Method for on-line control of yeast protein peptide preparation based on conductivity change rate threshold

Abstract

This invention provides an online control method for yeast protein peptide preparation based on a conductivity change rate threshold, comprising the following steps: S1: Real-time monitoring of the conductivity of the enzymatic hydrolysis reaction system; during the enzymatic hydrolysis reaction, the conductivity change of the hydrolysate is monitored in real time using an online conductivity sensor; S2: Calculation of the conductivity change rate; based on the real-time monitoring data, the conductivity change rate (dσ / dt) is calculated. As the enzymatic hydrolysis reaction proceeds, the concentration of mobile ions in the solution gradually increases, and the conductivity rises. The conductivity change rate is closely related to the progress of the enzymatic hydrolysis reaction and the degree of hydrolysis; S3: Setting at least one conductivity change rate threshold according to the target product; S4: When the conductivity change rate reaches or falls below the threshold, the enzymatic hydrolysis reaction is automatically terminated. This application solves the reaction control problem in the preparation of yeast protein peptides, not only improving production efficiency and product quality but also promoting the automation and intelligence of the production process.

Description

Technical Field

[0001] This invention relates to the field of yeast protein peptide preparation control methods, and in particular to an online control method for yeast protein peptide preparation based on a conductivity change rate threshold. Background Technology

[0002] Yeast protein peptides are products obtained by converting yeast cells into small peptide molecules through autolysis or exogenous enzymatic hydrolysis. These peptides are widely used in the food, health product, and cosmetic industries due to their high bioactivity and good solubility, especially in functional foods and beauty products. The main production steps include yeast cell disruption, enzymatic hydrolysis, separation and purification, and drying. Among these, enzymatic hydrolysis is the key step affecting the quality of yeast protein peptides. During enzymatic hydrolysis, the enzyme hydrolyzes proteins to produce peptide chains. The degree of hydrolysis and the molecular weight distribution of the peptides directly affect the bioactivity, solubility, flavor, and other functional properties of the final product. To obtain high-quality yeast protein peptides, it is necessary to precisely control the degree of hydrolysis and the termination point during the reaction.

[0003] In recent years, continuous improvements and optimizations in enzymatic hydrolysis technology have provided more possibilities for the production of yeast protein peptides. Traditional enzymatic hydrolysis processes mostly rely on manual control of reaction conditions (such as temperature, pH, and enzyme dosage). However, with the expansion of production scale and the increasing demands for product quality control, industrial production requires greater automation. The application of intelligent reaction monitoring and automated control technologies in yeast protein peptide production is gradually becoming a key technology for improving product quality, reducing costs, and increasing production efficiency.

[0004] Currently, the control of enzymatic hydrolysis reactions in the preparation of yeast protein peptides mainly relies on traditional empirical methods. For example, operators typically determine the degree of hydrolysis by terminating the reaction at set times or by periodically sampling and analyzing samples. Specific methods include: (1) Timed termination This method involves setting a fixed enzymatic hydrolysis time based on experience, and terminating the reaction when the predetermined time is reached. However, this method is susceptible to factors such as differences in yeast raw materials and fluctuations in enzyme activity, making it difficult to ensure the stability of each reaction.

[0005] (2) Periodic sampling and analysis Operators periodically take samples to determine the degree of hydrolysis or analyze the molecular weight distribution of peptides using methods such as chromatography. This method can more accurately reflect the reaction progress, but because the analytical process is relatively cumbersome and time-consuming, it is difficult to achieve real-time monitoring and control.

[0006] In addition, recent studies have proposed using pH monitoring, viscosity monitoring or online chromatography to control the enzymatic hydrolysis process. Although these methods can provide real-time feedback on the reaction to some extent, they usually have drawbacks such as slow response, complex equipment and unstable signals, and have not been widely used in large-scale industrial production.

