Polypeptide induction type biological fermentation culture medium optimization method and system

Abstract

The invention relates to the technical field of polypeptide induction, and discloses a polypeptide induction type biological fermentation culture medium optimization method and system.The method comprises the steps that ideal experimental environment conditions suitable for a biological fermentation culture medium are established, and polypeptide with the preset concentration is added to serve as signal molecules when cells enter a product induction period from a logarithmic 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 increased ratio after polypeptide is added to serve as a signal molecule on the original basis, and when the actual yield is smaller than the standard yield, a polypeptide concentration optimization strategy is executed. According to the method disclosed by the invention, the to-be-optimized window period with an unobvious reaction effect is accurately identified by detecting the concentration of the target product in real time and calculating the real-time obtaining rate by utilizing a method of adding polypeptide signals step by step in stages in the window period, so that the polypeptide adding condition is optimized aiming at the to-be-optimized window period.

Description

Technical Field

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

[0002] Peptides can act as signal molecules and trigger intracellular signal transduction cascades by binding to receptors in host cells or on the cell surface, thereby inducing gene expression or activation of metabolic pathways related to the synthesis of target products. By utilizing the signals induced by polypeptides, 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 purpose 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 purpose of increasing the yield of the target product, it is critical to scientifically and rationally design and optimize the conditions for peptide addition. Summary of the invention

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

[0005] In order to achieve the above-mentioned purpose, the present invention provides the following technical solution: a method for optimizing a polypeptide-induced biological fermentation medium, the method comprising the following steps: Establish 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; 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 less than the target yield, the peptide concentration optimization strategy is implemented; The peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of cells to the product induction phase into multiple window periods, adding the peptides in equal amounts in the multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; The real-time yield rate is obtained by the induction concentration of the target product in each window period, and the yield rate growth amount in each window period is compared with the yield rate improvement threshold. The window period with a yield rate growth amount 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.

[0006] 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 obtain the real-time yield 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 yield rate improvement threshold is to obtain the real-time yield rate Sn in the initial window period. 1 The rate difference Dc=1-Sn is obtained 1 And the rate improvement threshold Dt=Dc÷Cs is obtained by obtaining the rate difference Dc and the total number of window periods Cs.

[0007] In some embodiments, the specific method for 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 the 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 a corresponding response according to the comparison result.

[0008] In some embodiments, if the increase in the window period to be optimized is the largest when the preset concentration of polypeptide is added, the concentration of the added polypeptide is not adjusted; If the increase in the window period to be optimized is the largest when adding the diluted concentration peptide, 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 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.

[0009] 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 judgment should be made to improve the overall process.

[0010] In some embodiments, when the adjusted yield is less than the estimated target yield, a tolerance threshold should also be set, and an optimized phase difference value should be obtained, and 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 improvement is required is maintained; if the optimized phase difference value is less than or equal to the tolerance threshold, the polypeptide optimization effect judgment is performed.

[0011] 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 obtained to obtain the adjusted growth value Tz=Td-Sn 1 , and 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 The difference is obtained to obtain the growth value before adjustment Tu=Sn 5 -Sn 1 , the optimized improvement Fr=Tz÷Tu is obtained by adjusting the growth value Tz after adjustment and the growth value Tu before adjustment, and 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.

[0012] In some embodiments, when the adjusted yield fails to reach the estimated target yield and the optimized increase is greater than or equal to the increase threshold, the window period with the highest rate increase is set as the optimal window period, and when the polypeptide is added during the optimal window period, a method that allows the polypeptide to be sustained-released during the optimal window period is used to extend the actual induction time of the polypeptide during the optimal window period.

[0013] The present invention also provides the following technical solution: a polypeptide-induced biological fermentation medium optimization system, comprising: The environment building module builds the ideal experimental environment conditions for biological fermentation medium, and adds a preset concentration of peptides as signal molecules 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 less than the target yield, the peptide concentration optimization strategy is implemented; Strategy execution module: the peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of cells to the product induction phase into multiple window periods, adding peptides in equal amounts in multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; The condition optimization module obtains the real-time yield rate through the induction concentration of the target product in each window period, and compares the yield rate growth amount in each window period with the yield rate improvement threshold. The window period with the yield rate growth amount 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.

