CN108102906B

CN108102906B - Parallel reactor system and parallel control experiment method

Info

Publication number: CN108102906B
Application number: CN201710901063.8A
Authority: CN
Inventors: 张明; 唐寅; 王泽建; 曾梁平; 马文峰; 张鑫; 郭美锦; 庄英萍
Original assignee: Shanghai Guoqiang Biochemical Engineering Equipment Co ltd; East China University of Science and Technology
Current assignee: Shanghai Guoqiang Biochemical Engineering Equipment Co ltd; East China University of Science and Technology
Priority date: 2017-09-28
Filing date: 2017-09-28
Publication date: 2023-12-15
Anticipated expiration: 2037-09-28
Also published as: CN108102906A

Abstract

The invention discloses a parallel reactor system, which comprises a plurality of reactor tanks, wherein sensing units for measuring physicochemical parameters are arranged in the reactor tanks, and each reactor tank is provided with a first air inlet pipeline and a first air outlet pipeline; the seed stirring tank is used for culturing seeds and is respectively communicated with each reactor tank through an inoculation pipeline for inoculation, an inoculation pump is arranged on each inoculation pipeline, and a first stirring mechanism is arranged in the seed stirring tank to uniformly mix seed liquid in the seed stirring tank; the first exhaust pipelines of all the reactor tanks are communicated with the buffer bottles, the buffer bottles are additionally provided with main exhaust pipelines, and the main exhaust pipelines are provided with main exhaust regulating valves; a pressure detection mechanism is arranged on the buffer bottle or the main exhaust pipe; and a parallel sampling device in communication with each of the reactor tanks for sampling. The deviation of each parallel reactor is reduced so as to improve the reliability of research results in the culture process.

Description

Parallel reactor system and parallel control experiment method

Technical Field

The invention relates to the technical field of biology, in particular to a parallel reactor system and a parallel control experiment method.

Background

The culture process of organisms (microorganisms or animal and plant cells) in a bioreactor depends on the genetic metabolic characteristics of the organisms themselves and is also influenced by the culture environment in the reactor. The genetic characteristics of the biological culture process and the culture environment can interact and influence each other, so that the biological culture process is different from the general chemical reaction process, and the orientation and repetition cannot be realized, which is the characteristic of the biological culture process. The bioreactor not only needs to ensure the conventional sterile requirements to meet the basic requirements of pure culture of organisms, the reactor material does not influence the biological culture process and the like, but also accurately controls various parameters of the culture environment to meet the requirements of the biological culture process, and is a core problem for measuring the performance of the reactor. Due to the unrepeatability of the biological culture process, in the research process of optimizing the biological culture process by changing certain culture conditions, the variation amplitude of the culture result caused by the variation of the conditions is often difficult to distinguish from the deviation amplitude of the results of the biological culture processes of different batches under the same conditions, so that the reliability of the optimized research result is reduced, the experimental efficiency is affected, and even misguidance is generated. For this reason, the concept of parallel reactions is generally employed, in order to increase the reliability of the results of the culture process study by reducing the deviation (i.e., systematic deviation) of the individual parallel reactors from each other. To achieve this requirement, several reactors of identical specifications cannot simply be combined, but rather a specific targeted systematic design of parallel reactor performance is required from the accuracy requirement characteristics of the parallel culture studies themselves.

Disclosure of Invention

The invention aims to provide a parallel reactor system and a parallel control test method, which can reduce the system deviation as much as possible so as to improve the reliability of research results.

The technical scheme provided by the invention is as follows: a parallel reactor system comprising:

the reactor comprises a plurality of reactor tanks, a plurality of first air inlet pipelines and a plurality of first air outlet pipelines, wherein the reactor tanks are used for carrying out parallel control tests, sensing units for measuring physical and chemical parameters are arranged in the reactor tanks, and each reactor tank is provided with a first air inlet pipeline and a first air outlet pipeline;

the material beating tanks are used for culturing seeds and are respectively communicated with each reactor tank through an inoculation pipeline for inoculation, each inoculation pipeline is provided with a material beating pump, and a first stirring mechanism is arranged in each material beating tank to uniformly mix feed liquid in the material beating tanks;

the first exhaust pipelines of all the reactor tanks are communicated with the buffer bottles, the buffer bottles are additionally provided with main exhaust pipelines, and the main exhaust pipelines are provided with main exhaust regulating valves; a pressure detection mechanism is arranged on the buffer bottle or the main exhaust pipe;

and a parallel sampling device in communication with each of the reactor tanks for sampling.

According to the technical scheme, seed liquid input into each reactor tank of the parallel reactor system comes from the same stirring tank, the stirring tank can be used for culturing the seed liquid, and the seed liquid in the tank can be uniformly mixed through stirring of the stirring mechanism. The flow rate of each inoculation pipeline of the material can be regulated, so that the consistency of the quantity and quality of the initial seed liquid in each reactor can be realized, and particularly when the material is used for inoculation, the liquid quality and quantity of the initial seed liquid in each reactor can be consistent, and the reliability of the final research result is improved. The exhaust pipelines of the reactor tanks are communicated with the buffer bottle, a main exhaust pipeline is additionally led out of the buffer bottle, an exhaust regulating valve is arranged on the buffer bottle, and the pressure detection mechanism is matched with the exhaust regulating valve on the main exhaust pipeline to regulate the exhaust. The exhaust of each reactor tank is buffered, so that the fluctuation range of the gas pressure is reduced, the detection is facilitated, the control sensitivity is improved, and the overall control precision is improved. Meanwhile, the problem that fluctuation time is asynchronous in the process of independently controlling the tank pressure is solved, and the circulation interference on measurement and control of the tank pressure is caused by circulation sampling of exhaust gas by each reactor tank. Even if the fluctuation is slight, the consistency of the changes among the tanks can be ensured because the tanks synchronously fluctuate.

