WO2009121211A1 - 一种发酵控制系统及发酵控制方法 - Google Patents

一种发酵控制系统及发酵控制方法 Download PDF

Info

Publication number
WO2009121211A1
WO2009121211A1 PCT/CN2008/000688 CN2008000688W WO2009121211A1 WO 2009121211 A1 WO2009121211 A1 WO 2009121211A1 CN 2008000688 W CN2008000688 W CN 2008000688W WO 2009121211 A1 WO2009121211 A1 WO 2009121211A1
Authority
WO
WIPO (PCT)
Prior art keywords
culture
distribution
value
cell population
solution
Prior art date
Application number
PCT/CN2008/000688
Other languages
English (en)
French (fr)
Other versions
WO2009121211A8 (zh
Inventor
范顺杰
杨宏伟
孔兵
贺伯特·格里布
卓越
库特·贝腾豪森
Original Assignee
西门子公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 西门子公司 filed Critical 西门子公司
Priority to PCT/CN2008/000688 priority Critical patent/WO2009121211A1/zh
Publication of WO2009121211A1 publication Critical patent/WO2009121211A1/zh
Publication of WO2009121211A8 publication Critical patent/WO2009121211A8/zh

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/26Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH

Definitions

  • the invention relates to the field of industrial automation, in particular to a fermentation control system and a fermentation control method.
  • Fermentation (biological reaction) processes are important for the pharmaceutical, energy, food, beverage, environmental engineering and other industries.
  • the fermentation process is a method of cultivating a culture (microorganism, animal cell or plant cell) in a certain amount of nutrient solution (nutrient solution for growth of the culture), and currently three general fermentation processes include : Batch processing, fed-batch processing and continuous processing.
  • the culture and nutrient solution in the fermenter were not increased or decreased by external means during the batch process.
  • the nutrient solution is continuously replenished into the fermentor, and the culture solution (the liquid composed of the culture and the nutrient solution) is also continuously discharged from the fermentor.
  • the process of fed-batch processing is relatively complicated, which means that a certain amount of nutrient solution is added to the fermenter one or more times in the stage of culture growth and product formation on the basis of batch processing, wherein, The amount of nutrient solution is preset or calculated in real time, and during the process, the culture solution is not discharged until one fermentation is completed.
  • the amount of nutrient solution is preset or calculated in real time, and during the process, the culture solution is not discharged until one fermentation is completed.
  • the fermentation conditions can be optimized to obtain more and better products.
  • FIG. 1 is a logical structural diagram of a fermentation control system in the prior art.
  • the system performs multi-loop control of the fermentation process.
  • the pH of the culture solution is controlled by a closed loop, and the injection of the nutrient solution is controlled by an open loop.
  • the PH sensor sends the measured pH value of the culture solution to the PH controller; the PH controller compares the PH value measured by the pH sensor with a preset PH threshold value, according to different The comparison result controls the switch of the acid/base control valve (such as solenoid valve or control valve) to open for a certain period of time, so as to inject a certain amount of acid/base solution into the fermenter to control the culture.
  • the pH of the liquid such as solenoid valve or control valve
  • the pH sensor then measures the pH of the culture solution again and feeds the newly measured pH value back to the pH controller for the next round of control.
  • the nutrient solution feed controller is based on the nutrient solution feed curve shown on the left side of Figure 1, that is, the curve of the nutrient solution flow rate with time, and the switch of the nutrient solution feed valve is controlled to open for a specific time to achieve the fermentation tank. A certain amount of nutrient solution is injected to control the fermentation process.
  • the cell population in the fermenter is not a continuous uniform object, but a distributed cell population, that is, the properties of the culture cells in the cell population, such as cell size, vigor and age are different. .
  • the culture solution in the fermenter is also largely affected by changes in cell distribution. For example, as the fermentation reaction progresses, changes in the distribution of the culture caused by changes in the physiological state of cells in the fermentor cause the culture solution. Changes in temperature and pH affect the control of the fermentation process.
  • the control scheme in the prior art only treats the cell population as a continuous uniform object, that is, the cells in the fermenter are considered to be the same, and there is no difference in properties such as size, vitality or age, and the cells in the cell metabolism cycle are not considered. The influence of changes in the internal physiological state on the fermentation process, so that the fermentation process cannot be better controlled, and the efficiency of the fermentation process is low. .
  • a fermentation control system provided by the present invention includes: a PH value sensor for acquiring pH value data of a culture solution; the system further comprising: a culture distribution sensor and a culture distribution controller ;
  • the culture distribution sensor is configured to obtain culture distribution data, wherein the culture distribution data includes a cell population distribution width of the culture;
  • the culture distribution controller is configured to perform multivariate on the pH value of the culture solution and the cell population distribution width of the culture according to the culture liquid pH value data acquired by the pH value sensor and the culture distribution data acquired by the culture distribution sensor. control.
  • the culture distribution data obtained by the culture distribution sensor further includes a cell population average property of the culture
  • the culture distribution controller is further configured to control the average population of the cell population of the culture based on the culture distribution data acquired by the culture distribution sensor.
  • the system further comprises a pH controller for controlling the pH of the culture solution and the cell population distribution width of the culture under the control of the culture distribution controller. Take control.
  • the system further comprises a nutrient solution controller for controlling the cell population average property of the culture under the control of the culture distribution controller.
  • the culture distribution sensor is an online flow cytometer.
  • the culture distribution controller is a programmable logic controller.
  • a fermentation control method provided by the present invention includes:
  • the pH value of the culture solution and the cell population distribution width of the culture were subjected to multivariate control.
  • the multi-variable control is performed on the pH value of the culture solution and the cell population distribution width of the culture:
  • the pH of the culture solution is controlled, and the distribution of the cell population of the culture is controlled by the pH value of the culture solution.
  • the step of controlling the pH value of the culture solution and controlling the distribution width of the cell population of the culture using the pH value of the culture solution is:
  • the pH value of the culture solution is controlled so that the cell population distribution width of the culture satisfies the requirement of the width threshold value.
  • the step of controlling the pH value of the culture solution and controlling the distribution width of the cell population of the culture using the pH value of the culture solution For:
  • the obtained culture distribution data further includes a cell population average property of the culture; the method further comprising:
  • the average population of the cell population of the culture is controlled.
  • the control of the average attribute of the cell population of the culture is performed before the control of the distribution width of the cell population of the culture.
  • the average population attribute of the culture of the culture is controlled as follows:
  • the average population properties of the culture are controlled by controlling the feeding operation of the nutrient solution.
  • the step of controlling the average attribute of the cell population of the culture by controlling the feeding operation of the nutrient solution is:
  • each of the PH threshold value, the width threshold value, and the average property threshold value are determined in advance by an experimental or mathematical model.
  • the pH value of the culture solution in the fermenter and the culture distribution data are obtained, and the pH value of the culture solution and the culture in the culture solution are obtained based on the obtained pH value data of the culture solution and the culture distribution data.
  • the multivariate control of the distribution of substances can improve the production efficiency and product quality of the industrial fermentation process, and can be applied to a variety of fermentation processes, such as biopharmaceuticals, energy, food processing, and the like.
  • the present invention can be implemented on the basis of the existing automatic fermentation control system in the specific implementation, without major changes to the existing fermentation control system, and therefore has low cost, high efficiency and easy implementation. advantage.
  • FIG. 1 is a logical structural diagram of a fermentation control system in the prior art
  • FIG. 2 is a logical structural diagram of a fermentation control system according to an embodiment of the present invention.
  • FIG. 3 is an exemplary structural diagram of a fermentation control system in an embodiment of the present invention.
  • Figure 4 is a schematic view showing the structure and connection relationship of the nutrient solution measuring tank in the fermentation control system shown in Figure 3;
  • FIG. 5 is an exemplary flowchart of a fermentation control method according to an embodiment of the present invention.
  • Figure ⁇ is an exemplary flow chart of a fermentation control method in another embodiment of the present invention.
  • FIG. 2 is a logical block diagram of a fermentation control system in accordance with a preferred embodiment of the present invention.
  • the fermentation control system of this embodiment adds a culture distribution controller and a culture distribution sensor to the system shown in Fig. 1.
  • the culture distribution controller measures the PH value of the culture solution in the fermenter and the distribution of the culture through the pH sensor and the culture distribution sensor, and the measurement results control the fermentation process to achieve the purpose of optimizing the fermentation process.
  • the pH sensor measures the pH of the fermentor and feeds it back to the culture distribution controller.
  • the culture distribution sensor measures the distribution of the culture in the fermentor and feeds it back to the culture distribution controller.