[0014] The present invention further provides a computer-readable storage medium, wherein the computer-readable storage medium stores a computer program, and the computer program is executed by a processor to implement the above-mentioned polypeptide-induced biological fermentation medium optimization method and system.

[0015] Compared with the prior art, the technical solution provided by the present invention has the following beneficial effects: Firstly, the present invention utilizes a method of adding polypeptide signals in stages and steps within a window period, and detects the concentration of the target product 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.

[0016] 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 the 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.

[0017] Thirdly, the present invention realizes a dynamic evaluation of the induction effect of the polypeptide 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 used as subsequent production indicators.

[0018] Fourthly, the present invention uses the optimal window period as the key induction period and adopts a sustained release method during this period, which not only prolongs 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 the yield loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a schematic diagram of a process for optimizing a polypeptide-induced biological fermentation medium of the present invention; Figure 2 This is a module schematic diagram of a polypeptide-induced biological fermentation medium optimization system of the present invention. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. 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 creative work are within the scope of protection of the present invention.

[0021] 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 element may be multiple, and the term "one" should not be understood as a limitation on the quantity.

[0022] The present invention provides a method for optimizing a polypeptide-induced biological fermentation medium. Figure 1 As shown, including: Step 1: Establish ideal experimental environment conditions suitable for biological fermentation medium, including temperature control, pH value control and dissolved oxygen level control, and add a preset concentration of polypeptide as a signal molecule when the cells enter the product induction phase from the logarithmic growth phase. At this time, the cells are in an active metabolic state and are more sensitive to the induction signal. It should also be ensured that the polypeptide is fully dissolved in the medium to avoid weakening of activity due to excessive local concentration or precipitation; 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 polypeptide as a signal molecule on the original basis. For example, if the target product is protein, the protein yield is 1g / L on the original basis, and the protein yield is 1.5g / L after adding the polypeptide as a signal molecule on the original basis, then the yield is 150%. The actual yield is compared with the preset standard yield. When the actual yield is greater than or equal to the standard yield, it means that the expected target product yield can be obtained according to the current polypeptide addition conditions; and when the actual yield is less than the standard yield, it means that the expected target product yield cannot be obtained according to the current polypeptide addition conditions, then the polypeptide concentration optimization strategy is executed; Step 3, executing the peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of the cells to the product induction phase into multiple window periods, adding the peptide in equal amounts in the multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; Step 4: The real-time yield rate is obtained through the induction concentration of the target product in each window period, and the yield rate growth amount in each window period is compared with the yield rate improvement threshold. The window period in which the yield rate growth amount is less than the yield rate improvement threshold is set as the window period to be optimized, and the concentration of the polypeptide added in the window period to be optimized is adjusted.

[0023] 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 acquisition rate Dn, and then obtain the real-time acquisition rate Sn=Nr÷Yc by the estimated target yield Yc and the induced concentration Nr of the target product in the corresponding window period. The specific method of setting the acquisition rate enhancement threshold is to calculate the real-time acquisition rate Sn in the initial window period. 1 The rate difference Dc=1-Sn is obtained 1 And the rate improvement threshold Dt=Dc÷Cs is obtained by obtaining the rate difference Dc and the total number of window periods Cs.

[0024] 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 products 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 achievement rate is set to 200%. The estimated target yield is 2 g / L, and the achievement rate improvement threshold is 15%. The real-time achievement rates of the five window periods are 25%, 30%, 45%, 65% and 75%, respectively. The real-time achievement 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 gain 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 base value, with no gain rate increase. The gain rate increase in the second window period is 5%, the gain rate increase in the third window period is 15%, the gain rate increase in the fourth window period is 20%, and the gain rate increase in the fifth window period is 10%. If the gain rate increase in the second and fifth window periods is less than the gain rate increase threshold, the second and fifth window periods are set as the window periods to be optimized. The window period to be optimized can represent the period during the entire polypeptide induction process when the effect of adding polypeptides on the yield of the target product is not obvious.