Preferably, the parallel sampling device comprises: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; the first peristaltic pump is arranged on each sampling pipe and is positioned between the sampling end and the sample outlet; the second peristaltic pump is arranged on each sampling pipe and is positioned between the first peristaltic pump and the sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second peristaltic pump and the sample outlet; one end of the gas main pipeline is connected with an air filter, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the first peristaltic pump and the second peristaltic pump; the first pinch valve is arranged on the gas main pipeline and is positioned between the air filter and the gas branch pipeline.

Specifically, the parallel sampling device includes: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; the first peristaltic pump is arranged on each sampling pipe and is positioned between the sampling end and the sample outlet; a second pinch valve disposed on each of said sampling lines and positioned between said first peristaltic pump and said sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second pinch valve and the sample outlet; one end of the gas main pipeline is connected with an air filter, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the first peristaltic pump and the second pinch valve; the first pinch valve is arranged on the gas main pipeline and is positioned between the air filter and the gas branch pipeline;

Each gas branch pipeline is respectively provided with a one-way valve, and the one-way valves are used for conducting the gas branch pipelines in one way towards the direction of the sampling pipeline.

Specifically, the parallel sampling device includes: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; a third pinch valve disposed on each of the sampling lines and located between the reactor tank and the sample outlet; the second pinch valve is arranged on each sampling pipe and is positioned between the third pinch valve and the sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second pinch valve and the sample outlet; a fourth pinch valve and an air filter are sequentially arranged at one end of the gas main pipeline, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the first peristaltic pump and the second pinch valve; the injection pump is connected with the gas main pipeline, and a connecting point is positioned between the fourth pinch valve and the air filter; the buffer tanks are respectively arranged on each gas branch pipeline.

According to the technical scheme, the equipment cost and maintenance workload of the sampling device are reduced by integrating the sampling functions of the plurality of reactor tanks of the parallel reactor; based on the parallel culture experiment, the sampling operation time is shortened, the sampling limit, sterility and synchronous consistency can be ensured from the sampling link, and the systematic deviation of the experimental device is reduced.

Further, the parallel reactor system further comprises an electrode calibration device for calibrating the sensing unit; the electrode calibration device comprises:

the tank body is used for containing the liquid to be tested;

the tank cover can be covered on the tank body and is provided with a plurality of faucet openings, and the electrode to be tested is inserted into the faucet openings;

the third air inlet pipeline and the third air outlet pipeline are arranged on the tank body or the tank cover, the third air inlet pipeline comprises a plurality of air inlet branches which are arranged in parallel, each air inlet branch is provided with a gas flowmeter and a third air inlet regulating valve, and the outlet pipeline is provided with a pressure detection device and a third air outlet regulating valve;

the second stirring mechanism acts on the tested liquid in the tank body to uniformly mix the tested liquid;

The temperature electrode and the standard thermometer are inserted into the socket of the tank cover to measure the temperature;

a temperature adjusting mechanism for heating and cooling the tank body;

and the calibration mechanism is used for calibrating the electrode to be measured.

The sensing units mentioned herein refer to temperature electrodes, pH electrodes, DO electrodes and other electrodes that can be calibrated by the calibration device, such as dissolved carbon dioxide, ORP, etc. According to the technical scheme, the socket is designed on the tank cover of the calibration tank so as to insert the electrode to be measured, and the stirring mechanism is used for stirring the liquid to be measured in the tank body to ensure that the internal physical and chemical parameters of the liquid to be measured are uniform, so that the effect of calibrating a plurality of electrodes simultaneously under the same liquid environment to be measured is realized, and the calibration errors caused by fluctuation and change of a standard medium and uncertainty of external interference factors in the independent calibration process of each electrode can be solved, and one-key parallel calibration is realized. Meanwhile, through setting basic structures such as a tank body, a tank cover, a stirring mechanism, and gas inlet and exhaust, such as tank pressure measurement and control, temperature measurement and control, calibration of different types of electrodes, such as a temperature electrode, a pH electrode, a DO electrode, a carbon dioxide electrode, a methanol electrode and the like, can be realized.

In particular, the method comprises the steps of,

an auxiliary exhaust pipeline is further arranged on the buffer bottle, and a manual bypass valve is arranged on the auxiliary exhaust pipeline;

and/or;

the buffer bottle is also provided with a pressure relief pipeline, and a safety pressure relief valve is arranged on the pressure relief pipeline;

and/or;

a drain pipeline is further arranged on the buffer bottle, the drain pipeline is led out of the buffer bottle from the bottom of the buffer bottle, and a drain valve is arranged on the drain pipeline;

according to the technical scheme, the manual bypass valve on the auxiliary exhaust pipeline is used for manually adjusting when the main exhaust regulating valve fails or the flow is insufficient. The safety relief valve on the relief pipeline is used for avoiding the excessive high tank pressure. The sewage discharge pipeline is used for timely discharging condensate and the like.

Specifically, the sensing unit comprises one or more of a temperature sensor, a pH sensor, a dissolved oxygen sensor, a full tank weighing sensor, a tail gas analyzer, a speed measuring sensor, a pressure sensor, a defoaming sensor, a feeding weighing sensor, a cell microscopic online observer, a viable bacteria amount sensor, an oxidation-reduction potential ORP sensor, a bacteria concentration OD sensor and an air flow sensor.

