A method and system for optimizing polypeptide-induced biological fermentation medium

Abstract

The present invention relates to the field of polypeptide induction technology, and discloses a polypeptide-induced bio-fermentation culture medium optimization method and system, including building an ideal experimental environment condition suitable for the bio-fermentation culture medium, adding a preset concentration of polypeptide as a signal molecule between the time when the cells enter the product induction period from the logarithmic growth phase; detecting the actual yield of the target product in the culture medium after the product induction period, the actual yield refers to the ratio increased by adding the polypeptide as a signal molecule on the original basis, and when the actual yield is less than the standard yield, executing the polypeptide concentration optimization strategy. The present invention utilizes a method of adding polypeptide signals in stages and steps within a window period, and detects the target product concentration in real time and calculates the real-time yield rate, thereby accurately identifying the window period to be optimized where the reaction effect is not obvious, so as to optimize the polypeptide addition conditions for the window period to be optimized.

Description

Technical Field

[0001] The present invention relates to the technical field of polypeptide induction, in particular to a method and system for optimizing a polypeptide-induced biological fermentation medium. Background Art

[0002] Peptides can act as signal molecules, binding to receptors inside or on the surface of host cells to trigger intracellular signal transduction cascades, thereby inducing gene expression or activation of metabolic pathways related to the synthesis of target products. By utilizing the signals induced by peptides, the culture medium can enable cells to concentrate metabolic resources on the synthesis of target products (such as proteins, antibiotics, enzymes, metabolites, etc.) at the appropriate time point, thereby achieving the goal of increasing product yield.

[0003] In practical applications, when using peptides as signal molecules to induce the synthesis of target products, different cells may have large differences in sensitivity to signal molecules at different growth stages and under different environmental conditions. Changes in receptor expression levels will directly affect the effect of peptide induction. Therefore, in order to achieve the goal of increasing the yield of target products, it is critical to scientifically and rationally design and optimize the conditions for peptide addition. Summary of the Invention

[0004] In response to the problems existing in the above-mentioned prior art, the purpose of the present invention is to provide a method and system for optimizing polypeptide-induced bio-fermentation culture medium, so as to optimize the polypeptide addition conditions in combination with the expression level of culture medium receptors, so that the polypeptide-induced signal transmission can achieve the ideal application effect, thereby achieving the purpose of increasing the yield of the target product.

[0005] In order to achieve the above-mentioned object, the present invention provides the following technical solution: a method for optimizing a polypeptide-induced bio-fermentation medium, the method comprising the following steps:

[0006] Establish ideal experimental environment conditions for biofermentation culture medium, and add a preset concentration of peptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase;

[0007] After the product induction period, the actual yield of the target product in the culture medium is detected. The actual yield refers to the ratio increased by adding the peptide as a signal molecule on the original basis. When the actual yield is lower than the target yield, the peptide concentration optimization strategy is implemented;

[0008] The peptide concentration optimization strategy involves dividing the time period between the cells' logarithmic growth phase and the product induction phase into multiple window periods. The peptide is then added in equal portions according to the original dosage in multiple window periods, and the post-induction concentration of the target product in each window period is measured.

[0009] The real-time yield rate is obtained by the concentration of the target product after induction in each window period, and the yield rate growth in each window period is compared with the yield rate improvement threshold. The window period with a yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted;

[0010] After executing the peptide concentration optimization strategy, the adjusted yield of the target product in the culture medium is tested. When the adjusted yield is less than the estimated target yield, a tolerance threshold should be set and the optimized phase difference value should be obtained. The optimized phase difference value should be compared with the tolerance threshold. If the optimized phase difference value is greater than the tolerance threshold, the judgment that the overall process needs to be improved is maintained; if the optimized phase difference value is less than or equal to the tolerance threshold, the peptide optimization effect is judged.

[0011] When the adjusted yield fails to reach the estimated target yield, and the optimized improvement is greater than or equal to the improvement threshold, the window period with the highest rate increase is set as the optimal window period. When adding peptides during the optimal window period, a method that allows the peptides to be slowly released during the optimal window period is used to extend the actual induction time of the peptides during the optimal window period.