  • the culture distribution controller compares the PH value data measured by the pH sensor and the culture distribution data measured by the culture distribution sensor with the set threshold value, and can respectively be compared to the PH value controller according to the comparison result.
  • the nutrient solution feed controller sends acid/alkaline solution feed data and nutrient solution feed data. After receiving the above acid/base solution feed data, the pH controller can inject the acid/base solution into the fermenter by controlling the switch of the acid/sincere control valve. After receiving the nutrient solution feed data, the nutrient solution feed controller can inject nutrient solution into the fermenter by controlling the switch of the nutrient solution feed valve.
  • the culture distribution controller is the main controller, the PH value controller and the nutrient liquid feeding controller are slave controllers, and the main controller controls the alkali solution by controlling the slave controllers.
  • the feeding operation of the liquid and the feeding operation of the nutrient solution make it possible to implement the invention with minor modifications on the basis of the existing fermentation control system, reducing the cost and complexity of upgrading the existing system. If the problem of upgrading the existing fermentation control system is not considered, the fermentation control system of the present invention can directly control the feeding operation of the acid/alkaline solution and the feeding operation of the nutrient solution by using only the culture distribution controller.
  • Fig. 3 is an exemplary structural diagram of a fermentation control system based on the logical structure shown in Fig. 2.
  • the system comprises: fermenting tank 1, nutrient liquid feeding valve 2, acid/alkali control valve 3, acid/subduction metering tank 4, nutrient liquid measuring tank 5, pH controller 6, nutrient solution
  • the fermentor 1 operates in a fed-batch mode.
  • the fermentor 1 has an outer casing 16 that can be heated or cooled, and an agitator 15 for agitating the culture liquid 17.
  • the agitator 15 is used to provide a mixture between the culture and the nutrient solution in the fermentor 1. It should be noted that the agitator 15 should avoid excessive agitation to agitate the culture cells while stirring.
  • the nutrient solution feed valve 2 is used to open or close under the control of the nutrient solution feed controller 7, and when the nutrient solution feed valve 2 is opened, the nutrient solution metering tank 5 injects the nutrient solution into the fermentor 1, preferably,
  • the nutrient solution feed valve 2 can be a solenoid valve or a control valve.
  • the nutrient solution metering tank 5 is for storing a nutrient solution, such as a glucose solution, a nitrogen-containing solution, etc., and supplements the nutrient solution to the fermentor 1 through the nutrient solution feed valve 2.
  • the nutrient solution metering tank 5 may further include an input port valve for controlling the nutrient solution injection of the nutrient solution metering tank 5, and the inlet port valve may also be controlled by the nutrient solution feed controller 7.
  • FIG. 4 is a schematic view showing the structure of a nutrient solution measuring tank 5 and its connection relationship in the present embodiment.
  • the nutrient solution feeding controller 7 controls the nutrient solution measuring tank 5.
  • the input port valve (first solenoid valve 19) and the metering tank 5 flow to the valve of the fermentor 1 (the second solenoid valve 20, that is, the nutrient solution feed valve 2 in Fig. 3).
  • the nutrient solution metering tank 5 has two level electrodes 18, and when the nutrient solution is injected into the nutrient solution metering tank 5, the two electrode circuits are respectively turned on, the electrodes generating a signal representative of the liquid position of the nutrient solution metering tank.
  • the first electromagnetic valve 19 controls the injection of the nutrient solution into the nutrient solution metering tank 5, and the second solenoid valve 20 controls the injection of the nutrient solution from the nutrient solution metering tank 5 to the fermentor 1.
  • the second solenoid valve 20 connected to the fermentor 1 is closed under the control of the nutrient solution fed controller 7, and the first solenoid valve 19 connected to the nutrient solution metering tank 5 is opened to make nutrition
  • the liquid metering tank 5 is loaded with nutrient solution.
  • the second solenoid valve 20 is hit After a certain amount of nutrient solution (for example, >1 liter or 2 liters of nutrient solution) is injected into the fermenter 1, the second solenoid valve 20 is closed, waiting for the control command of the nutrient solution feeding controller 7 in the next feeding cycle. .
  • the acid tank 11 is for storing an acid solution; the acid solution pump 13 is for pumping the acid solution into the acid/base metering tank 4 under the control of the pH controller 6.
  • the alkali tank 12 is for storing an alkali solution; the alkaline solution pump 14 is for pumping the alkali solution into the acid/base metering tank 4 under the control of the pH controller 6.
  • the alkali metering tank 4 is used to supply the acid solution or the alkali solution to the fermentor 1 through the ⁇ /subtraction control valve 3.
  • the acid/base control valve 3 is used to open or close under the control of the pH controller 6.
  • the acid/base control valve 3 is opened, the acid/base metering tank 4 injects an acid solution or an alkali solution into the fermentor 1 through the acid/base control valve 3.
  • the acid/base control valve 3 may be a solenoid valve or a control valve.
  • the nutrient solution metering tank 5 and the alkali metering tank 4 may each be a metering tank with a flow rate measuring device, wherein the flow rate measuring device may be used to measure the flow rate of the nutrient solution flowing into the fermentor 1 or the acid/base solution flowing into the fermentor 1 Flow rate.
  • the flow rate measuring device can be a solenoid valve or a control valve.
  • the nutrient solution feeding controller 7 is used for controlling the nutrient solution feeding operation, and according to the nutrient solution feeding data of the culture distribution controller 8, the switch of the nutrient solution feeding valve 2 is controlled to open, and the nutrient solution measuring tank 5 is realized. The amount of nutrient solution is injected into the fermentor 1.
  • the pH controller 6 is configured to control the acid solution pump 13 or the alkaline solution pump 14 to pump the acid solution or the solution to the acid/antium metering tank 4 according to the acid/reduction solution feeding data of the culture distribution controller 8.
  • the switch of the acid/base control valve 3 is controlled to open, and a certain amount of the alkali/alkali solution in the alkali metering tank 4 is injected into the fermentor 1.
  • the pH sensor 9 is used to measure the pH value in the fermentor 1 and transmit the measured pH value data back to the culture distribution controller 8.
  • the pH sensor 9 can be a PH electrode 9 for real-time measurement of the pH in the fermentor 1.
  • the culture distribution sensor 10 is used to measure the culture distribution in the fermentor 1, and the measured culture distribution data is transmitted back to the culture distribution controller 8. Among them, the culture distribution data includes "average attribute of cell population” and “distribution width of cell population”.
  • the average property of the cell population refers to the average of the cell properties in the cell population and is used to reflect the overall trait of the culture cells. Targeting the size, vitality and age of the culture cells, the cell population can Has average properties such as size, vitality, and age.
  • the distribution width of the cell population is a parameter that reflects the heterogeneity of the culture cells.
  • the small distribution width of the cell population indicates that the culture cells have uniform properties and small differences, and the large distribution width of the cell population indicates that the culture cells have heterogeneous and large differences in properties.
  • the distribution width of the cell population is a dimensionless value, and the distribution width of the cell population can be calculated according to the properties of the size, vitality or age of the culture cells.
  • Distribution width of cell population D90 z D10
  • /) 90 represents a particle size value, 90% of the cells in the cell population have a diameter smaller than the particle size; D10 represents another particle size value, and 10% of the cells in the cell population have a diameter smaller than the particle size.
  • the culture distribution sensor 10 can be a variety of advanced biochemical sensors, for example, an online flow cytometer as described in U.S. Patent 6,555,360, or other suitable measuring instrument.
  • the culture distribution controller 8 serves as a main controller, and detects the pH value and the culture distribution of the culture liquid 17 in the fermentor 1 by the pH value sensor 9 and the culture distribution sensor 10, respectively, and detects the detected pH value data and the culture distribution data, respectively. Comparing with the set threshold value, the alkali solution feed data and the nutrient solution feed data are sent to the pH controller 6 and the nutrient solution feed controller 7 respectively according to the comparison result.
  • the threshold value of the pH data and the threshold value of the culture distribution data may be determined in advance by an experimental or mathematical model.
  • the culture distribution controller 8 is a multivariable controller that controls the pH value and culture distribution of the culture solution, that is, between the change in the distribution of the culture and the change in the pH of the culture solution in the control process. Interaction, based on the existing simple control of the pH of the culture solution, while using the pH value of the culture solution to control the distribution of the cell population distribution of the culture, and preferably, further using the amount of nutrient solution to feed Control the average properties of the cell population of the culture to optimize control of the fermentation process.
  • the culture distribution controller 8, the nutrient solution controller 7, and the pH controller 6 can be realized by an electronic device, for example, an industrial control computer, a programmable logic controller (PLC), Or a combination thereof. Communication between devices can be achieved via standard 4 to 20 mA (mA) electrical signals or electrical signals in accordance with fieldbus standards such as Profibus.
  • Both the nutrient solution controller 7 and the pH controller 6 can use a proportional-integral-derivative method (PID), on/off or
  • PID proportional-integral-derivative method
  • TPC time proportional control
  • the control method provided by TPC is relatively low in cost based on relatively low control complexity. During each feed cycle, the TPC changes the time ratio of the valve switch based on changes in the manipulated variable, for example, the amount of feed flow.