[0007] Existing yeast protein peptide preparation technologies have the following main drawbacks: (1) The hysteresis of the timed termination reaction Relying on a fixed time to terminate the reaction makes it impossible to accurately determine the actual degree of hydrolysis. Due to differences in substrate type, enzyme activity, and reaction conditions, traditional methods can easily lead to over-hydrolysis or under-hydrolysis, affecting the quality and function of the final peptide.

[0008] (2) Operational complexity of periodic sampling analysis Sampling and analysis require a certain amount of time, resulting in a lag between the reaction process and the actual analytical results, making it difficult to accurately determine the reaction endpoint. Furthermore, the analytical process relies on manual operation, which introduces certain errors, and real-time feedback control is not possible.

[0009] (3) Insufficient automation and real-time control Existing technologies struggle to automate the production process, particularly during enzymatic hydrolysis. While some methods attempt to monitor changes in pH or viscosity, these parameters are not directly correlated with the accuracy of the hydrolysis process and cannot adapt to variations in different batches of yeast feedstock. Consequently, manual intervention remains necessary, resulting in low production efficiency and poor product consistency.

[0010] (4) The process is difficult to standardize. Traditional technologies often lack unified standards, and reaction conditions and termination timing depend on the operator's experience, making it difficult to achieve efficient, stable, and standardized large-scale production.

[0011] Therefore, existing technologies still have significant shortcomings in yeast protein peptide production, especially in areas such as precise control of the reaction process, improving production automation, and reducing human intervention, where new technological breakthroughs are urgently needed. This invention solves these problems through an online monitoring and automatic control method based on the rate of change of conductivity. It can accurately and in real-time determine the progress of the enzymatic hydrolysis reaction, ensuring stable product quality and a high level of automation.

[0012] In the preparation of yeast protein peptides, controlling the enzymatic hydrolysis reaction has always been a crucial and challenging step in the production process. Existing technologies mostly rely on traditional methods such as timed reaction termination or periodic sampling and analysis of the degree of hydrolysis to determine the reaction progress. However, these methods have the following shortcomings: (1) Lag in control of reaction endpoint: Traditional timed termination methods are prone to over-hydrolysis or under-hydrolysis due to failure to monitor the degree of hydrolysis in real time, which affects the quality of the final peptide.

[0013] (2) The sampling and analysis process is cumbersome and delayed: It takes time to periodically sample and analyze the degree of hydrolysis or molecular weight distribution, and there are certain operational errors, making it impossible to achieve real-time feedback control.

[0014] (3) Production process is difficult to automate: Existing technologies lack precise online monitoring methods and still rely on human experience and intervention, which makes it difficult to meet the demand for high-precision and automated control in large-scale industrial production.

[0015] Therefore, the technical problem of this invention is: how to achieve real-time and precise control of the enzymatic hydrolysis reaction during the preparation of yeast protein peptides, and how to automatically determine the end point of enzymatic hydrolysis, thereby improving production efficiency, ensuring product quality consistency, and reducing human intervention. Summary of the Invention

[0016] The problem with the prior art that this application addresses is: Existing technologies still have significant shortcomings in the production of yeast protein peptides, especially in areas such as precise control of the reaction process, improvement of production automation, and reduction of human intervention, where new technological breakthroughs are urgently needed.

[0017] The solution to the technical problem of this invention is: An online control method for yeast protein peptide preparation based on a conductivity change rate threshold is provided, comprising the following steps: Real-time monitoring of the conductivity of the enzymatic hydrolysis reaction system; Calculate the rate of change of conductivity; At least one conductivity change rate threshold is set based on the target product; The enzymatic hydrolysis reaction is automatically terminated when the rate of change of conductivity reaches or falls below the threshold.

[0018] Preferably, the conductivity change rate threshold is determined experimentally by detecting the relationship between the conductivity change rate and the degree of protein hydrolysis.

[0019] Preferably, the threshold includes: The first threshold δ1 is used to control the preparation of peptides with a molecular weight >5000 Da; The second threshold δ2 is used to control the preparation of peptides with a molecular weight of 1000–5000 Da. The third threshold δ3 is used to control the preparation of peptides with a molecular weight <1000 Da.