[0012] In some embodiments, the specific method of setting the real-time yield rate is to obtain the estimated target yield Yc=Jc×Dn by the basic yield Jc of the target product and the target yield rate Dn, and to obtain the real-time yield rate Sn=Nr÷Yc by the estimated target yield Yc and the target product concentration after induction Nr in the corresponding window period; the specific method of setting the yield rate improvement threshold is to obtain the real-time yield rate Sn in the initial window period. 1 Obtain the rate difference Dc=1-Sn 1 The rate improvement threshold Dt=Dc÷Cs is obtained by obtaining the rate difference Dc and the total number of window periods Cs.

[0013] In some embodiments, the specific method of adjusting the window period to be optimized is: diluting and concentrating the polypeptide of the preset concentration according to the polypeptide concentration adjustment amount, and adding the polypeptide of the diluted concentration and concentrated concentration to replace the polypeptide of the preset concentration in the window period to be optimized for testing, obtaining the increase in the yield rate of the window period to be optimized under the conditions of using the preset concentration, diluted concentration and concentrated concentration polypeptide, comparing the increase in the yield rate of the window period to be optimized when adding the preset concentration, diluted concentration and concentrated concentration polypeptide, and making corresponding responses based on the comparison results.

[0014] In some embodiments, if the increase in the window period to be optimized is the largest when the predetermined concentration of polypeptide is added, the concentration of the added polypeptide is not adjusted;

[0015] If the increase in the window period to be optimized is the largest when adding the diluted concentration peptide, then the dilution concentration of the peptide is continued to be reduced according to the peptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained;

[0016] If the increase in the window period to be optimized is the largest when the concentrated concentration of the peptide is added, the dilution concentration of the peptide is increased incrementally according to the peptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained.

[0017] In some embodiments, after executing the polypeptide concentration optimization strategy, the adjusted yield of the target product in the culture medium is detected, and the adjusted yield is compared with the estimated target yield. When the adjusted yield is greater than or equal to the estimated target yield, the polypeptide concentration optimization strategy is used as a subsequent operation indicator; when the adjusted yield is less than the estimated target yield, a decision should be made to improve the overall process.

[0018] In some embodiments, the specific method for determining the effect of polypeptide optimization is to set a threshold value for the improvement, compare the adjusted acquisition rate Td with the real-time acquisition rate Sn in the initial window period before performing polypeptide concentration optimization, and 1 The difference is the adjusted growth value Tz=Td-Sn 1 The real-time acquisition rate Sn of the final window period before peptide concentration optimization is performed 5 The real-time rate Sn obtained during the initial window period before performing peptide concentration optimization 1 Take the difference to get the unadjusted growth value Tu=Sn 5 -Sn 1 The optimized improvement Fr=Tz÷Tu is obtained by dividing the adjusted growth value Tz by the growth value Tu before adjustment. The optimized improvement is compared with the improvement threshold. If the optimized improvement is greater than or equal to the improvement threshold, the peptide concentration optimization strategy is used as the subsequent operation indicator.

[0019] The present invention also provides the following technical solution: a polypeptide-induced bio-fermentation medium optimization system, comprising:

[0020] The environment construction module is used to build the ideal experimental environment conditions for biological fermentation medium, and add a preset concentration of peptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase;

[0021] The improvement comparison module detects the actual yield of the target product in the culture medium after the product induction period. The actual yield refers to the ratio increased by adding peptides as signal molecules on the original basis. When the actual yield is lower than the target yield, the peptide concentration optimization strategy is implemented;

[0022] Strategy execution module: The peptide concentration optimization strategy includes dividing the time period between the cells entering the logarithmic growth phase and the product induction phase into multiple window periods. The peptide is added in equal amounts in multiple window periods according to the original addition amount, and the concentration of the target product after induction in each window period is detected;

[0023] The condition optimization module obtains the real-time yield rate through the concentration of the target product after induction in each window period, and compares the yield rate growth in each window period with the yield rate improvement threshold. The window period with the yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted.

[0024] The present invention further provides a computer-readable storage medium storing a computer program, wherein the computer program is executed by a processor to implement the above-mentioned method and system for optimizing a polypeptide-induced biological fermentation medium.

[0025] Compared with the prior art, the technical solution provided by the present invention has the following beneficial effects:

[0026] First, the present invention utilizes a method of adding polypeptide signals in stages and steps within a window period. By real-time detection of the target product concentration and calculation of the real-time yield, the window period to be optimized where the reaction effect is not obvious can be accurately identified, so as to optimize the polypeptide addition conditions for the window period to be optimized.