[0025] The specific method for adjusting the window period to be optimized is as follows: dilute and concentrate the polypeptide of the preset concentration according to the polypeptide concentration adjustment amount, the polypeptide concentration adjustment amount can be 1-5%, and replace the polypeptide of the preset concentration in the window period to be optimized with the polypeptide of the diluted concentration and concentrated concentration for addition test, and obtain the increase of the yield rate in the window period to be optimized under the conditions of using the preset concentration, diluted concentration and concentrated concentration polypeptide, and compare the increase of the yield rate in the window period to be optimized under the conditions of adding the preset concentration, diluted concentration and concentrated concentration polypeptide. If the increase of the yield rate in the window period to be optimized is the largest when the preset concentration polypeptide is added, it means that the preset concentration polypeptide is already the most suitable production condition for the target product in this culture medium, and it is not correct. The concentration of the added polypeptide is adjusted; if the increase in the window period to be optimized is the largest when the diluted concentration polypeptide is added, it means that the concentration of the preset concentration polypeptide is high and is not suitable for the production conditions of the target product in this culture medium. The dilution concentration of the polypeptide 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 the concentrated concentration polypeptide is added, it means that the concentration of the preset concentration polypeptide is low and is not suitable for the production conditions of the target product in this culture medium. The dilution concentration of the polypeptide 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, in combination with the above embodiment, the polypeptide concentration adjustment amount is set to 5%. Since the increase in the yield rate in the fifth window period is 10%, it is the window period to be optimized. The preset concentration of the polypeptide is 100%. The polypeptides with a dilution concentration of 95% and a concentrated concentration of 105% are added in the fifth window period for testing. After testing, the increase in the yield rate of the polypeptide in the fifth window period at the preset concentration is 10%, the increase in the yield rate in the fifth window period at a dilution concentration of 95% is 11%, and the increase in the yield rate in the fifth window period at a concentrated concentration of 105% is 9%. Among them, the increase in the yield rate at the diluted concentration is the largest, and the concentration is continued to decrease according to the polypeptide concentration adjustment amount. The increase in the yield rate after adding the polypeptide at a concentration of 90% is 13%, the increase in the yield rate after adding the polypeptide at a concentration of 85% is 15%, and the increase in the yield rate after adding the polypeptide at a concentration of 80% is 12%. When the increase in the yield rate begins to decrease, the 85% concentration with the largest increase in the yield rate is obtained as the polypeptide concentration in the fifth window period.

[0026] 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 cell activation by the peptide 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 cells, 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 the peptide will take effect when the concentration is increased, 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.

[0027] After the implementation of the peptide concentration optimization strategy, the addition method of the peptide is changed to a step-by-step addition, which can achieve a more ideal purpose of maintaining the induction effect, and by setting the window period to be optimized, the peptide concentration can be adjusted for the time period 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 the adjusted yield is 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 subsequent operation indicators; and 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.

[0028] 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, and the optimized difference is obtained by dividing the adjusted difference by the estimated target yield. The optimized difference is compared with the tolerance threshold. If the optimized difference is greater than the tolerance threshold, it means that the gap between the adjusted yield and the estimated target yield is obvious, and the judgment that the overall process needs to be improved is maintained; if the optimized difference is less than or equal to the tolerance threshold, the peptide optimization effect is determined. Specifically, the adjusted yield rate Td is compared with the real-time yield rate Sn in the initial window period before the peptide concentration optimization is performed. 1 The difference is obtained to obtain the adjusted growth value Tz=Td-Sn 1 , and 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 The difference is obtained to obtain the growth value before adjustment Tu=Sn 5 -Sn 1, the optimized improvement Fr=Tz÷Tu is obtained by comparing the adjusted growth value Tz with the growth value Tu before adjustment, and the improvement threshold is set. The optimized improvement is compared with the improvement threshold. If the optimized improvement is less than the improvement threshold, it means that after the peptide concentration optimization strategy is implemented, the adjusted yield fails to reach the estimated target yield, and the target product can only increase a limited yield, indicating that the change in peptide conditions has little effect on cells in such culture media, and the judgment that overall process improvement is needed should be maintained; if the optimized improvement is greater than or equal to the improvement threshold, it means that after the peptide concentration optimization strategy is implemented, the target product can increase the yield significantly, indicating that the change in peptide conditions has a significant response to cells in such culture media, and the peptide optimization conditions designed above are used as subsequent operation indicators. This is because different production systems or cell lines may have large differences in sensitivity to peptides. By setting a tolerance threshold and quantifying the optimized improvement, the actual effect can be captured in dynamic adjustment. Only when the peptide optimization conditions do show a stable and significant positive effect, these conditions are considered suitable for subsequent production and are executed 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 the peptide concentration optimization is performed 1 When the adjusted yield is 25%, dividing the adjusted yield by the estimated target yield can get an adjusted yield of 90%, that is, the adjusted growth value is 65%, the growth value before adjustment is 50%, the optimized improvement is 1.3, and the improvement threshold is set to 1.2. Since the optimized improvement is greater than the improvement threshold, the peptide optimization conditions designed above are used as subsequent operation indicators.