[0020] Preferably, δ1 is 0.4 mS / cm·min, δ2 is 0.2 mS / cm·min, and δ3 is 0.05 mS / cm·min.

[0021] Preferably, the conductivity is acquired in real time by an online conductivity sensor.

[0022] Preferably, the method further includes the step of: establishing a mathematical model relating the rate of change of conductivity to the degree of hydrolysis and the molecular weight distribution of peptides.

[0023] Preferably, the automatic termination of the reaction includes sending a stop signal to the control system and initiating subsequent purification steps.

[0024] Preferably, the method is applicable to the enzymatic hydrolysis of yeast protein peptides, and can also be extended to other protein enzymatic hydrolysis processes.

[0025] Preferably, the online control method includes: an online conductivity sensor; a data acquisition and processing module; a threshold comparison and judgment module; and an automatic control execution module.

[0026] Preferably, it also includes a human-machine interface for real-time display of conductivity curves, setting thresholds, and alarm prompts.

[0027] The technical effects achieved by this application in solving the technical problem are as follows: Compared with existing technologies, the present invention provides an online control method for yeast protein peptide preparation based on a conductivity change rate threshold, which has the following advantages: (1) Real-time monitoring and precise control: By monitoring the rate of change in conductivity in real time, the progress of the enzymatic hydrolysis reaction can be accurately determined without sampling. This can avoid over-hydrolysis or under-hydrolysis, ensuring the consistency of product quality for each batch.

[0028] (2) Reduce human intervention and improve automation: Traditional methods rely on human experience for reactive control and are easily affected by human factors. This invention enables full-process online monitoring and control through an automated control system, which greatly reduces human intervention and improves the automation and repeatability of the production process.

[0029] (3) Improved production efficiency and product consistency: The lag in traditional methods makes the control of the enzymatic hydrolysis process insufficiently precise, leading to fluctuations in product quality. This invention can adjust the enzymatic hydrolysis process in real time, ensuring the accuracy and stability of each reaction, thereby improving product consistency.

[0030] (4) Simplified operation and reduced cost: Unlike traditional frequent sampling analysis, this invention provides a simple and efficient control method, which reduces the number of operation steps and the dependence on experimental equipment, thereby effectively reducing production costs.

[0031] (5) High adaptability and potential for industrial application: This method can not only be applied to the preparation of yeast protein peptides, but also extended to other protein hydrolysis processes, with wide applicability and good prospects for industrial application. Attached Figure Description

[0032] Figure 1 This is a schematic flowchart of an online control method for preparing yeast protein peptides based on a threshold of conductivity change rate, according to the present invention. Detailed Implementation

[0033] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0034] It should be understood that the specific embodiments described herein are merely for explaining the invention and are not intended to limit the invention.

[0035] Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] It should be noted that when a component is said to be "fixed" to another component, it can be directly on the other component or it can be in a middle component. When a component is said to be "connected" to another component, it can be directly connected to the other component or it may be in a middle component.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0038] The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0039] The following detailed description of some embodiments of the present invention is provided in conjunction with the accompanying drawings. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0040] Please see Figure 1 The present invention provides an online control method for the preparation of yeast protein peptides based on a threshold of conductivity change rate, comprising the following steps: Real-time monitoring of the conductivity of the enzymatic hydrolysis reaction system; Calculate the rate of change of conductivity; At least one conductivity change rate threshold is set based on the target product; The enzymatic hydrolysis reaction is automatically terminated when the rate of change of conductivity reaches or falls below the threshold.

[0041] In some other embodiments, the conductivity change rate threshold is determined by experimentally detecting the relationship between the conductivity change rate and the degree of protein hydrolysis.

[0042] The threshold includes: The first threshold δ1 is used to control the preparation of peptides with a molecular weight >5000 Da; The second threshold δ2 is used to control the preparation of peptides with a molecular weight of 1000–5000 Da. The third threshold δ3 is used to control the preparation of peptides with a molecular weight <1000 Da.