[0027] Secondly, the present invention adjusts the conditions for adding polypeptides within the optimization window period, taking into account the differences in sensitivity and tolerance of different production systems and cell lines to polypeptides. By gradually adjusting the concentration and based on experimental data, the optimal concentration is accurately found to ensure the best induction effect of the polypeptide within the target window period, thereby significantly improving the accumulation of the target product and the stability of the overall process.

[0028] Third, the present invention achieves a dynamic evaluation of the polypeptide induction effect by comparing the relationship between the adjusted yield and the estimated target yield, and setting tolerance thresholds and quantitative optimization indicators. It can accurately evaluate the improvement effect of the target product yield before and after polypeptide optimization, thereby ensuring that only when the polypeptide conditions do show a stable and obvious positive effect will they be implemented as subsequent production indicators.

[0029] Fourthly, the present invention uses the optimal window period as the key induction period and adopts a sustained-release method during this period. This not only extends the actual induction time of the polypeptide and maintains a relatively constant polypeptide concentration throughout the window period, but also specifically enhances the response of cells to polypeptide signals during the optimal window period. Although this method may slightly increase the production steps, if the adjusted yield does not reach the estimated target, this move can further narrow the gap and reduce the impact of yield loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Schematic diagram of a process for optimizing a polypeptide-induced bio-fermentation medium according to the present invention;

[0031] Figure 2 This is a module schematic diagram of a polypeptide-induced biological fermentation medium optimization system of the present invention. DETAILED DESCRIPTION

[0032] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0033] It is to be understood that the term "one" should be understood as "at least one" or "one or more", that is, in one embodiment, the number of an element may be one, while in another embodiment, the number of the elements may be multiple, and the term "one" should not be understood as a limitation on the quantity.

[0034] The present invention provides a method for optimizing a polypeptide-induced bio-fermentation medium. Figure 1 As shown, including:

[0035] Step 1: Establish ideal experimental environmental conditions suitable for biofermentation culture medium, including temperature control, pH control, and dissolved oxygen level control. Add a preset concentration of peptide as a signal molecule between the cells' transition from the logarithmic growth phase to the product induction phase. During this period, the cells are in an active metabolic state and are more sensitive to induction signals. Ensure that the peptide is fully dissolved in the culture medium to avoid weakening of activity due to localized excessive concentration or precipitation.

[0036] Step 2: After the product induction period, the actual yield of the target product in the culture medium is detected. The actual yield refers to the ratio increased by adding the peptide as a signal molecule on the original basis. For example, if the target product is a protein, the protein yield on the original basis is 1 g / L, and the protein yield after adding the peptide as a signal molecule is 1.5 g / L, then the yield is 150%. The actual yield is compared with the preset target yield. When the actual yield is greater than or equal to the target yield, it means that the expected target product yield can be obtained based on the current peptide addition conditions. When the actual yield is less than the target yield, it means that the expected target product yield cannot be obtained based on the current peptide addition conditions, and the peptide concentration optimization strategy is executed.

[0037] Step 3: Executing a peptide concentration optimization strategy involves dividing the time period between the cells entering the logarithmic growth phase and the product induction phase into multiple window periods, adding the peptide in equal portions according to the original addition amount in multiple window periods, and measuring the post-induction concentration of the target product in each window period;

[0038] Step 4: The real-time yield rate is obtained by the concentration of the target product after induction in each window period, and the yield rate growth in each window period is compared with the yield rate improvement threshold. The window period with a yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted.

[0039] The specific method of obtaining the real-time acquisition rate is to obtain the estimated target yield Yc=Jc×Dn by the basic yield Jc of the target product and the target yield rate Dn, and then obtain the real-time acquisition rate Sn=Nr÷Yc by the estimated target yield Yc and the target product induced concentration Nr in the corresponding window period. The specific method of setting the acquisition rate improvement threshold is to use the real-time acquisition rate Sn in the initial window period. 1 Obtain the rate difference Dc=1-Sn 1 The rate improvement threshold Dt=Dc÷Cs is obtained by obtaining the rate difference Dc and the total number of window periods Cs.