[0029] Furthermore, when the adjusted yield fails to reach the estimated target yield, but the optimized increase is greater than or equal to the increase 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 polypeptide 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 polypeptides can bring about the maximum increase in product concentration. When adding polypeptides during the optimal window period, a method is designed to allow the polypeptide to be slowly released during the optimal window period, such as using microcapsules, liposomes and other carriers. The peptide is encapsulated into a sustained-release preparation so that the duration of the peptide action can be extended throughout the optimal window period, and the actual induction time of the peptide in the optimal window period is extended. Taking the optimal window period as the key induction period and adopting a sustained-release method can help reduce the volatility between batches and make the fermentation process more stable and controllable. This is different from directly adding the peptide once in the optimal window period, which can maintain a relatively constant peptide concentration throughout the optimal window period. Although it increases the production steps, when there is a gap between the adjusted yield and the estimated target yield, this method can further reduce the gap between the adjusted yield and the estimated target yield. In addition, the optimal window period is the time period when cells are most sensitive and efficient in responding to peptide signals. At this time, the metabolic state and signal pathways of the cells are in the activation stage, and the sustained-release peptide signals can be fully utilized to maximize the synthesis of the target product. Cells in other window periods are less sensitive to peptide signals, and the sustained-release effect may be difficult to achieve the expected induction effect. And targeted sustained-release in the optimal window period can simplify process control and avoid introducing unnecessary variables in other window periods where cell responses are weak.

[0030] In general, the present invention aims to design a method for optimizing a polypeptide-induced biological fermentation medium. For the purpose of optimizing the polypeptide addition conditions in combination with the expression level of the medium receptor, the present invention uses 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. And the adjustment of the polypeptide addition conditions within the window period to be optimized takes into account the differences in sensitivity and tolerance of different production systems and cell strains 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 it will only be used as a subsequent production indicator when the peptide conditions do show a stable and obvious positive effect. By taking the optimal window period as the key induction period and adopting a slow-release method during this period, not only the actual induction time of the peptide is extended, the relatively constant peptide concentration is maintained throughout the window period, but also the cells in the optimal window period are specifically enhanced to respond to the peptide signal. 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 the yield loss.

[0031] The present invention provides a polypeptide-induced bio-fermentation medium optimization system, such as Figure 2 As shown, including: The environment building module builds the ideal experimental environment conditions for biological fermentation medium, and adds a preset concentration of peptides as signal molecules 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 less than the target yield, the peptide concentration optimization strategy is implemented; Strategy execution module: the peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of cells to the product induction phase into multiple window periods, adding peptides in equal amounts in multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; The condition optimization module obtains the real-time yield rate through the induction concentration of the target product in each window period, and compares the yield rate growth amount in each window period with the yield rate improvement threshold. The window period with the yield rate growth amount 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.

[0032] The embodiments disclosed in the present invention, the process described above with reference to the flowchart can be implemented as a computer software program. The embodiments disclosed in the present invention include a computer program product, which includes a computer program carried on a computer-readable medium, and the computer program contains a program code for executing the method shown in the flowchart. In such an embodiment, the computer program can be downloaded and installed from the network through the communication part, and / or installed from a removable medium. When the computer program is executed by the central processing unit, the above functions defined in the method of the present application are executed. It should be noted that the computer-readable medium mentioned above in the present application can be a computer-readable signal medium or a computer-readable storage medium or any combination of the above two. The computer-readable storage medium can be, for example, but not limited to, a system, device or device of an electrical, magnetic, optical, electromagnetic, infrared segment, or semiconductor, or any combination of the above. More specific examples of computer-readable storage media can include, but are not limited to: an electrical connection with one or more wire segments, a portable computer disk, a hard disk, a random access memory, a read-only memory, an erasable programmable read-only memory, an optical fiber, a portable compact disk read-only memory, an optical storage device, a magnetic storage device, or any suitable combination of the above. In the present application, a computer-readable storage medium may be any tangible medium containing or storing a program that can be used by or in combination with an instruction execution system, apparatus, or device. In the present application, a computer-readable signal medium may include a data signal propagated in a baseband or as part of a carrier wave, which carries a computer-readable program code. Such propagated data signals may take a variety of forms, including but not limited to electromagnetic signals, optical signals, or any suitable combination of the above. A computer-readable signal medium may also be any computer-readable medium other than a computer-readable storage medium, which may send, propagate, or transmit a program for use by or in combination 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 segments, wire segments, optical cables, RF, etc., or any suitable combination of the above.