  • FIG. 5 Based on the system shown in Fig. 3, an exemplary flow chart of the fermentation control method in the embodiment of the present invention is shown in Fig. 5.
  • the process includes the following steps:
  • Step 501 Obtain PH data of the culture solution.
  • the PH value data of the culture solution can be obtained by the PH value sensor 9.
  • Step 502 Determine whether the PH value of the culture solution exceeds the set PH threshold. If yes, go to step 503; otherwise, go to step 504.
  • the culture distribution controller 8 compares the PH value obtained by the pH sensor 9 with the set PH threshold value, and determines whether the PH value of the culture solution satisfies the set PH threshold value, and Select the next step based on the comparison result.
  • Step 503 Control the PH value of the culture solution to meet the set PH threshold requirement, and then perform step 504.
  • the culture distribution controller 8 controls the pH of the culture solution to meet the set PH threshold.
  • the acid distribution controller 6 sends acid/base solution feed data to the pH controller 6, and the pH controller 6 controls the acid/base control valve 3 based on the ⁇ /base solution feed data.
  • the pH of the culture solution satisfies the set pH threshold.
  • Step 504 obtaining culture distribution data.
  • the culture distribution data of the culture solution can be obtained by the culture distribution sensor 10, wherein the culture distribution data includes the distribution width of the cell population of the culture.
  • Step 505 Determine whether the cell population distribution width of the culture is greater than a set width threshold. If yes, execute step 506, otherwise end the control process.
  • the culture distribution controller 8 compares the distribution width of the cell population acquired by the culture distribution sensor 10 with the set width threshold value, and determines whether the cell population distribution width of the culture satisfies the set width gate P. Depreciate the requirements and select the next steps based on the comparison results. Step 506, controlling the distribution width of the cell population of the culture to meet the set width threshold requirement and ending the control process.
  • the culture distribution controller 8 further utilizes the pH value of the culture solution to control the distribution of the cell population distribution of the culture to meet the set width threshold.
  • the culture distribution controller 8 sends the / alkali solution feed data to the pH controller 6, and the pH controller 6 controls the acid/base control valve 3 based on the ⁇ /base solution feed data.
  • the culture cells having undesired properties are reduced, and the culture cells having the desired properties are increased, thereby reducing the difference between the properties of the culture cells, and making the cell population of the culture
  • the distribution width satisfies the requirements of the set width threshold.
  • the culture distribution data extracted in step 504 further includes the average group attribute of the culture, so that after step 504, the distribution width of the cell population of the culture is controlled.
  • the culture distribution controller 8 can compare the average group attribute obtained by the culture distribution sensor 10 with a set average attribute threshold, and the average attribute of the cell group does not satisfy the threshold.
  • the nutrient solution feed data is sent to the nutrient solution feed controller 7 to control the feeding operation of the nutrient solution, so that the average attribute of the cell population meets the threshold value, that is, the culture cells are The overall trait first satisfies the set requirements, thus further optimizing the fermentation control process.
  • the vertical axis represents the pH value
  • the pH value gradually increases in the direction of the arrow
  • the horizontal axis represents the fermentation time.
  • the black solid line in the figure is a plot of the pH of the culture solution over time during the fermentation.
  • the two dotted lines corresponding to a maximum PH value and a minimum PH value in the figure are local control lines (LCL, Local Control Limit), which indicate the maximum alkaline pH and the maximum acidic pH value that the culture cells can withstand, respectively. That is, the upper and lower thresholds of the set PH value.
  • LCL Local Control Limit
  • the pH of the culture medium exceeds the pH range indicated by the two LCLs, for example, the pH of the culture solution is less than the set maximum acidic pH value, or greater than the set maximum alkaline pH value, the culture cells The growth will be hindered, making the fermentation process impossible.
  • the dotted line immediately below the minimum PH value LCL is a fine control line (FCL, Fine Control Limit), which indicates the cell to a certain attribute.
  • FCL Fine Control Limit
  • the physiological cycle of the viable culture cells will be significantly shortened, but the physiological cycle of the viable culture cells will not change significantly.
  • another FCL (not shown in Figure 6 for clarity) can be set immediately below the maximum PH value LCL, indicating a threshold value for an alkaline growth environment.
  • the present invention can set the upper and lower limits of pH value LCL and the acidic growth environment FCL, or the upper and lower limit of pH value LCL and the alkaline growth environment FCL.
  • a dotted line is also displayed on the FCL, and the corresponding PH value is the pH value of the most suitable growth environment.
  • the culture liquid having the pH value of the optimum growth environment is suitable for the growth of all cells in the cell population.
  • FIG. 7 Based on the system shown in Fig. 3 and the pH control curve shown in Fig. 6, an exemplary flowchart of the fermentation control method in the present embodiment is shown in Fig. 7.
  • the process includes the following steps:
  • Step 701 Obtain a PH value of the culture solution.
  • the culture distribution controller 8 can obtain the PH value data of the culture solution in real time through the pH value sensor 9.
  • step 702 it is determined whether the PH value of the culture solution exceeds the PH value range indicated by the two LCLs. If yes, step 703 is performed; otherwise, step 704 is performed.
  • the culture distribution controller 8 compares the PH value obtained by the pH sensor 9 with the upper and lower limit PH thresholds indicated by the LCL, and determines whether the pH of the culture solution exceeds the upper and lower limits PH.
  • the range of values defined by the threshold value, and the subsequent steps are selected based on the comparison result.
  • Step 703 Control the PH value of the culture solution to be within the range of PH values indicated by the two LCLs, and then perform step 704.
  • the culture distribution controller 8 controls the pH of the culture solution, for example, the acid distribution controller 6 sends the acid/alkali solution feed data to the pH controller 6, and the pH controller 6 according to the The acid/base solution feed data controls the acid/base control valve 3, and the acid/base solution is injected into the culture solution.
  • the pH of the culture solution is within the pH range indicated by the two LCLs to meet the minimum requirements for cell growth of the culture.
  • Step 704 obtaining culture distribution data.
  • the pH of the culture solution is within the range of pH values indicated by the two LCLs, and then the culture distribution data of the culture solution can be obtained by the culture distribution sensor 10, wherein the culture distribution data includes the culture.
  • the distribution of cell population width is the reason for the culture solution.
  • Step 705 Determine whether the cell population distribution width of the culture is greater than a set width threshold. If yes, go to step 706. Otherwise, go to step 707.
  • Step 706 controlling the distribution width of the cell population of the culture to meet the set width threshold requirement, and performing step 707.
  • the culture distribution controller 8 sends the acid/alkali solution feed data to the pH controller 6 according to the pH control curve, and the pH controller 6 according to the alkali solution feed data to the acid/machine
  • the control valve 3 is controlled, for example, in the time t1 to t2 shown in Fig. 6, the acid/base solution is injected into the culture solution so that the pH of the culture solution is between the FCL and the minimum PH value LCL, so that it will have It is desirable to reduce the culture of the culture cells, such as reducing the culture cells with weak vigor, and increasing the number of culture cells with desired properties, such as increasing the number of culture cells with strong vigor, and then making the distribution width of the cell population meet the set
  • the width gate P ⁇ value is required.
  • step 707 the PH value of the culture solution is further controlled to maintain the pH value of the optimum growth environment, and then step 708 is performed.
  • the pH of the culture solution in order to promote the growth of the culture cells, can be further controlled to maintain the pH value of the optimum growth environment, so that the culture cells having the desired properties can grow faster and better. .
  • step 708 it is judged whether the fermentation production process is finished, and if so, the control process is ended, otherwise the process returns to step 704.
  • the culture distribution data obtained in step 704 also includes the average population of the cell population of the culture.
  • the cell group average attribute acquired by the culture distribution sensor 10 is first compared with a set average attribute threshold value, and when the average attribute of the cell group does not satisfy the requirement of the gate P ⁇ value, The nutrient solution feeding controller 7 controls the feeding operation of the nutrient solution according to the nutrient solution feeding data sent from the culture distribution controller 8 The average properties of the cell population are made to meet the threshold requirements.
  • the feed controller 7 controls the injection of a certain amount of nutrient solution into the fermenter based on the nutrient solution feed data output from the culture distribution controller 8, to promote the growth of the culture cells.
  • the present invention considers the distribution of the culture and its interaction with the pH of the culture solution in the control process, and controls the distribution of the cell population of the culture by controlling the pH value of the culture solution.
  • the width allows the production efficiency and product quality of the fermentation process to be improved.
  • the average population property of the culture can be controlled by controlling the feeding operation of the nutrient solution, so that the overall trait of the culture cell first satisfies the set requirements, thereby further improving the production efficiency of the fermentation process. And product quality.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Description