[0040] For example, the time period from the logarithmic growth phase to the product induction phase of the cells is divided into five window periods. The post-induction concentrations of the target product detected in the five window periods are 0.5 g / L, 0.6 g / L, 0.9 g / L, 1.3 g / L and 1.5 g / L, respectively. The basic yield of the target product is set to 1 g / L, and the compliance rate is set to 200%. The estimated target yield is 2 g / L, and the rate improvement threshold is 15%. The real-time rates of the five window periods are 25%, 30%, 45%, 65% and 75%, respectively. The real-time rate refers to the value of the post-induction concentration of the target product detected in the corresponding window period divided by the estimated target yield. The yield rate increase in a window period is the difference between the corresponding window period and the previous window period. Therefore, the first window period is the baseline value with no yield rate increase, the yield rate increase in the second window period is 5%, the yield rate increase in the third window period is 15%, the yield rate increase in the fourth window period is 20%, and the yield rate increase in the fifth window period is 10%. If the yield rate increase in the second and fifth window periods is less than the yield rate improvement threshold, the second and fifth window periods are set as the window period to be optimized. The window period to be optimized can represent the period during the entire peptide induction process when the addition of peptide does not significantly increase the yield of the target product.

[0041] The specific method for adjusting the window period to be optimized is: dilute and concentrate the preset concentration of polypeptide according to the polypeptide concentration adjustment amount, the polypeptide concentration adjustment amount can be 1-5%, and replace the preset concentration of polypeptide with diluted concentration and concentrated concentration in the window period to be optimized for addition testing, and obtain the increase in the yield rate of the window period to be optimized under the conditions of using the preset concentration, diluted concentration and concentrated concentration of polypeptide, and compare the increase in the yield rate of the window period to be optimized under the conditions of adding the preset concentration, diluted concentration and concentrated concentration of polypeptide. If the increase in the yield rate of the window period to be optimized is the largest when the preset concentration of polypeptide is added, it means that the preset concentration of polypeptide is already the most suitable production condition for the target product in this culture medium, and it is not correct. Adjustments are made by adding polypeptide concentrations; if the increase in the window period to be optimized is the largest when adding a diluted concentration polypeptide, it means that the concentration of the polypeptide at the preset concentration is too high and is not suitable for the production conditions of the target product in this culture medium. The polypeptide dilution concentration is continuously decreased according to the polypeptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained; if the increase in the window period to be optimized is the largest when adding a concentrated concentration polypeptide, it means that the concentration of the polypeptide at the preset concentration is too low and is not suitable for the production conditions of the target product in this culture medium. The polypeptide dilution concentration is continuously increased according to the polypeptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained. For example, the polypeptide concentration adjustment amount is set to 5%. Since the yield increase in the fifth window period is 10%, it is the window period to be optimized. The preset concentration of the polypeptide is 100%. The polypeptide is added in the fifth window period at a 95% dilution concentration and a 105% concentrated concentration. After testing, the yield increase in the fifth window period at the preset concentration is 10%, the yield increase in the fifth window period at a 95% dilution concentration is 11%, and the yield increase in the fifth window period at a 105% concentrated concentration is 9%. Among them, the yield increase at the diluted concentration is the largest. Then, the concentration is continued to decrease according to the polypeptide concentration adjustment amount. After adding the polypeptide at a 90% concentration, the yield increase is 13%, after adding the polypeptide at an 85% concentration, the yield increase is 15%, and after adding the polypeptide at an 80% concentration, the yield increase is 12%. When the yield increase begins to decrease, the 85% concentration with the largest yield increase is obtained as the polypeptide concentration for the fifth window period.

[0042] The reason for adjusting the peptide concentration is that within a certain concentration range, the peptide can effectively activate the target metabolic pathway. If the peptide concentration is insufficient, the level of peptide activation on the cell will be less than ideal. If the concentration is excessive, high concentrations of active peptides may cause cell stress responses or side effects, disrupting the normal metabolic balance of the cell, thereby reducing the synthesis efficiency of the target product. In addition, different production systems and cell lines have different sensitivities and tolerances to peptides. Sometimes, increasing the concentration of the peptide will be effective, but in fact, due to differences in cell characteristics, reducing the concentration can bring better induction effects. Therefore, it is key to adopt a flexible adjustment plan based on the above design and compare the performance at different concentrations.