Specifically, the system further comprises a monitoring system and an analysis system; the monitoring system is communicated with the sensing unit, the parallel sampling device, the electrode calibration device, the main exhaust regulating valve and the pressure detection mechanism for data acquisition and control, and the analysis system analyzes the data acquired by the monitoring system.

Specifically, the first stirring mechanism and the second stirring mechanism both comprise stirring paddles and stirring motors for driving the stirring paddles.

The invention also discloses a parallel control test method, which uses the parallel reactor system and comprises the following steps:

s100, calibrating the sensing unit through the electrode calibration device;

s200, adding a seed culture medium into the stirring tank, and sterilizing, inoculating and culturing;

s300, adding a culture medium into the reactor tank, and sterilizing;

s400, inoculating the seeds cultured in the material beating tanks into each reactor tank through a material beating device;

s500, controlling the pressure of each reactor tank through the buffer bottle by matching the pressure detection mechanism and the main exhaust regulating valve, and fermenting;

s600, sampling and analyzing each reactor tank through the parallel sampling device, and inputting analysis obtained data into the analysis system;

and S700, performing relevant analysis on the data obtained by the monitoring system and the data obtained in the step S600 through the analysis system, and determining the optimal control parameters in the fermentation process.

The parallel reactor system and the parallel control test method provided by the invention can bring at least one of the following beneficial effects:

during inoculation, seed liquid input into each reactor tank of the parallel reactor system comes from the same beating tank for parallel inoculation; when fermentation is carried out, the exhaust pipelines of all the reactor tanks are communicated through one buffer tank, and the parallel control of the tank is carried out by matching with a main exhaust pipeline further led out on the buffer tank; and parallel sampling is carried out through an integrated pipeline during sampling. In addition, at the beginning, the parallel calibration device is used for carrying out one-key parallel calibration on the test electrodes used by each reactor tank based on the tested seed liquid in one tank body. By a series of device designs, the mutual deviation (namely, systematic deviation) of each parallel reactor is reduced, so that the reliability of research results of the culture process is improved.

Drawings

The above features, technical features, advantages and implementation of the inverted metered dose aerosol valve will be further described in the following description of preferred embodiments in a clearly understood manner with reference to the accompanying drawings.

Fig. 1 is a schematic view of a parallel feed apparatus.

Fig. 2 is a schematic diagram of a tank pressure control device.

FIG. 3-a is a schematic diagram of one form of a parallel sampling device.

FIG. 3-b is a schematic illustration of another form of parallel sampling device.

Fig. 3-c is a schematic illustration of yet another form of composition of the parallel sampling device.

FIG. 4 is a schematic diagram of an electrode calibration device.

Reference numerals illustrate:

100. the device comprises a material beating tank body 110, a material beating tank cover 120, a first stirring mechanism 130, a liquid outlet pipe 131, a liquid outlet pump 140, a material supplementing pipe 141, a material supplementing pump 150, a control system 160, a pH electrode 161, a pH adjusting pipeline 162, an acid-base peristaltic pump 170, a temperature electrode 171, a temperature control system 180, a DO electrode 181, an air inlet pipeline 182 and an exhaust pipeline;

200. a reactor tank 210, a first air inlet pipeline 211, a mass flowmeter 212, a rotameter 213, a one-way valve 220, a stirring motor 221, a stirring paddle 222, a rotation speed detection device 230, a first air outlet pipeline 240, a buffer bottle 250, a main air outlet pipeline 251, a self-control air outlet regulating valve 252, a pressure sensor 253, a mechanical back pressure valve 254, a manual bypass valve 255, a safety relief valve 260 and a sewage pipeline;

300. sampling tubing, 301, sample outlet, 302, first peristaltic pump, 303, second peristaltic pump, 304, heating cooler, 310, main tubing, 311, air filter, 312, gas manifold, 320, first pinch valve, 321, second pinch valve, 322, third pinch valve, 323, fourth pinch valve, 324, one-way valve, 325, buffer tank, 326, syringe pump;

400. The device comprises a calibration tank body 410, a calibration tank cover 411, a socket, 420, a third air inlet pipeline, 421, a gas flowmeter, 430, a third air outlet pipeline, 431, a third air outlet regulating valve, 432, a pressure detection device, 440, a temperature electrode, 441, a standard thermometer, 442, a heating source, 443, a cooling pipe, 450, an electrode to be tested, 460, a calibration mechanism, 470 and a control system;

Detailed Description

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following description will explain the specific embodiments of the present invention with reference to the accompanying drawings. It is evident that the drawings in the following description are only examples of the invention, from which other drawings and other embodiments can be obtained by a person skilled in the art without inventive effort.

For simplicity of the drawing, only the parts relevant to the present invention are schematically shown in each drawing, and they do not represent the actual structure thereof as a product.

Device embodiment 1

This example discloses a parallel reactor system comprising: a plurality of reactor tanks 200 for parallel control tests, in which a sensing unit for measuring physicochemical parameters is provided,