[0043] The values ​​of δ1 are 0.4 mS / cm·min, δ2 are 0.2 mS / cm·min, and δ3 are 0.05 mS / cm·min.

[0044] The conductivity is collected in real time by an online conductivity sensor.

[0045] It also includes the step of establishing a mathematical model relating the rate of change in conductivity to the degree of hydrolysis and the molecular weight distribution of peptides.

[0046] The automatic termination of the reaction includes sending a stop signal to the control system and initiating subsequent purification steps.

[0047] The method is applicable to the enzymatic hydrolysis of yeast protein peptides and can also be extended to other protein hydrolysis processes.

[0048] The online control method includes: an online conductivity sensor; a data acquisition and processing module; a threshold comparison and judgment module; and an automatic control execution module.

[0049] It also includes a human-computer interaction interface for real-time display of conductivity curves, setting thresholds, and alarm prompts.

[0050] The technical solution of this application is used to accurately determine the enzymatic hydrolysis process in the preparation of yeast protein peptides and to control the termination time of the reaction through multiple thresholds, thereby obtaining small peptide products of different molecular weights. This method mainly includes the following steps: (1) Real-time conductivity monitoring: During the enzymatic hydrolysis process, the conductivity changes of the enzymatic hydrolysate are monitored in real time by an online conductivity sensor.

[0051] (2) Calculation of conductivity change rate: The conductivity change rate (dσ / dt) is calculated based on real-time monitoring data. As the enzymatic reaction proceeds, the concentration of mobile ions in the solution gradually increases, and the conductivity rises. The conductivity change rate is closely related to the progress of the enzymatic reaction and the degree of hydrolysis.

[0052] (3) Establishment of the relationship between the rate of change of conductivity and the degree of hydrolysis: Through experimental calibration, a mathematical relationship (functional model) between the rate of change of conductivity (dσ / dt) and the degree of hydrolysis (such as degree of hydrolysis DH, molecular weight distribution, etc.) is established. This relationship can accurately reflect the nonlinear correlation between the rate of change of conductivity and the hydrolysis process, and provide a theoretical basis for subsequent control.

[0053] (4) Setting and controlling multiple thresholds: By using different conductivity change rate thresholds (δ1, δ2, δ3, etc.), the reaction progress is precisely controlled at different hydrolysis stages. Specifically, based on the required degree of hydrolysis, appropriate thresholds are selected to determine whether the reaction needs to continue or be terminated. Low molecular weight peptide control: When the rate of change of conductivity approaches a certain high threshold (e.g., δ1), it is determined that the reaction has entered the early stage. At this time, less small peptides are generated, which is suitable for peptide products with larger molecular weight that require shorter hydrolysis time.

[0054] Medium molecular weight peptide control: When the rate of change of conductivity drops to another threshold (e.g., δ2), the degree of hydrolysis is moderate, and the product is mainly medium molecular weight peptides.

[0055] High degree of hydrolysis control: When the rate of change of conductivity further decreases and approaches the low threshold (e.g., δ3), the enzymatic hydrolysis reaction enters the final stage, with a high degree of hydrolysis and a high content of small molecule peptides, which is suitable for applications that require low molecular weight peptides.

[0056] By flexibly adjusting these thresholds, the enzymatic hydrolysis process can be precisely controlled to form peptide products with different molecular weight distributions, thereby meeting the peptide requirements of different application scenarios.

[0057] (5) Automated control system: Based on the comparison between the real-time rate of change of conductivity and the set threshold, the automated control system determines whether the reaction should continue or terminate. When the rate of change of conductivity is lower than the set threshold, the system automatically issues a command to terminate the enzymatic hydrolysis and initiates subsequent separation and purification steps.

[0058] This multi-threshold control-based approach not only precisely controls the enzymatic hydrolysis process but also generates peptides of varying molecular weights, offering greater flexibility and adaptability. This technology ensures precise control at every stage of the production process and reduces human intervention through intelligent feedback, thereby improving automation levels and product consistency.