[0043] By implementing the peptide concentration optimization strategy, the peptide addition method is changed to a step-by-step addition, which can achieve the goal of more ideally maintaining the induction effect. By setting the window period to be optimized, the peptide concentration can be adjusted for the period of time when the induction effect is not ideal. After the above adjustment, the adjusted yield of the target product in the culture medium is detected and compared with the estimated target yield. When the adjusted yield is greater than or equal to the estimated target yield, it means that the expected target product yield can be obtained according to the above peptide concentration optimization method, and the above peptide optimization conditions are used as the subsequent operation indicators; when the adjusted yield is less than the estimated target yield, it means that the expected target product yield cannot be obtained according to the above peptide concentration optimization method, and the overall process improvement should be considered, such as adjusting the culture time, cell seeding density, or adding other nutrients.

[0044] When the adjusted yield is less than the estimated target yield, a tolerance threshold should also be set. Specifically, the estimated target yield is subtracted from the adjusted yield to obtain the adjusted difference value, and the optimized phase difference value is obtained by dividing the adjusted difference value by the estimated target yield. The optimized phase difference value is compared with the tolerance threshold. If the optimized phase difference value is greater than the tolerance threshold, it means that the gap between the adjusted yield and the estimated target yield is more obvious, and the judgment of overall process improvement is maintained; if the optimized phase difference value is less than or equal to the tolerance threshold, the peptide optimization effect is judged. Specifically, the adjusted yield rate Td is compared with the real-time yield rate Sn in the initial window period before peptide concentration optimization. 1 The difference is the adjusted growth value Tz=Td-Sn 1 The real-time acquisition rate Sn of the final window period before peptide concentration optimization is performed 5 The real-time rate Sn obtained during the initial window period before performing peptide concentration optimization 1 Take the difference to get the unadjusted growth value Tu=Sn 5 -Sn 1The optimized improvement Fr = Tz ÷ Tu is calculated by comparing the adjusted growth value Tz with the unadjusted growth value Tu. A threshold for the improvement is set and the optimized improvement is compared to the threshold. If the optimized improvement is less than the threshold, it means that after implementing the peptide concentration optimization strategy, the adjusted yield has failed to reach the estimated target yield, and the yield of the target product can be increased only slightly. This indicates that the change in peptide conditions has little effect on cells in this culture medium, and the judgment that overall process improvement is necessary should be maintained. If the optimized improvement is greater than or equal to the threshold, it means that after implementing the peptide concentration optimization strategy, the yield of the target product can be significantly increased, indicating that the change in peptide conditions has a significant response to cells in this culture medium. The peptide optimization conditions designed above are then used as indicators for subsequent operations. This is because different production systems or cell lines may have significant differences in sensitivity to peptides. By setting a tolerance threshold and quantifying the optimized improvement, the actual effect can be captured during dynamic adjustments. Only when the peptide optimization conditions truly demonstrate stable and significant positive effects are they considered suitable for subsequent production and implemented as indicators in subsequent operations. For example, in combination with the above embodiment, the adjusted yield is 1.8 g / L, which is less than the estimated target yield of 2.0 g / L. The adjusted difference is 0.2 g / L, while the optimized difference is 0.1. The tolerance threshold is set to 0.2. Since the optimized difference is less than the tolerance threshold, the peptide optimization effect is determined: the real-time yield rate Sn in the initial window period before performing peptide concentration optimization is 1 When the yield is 25%, the adjusted yield is divided by the estimated target yield to obtain an adjusted yield of 90%, that is, the adjusted growth value is 65%, the growth value before adjustment is 50%, the optimization improvement is 1.3, and the improvement threshold is set to 1.2. Since the optimization improvement is greater than the improvement threshold, the peptide optimization conditions designed above are used as subsequent operation indicators.

