CN-122012660-A - Method for producing and preparing functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal

CN122012660ACN 122012660 ACN122012660 ACN 122012660ACN-122012660-A

Abstract

The invention discloses a method for producing and preparing functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal, belonging to the technical field of bioengineering. The invention solves the problems of unstable enzymolysis process and large fluctuation of polypeptide yield and activity index caused by the lack of causal inference and real-time dynamic adjustment mechanism due to the dependence analysis of the prior method and the adoption of fixed parameters for controlling the enzymolysis process, increases the contact area of enzyme and substrate by a two-stage crushing mechanism, enhances the contact of enzyme and substrate by combining the ultrasonic auxiliary enzymolysis technology, and utilizes gradient ethanol-supercritical The combined extraction technology can effectively remove fat and fat-soluble anti-nutritional factors and improve the purity of raw materials, the multistage filtration mechanism and the nano filtration technology can effectively remove impurities and improve the purity of polypeptide, and the microwave-assisted enzyme deactivation treatment can rapidly inactivate the activity of enzyme and reduce the potential influence of high temperature on the structure and activity of polypeptide.

Inventors

YU MIAO
ZHANG LIANGCHEN
XIE MENGXI
TIAN YUAN
SUN LIN

Assignees

辽宁省农业科学院

Dates

Publication Date: 20260512
Application Date: 20260304

Claims (10)

1. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of the soybean meal and the peanut meal is characterized by comprising the following steps: s1, respectively crushing soybean meal and peanut meal, and sieving with a 80-mesh sieve; degreasing the crushed soybean meal and peanut meal respectively, and adopting gradient ethanol-supercritical Combined extraction, soaking in 60% ethanol solution for 12 hr to remove free fat; Then supercritical is carried out under the condition of 30MPa and 35 DEG C Extracting for 5 hours, selectively removing the combined fat and fat-soluble anti-nutritional factors, and drying after filtering; Mixing defatted soybean meal and peanut meal according to a mass ratio of 1:2, and adding a proper amount of water to enable the water content of the mixed material to reach 55%; S2, adding a compound enzyme preparation into the mixed material, adjusting the pH value of the mixed material to 8.0, controlling the temperature to 50 ℃, stirring and carrying out enzymolysis for 3 hours; S3, heating the material subjected to enzymolysis to 92 ℃ and keeping the temperature for 12 minutes for enzyme deactivation treatment, centrifuging the material subjected to enzyme deactivation, adopting a variable frequency centrifugation technology, automatically adjusting the centrifugation speed according to the density and viscosity of the material, and centrifuging for 12 minutes at the rotation speed of 4500 rpm; filtering the supernatant by using a functional membrane element with a molecular weight of 5kDa, introducing a nanofiltration technology in the separation and purification process, and effectively removing impurities by using a nanofiltration membrane; and S4, spray drying the filtered polypeptide solution to obtain functional polypeptide powder, and detecting the quality of a final product, wherein the quality comprises the content, purity, antioxidant activity and immunoregulatory activity indexes of the polypeptide.
2. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of the soybean meal and the peanut meal according to claim 1, wherein in the step S1, the soybean meal and the peanut meal are respectively crushed and pass through a 80-mesh sieve, and the method comprises the following steps: constructing a two-stage crushing mechanism, wherein the first stage adopts molar-simulated low-temperature shearing crushing to simulate a human body chewing mechanism, and crushing the raw materials to 40 meshes at a temperature of below 15 ℃; the second stage adopts simulated enamel high-frequency vibration grinding, and the material is further refined to 80 meshes through a vibration field with specific frequency, and simultaneously nanoscale cracks are generated, so that the enzyme contact area is increased.
3. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of soybean meal and peanut meal according to claim 1, wherein in the step S3, supernatant fluid is taken and filtered by using a functional membrane element with a molecular weight of 5kDa, and the method comprises the following steps of: Adopting a multistage filtration mechanism, firstly performing coarse filtration by using a membrane element with a molecular weight of 10kDa to remove macromolecular impurities, and then performing fine filtration by using a membrane element with a molecular weight of 5 kDa; Adding bioactive factor including coenzyme Q10 into the filtered polypeptide solution, and dispersing coenzyme Q10 homogeneously into the polypeptide solution via nanometer emulsification to combine with the polypeptide solution.
4. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of soybean meal and peanut meal according to claim 1, wherein the compound enzyme preparation consists of alkaline protease, papain and flavourzyme according to the enzyme activity ratio of 1:2:1.
5. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of soybean meal and peanut meal according to claim 1, which is characterized by further comprising the following steps: S5, establishing a quality tracing system based on the RIFD radio frequency technology, and recording production process data and quality detection results of each batch of products; and S6, introducing a self-adaptive control strategy, and dynamically adjusting enzymolysis parameters by monitoring the pH value, the temperature and the enzyme activity of the enzymolysis liquid in real time through an online sensor.
6. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of the soybean meal and the peanut meal according to claim 5, wherein in the step S6, an adaptive control strategy is introduced, and enzymolysis parameters are dynamically adjusted, and the method comprises the following steps: The method comprises the steps of rapidly detecting each batch of soybean meal/peanut meal raw materials to obtain basic data of the soybean meal and the peanut meal in different batches, wherein the basic data comprise protein content, fat content and ash content, recording various parameters in the enzymolysis process, including temperature, pH value, enzyme dosage and enzymolysis time, and measuring various indexes after enzymolysis, including yield, purity, antioxidant activity and immunoregulation activity of polypeptides; A near infrared spectrum analysis technology is introduced to rapidly detect protein structures of the soybean meal and the peanut meal, and the enzymolysis efficiency of the protein and the potential activity of the polypeptide are predicted; analyzing all collected data based on a decision tree model, taking basic data and enzymolysis parameters as input variables, and taking yield, purity and activity indexes of the polypeptide as output variables; and (3) calculating correlation coefficients between each input variable and each output variable through training of a decision tree model, and determining key parameters and influence degree of the key parameters on the polypeptide preparation effect.
7. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of soybean meal and peanut meal according to claim 5, wherein the method is characterized by calculating the correlation coefficient between each input variable and each output variable through training of a decision tree model and comprises the following steps: introducing causal inference analysis, and constructing a plurality of factor trees for comprehensively analyzing causal effects of each input variable to determine causal effects of the input variables on the output variables; Calculating, for each input variable, its causal effect on each output variable; evaluating the influence of an input variable on the polypeptide preparation effect by combining the correlation coefficient and the causal effect; determining key parameters and the influence degree of the key parameters on the polypeptide preparation effect according to the correlation coefficient and the causal effect; For each key parameter, evaluating the influence degree of the key parameter on different output variables; Based on the evaluation result, generating a characteristic importance list, displaying the relative importance of each input variable to the output variable, and normalizing the importance value to form a correlation coefficient.
8. The method for producing and preparing the functional polypeptide by enzymatic hydrolysis of soybean meal and peanut meal according to claim 7, wherein after generating the feature importance list based on the evaluation result, the method comprises the following steps: acquiring a feature importance list generated by a decision tree model, and acquiring the influence degree of each input variable on the output variable; According to the real-time data and the prediction result of the decision tree model, evaluating the state of the current enzymolysis process; Formulating a parameter adjustment rule according to the prediction result of the decision tree model; And determining the adjustment range and step length of each parameter, and determining the frequency of parameter adjustment according to the dynamic characteristics and production requirements of the enzymolysis process.
9. The method for producing and preparing the functional polypeptide by enzymatic hydrolysis of soybean meal and peanut meal according to claim 8, further comprising the steps of, after generating the feature importance list based on the evaluation result: setting an evaluation period according to the actual production process; Continuously collecting various data in the enzymolysis process in an evaluation period, analyzing the data, evaluating whether the self-adaptive control strategy reaches an expected target or not, comparing actual production data with a prediction result of a decision tree model, and analyzing the reason of deviation; Setting evaluation indexes, namely polypeptide yield improvement rate, antioxidant activity standard reaching rate, parameter adjustment stability and production cost change; updating the decision tree model according to the evaluation result, re-evaluating the influence degree of each input variable on the output variable, and updating the feature importance list; optimizing parameter adjustment rules according to the model updating result and actual production data, wherein the parameter adjustment rules comprise redetermining the adjustment range and step length of each parameter according to the new model prediction result; the frequency of parameter adjustment is redetermined according to the dynamic characteristics and production requirements of the enzymolysis process; and adjusting the self-adaptive control strategy according to the evaluation result.
10. The method for producing and preparing the functional polypeptide by utilizing the enzymolysis of soybean meal and peanut meal according to claim 8, wherein the parameter adjustment rule is formulated according to the prediction result of the decision tree model, and the method comprises the following steps: comparing the real-time monitoring data with the prediction result of the decision tree model, and evaluating the state of the current enzymolysis process; According to the prediction result of the decision tree model and the deviation of real-time data, formulating parameter adjustment directions and amplitudes comprises the following steps: according to the prediction result of the decision tree model, maintaining the temperature of the enzymolysis liquid within the range of the prediction result, and optimizing the yield of the polypeptide; According to the prediction result of the decision tree model, maintaining the pH value of the enzymolysis liquid within the range of the prediction result, and optimizing the antioxidant activity of the polypeptide; According to the change of the enzyme activity, the enzyme dosage is dynamically adjusted, so that the efficient performance of the enzymolysis reaction is ensured; according to the change of the polypeptide yield and the antioxidant activity, the enzymolysis time is dynamically adjusted, and the quality of the final product is optimized.