[0059] The online control method based on the conductivity change rate threshold proposed in this invention has the following significant advantages compared to traditional reaction control methods: (1) Real-time monitoring and precise control: By monitoring the rate of change in conductivity in real time, the progress of the enzymatic hydrolysis reaction can be accurately determined without sampling. This can avoid over-hydrolysis or under-hydrolysis, ensuring the consistency of product quality for each batch.

[0060] (2) Reduce human intervention and improve automation: Traditional methods rely on human experience for reactive control and are easily affected by human factors. This invention enables full-process online monitoring and control through an automated control system, which greatly reduces human intervention and improves the automation and repeatability of the production process.

[0061] (3) Improved production efficiency and product consistency: The lag in traditional methods makes the control of the enzymatic hydrolysis process insufficiently precise, leading to fluctuations in product quality. This invention can adjust the enzymatic hydrolysis process in real time, ensuring the accuracy and stability of each reaction, thereby improving product consistency.

[0062] (4) Simplified operation and reduced cost: Unlike traditional frequent sampling analysis, this invention provides a simple and efficient control method, which reduces the number of operation steps and the dependence on experimental equipment, thereby effectively reducing production costs.

[0063] (5) High adaptability and potential for industrial application: This method can not only be applied to the preparation of yeast protein peptides, but also extended to other protein hydrolysis processes, with wide applicability and good prospects for industrial application.

[0064] In summary, this invention solves the reaction control problem in the preparation of yeast protein peptides through an innovative online monitoring and control method, which not only improves production efficiency and product quality, but also promotes the automation and intelligence of the production process.

[0065] Example 1: Preparation of mid-molecular-weight peptides from yeast protein peptides based on conductivity change rate control.

[0066] (1) Yeast pretreatment Take 5000 mL of freshly cultured yeast fermentation broth and remove the culture medium by centrifugation (5000 rpm, 10 min); after washing the precipitate, resuspend the fermentation broth in 1 / 10 volume of deionized water to obtain a uniform yeast cell emulsion.

[0067] (2) Homogenization and cell wall disruption The yeast suspension was fed into a high-pressure homogenizer, set to 50 MPa, and homogenized 5 times. Centrifuged at 5000 rpm for 10 min, filtered to remove the precipitate, and the supernatant was kept for later use.

[0068] (3) Enzymatic hydrolysis reaction Add trypsin to the yeast supernatant after cell wall disruption to a final concentration of 1 U / ml, adjust the pH to 7.0, set the temperature to 40℃, start the enzymatic hydrolysis reaction and monitor the conductivity in real time.

[0069] (4) Calculation of conductivity change rate and threshold setting: Based on the relationship between the rate of change of conductivity and the degree of hydrolysis obtained from previous experiments, the following threshold value for the rate of change of conductivity, δ, is set: δ1 = 0.4 mS / cm·min (early enzymatic hydrolysis stage, suitable for large molecular weight peptides, molecular weight > 5000 Da) δ2 = 0.2 mS / cm·min (mid-stage enzymatic hydrolysis, suitable for medium molecular weight peptides, molecular weight 1000-5000 Da) δ3 = 0.05 mS / cm·min (final enzymatic hydrolysis stage, suitable for generating low molecular weight peptides, molecular weight <1000 Da) The change in conductivity σ of the enzymatic hydrolysate is monitored by a real-time online conductivity sensor, and the rate of change of conductivity (dσ / dt) is calculated.

[0070] (5) Automated control: When the rate of change of conductivity drops to δ2 = 0.2 mS / cm·min, the system automatically issues a command to stop the enzymatic hydrolysis reaction.

[0071] (6) Separation, purification and drying Impurities such as nucleic acids and polysaccharides in the enzymatic hydrolysate were removed using conventional methods, and then medium-molecular-weight brewer's yeast protein peptides were obtained by spray drying.

[0072] Example 2: Preparation of yeast protein peptides with small molecular weight peptides based on conductivity change rate control.