The parallel reactor system further comprises a parallel feed device for inoculating each reactor tank for feeding seed liquid into the parallel reaction reactor tank 200. As shown in fig. 1, the parallel feeding device includes: the stirring device comprises a stirring tank, a first stirring mechanism 120, a liquid outlet pipe 130 and a liquid outlet pump 131. The material beating pot is composed of a material beating pot body 100 and a material beating pot cover 110. The material beating pot body 100 is used for containing seed liquid, and the material beating pot cover 110 is covered on the material beating pot body 100 to seal the material beating pot body 100. The first stirring mechanism 120 acts on the seed liquid in the material stirring tank body 100 to uniformly mix the seed liquid, and as shown in the drawing, the first stirring mechanism 120 may be a stirring paddle structure driven by a motor. One end of the liquid outlet pipe 130 is divided into a plurality of branches of the liquid outlet pipe 130, and each branch of the liquid outlet pipe 130 can be connected into one reactor. And the other end extends to the bottom of the can body 100 to extract the seed liquid in the can body 100. The liquid outlet pump 131 is arranged on a branch of the liquid outlet pipe 130 to enable seed liquid to enter the reactor through the liquid outlet pipe 130. That is, under the power support of the liquid outlet pump 310, the seed liquid in the material-filling tank body 100 is led out through the liquid outlet pipe 130, and then is split through a plurality of branches of the liquid outlet pipe 130, and meanwhile, the liquid outlet pump 131 controls the flow in the corresponding branch, and the action can also be controlled by the control system 150 to adjust the same amount of the seed liquid entering each reactor by controlling a plurality of liquid outlet pumps 131. In addition to the discharge amount controlled by the liquid pump 131 and the control mechanism, a pinch valve (not shown in the figure) may be disposed on the branch of the liquid outlet pipe 130 to control the discharge amount, and the discharge amount is opened and closed by using the tank pressure as power. The parallel feeding device further includes a feeding pipe 140, and a feeding pump 141 is disposed on the feeding pipe. Feed liquid can be fed through the feed pipe 140 to be fed to the feed tank body 100, and peristaltic pumps or syringe pumps can be used as the feed pump 131 and the feed pump 141, for example.

The parallel feeding device further comprises an air pressure adjusting device, a temperature adjusting device and a pH adjusting device which act on the feeding tank body 100. The tank cover 110 of the material tank is provided with an interface for installing a temperature electrode 170, a pH electrode 160 and a DO electrode 180, which are respectively used for measuring the temperature, the pH value, the dissolved oxygen and the pressure of the material liquid in the tank body, the indication of the temperature electrode 170 is analyzed by a control system, the action of a final control temperature control mechanism 171 is controlled by the analysis of the control system, the temperature is controlled according to the preset parameters, and the pH electrode 160 is analyzed by the control system, the action of an acid-base peristaltic pump 162 is finally controlled by the analysis of the control system, so that the reagent introduced by a pH adjusting pipeline 161 is controlled, and the pH value of the material liquid in the tank body is adjusted. The air inlet pipeline 181 and the air outlet pipeline 182 are also inserted on the can cover 110 of the can, and the air inlet and the air outlet of the can are regulated and controlled by the control system so as to maintain the required can pressure.

The parallel reactor system further includes tank pressure control means for controlling the tank pressure of each reactor tank, as shown in fig. 2, including a first inlet line 210 and an outlet line 230 communicating with the reactor tank 200, a buffer bottle 240, a pressure detecting mechanism, and an outlet regulating valve.

The reactor tanks 200 are provided with a first air inlet pipeline 210 and an air outlet pipeline 230 for air inlet and air outlet, each reactor tank 200 is provided with at least one group of air inlet and air outlet pipelines, wherein all the air outlet pipelines 230 are communicated with the buffer bottle 240, the pressure detection mechanism detects the pressure in the buffer bottle 240, a main air outlet pipeline 250 is led out of the buffer bottle 240, and an air outlet regulating valve is arranged on the main air outlet pipeline 250 to regulate the pressure in all the reactor tanks 200 through the buffer bottle 240. The action of the exhaust gas regulating valve is controlled by the action of a human hand or a control mechanism, namely, a self-control exhaust gas regulating valve 251 or a mechanical back pressure valve 253 is selected as shown in the figure. Illustratively, the pressure sensing mechanism may be a mechanical pressure gauge or a pressure sensor 252.

Further preferably, after the self-controlled exhaust gas regulating valve 251 or the mechanical back pressure valve 253 is selected, one or more of the following pipeline structures may be added.

(1) A secondary exhaust line is led out of the buffer bottle 240, and a manual bypass valve 254 is arranged on the secondary exhaust line. Is manually adjusted by a manual bypass valve 254 when the exhaust flow is insufficient or the control of the self-controlled exhaust gas adjustment valve 251 fails.

(2) The buffer bottle 240 is further led out of a pressure relief pipeline, and a safety pressure relief valve 255 is arranged on the pressure relief pipeline. This avoids the risk of tank pressure of the tank pressure control device of the parallel reactor being too high.

(3) A drain 260 is further led out of the buffer bottle 240, and is led out from the bottom of the buffer bottle 240 to drain the waste liquid.

An intake flow meter and a check valve 213 are provided in the first intake pipe 210. Among them, the intake flowmeter may be a mass flowmeter 211 and/or a rotameter 212. The check valve 213 is designed to avoid reverse channeling of the gas.

Device example two

Based on the first embodiment of the device, this embodiment discloses three specific composition forms of the parallel sampling device.

Composition form 1

Referring to fig. 3-a, a parallel sampling device for sampling reactants in a reactor tank 200, comprising: a plurality of sampling pipes 300, one ends of which are respectively positioned in different reactor tanks 200, and the other ends of which are sample outlets 301; a first peristaltic pump 302 disposed on each of the sampling tubes 300 and positioned between the reactor tank 200 and the sample outlet 301; a second peristaltic pump 303 disposed on each of the sampling tubes 300 and positioned between the first peristaltic pump 302 and the sample outlet 301; a heating cooler 304 provided on each sampling tube 300, respectively, and located between the second peristaltic pump 303 and the sample outlet 301; a main pipe 310, one end of which is connected to the air filter 311, and the other end of which is provided with a plurality of gas sub-pipes 312, each gas sub-pipe 312 is respectively communicated with one sampling pipe 300, and the connection part is positioned between the first peristaltic pump 302 and the second peristaltic pump 303; a first pinch valve 320 is disposed on the main pipe 310 and is located between the air filter 311 and the gas branch pipe 312. The heating cooler 304 is used for maintaining high temperature when not sampling to avoid contamination of the culture solution by mixed bacteria along the pipeline, and sampling after cooling when sampling, so as to avoid influence of high temperature on the form of microorganisms in the sampling solution. Specifically, in the embodiment of the invention, three-channel automatic quantitative sterile sampling devices are used for explanation.