一种发酵控制系统及发酵控制方法 技术领域
本发明涉及工业自动化领域, 特别涉及一种发酵控制系统及发酵控制 方法。
背景技术
发酵 (生物反应) 处理过程对于制药, 能源, 食品、 饮料, 环境工程 等行业来说都是很重要的。 发酵处理过程是将培养物 (微生物、 动物细胞 或植物细胞) 在一定量的营养液(供培养物生长的营养物溶液) 中进行培 养的一种方法, 目前比较通用的三种发酵处理过程包括: 分批处理、 补料 分批处理和连续处理。 在分批处理过程中, 发酵罐中的培养物和营养液没 有通过外部手段增加或减少。 在连续处理过程中, 营养液被不断的补充入 发酵罐, 培养液(由培养物和营养液构成的液体)也被不断的排出发酵罐。 而补料分批处理的过程相对复杂一些, 是指在分批处理的基础上, 在培养 物生长和产物形成阶段中, 一次或多次向发酵罐中补充一定量的营养液, 其中, 添加的营养液量是预先设定的或者是实时计算获得的, 在该处理过 程中, 直到一次发酵完成时才排出培养液。 通过该过程可优化发酵条件, 进而获得更多、 更好的产物。
培养物细胞的生长需要一个合适的 PH值范围。 而在发酵过程中, 细胞 代谢物的积累, 例如有机酸或氨基酸, 会导致培养液的 PH值降低。 此外, 细胞在缺乏营养液的情况下, 会进入另一种代谢状态, 造成胞内氮源过剩, 并以氨离子的形式释放到培养液中,从而使培养液的 PH值急剧上升。 无论 PH值降低还是上升都不利于发酵过程, 实际生产中, 可采用补料分批处理 模式, 通过向发酵罐中注入酸或碱溶液以控制 PH值, 和 /或通过向发酵罐 注入营养液以控制发酵过程。 图 1 为现有技术中一种发酵控制系统的逻辑 结构图, 如图 1 所示, 该系统对发酵过程进行多回路控制。 其中, 培养液 的 PH值通过一个闭环回路进行控制,营养液补料的注入通过一个开环回路 进行控制。 该系统中, PH值传感器将测得的培养液的 PH值发送给 PH值 控制器; PH值控制器将 PH值传感器测量的 PH值与预先设定的 PH门限值 相比较, 根据不同的比较结果控制酸 /碱控制阀 (例如电磁阀或控制阀) 的 开关打开特定的时间, 实现向发酵罐中注入一定量的酸 /碱溶液以控制培养 液的 PH值。然后 PH值传感器再次测量培养液的 PH值,并将新测得的 PH 值又反馈给 PH值控制器, 从而进行下一轮的控制过程。 营养液补料控制器 才艮据图 1 左侧所示的营养液补料曲线, 即营养液流量随时间的变化曲线, 控制营养液补料阀的开关打开特定的时间, 实现向发酵罐中注入一定量的 营养液以控制发酵过程。
实际应用中, 发酵罐中的细胞群并不是一个连续均一的对象, 而是一 个分布式的细胞群体, 即, 细胞群中培养物细胞的属性, 例如细胞的大小、 活力及年龄都是不同的。 而且发酵罐中的培养液很大程度上也受到细胞分 布变化的影响, 例如随着发酵反应的进行, 发酵罐中细胞发生的生理状态 的变化所引起的培养物分布的变化会引起培养液的温度和 PH值的变化, 从 而影响对发酵过程的控制。 而现有技术中的控制方案只是把细胞群作为连 续均一的对象, 即认为发酵罐中的细胞都是一样的, 没有大小、 活力或年 龄等属性上的区别, 并没有考虑细胞新陈代谢周期中细胞内部生理状态的 变化对发酵过程的影响, 从而无法更好的控制发酵过程, 使得发酵过程的 效率较低。 .
发明内容
为了解决以上问题, 本发明的一个目的是提供一种发酵控制系统, 另 一目的是提供一种发酵控制方法。
为了实现本发明的上述目的, 本发明所提供的一种发酵控制系统, 包 括, PH值传感器, 用于获取培养液的 PH值数据; 该系统还包括, 培养物 分布传感器和培养物分布控制器;
所述培养物分布传感器用于获取培养物分布数据, 其中, 所述培养物 分布数据包括培养物的细胞群分布宽度;
所述培养物分布控制器用于根据所述 PH值传感器获取的培养液 PH值 数据和培养物分布传感器获取的培养物分布数据,对培养液的 PH值及培养 物的细胞群分布宽度进行多变量控制。
根据本发明所提供的发酵控制系统, 优选的, 所述培养物分布传感器 所获取的培养物分布数据中还包括培养物的细胞群平均属性;
所述培养物分布控制器进一步用于根据所述培养物分布传感器获取的 培养物分布数据, 对培养物的细胞群平均属性进行控制。 根据本发明所提供的发酵控制系统, 优选的, 该系统进一步包括 PH值 控制器,用于在所述培养物分布控制器的控制之下对培养液的 PH值及培养 物的细胞群分布宽度进行控制。
根据本发明所提供的发酵控制系统, 优选的, 该系统进一步包括营养 液补料控制器, 用于在所述培养物分布控制器的控制之下对培养物的细胞 群平均属性进行控制。
根据本发明所提供的发酵控制系统, 优选的, 所述培养物分布传感器 为在线流式细胞仪。
根据本发明所提供的发酵控制系统, 优选的, 所述培养物分布控制器 为可编程逻辑控制器。
为了实现本发明的上述目的, 本发明所提供的一种发酵控制方法, 包 括:
获取培养液的 PH值数据和培养物分布数据, 其中, 所述培养物分布数 据包括培养物的细胞群分布宽度;
根据所述培养液的 PH值数据和培养物分布数据, 对培养液的 PH值及 培养物的细胞群分布宽度进行多变量控制。
根据本发明所提供的发酵控制方法, 优选的, 所述对培养液的 PH值及 培养物的细胞群分布宽度进行多变量控制为:
对培养液的 PH值进行控制, 并利用培养液的 PH值对培养物的细胞群 分布宽度进行控制。
根据本发明所提供的发酵控制方法, 优选的, 所述对培养液的 PH值进 行控制,并利用培养液的 PH值对培养物的细胞群分布宽度进行控制的步骤 为:
判断培养液的 PH值是否超出.设定的 PH门限值, 如果超出, 则对培养 液的 PH值进行控制, 使其满足所述 PH门限值的要求;
判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果大于, 则对培养液的 PH值进行控制,以使培养物的细胞群分布宽度满足所述宽度 门限值的要求。
根据本发明所提供的发酵控制方法, 优选的, 所述对培养液的 PH值进 行控制,并利用培养液的 PH值对培养物的细胞群分布宽度进行控制的步骤 为:
判断培养液的 PH值是否超出 PH值控制曲线中的局部控制线所设定的 PH门限值范围, 如果超出, 则将培养液的 PH值控制在所述局部控制线所 设定的 PH门限值范围内;
判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果大于, 则对培养液的 PH值进行控制, 使其满足 PH值控制曲线中的精细控制线所 设定的 PH门限值的要求,以使培养物的细胞群分布宽度满足所述宽度门限 值的要求。
根据本发明所提供的发酵控制方法, 优选的, 所获取的培养物分布数 据中还包括培养物的细胞群平均属性; 该方法进一步包括:
根据所述培养物分布数据, 对培养物的细胞群平均属性进行控制。 根据本发明所提供的发酵控制方法, 优选的, 所述对培养物的细胞群 平均属性的控制在所述对培养物的细胞群分布宽度的控制之前进行。