Description

Method for producing and preparing functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal Technical Field The invention relates to the technical field of bioengineering, in particular to a production and preparation method for forming functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal. Background The bean pulp and the peanut meal are main byproducts of grain and oil processing, have rich protein content, and are excellent raw materials for obtaining plant-derived protein peptides. In the prior art, single enzyme or simple complex enzyme is adopted for primary enzymolysis under fixed conditions, but the following defects exist: The traditional crushing method can not fully damage the cell walls of the bean pulp and the peanut pulp, so that the contact area of enzyme and substrate is limited, the enzymolysis efficiency is low, the combined fat and fat-soluble anti-nutritional factors are difficult to effectively remove by adopting a degreasing mode of single ethanol extraction, the purity of the raw materials is insufficient, the polypeptide structure and activity can be damaged due to conventional heating and enzyme deactivation, and the product quality is reduced. The traditional enzymolysis process is generally controlled by adopting fixed parameters, lacks of a real-time monitoring and dynamic adjusting mechanism, and cannot dynamically adjust enzymolysis parameters according to real-time data, so that the enzymolysis process is unstable, and the fluctuation of polypeptide yield and activity index is large. Therefore, in order to meet the existing requirements, a production and preparation method for forming functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal is provided. ‌ A Disclosure of Invention The invention aims to provide a method for producing and preparing functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal, which increases the contact area of enzyme and substrate by a two-stage crushing mechanism, enhances the contact of enzyme and substrate by combining an ultrasonic auxiliary enzymolysis technology and utilizes gradient ethanol-supercriticalThe combined extraction technology can effectively remove fat and fat-soluble anti-nutritional factors and improve the purity of raw materials, the multistage filtration mechanism and the nano filtration technology can effectively remove impurities and improve the purity of polypeptide, the microwave-assisted enzyme deactivation treatment can rapidly inactivate the activity of enzyme, the potential influence of high temperature on the structure and activity of polypeptide is reduced, and the problems in the background technology are solved. In order to achieve the above purpose, the present invention provides the following technical solutions: a method for producing and preparing functional polypeptide by utilizing enzymolysis of soybean meal and peanut meal comprises the following steps: s1, respectively crushing soybean meal and peanut meal, and sieving with a 80-mesh sieve; degreasing the crushed soybean meal and peanut meal respectively, and adopting gradient ethanol-supercritical Combined extraction, soaking in 60% ethanol solution for 12 hr to remove free fat; Then supercritical is carried out under the condition of 30MPa and 35 DEG C Extracting for 5 hours, selectively removing the combined fat and fat-soluble anti-nutritional factors, and drying after filtering; Mixing defatted soybean meal and peanut meal according to a mass ratio of 1:2, and adding a proper amount of water to enable the water content of the mixed material to reach 55%; S2, adding a compound enzyme preparation into the mixed material, adjusting the pH value of the mixed material to 8.0, controlling the temperature to 50 ℃, stirring and carrying out enzymolysis for 3 hours; S3, heating the material subjected to enzymolysis to 92 ℃ and keeping the temperature for 12 minutes for enzyme deactivation treatment, centrifuging the material subjected to enzyme deactivation, adopting a variable frequency centrifugation technology, automatically adjusting the centrifugation speed according to the density and viscosity of the material, and centrifuging for 12 minutes at the rotation speed of 4500 rpm; filtering the supernatant by using a functional membrane element with a molecular weight of 5kDa, introducing a nanofiltration technology in the separation and purification process, and effectively removing impurities by using a nanofiltration membrane; and S4, spray drying the filtered polypeptide solution to obtain functional polypeptide powder, and detecting the quality of a final product, wherein the quality comprises the content, purity, antioxidant activity and immunoregulatory activity indexes of the polypeptide. Further, in S1, respectively crushing soybean meal and peanut meal, and sieving with an 80-mesh sieve, wherein the method comprises the following steps: Constructing a two