[0033] The flow chart and block diagram in the accompanying drawings illustrate the possible architecture, function and operation of the system, method and computer program product according to various embodiments of the present invention. In this regard, each square box in the flow chart or block diagram can represent a module, a program segment or a part of a code, and the module, the program segment or a part of the 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 square box can also occur in a sequence different from that marked in the accompanying drawings. For example, two square 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 square box in the block diagram and / or flow chart, and the combination of the square 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.

[0034] Those skilled in the art should understand that the above description is only a specific implementation mode of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed in the present application, which should be covered by the protection scope of the present application.

Claims

1. A method for optimizing a polypeptide-induced biological fermentation medium, characterized in that: The method comprises the following steps: Establish 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; 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 less than the target yield, the peptide concentration optimization strategy is implemented; The peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of cells to the product induction phase into multiple window periods, adding the peptides in equal amounts in the multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; The real-time yield rate is obtained by the induction concentration of the target product in each window period, and the yield rate growth amount in each window period is compared with the yield rate improvement threshold. The window period with a yield rate growth amount 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.

2. The method for optimizing a polypeptide-induced biological fermentation medium according to claim 1, characterized in that: 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 acquisition rate Dn, and obtain the real-time acquisition rate Sn=Nr÷Yc by the estimated target yield Yc and the induced concentration Nr of the target product 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 The rate difference Dc=1-Sn is obtained 1 And 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 biological fermentation medium according to claim 2, characterized in that: 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 replace the preset concentration of polypeptide with the diluted concentration and concentrated concentration of polypeptide in the window period to be optimized for addition test, 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 biological fermentation medium according to claim 3, characterized in that: If the increase in the window period 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, 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 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 biological fermentation medium according to claim 4, characterized in that: 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 judgment should be made to improve the overall process.

6. The method for optimizing a polypeptide-induced biological fermentation medium according to claim 5, characterized in that: 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, and 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 judgment is performed.

7. The method for optimizing a polypeptide-induced biological fermentation medium according to claim 6, characterized in that: The specific method for determining the peptide optimization effect is: setting the improvement threshold, comparing 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 obtained to obtain the adjusted growth value Tz=Td-Sn 1 , and 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 The difference is obtained to obtain the growth value before adjustment Tu=Sn 5 -Sn 1 , the optimized improvement Fr=Tz÷Tu is obtained by adjusting the growth value Tz after adjustment and the growth value Tu before adjustment, and 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.

8. The method for optimizing a polypeptide-induced biological fermentation medium according to claim 7, characterized in that: When the adjusted yield fails to reach the estimated target yield, and the optimized increase is greater than or equal to the increase threshold, the window period with the highest rate increase is set as the optimal window period. When adding polypeptides during the optimal window period, a method that allows the polypeptides to be slowly released during the optimal window period is used to extend the actual induction time of the polypeptides during the optimal window period.

9. A polypeptide-induced biological fermentation medium optimization system, characterized in that: The method for optimizing a polypeptide-induced biofermentation medium according to any one of claims 1 to 8 comprises: The environment building module builds the ideal experimental environment conditions for biological fermentation medium, and adds a preset concentration of peptides as signal molecules 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 less than the target yield, the peptide concentration optimization strategy is implemented; Strategy execution module: the peptide concentration optimization strategy includes dividing the time period from the logarithmic growth phase of cells to the product induction phase into multiple window periods, adding peptides in equal amounts in multiple window periods in turn according to the original addition amount, and detecting the concentration of the target product after induction in each window period; The condition optimization module obtains the real-time yield rate through the induction concentration of the target product in each window period, and compares the yield rate growth amount in each window period with the yield rate improvement threshold. The window period with the yield rate growth amount 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.

10. 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 biological fermentation medium as described in any one of claims 1 to 8.

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