The three-channel automatic quantitative sterile sampling device can simultaneously or respectively sample three reactor tanks 200, integrates the work originally needing three sets of sampling devices into one set of device, and reduces the equipment cost and maintenance workload of the sampling devices; and simultaneously, the connection use of the connection pipeline is simplified, the three reactor tanks 200 are conveniently sampled in a short time, and the risk of experimental failure caused by the pipeline system is reduced. In addition, since the set parameters and the environmental parameters of all the sampling pipes 300 are kept consistent, the system deviation can be well eliminated.

The specific operation steps of sampling by using the parallel sampling device with the composition are as follows:

1. control of contamination prevention of sampling operations:

1. all devices on the main pipe 310, the gas branch pipe 312 and the sampling pipe 300 of the device adopt a pipe clamping mode, for example: the pinch valve, peristaltic pump and heating cooler 304 are not in direct contact with the sampled device, so that secondary pollution can be avoided.

2. The main pipeline 310, the gas branch pipeline 312 and the sampling pipeline 300 adopt flexible silicone tubes, an air filter 311 is arranged on the main pipeline 310, the sampling pipeline 300 and the reactor tank 200 are connected in place, and the main pipeline 310 and the sampling pipeline 300 are clamped or aseptically wrapped with an external interface. Before use, the reactor tank 200 and the reactor tank are put into an autoclave for sterilization, and after sterilization, the reactor tank is installed and connected with each reactor, corresponding valves, peristaltic pumps and the like as shown in fig. 1. When in use, the pipeline is clamped into the pinch valve, the peristaltic pump and the heating cooler 304 respectively only by operating under the environment conditions required by the culture process. Because the first peristaltic pump 302 and the second peristaltic pump 303 clamp the sampling tube 300, the first pinch valve 320 clamps the main tube 310, and the main tube 310, the gas branch tube 312 and the sampling tube 300 cannot be directly communicated with the reactor tank 200, and at this time, when the sampling tube 300 is connected with an external interface, the pollution of the culture solution caused by the connection process can not be generated.

3. The sampling pipeline 300 provided with the first peristaltic pump 302 and the second peristaltic pump 303 is connected with an external non-sterile pipeline, so that mixed bacteria possibly pass through the first peristaltic pump 302 and the second peristaltic pump 303 to pollute the reactor tank 200 in the long-time culture process, sterile air (compressed air is filtered by the air filter 311) is adopted to perform sterile pressure maintaining on the system, the first pinch valve 320 is opened after sampling is finished, and the compressed air enters each sampling pipeline 300 from the main pipeline 310 through each gas branch pipeline 312 respectively to blow off liquid drops adhered to the pipeline wall; meanwhile, according to the principle of bus disinfection, the heating and cooling devices 304 arranged on the sampling pipelines 300 are utilized to carry out heat preservation on the sampling pipelines 300 so as to prevent the reverse propagation of mixed bacteria along the pipelines and pollute the inside of the reactor tank 200. If the sample is taken out and passed through the section of high-temperature pipeline, components, microscopic appearance and the like can be influenced, and the pipeline can be cooled by switching a heating mode into a cooling mode during sampling.

2. And (3) controlling the discharge amount of waste liquid in the sampling process:

1. due to the above mentioned sterility control measures, the sterility of the interior of the tubing is ensured by opening the first pinch valve 320 prior to sampling and reversing, e.g. counter-clockwise rotation, the first peristaltic pump 302 to pump the residual liquid in the sampling tubing 300 in the reactor tank 200 back into the sampling device. The first peristaltic pump 302 and the second peristaltic pump 303 are rotated in the forward direction, such as clockwise rotation, so that the sample is taken out in real time without discharging waste liquid.

2. According to the sampling amount requirement, the corresponding peristaltic pump flow is calibrated under the conditions of the peristaltic pump rotating speed, the pipeline inner diameter, the pressure difference at two ends of the peristaltic pump and the like, and the sampling amount can be controlled through setting the running time of the peristaltic pump. When the desired sample size is reached, the first peristaltic pump 302 is rotated in reverse, e.g., counter-clockwise, and the second peristaltic pump 303 continues to rotate in forward, e.g., clockwise.

At this time, with the access point of the sterile air pipeline as a demarcation point, the residual liquid near the end of the first peristaltic pump 302 is completely pressed into the reactor tank 200, the sample taken out near the end of the second peristaltic pump 303 completely flows into an external sampling device or an analyzer under the pushing of the sterile air, and excessive sampling and waste liquid cannot be generated in the whole process.

Composition form 2

Referring to fig. 3-b, a schematic diagram of another parallel sampling device according to the present embodiment is shown.

