through the feed ports at the top of the mixing tank. The motor is started, and the stirring paddle stirs at a speed of 500 revolutions per minute for 30 minutes to fully mix the two raw materials. The S3 reaction treatment includes a reaction kettle. A temperature sensor and a pH sensor are installed inside the reaction kettle. The temperature sensor is preferably of the tc-05b type, and the pH sensor is preferably of the RS232 type. The temperature sensor and the pH sensor are respectively used to monitor the reaction temperature and pH value in real time. The outside of the reaction kettle is wrapped with a heating jacket and a cooling pipe. The reaction temperature is controlled by the heating jacket and the cooling pipe. Moreover, the reaction kettle is also provided with an acid-base addition port. According to the monitoring result of the pH sensor, acid-base solution can be automatically added to adjust the pH value of the reaction system. The reaction kettle is kept connected to the mixing tank through a pipeline. The mixed solution is transported to the reaction kettle. The temperature sensor and the pH sensor are turned on. The reaction temperature is controlled at 37°C - 40°C through the heating jacket or the cooling pipe. When the pH sensor monitors that the pH value of the reaction system deviates from the predetermined range, acid-base solution is added through the acid-base addition port to adjust the pH value to 7.0 - 7.5, and the raw material solution reacts under this condition for 5 hours. The S4 separation and collection include a centrifuge separator and a collection tank. The centrifuge separator is connected to the reaction kettle through a pipeline, and the centrifuge separator is connected to the collection tank through a pipeline. After the reaction is completed, the mixed liquid inside the reaction kettle is transported to the centrifuge separator and centrifuged at a speed of 3000 revolutions per minute for 20 minutes for solid-liquid separation. The separated liquid flows into the collection tank equipped with a filtering device. After further removing impurities, a composite biological enzyme product is obtained.

[0016] S5 Sampling and detection: Quantitatively take out the composite bio-enzyme product in the collection tank multiple times, and separate and quantitatively analyze each component in the composite bio-enzyme by a high-performance liquid chromatograph. According to the chemical properties of different bio-enzymes, select a suitable chromatographic column and mobile phase, and determine the types and contents of each component in the composite bio-enzyme by comparing the retention time and peak area with the standard product to ensure that it meets the predetermined component ratio. Appropriately process the prepared composite bio-enzyme sample, centrifuge it at a speed of 10,000 - 15,000 revolutions per minute for 10 - 15 minutes to remove insoluble impurities, then take the supernatant and filter it with a 0.22μm or 0.45μm filter membrane to ensure that there are no fine particles in the sample and prevent clogging of the chromatographic column. Inject the processed sample into the high-performance liquid chromatograph, set appropriate instrument parameters, set the flow rate at 0.8 - 1.2 mL / min. After setting the parameters, start the instrument for analysis. Each component in the composite bio-enzyme has different interactions with the stationary phase and the mobile phase in the chromatographic column, resulting in different retention times in the column, so they flow out of the chromatographic column in sequence and are detected by the detector. The detector converts the concentration signal of each component into an electrical signal and records it to form a chromatogram. By comparing the retention time with the standard product, the types of each component in the composite bio-enzyme can be determined. Through detection, it is determined that the component ratio of the composite bio-enzyme product meets the expectation and the bio-enzyme activity is relatively high.

[0017] In the description of this specification, the description with reference to terms such as "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0018] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art of this industry should understand that the present invention is not limited by the above embodiments. The above embodiments and the descriptions in the specification only illustrate the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed.

Claims

1. A composite bio-enzyme preparation device and its preparation method, characterized in that: It includes S1 raw material preparation; S2 mixing and processing; S3 reaction treatment; S4 separation and collection; S5 sampling and detection; the S1 raw material preparation includes: preparing amylase, protease, and lipase, weighing a certain mass of each raw material respectively using a high-precision electronic balance, dissolving the amylase in an appropriate amount of phosphate buffer, then dissolving the protease in an appropriate amount of tris(hydroxymethyl)aminomethane buffer, and then dissolving the lipase in an appropriate amount of sodium acetate buffer to make raw material solutions with appropriate concentrations.

2. The composite bio-enzyme preparation device and its preparation method according to claim 1, characterized in that: The S2 mixing and processing includes a mixing tank. The top of the mixing tank is provided with a plurality of feed inlets for inputting different bio-enzyme raw material solutions respectively. Inside the mixing tank, there is a stirring paddle, and the stirring paddle is connected to a motor; input the amylase raw material solution, protease raw material solution, and lipase raw material solution into the mixing tank through the feed inlets at the top of the mixing tank, start the motor, and stir at a speed of 500 revolutions per minute for 30 minutes through the stirring paddle to fully mix the two raw materials.

3. A composite bioenzyme preparation device and its preparation method according to claim 2, characterized in that: The S3 reaction treatment includes a reaction kettle. A temperature sensor and a pH value sensor are installed inside the reaction kettle for real-time monitoring of the reaction temperature and pH value respectively. The outside of the reaction kettle is wrapped with a heating jacket and a cooling pipe to control the reaction temperature through the heating jacket and the cooling pipe. And the reaction kettle is also provided with an acid-base addition port, and the acid-base solution can be automatically added according to the monitoring result of the pH value sensor to adjust the pH value of the reaction system. The reaction kettle is kept connected to the mixing tank through a pipeline; transport the mixed solution to the reaction kettle, turn on the temperature sensor and the pH value sensor, control the reaction temperature at 37°C - 40°C through the heating jacket or the cooling pipe. When the pH value sensor monitors that the pH value of the reaction system deviates from the predetermined range, add the acid-base solution through the acid-base addition port to adjust the pH value to 7.0 - 7.5, and make the raw material solution react under this condition for 5 hours.

4. A composite bio-enzyme preparation device and its preparation method according to claim 3, characterized in that: The S4 separation and collection includes a centrifuge and a collection tank. The centrifuge is connected to the reaction kettle through a pipeline, and the centrifuge is connected to the collection tank through a pipeline. After the reaction is completed, transport the mixed liquid inside the reaction kettle to the centrifuge and centrifuge at a speed of 3000 revolutions per minute for 20 minutes for solid-liquid separation. The separated liquid flows into the collection tank equipped with a filtering device, and after further removing impurities, a composite bio-enzyme product is obtained.

5. A composite bio-enzyme preparation device and its preparation method according to claim 4, characterized in that: For the S5 sampling and detection, quantitatively take out the composite bio-enzyme product in the collection tank multiple times, and separate and quantitatively analyze the components in the composite bio-enzyme through a high-performance liquid chromatograph; according to the chemical properties of different bio-enzymes, select a suitable chromatographic column and mobile phase, and determine the types and contents of the components in the composite bio-enzyme by comparing with the retention time and peak area of the standard product to ensure that it meets the predetermined component ratio. Through detection, it is determined that the component ratio in the composite bio-enzyme product meets the expectation and the bio-enzyme activity is relatively high.

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