[0045] Furthermore, when the adjusted yield fails to reach the estimated target yield, but the optimized improvement is greater than or equal to the improvement threshold, the window period with the highest rate increase should be set as the optimal window period. The optimal window period represents the time period when the cell is most sensitive and efficient in responding to the peptide signal when entering the product induction stage. During this window period, the metabolic state and physiological conditions of the cell are at a stage that is most suitable for receiving signals and responding quickly, so that the addition of the peptide can bring about the maximum increase in product concentration. When adding the peptide during the optimal window period, a method is designed to allow the peptide to be slowly released during the optimal window period, such as using microcapsules or liposomes as carriers. By encapsulating the peptide into a sustained-release formulation, the peptide's effect can be extended throughout the optimal window, thereby extending the actual induction duration of the peptide during the optimal window. Focusing the optimal window as the key induction period and employing a sustained-release formulation can help reduce batch-to-batch variability and make the fermentation process more stable and controllable. This approach, compared to directly adding the peptide all at once during the optimal window, can maintain a relatively constant peptide concentration throughout the entire optimal window. While this increases the number of production steps, it can further reduce the gap between the adjusted yield and the estimated target yield when there is a discrepancy between the adjusted yield and the estimated target yield. Furthermore, the optimal window is the time period when cells are most sensitive and responsive to peptide signals. During this time, the cell's metabolic state and signaling pathways are activated, making full use of the sustained-release peptide signal to maximize the synthesis of the target product. During other windows, cells are less sensitive to peptide signals, and sustained-release may not achieve the desired induction effect. Targeting sustained-release during the optimal window simplifies process control and avoids introducing unnecessary variables during other windows, when the cell response is weaker.

[0046] In general, the present invention aims to design a method for optimizing a polypeptide-induced biofermentation culture medium. For the purpose of optimizing the polypeptide addition conditions in combination with the expression level of the culture medium receptor, the present invention utilizes a method of adding polypeptide signals in stages and steps within the window period, and detects the concentration of the target product in real time and calculates the real-time rate, thereby accurately identifying the window period to be optimized where the reaction effect is not obvious. In addition, the adjustment of the polypeptide addition conditions within the optimization window period takes into account the differences in sensitivity and tolerance of different production systems and cell lines to polypeptides. By gradually adjusting the concentration and taking the experimental data as the basis, the optimal concentration is accurately found to ensure that the polypeptide has the best induction effect within the target window period, thereby significantly improving the accumulation of the target product and the stability of the overall process. This dynamic, step-by-step adjustment optimization strategy not only effectively improves the accumulation level of the target product, but also reduces the risk of over-stimulation of the cells, ensuring that the cells are always in the best state during the entire induction process. By comparing the relationship between the adjusted yield and the estimated target yield, and setting tolerance thresholds and quantitative optimization indicators, a dynamic evaluation of the peptide induction effect is achieved, which can accurately evaluate the improvement effect of the target product yield before and after peptide optimization, thereby ensuring that only when the peptide conditions truly show a stable and obvious positive effect will they be used as subsequent production indicators. By using the optimal window period as the key induction period and adopting a sustained-release method during this period, not only is the actual induction time of the peptide extended and a relatively constant peptide concentration maintained throughout the window period, but the response of cells to peptide signals during the optimal window period is also specifically enhanced. Although this approach may slightly increase the number of production steps, if the adjusted yield does not reach the estimated target, this move can further narrow the gap and reduce the impact of the yield shortfall.

[0047] The present invention provides a polypeptide-induced bio-fermentation medium optimization system, such as Figure 2 As shown, including:

[0048] The environment construction module is used to build the ideal experimental environment conditions for biological fermentation medium, and add a preset concentration of peptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase;

[0049] The improvement comparison module detects the actual yield of the target product in the culture medium after the product induction period. The actual yield refers to the ratio increased by adding peptides as signal molecules on the original basis. When the actual yield is lower than the target yield, the peptide concentration optimization strategy is implemented;

[0050] Strategy execution module: The peptide concentration optimization strategy includes dividing the time period between the cells entering the logarithmic growth phase and the product induction phase into multiple window periods. The peptide is added in equal amounts in multiple window periods according to the original addition amount, and the concentration of the target product after induction in each window period is detected;

[0051] The condition optimization module obtains the real-time yield rate through the concentration of the target product after induction in each window period, and compares the yield rate growth in each window period with the yield rate improvement threshold. The window period with the yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted.