The parallel sampling device includes: a plurality of sampling pipes 300, one ends of which are respectively positioned in different reactor tanks 200, and the other ends of which are sample outlets 301; a first peristaltic pump 302 disposed on each of the sampling tubes 300 and positioned between the reactor tank 200 and the sample outlet 301; a second pinch valve 321 disposed on each of the sampling tubes 300 and positioned between the first peristaltic pump 302 and the sample outlet 301; a heating cooler 304 provided on each sampling pipe 300, respectively, and located between the second pinch valve 321 and the sample outlet 301; a main pipe 310, one end of which is connected to the air filter 311, and the other end of which is provided with a plurality of gas sub-pipes 312, each gas sub-pipe 312 is respectively communicated with one sampling pipe 300, and the connection part is positioned between the first peristaltic pump 302 and the second pinch valve 321; a first pinch valve 320 is disposed on the main pipe 310 and is located between the air filter 311 and the gas branch pipe 312. The heating cooler 304 is used for maintaining high temperature when not sampling to avoid contamination of the culture solution by mixed bacteria along the pipeline, and sampling after cooling when sampling, so as to avoid influence of high temperature on the form of microorganisms in the sampling solution. Specifically, in the embodiment of the invention, three-channel automatic quantitative sterile sampling devices are used for explanation.

The second peristaltic pump 303 in the first embodiment is used to withdraw residual sample from the tubing between the first peristaltic pump 302 and the second peristaltic pump 303 after sampling is completed out of the sampling tubing 300 and to seal the sampling tubing 300 from contamination spreading from the free port of the sampling tubing 300 to the reactor tank 200. Since the sterile air entering the sampling tube 300 from the gas sub-tube 312 can blow out the residual sample in the tube between the first peristaltic pump 302 and the second peristaltic pump 303 out of the sampling tube 300, in order to simplify the system structure, in this embodiment, the second peristaltic pump 303 in the first embodiment is replaced by the second pinch valve 321, and in order to avoid the interference of the mutual flow of the liquid through the main tube 310 when the sampling channel samples from the reactor, a one-way valve 324 is disposed on each of the three gas sub-tubes 312, and the reverse flow of the sterile air is avoided by virtue of the one-way valve 324, so as to prevent the cross flow of the liquid after the liquid enters the main tube 310 through the gas sub-tube 312.

Form 3

Referring to fig. 3-c, a schematic diagram of another parallel sampling device according to the present embodiment is shown.

In this embodiment, the parallel sampling device for sampling the reactants in the reactor tank 200 comprises: a plurality of sampling pipes 300, one ends of which are respectively positioned in different reactor tanks 200, and the other ends of which are sample outlets 301; a third pinch valve 322 disposed on each of the sampling pipes 300 and located between the reactor tank 200 and the sample outlet 301; a second pinch valve 321 disposed on each of the sampling pipes 300 and located between the third pinch valve 322 and the sample outlet 301; a heating cooler 304 provided on each sampling pipe 300, respectively, and located between the second pinch valve 321 and the sample outlet 301; a main pipe 310, one end of which is sequentially provided with a fourth pinch valve 323 and an air filter 311, the other end of which is provided with a plurality of gas sub-pipes 312, each gas sub-pipe 312 is respectively communicated with one sampling pipe 300, the joint is positioned between the first peristaltic pump 302 and the second pinch valve 321, and an injection pump 326 is connected between the fourth pinch valve 323 and the air filter 311; buffer tanks 325 are respectively provided on each of the gas sub-pipes 312. The heating cooler 304 is used for maintaining high temperature when not sampling to avoid contamination of the culture solution by mixed bacteria along the pipeline, and sampling after cooling when sampling, so as to avoid influence of high temperature on the form of microorganisms in the sampling solution. Specifically, in the embodiment of the invention, three-channel automatic quantitative sterile sampling devices are used for explanation.

The specific flow of the sampling operation performed sequentially for each channel of the parallel sampling device provided by composition form 3 is as follows,

1. the culture solution in the reactor tank 200 is pumped into the buffer tank 325 on the gas distribution pipe 312:

closing the fourth pinch valve 323 on the main line 310 and the second pinch valve 321 on the sampling line 300; the third pinch valve 322 on the sampling tube 300 is opened and then the syringe pump 326 is activated to pump the sample into the buffer tank 325.

2. Sample preparation:

the third pinch valve 322 is closed, the second pinch valve 321 is opened, the syringe pump 326 is compressed to press the culture medium temporarily stored in the buffer tank 325 into the sample container through the sampling pipe 300, and then the second pinch valve 321 is closed.

And (3) sequentially repeating the operations of the step (1) and the step (2) to finish the sampling operation of each sampling channel.

3. Blowing:

the fourth pinch valve 323 on the main pipe 310 is opened, and then the second pinch valve 321 on each sampling pipe 300 is sequentially or simultaneously opened, so that the buffer tank 325 of each sampling passage and the residual culture solution in the sampling pipe 300 are blown into the sample container.

In the foregoing embodiments, in an embodiment of the present invention, the sampling device further includes a controller, where the controller includes: an input/output device; the central controller is connected with the input and output device; a multi-path relay connected to the control part of each sampling channel, such as pinch valve, peristaltic pump and syringe pump 326; and the power supply module is connected with the multipath relay. Each relay is connected with the central controller in parallel, namely each relay independently controls one sampling channel, and the sampling operation process of each sampling channel can be independently set; that is, each sampling channel may be simultaneously or separately sampled.

Device example III

On the basis of the first device embodiment or the second device embodiment, as shown in fig. 4, the parallel reactor system further includes an electrode calibration device for calibrating the sensing unit, where the electrode calibration device includes: a calibration tank, a third air inlet pipeline 420, a third air outlet pipeline 430, a second stirring mechanism, a temperature electrode 440, a temperature adjusting mechanism and a calibration mechanism 9. Wherein the calibration tank is composed of a calibration tank body 400 and a calibration tank cover 410. The second stirring mechanism acts on the liquid in the calibration tank body 400 to enable the liquid to be mixed more uniformly, so that the physical and chemical parameters of the liquid in the calibration tank body 400 are distributed uniformly. The second stirring mechanism used by the electrode calibration device is similar in structural composition to the parallel feed device and the stirring mechanism used in the reactor tank, and comprises a stirring motor 220, a stirring paddle 221 and a rotation speed detection device 222 as shown in fig. 2. The temperature adjusting mechanism can be heated by a heating source 442 positioned outside the calibration tank 400, and is cooled by a cooling pipe 443 positioned inside the calibration tank 400.