[0073] (1) Yeast pretreatment Take 5000 mL of freshly cultured yeast fermentation broth and remove the culture medium by centrifugation (5000 rpm, 10 min); after washing the precipitate, resuspend the fermentation broth in 1 / 10 volume of deionized water to obtain a uniform yeast cell emulsion.

[0074] (2) Homogenization and cell wall disruption The yeast suspension was fed into a high-pressure homogenizer, set to 50 MPa, and homogenized 5 times. Centrifuged at 5000 rpm for 10 min, filtered to remove the precipitate, and the supernatant was kept for later use.

[0075] (3) Enzymatic hydrolysis reaction Add trypsin to the yeast supernatant after cell wall disruption to a final concentration of 1 U / ml, adjust the pH to 7.0, set the temperature to 40℃, start the enzymatic hydrolysis reaction and monitor the conductivity in real time.

[0076] (4) Calculation of conductivity change rate and threshold setting: Based on the relationship between the rate of change of conductivity and the degree of hydrolysis obtained from previous experiments, the following threshold value for the rate of change of conductivity, δ, is set: δ1 = 0.4 mS / cm·min (early enzymatic hydrolysis stage, suitable for large molecular weight peptides, molecular weight > 5000 Da) δ2 = 0.2 mS / cm·min (mid-stage enzymatic hydrolysis, suitable for medium molecular weight peptides, molecular weight 1000-5000 Da) δ3 = 0.05 mS / cm·min (final enzymatic hydrolysis stage, suitable for generating low molecular weight peptides, molecular weight <1000 Da) The change in conductivity σ of the enzymatic hydrolysate is monitored by a real-time online conductivity sensor, and the rate of change of conductivity (dσ / dt) is calculated.

[0077] (5) Automated control: When the rate of change of conductivity drops to δ2 = 0.05 mS / cm·min, the system automatically issues a command to stop the enzymatic hydrolysis reaction.

[0078] (6) Separation, purification and drying Impurities such as nucleic acids and polysaccharides in the enzymatic hydrolysate were removed using conventional methods, and finally, small molecular weight brewer's yeast protein peptides were obtained by spray drying.

[0079] Compared with existing technologies, the present invention provides an online control method for yeast protein peptide preparation based on a conductivity change rate threshold. The embodiments of the present invention described above do not constitute a limitation on the scope of protection of the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A method for on-line control of the preparation of yeast protein peptides based on the threshold rate of change of conductivity, characterized in that: The method comprises the following steps: S1: Real-time monitoring of the conductivity of the enzymatic reaction system; S2: Calculating the conductivity change rate; S3: Setting at least one conductivity change rate threshold according to the target product; S4: Automatically terminating the enzymatic reaction when the conductivity change rate reaches or is lower than the threshold.

2. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: The conductivity change rate threshold is determined by experimental detection of the relationship between the conductivity change rate and the degree of proteolysis.

3. The online control method of yeast protein peptide preparation based on the threshold of the rate of change of electrical conductivity according to claim 1 or 2, characterized in that: The threshold includes: The first threshold δ1 is 0.4 mS / cm·min, the second threshold δ2 is 0.2 mS / cm·min, and the third threshold δ3 is 0.05 mS / cm·min. The conductivity is collected in real time by an online conductivity sensor. It also includes the step of establishing a mathematical model between the conductivity change rate and the degree of hydrolysis and the distribution of peptide molecular weight.

4. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 3, characterized in that: The automatic termination of the reaction includes sending a stop signal to the control system and starting the subsequent purification process.

5. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: The method is suitable for the enzymatic reaction of yeast protein peptides and can be extended to other protein enzymatic processes.

6. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: The online control method includes an online conductivity sensor, a data acquisition and processing module, a threshold comparison and judgment module, and an automatic control execution module.

7. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: It also includes a human-computer interaction interface for real-time display of the conductivity curve, threshold setting, and alarm prompt.

8. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: ​ 9. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 1, characterized in that: ​ 10. The online control method of yeast protein peptide preparation based on the change rate threshold of electrical conductivity according to claim 9, characterized in that: ​

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