[0052] In the embodiments disclosed herein, the processes described above with reference to the flowcharts can be implemented as computer software programs. The embodiments disclosed herein include a computer program product comprising a computer program carried on a computer-readable medium, the computer program containing program code for executing the method illustrated in the flowchart. In such an embodiment, the computer program can be downloaded and installed from a network via a communication component and / or installed from removable media. When the computer program is executed by a central processing unit, the functions defined in the methods of this application are performed. It should be noted that the computer-readable medium referred to herein can be a computer-readable signal medium or a computer-readable storage medium, or any combination thereof. Computer-readable storage media can be, for example, but not limited to, electrical, magnetic, optical, electromagnetic, infrared, or semiconductor systems, devices, or components, or any combination thereof. More specific examples of computer-readable storage media can include, but are not limited to, an electrical connection having one or more wire segments, a portable computer disk, a hard disk, random access memory, read-only memory, erasable programmable read-only memory, optical fiber, a portable compact disk read-only memory, an optical storage device, a magnetic storage device, or any suitable combination thereof. In this application, a computer-readable storage medium may be any tangible medium that contains or stores a program that can be used by or in conjunction with an instruction execution system, apparatus, or device. Furthermore, in this application, a computer-readable signal medium may include a data signal propagated in baseband or as part of a carrier wave, which carries computer-readable program code. This propagated data signal may take a variety of forms, including but not limited to electromagnetic signals, optical signals, or any suitable combination thereof. A computer-readable signal medium may also be any computer-readable medium other than a computer-readable storage medium that can transmit, propagate, or transfer a program for use by or in conjunction with an instruction execution system, apparatus, or device. The program code contained on the computer-readable medium may be transmitted using any suitable medium, including but not limited to wireless, electrical, optical, RF, or any suitable combination thereof.

[0053] The flow charts and block diagrams in the accompanying drawings illustrate the possible implementation architecture, functions and operations of the system, method and computer program product according to various embodiments of the present invention. In this regard, each box in the flow chart or block diagram can represent a module, program segment, or a part of code, and the module, program segment, or a part of code contains one or more executable instructions for realizing the specified logical function. It should also be noted that in some alternative implementations, the functions marked in the box can also occur in a different order than that marked in the accompanying drawings. For example, two boxes represented in succession can actually be executed substantially in parallel, and they can sometimes be executed in the opposite order, depending on the functions involved. It should also be noted that each box in the block diagram and / or flow chart, and the combination of the boxes in the block diagram and / or flow chart, can be implemented with a dedicated hardware-based system that performs the specified function or operation, or can be implemented with a combination of dedicated hardware and computer instructions.

[0054] Those skilled in the art should understand that the above is only a specific implementation method of the present application, but the scope of protection of the present application is not limited thereto. Any technician familiar with this technical field can easily think of changes or replacements within the technical scope disclosed in the present application, which should be covered by the scope of protection of the present application.

Claims

1. A method for optimizing a polypeptide-induced biofermentation medium, characterized in that: The method comprises the following steps: Establish ideal experimental environment conditions for biofermentation culture medium, and add a preset concentration of peptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase; After the product induction period, the actual yield of the target product in the culture medium is detected. The actual yield refers to the ratio increased by adding the peptide as a signal molecule on the original basis. When the actual yield is lower than the target yield, the peptide concentration optimization strategy is implemented; The peptide concentration optimization strategy involves dividing the time period between the cells' logarithmic growth phase and the product induction phase into multiple window periods. The peptide is then added in equal portions according to the original dosage in multiple window periods, and the post-induction concentration of the target product in each window period is measured. The real-time yield rate is obtained by the concentration of the target product after induction in each window period, and the yield rate growth in each window period is compared with the yield rate improvement threshold. The window period with a yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted; After executing the peptide concentration optimization strategy, the adjusted yield of the target product in the culture medium is tested. When the adjusted yield is less than the estimated target yield, a tolerance threshold should be set and the optimized phase difference value should be obtained. The optimized phase difference value should be compared with the tolerance threshold. If the optimized phase difference value is greater than the tolerance threshold, the judgment that the overall process needs to be improved is maintained; if the optimized phase difference value is less than or equal to the tolerance threshold, the peptide optimization effect is judged. When the adjusted yield fails to reach the estimated target yield, and the optimized improvement is greater than or equal to the improvement threshold, the window period with the highest rate increase is set as the optimal window period. When adding peptides during the optimal window period, a method that allows the peptides to be slowly released during the optimal window period is used to extend the actual induction time of the peptides during the optimal window period.