The calibration tank cover 410 is covered at the tank opening of the calibration tank body 400, the liquid to be measured is contained in the calibration tank body 400, and a plurality of plug connectors 411 are designed on the calibration tank cover 410. In this embodiment, three electrodes 450 to be measured can be inserted into the left 3 insertion ports 411, and a standard thermometer 441 and a temperature electrode 440 are disposed on the right side. The temperature adjusting mechanism and the calibration mechanism 460 can be integrated in a control cabinet of the parallel reactor, and can also be independently integrated in an electrode calibration device, and a main measurement and control system of the reactor is connected through communication to complete the calibration step.

The temperature electrode 440 is used to measure the temperature of the liquid in the calibration tank 400 so that the temperature adjustment structure can be controlled to adjust the temperature of the liquid in the calibration tank 400. The third air inlet pipeline 420 and the third air outlet pipeline 430 are arranged on the calibration tank body 400 or the calibration tank cover 410 for air inlet and air outlet, wherein the third air inlet pipeline 420 is provided with a gas flow meter 421 and an air inlet regulating valve, and the air outlet pipeline is provided with a pressure detection device 42 and an air outlet regulating valve 41. Meanwhile, the third air inlet pipeline 420 includes a plurality of air inlet branches arranged in parallel, and the air flow meter 421 is disposed on the air inlet branches. The device can comprise 2 air inlet branches, and mixed gas of nitrogen and oxygen is respectively introduced, so that liquid to be detected with certain oxygen dissolution concentration can be formed by controlling the gas phase partial pressure of the nitrogen and the oxygen, the proportion and the tank pressure, and the value of the oxygen dissolution concentration can be used as a calibration standard value of the DO electrode. And the oxygen is changed into carbon dioxide or methanol, so that the calibration of the carbon dioxide electrode or the methanol electrode can be realized.

The two adjustment actions of temperature regulation and air intake and exhaust regulation can be realized manually or automatically by the control system 470.

The electrode calibration device is used for calibrating the temperature electrode in parallel and comprises the following steps:

s100, inserting the temperature electrode to be measured of the parallel reactor into an inserting port 411;

s200, injecting liquid into the calibration tank body 400, placing the calibration tank cover 410 on the calibration tank body 400 for fixing, starting the stirring mechanism, controlling the temperature through the temperature electrode 440 and the temperature regulating mechanism to enable the measured liquid to reach a preset temperature value, and taking the indication value of the standard thermometer 441 as a calibration standard value;

s300, parallel calibration of the temperature electrode to be measured is carried out through the calibration mechanism 460.

The invention also discloses a parallel calibration method of the pH electrode, which uses the parallel reactor electrode calibration device and comprises the following steps:

s100, inserting a pH electrode to be tested of the parallel reactor into the plug-in port 411;

s200, filling a buffer solution with a required pH value into the calibration tank body 400, placing the calibration tank cover 410 on the calibration tank body 400 for fixing, and setting the pH value of the buffer solution as a standard value for calibrating the pH electrode;

s300, parallel calibration of the pH electrode is carried out through the calibration mechanism 460.

Of course, the ORP electrode is inserted into the interface, and ORP standard liquid is injected into the tank body, and the parallel calibration method is also suitable for measuring the calibration of the corresponding OPR electrode. In addition, the method of this embodiment may be used for calibration as long as it is an electrode calibrated with a standard solution, such as ORP electrode, conductivity electrode, fluoride electrode.

The invention also discloses a parallel calibration method of the DO electrode, which uses the parallel reactor electrode calibration device and comprises the following steps:

s100, inserting DO electrodes to be tested of the parallel reactor into the plug-in interface 411;

s200, injecting proper liquid into the tank body, and placing the calibration tank cover 410 on the calibration tank body 400 for fixing; forming liquid with required oxygen dissolved concentration by controlling the proportion of partial pressure of air inlet and tank pressure, and setting the oxygen dissolved concentration as the standard value of DO electrode calibration; and a reference electrode with higher precision can be further inserted into the connector 21, so that the calculated gas dissolution concentration formed by the gas components controlled by the flowmeter can be compared with the measured concentration of the reference electrode, and the trusted result is input into a calibration interface of the parallel reactor as a standard value for DO electrode calibration.

S300, submerging the electrode to be measured and the DO electrode, and carrying out parallel calibration of the DO electrode through the calibration mechanism 460. The ratio of oxygen to nitrogen components and/or the total pressure of gas introduced into the calibration tank 400 can be changed according to the calibration requirement, the balance state is accelerated by stirring, and the DO electrode of the parallel reactor can be calibrated in parallel after stabilization, so that the linearity of the calibration result or the parallelism among the calibration results under different calibration values in the calibration interval can be compared.

In this embodiment, the oxygen introduced in the above is changed into other gases (such as carbon dioxide, etc.), and the calibration of the electrode with the solubility of the gases (such as dissolved carbon dioxide, etc.) can be achieved by operating according to the same flow steps.

It should be noted that, in the foregoing embodiment, the parallel sampling device, the tank pressure control device, the electrode calibration device, the control component of the parallel feeding device, the corresponding software and the system may be integrated in the control cabinet of the parallel reactor, or may be separately arranged as a control structure independent of the control cabinet of the parallel reactor, and connected with the parallel reactor through the communication device in a communication manner, so as to complete the corresponding action.