根据本发明所提供的发酵控制方法, 优选的, 所述对培养物的细胞群 平均属性进行控制为:
通过控制营养液的补料操作来对培养物的细胞群平均属性进行控制。 根据本发明所提供的发酵控制方法, 优选的, 所述通过控制营养液的 补料操作来对培养物的细胞群平均属性进行控制的步骤为:
判断培养物的细胞群平均属性是否满足设定的平均属性门限值的要 求, 如果不满足, 则对营养液的补料操作进行控制, 以使培养物的细胞群 平均属性满足所述平均属性门限值的要求。
根据本发明所提供的发酵控制方法, 优选的, 所述的各 PH门限值、 宽 度门限值和平均属性门 P艮值预先通过实验或数学模型确定。
在本发明中, 通过获取发酵罐中培养液的 PH值数据和培养物分布数 据, 并根据所获取的培养液 PH值数据和培养物分布数据, 对培养液的 PH 值及培养液中的培养物分布进行多变量控制, 可以提高工业发酵过程的生 产效率和产品质量, 并且可以被应用到多种多样的发酵过程中, 例如, 生 物制药、 能源、 食品加工等。 此外, 本发明在具体实施时, 还可以在现有 的自动化发酵控制系统的基础上实现, 无需对现有的发酵控制系统进行很 大的改动, 因此还具有成本低、 效率高和易于实施的优点。 附图说明
下面将通过参照附图详细描述本发明的示例性实施例, 使本领域的普 通技术人员更清楚本发明的上述及其他特征和优点, 附图中:
图 1为现有技术中发酵控制系统的逻辑结构图;
图 2为本发明实施例中发酵控制系统的逻辑结构图;
图 3为本发明实施例中发酵控制系统的示例性结构图;
图 4为图 3所示发酵控制系统中营养液计量罐的结构及连接关系示意 图;
图 5为本发明实施例中发酵控制方法的示例性流程图;
图 6为本发明另一实施例中的 PH值控制曲线;
图 Ί为本发明另一实施例中发酵控制方法的示例性流程图。
具体实施方式
为使本发明的目的、 技术方案及优点更加清楚明白, 以下参照附图并 举实施例, 对本发明进一步详细说明。
图 2为本发明的一个优选实施例中发酵控制系统的逻辑结构图。
如图 2所示, 本实施例中的发酵控制系统在图 1所示系统基础上增加了 培养物分布控制器和培养物分布传感器。 其中, 培养物分布控制器通过 PH 值传感器和培养物分布传感器测量发酵罐中培养液的 PH值和培养物的分 布, 并 居这些测量结果控制发酵过程, 以达到优化发酵过程的目的。
根据图 2所示的逻辑结构, PH值传感器测量发酵罐中的 PH值状况, 并 反馈给培养物分布控制器。 培养物分布传感器测量发酵罐中的培养物分布 状况, 并反馈给培养物分布控制器。 培养物分布控制器将 PH值传感器测得 的 PH值数据和培养物分布传感器测得的培养物分布数据分别与设定的门限 值进行比较, 并可根据比较结杲分别向 PH值控制器和营养液补料控制器发 送酸 /碱溶液补料数据和营养液补料数据。 PH值控制器接收到上述酸 /碱溶液 补料数据后, 可通过控制酸 /诚控制阀的开关, 向发酵罐中注入酸 /碱溶液。 营养液补料控制器接收到上述营养液补料数据后, 可通过控制营养液补料 阀的开关, 向发酵罐中注入营养液。
在本实施例中, 培养物分布控制器为主控制器, PH值控制器和营养液 补料控制器为从属控制器, 主控制器通过控制从属控制器进而控制 碱溶 液的补料操作和营养液的补料操作, 从而可以在现有发酵控制系统的基础 上通过较小的改动实施本发明, 减小了对现有系统进行升级的成本和复杂 度。 如果不考虑对现有发酵控制系统升级的问题, 本发明的发酵控制系统 在具体实施时可以只采用培养分布控制器来直接控制酸 /碱溶液的补料操作 和营养液的补料操作。
图 3为基于图 2所示逻辑结构的发酵控制系统的示例性结构图。
如图 3所示, 该系统包括: 发酵罐 1、 营养液补料阀 2、 酸 /碱控制阀 3、 酸 /减计量罐 4、 营养液计量罐 5、 PH值控制器 6、 营养液补料控制器 7、 培 养物分布控制器 8、 PH值传感器 9、 培养物分布传感器 10、 酸罐 11、 碱罐 12、 酸溶液泵 13和碱溶液泵 14。
本实施例中, 发酵罐 1工作在补料分批处理模式下。 优选地, 发酵罐 1 具有可以加热或制冷的外壳 16, 以及用于搅拌培养液 17的搅拌器 15。 搅 拌器 15用于提供发酵罐 1 内培养物和营养液之间的混合, 应当注意的是, 搅拌器 15在搅拌时应当避免过度的搅拌而将培养物细胞打碎。
营养液补料阀 2用于在营养液补料控制器 7的控制下打开或者关闭, 当营养液补料阀 2打开时, 营养液计量罐 5向发酵罐 1 中注入营养液, 优 选地, 该营养液补料阀 2可以为电磁阀或者为控制阀。
营养液计量罐 5 用于存储营养液, 例如葡萄糖溶液、 含氮溶液等, 并 通过营养液补料阀 2向发酵罐 1 中补充营养液。 具体实现时, 该营养液计 量罐 5还可包括一个输入口阀, 用于控制该营养液计量罐 5的营养液注入, 且该输入口阀也可由营养液补料控制器 7控制。 如图 4所示, 图 4示出了 本实施例中一种营养液计量罐 5的结构及其连接关系示意图, 如图 4所示, 营养液补料控制器 7控制营养液计量罐 5的输入口阀 (第一电磁阀 19 ) 和 该计量罐 5流向发酵罐 1的阀 (第二电磁阀 20, 也即图 3中的营养液补料 阀 2 )。 营养液计量罐 5具有两个液位电极 18, 当向营养液计量罐 5注入营 养液时, 两个电极电路被分别接通, 该电极产生代表营养液计量罐液体位 置的信号。 第一电磁阀 19控制着营养液计量罐 5的营养液注入, 第二电磁 阀 20控制着从营养液计量罐 5向发酵罐 1的营养液注入。在装载营养液时, 在营养液补料控制器 7的控制下,与发酵罐 1相连接的第二电磁阀 20关闭, 与营养液计量罐 5相连接的第一电磁阀 19打开, 使营养液计量罐 5装载营 养液。 在补料周期中, 在营养液补料控制器 7的控制下, 第二电磁阀 20打 开,一定量的营养液(例如 > 1升或者 2升的营养液)被注入到发酵罐 1后, 第二电磁阀 20关闭, 等待营养液补料控制器 7在下一个补料周期的控制命 令。
酸罐 11用于存储酸溶液; 酸溶液泵 13, 用于在 PH值控制器 6的控制 下将酸溶液泵入酸 /碱计量罐 4中。
碱罐 12用于存储碱溶液; 碱溶液泵 14, 用于在 PH值控制器 6的控制 下将碱溶液泵入酸 /碱计量罐 4中。
碱计量罐 4用于通过 ^/减控制阀 3向发酵罐 1中补充酸溶液或碱溶 液。
酸 /碱控制阀 3用于在 PH值控制器 6的控制下打开或者关闭。 当酸 /碱 控制阀 3打开时, 酸 /碱计量罐 4通过酸 /碱控制阀 3向发酵罐 1中注入酸溶 液或者碱溶液。 优选地, 该酸 /碱控制阀 3可以为电磁阀或者为控制阀。
营养液计量罐 5和^/碱计量罐 4均可以是带有流速测量装置的计量罐, 其中流速测量装置可以用于测量营养液流入发酵罐 1的流速或酸 /碱溶液流 入发酵罐 1的流速。 流速测量装置可以是电磁阀也可以是控制阀。