2. The method for optimizing a polypeptide-induced biofermentation medium according to claim 1, wherein: The specific method of setting the real-time acquisition rate is to obtain the estimated target yield Yc=Jc×Dn by the basic yield Jc of the target product and the target yield rate Dn, and to obtain the real-time acquisition rate Sn=Nr÷Yc by the estimated target yield Yc and the target product induced concentration Nr in the corresponding window period; the specific method of setting the acquisition rate improvement threshold is to obtain the real-time acquisition rate Sn in the initial window period. 1 Obtain the rate difference Dc=1-Sn 1 The rate improvement threshold Dt=Dc÷Cs is obtained by obtaining the rate difference Dc and the total number of window periods Cs.

3. The method for optimizing a polypeptide-induced biofermentation medium according to claim 2, wherein: The specific method for adjusting the window period to be optimized is: dilute and concentrate the preset concentration of polypeptide according to the polypeptide concentration adjustment amount, and add the diluted concentration and concentrated concentration of polypeptide to replace the preset concentration of polypeptide in the window period to be optimized for testing, obtain the increase in the yield rate of the window period to be optimized under the conditions of using the preset concentration, diluted concentration and concentrated concentration of polypeptide, compare the increase in the yield rate of the window period to be optimized when adding the preset concentration, diluted concentration and concentrated concentration of polypeptide, and make corresponding responses according to the comparison results.

4. The method for optimizing a polypeptide-induced biofermentation medium according to claim 3, wherein: If the increase in the window to be optimized is the largest when adding the peptide at the preset concentration, no adjustment is made to the added peptide concentration; If the increase in the window period to be optimized is the largest when adding the diluted concentration peptide, then the dilution concentration of the peptide is continued to be reduced according to the peptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained; If the increase in the window period to be optimized is the largest when the concentrated concentration of the peptide is added, the dilution concentration of the peptide is increased incrementally according to the peptide concentration adjustment amount until the maximum increase in the window period to be optimized is obtained.

5. The method for optimizing a polypeptide-induced biofermentation medium according to claim 4, wherein: After executing the peptide concentration optimization strategy, detect the adjusted yield of the target product in the culture medium and compare the adjusted yield with the estimated target yield. When the adjusted yield is greater than or equal to the estimated target yield, the peptide concentration optimization strategy is used as the subsequent operation indicator; when the adjusted yield is less than the estimated target yield, a decision should be made to improve the overall process.

6. The method for optimizing a polypeptide-induced biofermentation medium according to claim 5, wherein: The specific method for determining the effect of peptide optimization is to set the improvement threshold, compare the adjusted acquisition rate Td with the real-time acquisition rate Sn in the initial window period before peptide concentration optimization. 1 The difference is the adjusted growth value Tz=Td-Sn 1 The real-time acquisition rate Sn of the final window period before peptide concentration optimization is performed 5 The real-time rate Sn obtained during the initial window period before performing peptide concentration optimization 1 Take the difference to get the unadjusted growth value Tu=Sn 5 -Sn 1 The optimized improvement Fr=Tz÷Tu is obtained by dividing the adjusted growth value Tz by the growth value Tu before adjustment. The optimized improvement is compared with the improvement threshold. If the optimized improvement is greater than or equal to the improvement threshold, the peptide concentration optimization strategy is used as the subsequent operation indicator.

7. A polypeptide-induced bio-fermentation medium optimization system, characterized in that: The method for optimizing a polypeptide-induced biofermentation medium according to any one of claims 1 to 6, comprising: The environment construction module is used to build the ideal experimental environment conditions for biological fermentation medium, and add a preset concentration of peptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase; The improvement comparison module detects the actual yield of the target product in the culture medium after the product induction period. The actual yield refers to the ratio increased by adding peptides as signal molecules on the original basis. When the actual yield is lower than the target yield, the peptide concentration optimization strategy is implemented; Strategy execution module: The peptide concentration optimization strategy includes dividing the time period between the cells entering the logarithmic growth phase and the product induction phase into multiple window periods. The peptide is added in equal amounts in multiple window periods according to the original addition amount, and the concentration of the target product after induction in each window period is detected; The condition optimization module obtains the real-time yield rate through the concentration of the target product after induction in each window period, and compares the yield rate growth in each window period with the yield rate improvement threshold. The window period with the yield rate growth less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the peptide added in the window period to be optimized is adjusted.

8. A computer-readable storage medium, characterized in that The computer-readable storage medium stores a computer program, and the computer program is executed by a processor to implement the method for optimizing a polypeptide-induced bio-fermentation medium according to any one of claims 1 to 6.

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