营养液补料控制器 7 用于控制营养液补料操作, 根据培养物分布控制 器 8的营养液补料数据, 控制营养液补料阀 2的开关打开, 实现将营养液 计量罐 5中一定量的营养液注入发酵罐 1中。
PH值控制器 6用于根据培养物分布控制器 8的酸 /减溶液补料数据,控 制酸溶液泵 13或碱溶液泵 14将酸溶液或誠溶液泵出至酸 /緘计量罐 4, 并 控制酸 /碱控制阀 3的开关打开,实现将 ^/碱计量罐 4中一定量的^ /碱溶液 注入发酵罐 1中。
PH值传感器 9用于对发酵罐 1中的 PH值进行测量, 将测量的 PH值 数据传送回培养物分布控制器 8。 优选地, 该 PH值传感器 9可以为 PH电 极 9, 用于对发酵罐 1中的 PH值进行实时测量。
培养物分布传感器 10用于对发酵罐 1中的培养物分布进行测量, 将测 量的培养物分布数据传送回培养物分布控制器 8。 其中, 培养物分布数据包 括"细胞群的平均属性"和"细胞群的分布宽度"。
细胞群的平均属性指细胞群中细胞属性的平均值, 用于反映培养物细 胞的总体性状。 针对培养物细胞的大小、 活力和年龄等属性, 细胞群可以 具有大小、 活力以及年龄等平均属性。 细胞群的分布宽度是一个反映培养 物细胞异质性的参数。 细胞群的分布宽度小表明培养物细胞的属性均一、 差异性小, 细胞群的分布宽度大表明培养物细胞的属性不均一、 差异性大。 细胞群的分布宽度是一个无量纲数值, 可以根据培养物细胞的大小、 活力 或年龄等属性来相应计算细胞群的分布宽度。 针对培养物细胞的大小这一 属性, 以培养物细胞的大小分布为例, 则细胞群的平均属性和细胞群的分 布宽度可分别采用以下的公式进行计算: 细胞群的平均属性 5 = 粒径 ^ 数
> 细胞总数
细胞群的分布宽度 = D90zD10
D
其中, /)90表示一个粒径值, 细胞群中 90%的细胞的直径小于此粒径值; D10表示另一个粒径值, 细胞群中 10%的细胞的直径小于此粒径值。
培养物分布传感器 10可以是各种先进的生物化学传感器, 例如, 可以 为美国专利 6,555,360中介绍的在线流式细胞仪, 或其它适用的测量仪器。
培养物分布控制器 8作为主控制器,通过 PH值传感器 9和培养物分布 传感器 10检测发酵罐 1中培养液 17的 PH值和培养物分布, 将检测的 PH 值数据和培养物分布数据分别与设定的门限值进行比较, 根据比较结果分 别向 PH值控制器 6和营养液补料控制器 7发送 碱溶液补料数据和营养 液补料数据。 其中, PH值数据的门限值和培养物分布数据的门限值可预先 通过实验或数学模型确定。
培养物分布控制器 8是一种针对培养液的 PH值和培养物分布进行控制 的多变量控制器, 即在控制过程中考虑培养物分布的变化与培养液 PH值的 变化两者之间的相互作用, 在现有的单纯对培养液的 PH值进行控制的基础 上, 同时利用培养液的 PH值来控制培养物的细胞群分布宽度, 并且优选的, 进一步利用营养液补料的多少来控制培养物的细胞群平均属性, 从而优化 对发酵过程的控制。
在本实施例的发酵控制系统中, 培养物分布控制器 8、 营养液补料控制 器 7和 PH值控制器 6可由电子设备实现, 例如, 工业控制计算机、 可编程逻 辑控制器(PLC ), 或其组合。 各设备之间的通信可通过标准的 4~20mA (毫 安) 电信号或者符合现场总线标准 (如 Profibus ) 的电信号实现。 营养液补 料控制器 7和 PH值控制器 6都可以采用比例-积分-微分方法 (PID )、 开 /关或 时间比例控制 (TPC ) 方法分别对营养液补料阀 2和酸 /碱控制阀 3进行调节 控制, 使用什么样的控制方法依赖于阀门的形式, 如控制阀或电磁阀。 TPC 提供的控制方式, 在相对较低的控制复杂度基础上成本相对较低。 在每次 补料周期内, TPC根据操作变量的变化, 例如, 补料流量的多少, 改变阀门 开关的时间比例。
基于图 3所示系统, 图 5中示出了本发明实施例中发酵控制方法的示例 性流程图。
如图 5所示, 该流程包括如下步骤:
步骤 501 , 获取培养液的 PH值数据。
具体实现时, 可通过 PH值传感器 9获取培养液的 PH值数据。
步骤 502, 判断培养液的 PH值是否超出设定的 PH门限值, 如果超出, 则执行步骤 503; 否则, 执行步骤 504。
本步骤中,培养物分布控制器 8将 PH值传感器 9获取的 PH值与设定的 PH 门限值相比较, 判断培养液的 PH值是否满足所设定的 PH门限值的要求, 并 根据比较结果选择后续步骤。
步骤 503 , 对培养液的 PH值进行控制, 使其满足所设定的 PH门限值的 要求, 然后执行步骤 504。
本步骤中, 培养物分布控制器 8对培养液的 PH值进行控制, 使其满足所 设定的 PH门限值的要求。 例如, 由培养物分布控制器 8向 PH值控制器 6发送 酸 /碱溶液补料数据, PH值控制器 6根据该 ^/碱溶液补料数据对酸 /碱控制阀 3进行开关控制。 通过将酸 /碱溶液注入培养液中, 使得培养液的 PH值满足 所设定的 PH门限值的要求。
步骤 504, 获取培养物分布数据。
本步骤中, 可通过培养物分布传感器 10获取培养液的培养物分布数据, 其中, 培养物分布数据包括培养物的细胞群的分布宽度。
步骤 505, 判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果是, 则执行步骤 506, 否则结束控制过程。
本步骤中, 培养物分布控制器 8将培养物分布传感器 10获取的细胞群分 布宽度与设定的宽度门限值相比较, 判断培养物的细胞群分布宽度是否满 足所设定的宽度门 P艮值的要求, 并根据比较结果选择后续步骤。 步骤 506, 对培养物的细胞群分布宽度进行控制, 使其满足所设定的宽 度门限值的要求, 并结束控制过程。
本步骤中,培养物分布控制器 8将进一步利用培养液的 PH值来对培养物 的细胞群分布宽度进行控制, 使其满足所设定的宽度门限值的要求。 例如, 由培养物分布控制器 8向 PH值控制器 6发送^ /碱溶液补料数据, PH值控制器 6根据该 ^/碱溶液补料数据对酸 /碱控制阀 3进行开关控制。通过将 碱溶液 注入培养液中, 使得具有不期望属性的培养物细胞減少, 而具有期望属性 的培养物细胞增加, 从而减小培养物细胞的属性之间的差异性, 使培养物 的细胞群分布宽度满足所设定的宽度门限值的要求。
至此, 培养物分布控制器 8的一次控制过程结束。
此外, 在本实施例中, 优选地, 步骤 504中所荻取的培养物分布数据中 还包括培养物的细胞群平均属性, 这样在步骤 504之后, 在对培养物的细胞 群分布宽度进行控制之前, 还可以首先通过营养液的补料操作来对培养物 的细胞群平均属性进行控制。 具体实现时, 培养物分布控制器 8可以将培养 物分布传感器 10获取的细胞群平均属性与一个设定的平均属性门限值相比 较, 并在细胞群的平均属性不满足该门限值的要求时, 通过向营养液补料 控制器 7发送营养液补料数据来控制营养液的补料操作, 从而使得细胞群的 平均属性符合所述门限值的要求, 也即使得培养物细胞的总体性状首先满 足设定的要求, 因而进一步优化了发酵的控制过程。
在本发明的具体实施过程中, 优选还可以根据 PH值控制曲线对培养液 的 PH值和培养物的细胞群分布宽度的进行控制,图 6中示出了本发明发酵控 制方法的另一实施例中的 PH值控制曲线。
如图 6所示, 纵轴代表 PH值, 按箭头方向 PH值逐渐变大, 横轴代表发 酵时间。图中的黑色粗实线为发酵过程中培养液的 PH值随时间变化的曲线。 图中分别对应一个最大 PH值和一个最小 PH值的两条虛线为局部控制线 ( LCL, Local Control Limit ), 分别表示培养物细胞能够承受的最大碱性 PH 值和最大酸性 PH值, 也即设定的 PH值上、 下限门限值。 如杲培养液的 PH 值超出了两条 LCL所表示的 PH值范围, 例如培养液的 PH值小于所设定的最 大酸性 PH值, 或大于所设定的最大碱性 PH值, 培养物细胞的生长将受到阻 碍, 从而使得发酵过程无法正常进行。 图中紧接着最小 PH值 LCL上面的虛 线为一条精细控制线( FCL, Fine Control Limit ), 表示对某种属性的细胞的 生长影响显著的一个 PH门限值。 当培养液的 PH值处于该 FCL与最小 PH值 LCL之间时,培养物细胞的生长环境呈偏酸性,这将显著影响某种属性的细 胞的生长, 而对其它属性的细胞的生长无明显影响。 例如, 当培养液的 PH 值处于该 FCL与最小 PH值 LCL之间时, 活力弱的培养物细胞的生理周期将 被显著缩短, 但活力强的培养物细胞的生理周期将不会有明显改变。 与之 相对应, 在实际应用中, 紧接着最大 PH值 LCL下面也可以设定另一条 FCL (为清楚起见, 图 6中未示出), 表示一种偏碱性生长环境的 ΡΗΠ限值, 当 培养液的 PH值处于该 FCL与最大 PH值 LCL之间时, 培养液呈偏减性, 这也 将显著影响某种属性的细胞的生长, 而对其它属性的细胞的生长无明显影 响。 根据不同发酵过程的控制需求, 本发明在具体实施时可设定 PH值上、 下限 LCL以及偏酸性生长环境 FCL,或者 PH值上、下限 LCL以及偏碱性生长 环境 FCL。 图中紧接着 FCL上面还显示了一条虛线, 其所对应的 PH值为最 适宜生长环境 PH值, 具有该最适宜生长环境 PH值的培养液适合细胞群中所 有细胞的生长。
基于图 3所示系统以及图 6所示 PH值控制曲线, 图 7中示出了本实施例中 发酵控制方法的示例性流程图。
如图 7所示, 该流程包括如下步骤:
步骤 701 , 获取培养液的 PH值数椐。
具体实现时, 培养物分布控制器 8可通过 PH值传感器 9实时地获取培养 液的 PH值数据。
步骤 702,判断培养液的 PH值是否超出两条 LCL所表示的 PH值范围,如 果超出, 则执行步骤 703; 否则, 执行步骤 704。
本步骤中, 培养物分布控制器 8将 PH值传感器 9获取的 PH值与两奈 LCL 所表示的上、 下限 PH门限值相比较, 判断培养液的 PH值是否超出所述上、 下限 PH门限值所限定的取值范围, 并根据比较结果选择后续步骤。
步骤 703 , 对培养液的 PH值进行控制, 使其处于两条 LCL所表示的 PH 值范围之内, 然后执行步骤 704。
本步骤中, 培养物分布控制器 8对培养液的 PH值进行控制, 例如, 由培 养物分布控制器 8向 PH值控制器 6发送酸 /碱溶液补料数据, PH值控制器 6根 据该酸 /碱溶液补料数据对酸 /碱控制阀 3进行控制, 将酸 /碱溶液注入培养液 中, 使得培养液的 PH值处于两条 LCL所表示的 PH值范围之内, 以满足培养 物细胞生长的最低要求。
步骤 704, 获取培养物分布数据。
本步骤中, 培养液的 PH值已在两条 LCL所表示的 PH值范围之内, 则进 而可通过培养物分布传感器 10获取培养液的培养物分布数据, 其中, 培养 物分布数据包括培养物的细胞群分布宽度。
步骤 705, 判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果是, 则执行步骤 706, 否则, 执行步骤 707。
步骤 706, 对培养物的细胞群分布宽度进行控制, 使其满足所设定的宽 度门限值的要求, 并执行步骤 707。
本步骤中, 培养物分布控制器 8将根据所述 PH值控制曲线向 PH值控制 器 6发送酸 /碱溶液补料数据,由 PH值控制器 6根据该 碱溶液补料数据对酸 /械控制阀 3进行控制, 例如在图 6中所示的 tl到 t2时间内, 将酸 /碱溶液注入 培养液中, 使得培养液的 PH值处于 FCL与最小 PH值 LCL之间, 从而将具有 不期望属性的培养物细胞减少, 如将活力弱的培养物细胞减少, 而将具有 期望属性的培养物细胞增多, 如将活力强的培养物细胞增多, 进而使细胞 群分布宽度满足所设定的宽度门 P艮值的要求。
步骤 707, 对培养液的 PH值进一步进行控制, 使其保持为所述最适宜生 长环境 PH值, 然后执行步骤 708。
本步骤中, 为了促进培养物细胞的生长, 可对培养液的 PH值进一步进 行控制, 使其保持为最适宜生长环境 PH值, 这样具有期望属性的培养物细 胞就可以更快更好的生长。
步骤 708, 判断本次发酵生产过程是否结束, 如果是, 则结束控制过程, 否则返回步骤 704。
在本实施例中, 为了进一步优化发酵的控制过程, 同样也可以在对培 养物的细胞群分布宽度进行控制之前, 首先通过营养液的补料操作来对培 养物的细胞群平均属性进行控制。 这样, 步骤 704中所获取的培养物分布数 据还包括培养物的细胞群平均属性。 在步骤 704之后, 首先将培养物分布传 感器 10获取的细胞群平均属性与一个设定的平均属性门限值相比较, 并可 在细胞群的平均属性不满足该门 P艮值的要求时, 由营养液补料控制器 7根据 培养物分布控制器 8发送的营养液补料数据来控制营养液的补料操作, 从而 使得细胞群的平均属性符合所述门限值的要求。 例如, 假设所考察的细胞 群平均属性为细胞大小, 且设定的平均大小门限值为 5um, 若培养物分布传 感器 10测得的细胞群平均大小比所述门限值低, 则营养液补料控制器 7将根 据培养物分布控制器 8输出的营养液补料数据, 控制向发酵罐中注入一定量 的营养液, 以促进培养物细胞生长。
根据上述的具体实施例可以看出, 本发明在控制过程中考虑了培养物 的分布及其与培养液 PH值之间的相互作用, 通过控制培养液的 PH值来控制 培养物的细胞群分布宽度, 使得发酵过程的生产效率和产品质量得以提高。 在此基础上, 优选的, 还可通过控制营养液的补料操作来控制培养物的细 胞群平均属性, 使得培养物细胞的总体性状首先满足设定的要求, 从而进 一步提高发酵过程的生产效率和产品质量。
以上所述仅为本发明的较佳实施例而已, 并非用于限定本发明的保护 范围。 凡在本发明的精神和原则之内, 所作的任何修改、 等同替换以及改 进等, 均应包含在本发明的保护范围之内。

Claims

权利要求
1. 一种发酵控制系统, 包括, PH值传感器 (9 ), 用于获取培养液的 PH值数据; 其特征在于, 该系统还包括, 培养物分布传感器( 10 )和培养 物分布控制器 (8 );
所述培养物分布传感器 (10 ) 用于获取培养物分布数据, 其中, 所述 培养物分布数据包括培养物的细胞群分布宽度;
所述培养物分布控制器(8 )用于 居所述 PH值传感器(9 )获取的培 养液 PH值数据和培养物分布传感器( 10 )获取的培养物分布数据, 对培养 液的 PH值及培养物的细胞群分布宽度进行多变量控制。
2. 根据权利要求 1所述的发酵控制系统, 其特征在于, 所述培养物分 布传感器 ( 10 ) 所获取的培养物分布数据中还包括培养物的细胞群平均属 性;
所述培养物分布控制器 (8 )进一步用于根据所述培养物分布传感器 ( 10 ) 获取的培养物分布数据, 对培养物的细胞群平均属性进行控制。
3. 根据权利要求 1或 2所述的发酵控制系统, 其特征在于, 该系统进 一步包括 PH值控制器 ( 6 ), 用于在所述培养物分布控制器 ( 8 ) 的控制之 下对培养液的 PH值及培养物的细胞群分布宽度进行控制。
4. 根据权利要求 2所述的发酵控制系统, 其特征在于, 该系统进一步 包括营养液补料控制器(7 ), 用于在所述培养物分布控制器(8 ) 的控制之 下对培养物的细胞群平均属性进行控制。
5. 根据权利要求 1至 4中任一所述的发酵控制系统, 其特征在于, 所 述培养物分布传感器 ( 10 ) 为在线流式细胞仪。
6. 根据权利要求 5所述的发酵控制系统, 其特征在于, 所述培养物分 布控制器(8 ) 为可编程逻辑控制器。
7. 一种发酵控制方法, 该方法包括:
获取培养液的 PH值数据和培养物分布数据, 其中, 所述培养物分布数 据包括培养物的细胞群分布宽度;
根据所述培养液的 PH值数据和培养物分布数据, 对培养液的 PH值及 培养物的细胞群分布宽度进行多变量控制。
8. 根据权利要求 7所述的发酵控制方法, 其特征在于, 所述对培养液 的 PH值及培养物的细胞群分布宽度进行多变量控制为:
对培养液的 PH值进行控制, 并利用培养液的 PH值对培养物的细胞群 分布宽度进行控制。
9. 根椐权利要求 8所述的发酵控制方法, 其特征在于, 所述对培养液 的 PH值进行控制, 并利用培养液的 PH值对培养物的细胞群分布宽度进行 控制的步骤为:
判断培养液的 PH值是否超出设定的 PH门限值, 如果超出, 则对培养 液的 PH值进行控制, 使其满足所述 PH门限值的要求;
判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果大于, 则对培养液的 PH值进行控制,以使培养物的细胞群分布宽度满足所述宽度 门限值的要求。
10. 根据权利要求 8 所述的发酵控制方法, 其特征在于, 所述对培养 液的 PH值进行控制, 并利用培养液的 PH值对培养物的细胞群分布宽度进 行控制的步骤为:
判断培养液的 PH值是否超出 PH值控制曲线中的局部控制线所设定的 PH门限值范围, 如果超出, 则将培养液的 PH值控制在所述局部控制线所 设定的 PH门限值范围内;
判断培养物的细胞群分布宽度是否大于设定的宽度门限值, 如果大于, 则对培养液的 PH值进行控制, 使其满足 PH值控制曲线中的精细控制线所 设定的 PH门限值的要求,以使培养物的细胞群分布宽度满足所述宽度门限 值的要求。
11. 根据权利要求 7至 10中任一所述的发酵控制方法, 其特征在于, 所获取的培养物分布数据中还包括培养物的细胞群平均属性; 该方法进一 步包括:
根据所述培养物分布数据, 对培养物的细胞群平均属性进行控制。
12. 根据权利要求 11所述的发酵控制方法', 其特征在于, 所述对培养 物的细胞群平均属性的控制在所述对培养物的细胞群分布宽度的控制之前 进行。
13. 根据权利要求 11所述的发酵控制方法, 其特征在于, 所述对培养 物的细胞群平均属性进行控制为:
通过控制营养液的补料操作来对培养物的细胞群平均属性进行控制。
14. 根据权利要求 13所述的发酵控制方法, 其特征在于, 所述通过控 制营养液的补料操作来对培养物的细胞群平均属性进行控制的步骤为: 判断培养物的细胞群平均属性是否满足设定的平均属性门限值的要 求, 如果不满足, 则对营养液的补料操作进行控制, 以使培养物的细胞群 平均属性满足所述平均属性门限值的要求。
15. 根据权利要求 9、 10或 14中任一所述的发酵控制方法, 其特征在 于, 所述的各 PH门限值、 宽度门限值和平均属性门限值预先通过实验或数 学模型确定。
PCT/CN2008/000688 2008-04-03 2008-04-03 一种发酵控制系统及发酵控制方法 WO2009121211A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2008/000688 WO2009121211A1 (zh) 2008-04-03 2008-04-03 一种发酵控制系统及发酵控制方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2008/000688 WO2009121211A1 (zh) 2008-04-03 2008-04-03 一种发酵控制系统及发酵控制方法

Publications (2)

Publication Number Publication Date
WO2009121211A1 true WO2009121211A1 (zh) 2009-10-08
WO2009121211A8 WO2009121211A8 (zh) 2009-12-10

Family

ID=41134799

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2008/000688 WO2009121211A1 (zh) 2008-04-03 2008-04-03 一种发酵控制系统及发酵控制方法

Country Status (1)

Country Link
WO (1) WO2009121211A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110669639A (zh) * 2019-10-08 2020-01-10 浙江中控技术股份有限公司 一种多发酵罐自动流加补料装置及方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0144474A1 (en) * 1983-12-09 1985-06-19 Fabriques De Tabac Reunies S.A. Continuous fermentation process
CN1177007A (zh) * 1996-09-19 1998-03-25 中国科学院化工冶金研究所 需氧微生物发酵过程控制菌体代谢节律的培养方法
US6492135B1 (en) * 1999-05-18 2002-12-10 Ebbe Busch Larsen U-shape and/or nozzle u-loop fermentor and method of carrying out a fermentation process
US6555360B1 (en) * 1998-03-30 2003-04-29 Friedrich Srienc Flow injection flow cytometry system for on-line monitoring of biroreactors and method for monitoring
CN1727468A (zh) * 2004-07-28 2006-02-01 薛宁 一种微生物发酵条件的控制方法
WO2006086489A1 (en) * 2005-02-10 2006-08-17 Broadley-James Corporation Integrated bio-reactor monitor and control system

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0144474A1 (en) * 1983-12-09 1985-06-19 Fabriques De Tabac Reunies S.A. Continuous fermentation process
CN1177007A (zh) * 1996-09-19 1998-03-25 中国科学院化工冶金研究所 需氧微生物发酵过程控制菌体代谢节律的培养方法
US6555360B1 (en) * 1998-03-30 2003-04-29 Friedrich Srienc Flow injection flow cytometry system for on-line monitoring of biroreactors and method for monitoring
US6492135B1 (en) * 1999-05-18 2002-12-10 Ebbe Busch Larsen U-shape and/or nozzle u-loop fermentor and method of carrying out a fermentation process
CN1727468A (zh) * 2004-07-28 2006-02-01 薛宁 一种微生物发酵条件的控制方法
WO2006086489A1 (en) * 2005-02-10 2006-08-17 Broadley-James Corporation Integrated bio-reactor monitor and control system

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110669639A (zh) * 2019-10-08 2020-01-10 浙江中控技术股份有限公司 一种多发酵罐自动流加补料装置及方法

Also Published As

Publication number Publication date
WO2009121211A8 (zh) 2009-12-10

Similar Documents

Publication Publication Date Title
CN102351945B (zh) 自动配酸装置和自动管道点酸装置及自动配酸点酸方法
CN206396227U (zh) 一种透明质酸生产用发酵罐
CN103698541A (zh) 谷氨酸发酵在线检测系统及采用该系统在线检测的方法
CN108315251A (zh) 一种菌种发酵控制系统
CN104028129A (zh) 一种干酪素生产自动溶解装置以及自动溶解的方法
JPWO2007032265A1 (ja) アルコール生産細菌の連続培養装置及びその方法
US11603517B2 (en) Method for monitoring a biotechnological process
CN101412952A (zh) 麦汁充氧设备系统及其控制方法
WO2009121211A1 (zh) 一种发酵控制系统及发酵控制方法
US10892033B2 (en) Method for monitoring bioprocesses
CN110656044A (zh) 新型智能微生物发酵装置
CN205088239U (zh) 一种餐厨垃圾厌氧发酵产沼气的快速启动装置
CN206289257U (zh) 一种全自动连续生产葡萄糖液系统
CN207204090U (zh) 一种带有自动传感器的聚羧酸减水剂合成装置
CN213376341U (zh) 一种半自动化全封闭的细胞转染试剂配制装置
CN105087355A (zh) 用于培养藻类的光合生物反应器系统
CN109321456A (zh) 一种微流控芯片细胞培养控制装置与方法
CN106155123B (zh) 一种发酵过程残糖浓度的在线控制方法
CN104946501B (zh) 一种醋酸发酵扩培酸蒸工艺及系统
CN210237506U (zh) 一种糖醛酸自动还原装置
CN211051415U (zh) 一种外加剂母料智能配料系统
CN206751833U (zh) 一种大米蛋白料液在线pH值调整装置
CN203355629U (zh) 一种干酪素生产自动溶解装置
CN209178397U (zh) 一种用于氨基酸生产菌种扩大培养的种子罐系统
CN205040612U (zh) 一种基于plc的蔬菜发酵系统

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08733902

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08733902

Country of ref document: EP

Kind code of ref document: A1