Method embodiment

s100, calibrating the sensing unit through the electrode calibration device;

s300, adding a culture medium into the reactor tank, and sterilizing;

In summary, to reduce the systematic deviation, the improvement of the detection accuracy of the sensor is mainly involved in addition to the geometric dimension of the reactor tank, secondary installation dimensions including the pitch of the stirring paddles, etc.

Common sensors for bioreactor designs include: temperature T, pH value, dissolved oxygen DO, stirring rotation speed AG, tank pressure P, inlet air flow F, feeding amount, defoaming AF, and other dissolved carbon dioxide, ORP, tail gas concentration and the like according to experimental requirements. If the accuracy requirement is high, the air inlet flow can adopt a thermal mass flowmeter, the stirring rotating speed can adopt a variable frequency motor or a servo motor, the feeding quantity can adopt a peristaltic pump, a weighing or electromagnetic flowmeter or a syringe pump for indirectly calculating the flow, and the like. From the control point of view, the parameters can be independently measured and controlled, and the method belongs to the conventional prior art.

Some parameters critical to the culture process can be optimized from the measurement and control technical scheme to eliminate or reduce the system deviation, and some parameters cannot be directly measured and must be optimally designed from the whole system scheme according to the principle. These parameters include:

■ Culture temperature T: directly influencing the kinetic constants of various non-reactions in cells during the culture of microorganisms

■ pH of the culture solution: can cause change of cell membrane charge, thereby affecting nutrient absorption by microorganism and enzyme activity in metabolic process

■ Dissolved oxygen DO: macroscopically reflecting the balance of oxygen supply and oxygen consumption in the culture process, and further improving the characteristics of fluid decomposition, DO variation, critical oxygen range, half-starvation state, etc. by matching with other parameters

■ Reactor pot pressure: affecting gas dissolution. The importance is the same as the stirring rotation speed and the air flow, but can be further optimized so as to improve the control consistency and reduce the measurement and control cost

■ Grafting ages: the microorganism is an active living body, if the inoculation process has a certain difference in the age of the seed, the further difference of the culture result can be caused, and the difference can not be quantified

■ Sampling: the culture process also has time-varying characteristics, and the difference of sampling time can interfere detection; meanwhile, for smaller-scale reactors, the sampling amount not only affects the limitation of sampling times, but also causes unquantifiable interference to the culture process and results due to the difference of the residual culture solution volumes of the reactors

Therefore, the invention is innovated and optimally designed from the parallel reactor system aiming at the characteristics, so as to reduce the system deviation of the parallel reactor to the maximum extent and improve the reliability and reliability of experimental results.

It should be noted that the above embodiments can be freely combined as needed. The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. A parallel reactor system, comprising:

The material beating tanks are respectively communicated with each reactor tank through an inoculation pipeline for inoculation, each inoculation pipeline is provided with an inoculation pump, and a first stirring mechanism is arranged in each material beating tank to uniformly mix seed liquid in each material beating tank;

one end of the liquid outlet pipe is divided into a plurality of branches of the liquid outlet pipe, each branch of the liquid outlet pipe is connected into one reactor, and the other end of the liquid outlet pipe extends to the bottom of the material beating tank body so as to extract seed liquid in the material beating tank body;

the liquid outlet pumps are respectively arranged on each branch of the liquid outlet pipe;

2. The parallel reactor system according to claim 1, wherein,

the parallel sampling device comprises: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; the first peristaltic pump is arranged on each sampling pipe and is positioned between the sampling end and the sample outlet; the second peristaltic pump is arranged on each sampling pipe and is positioned between the first peristaltic pump and the sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second peristaltic pump and the sample outlet; one end of the gas main pipeline is connected with an air filter, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the first peristaltic pump and the second peristaltic pump; the first pinch valve is arranged on the gas main pipeline and is positioned between the air filter and the gas branch pipeline.

3. The parallel reactor system according to claim 1, wherein,

the parallel sampling device comprises: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; the first peristaltic pump is arranged on each sampling pipe and is positioned between the sampling end and the sample outlet; a second pinch valve disposed on each of said sampling lines and positioned between said first peristaltic pump and said sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second pinch valve and the sample outlet; one end of the gas main pipeline is connected with an air filter, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the first peristaltic pump and the second pinch valve; the first pinch valve is arranged on the gas main pipeline and is positioned between the air filter and the gas branch pipeline;

4. The parallel reactor system according to claim 1, wherein,

the parallel sampling device comprises: the sampling ends of the sampling pipelines are respectively positioned in different reactor tanks, and the other ends of the sampling pipelines are sample outlets; a third pinch valve disposed on each of the sampling lines and located between the reactor tank and the sample outlet; the second pinch valve is arranged on each sampling pipe and is positioned between the third pinch valve and the sample outlet; the first heating cooler is respectively arranged on each sampling pipe and is positioned between the second pinch valve and the sample outlet; a fourth pinch valve and an air filter are sequentially arranged at one end of the gas main pipeline, the other end of the gas main pipeline is branched into a plurality of gas branch pipelines, each gas branch pipeline is respectively communicated with one sampling pipeline, and the joint is positioned between the third pinch valve and the second pinch valve; a syringe pump is connected with the gas main pipeline, and a connecting point is positioned between the fourth pinch valve and the air filter; the buffer tanks are respectively arranged on each gas branch pipeline.

5. The parallel reactor system according to any one of claims 1-4, further comprising electrode calibration means for calibrating the sensing unit; the electrode calibration device comprises:

The tank body is used for containing the liquid to be tested;

the tank cover can be covered on the tank body and is provided with a plurality of faucet ports, and electrodes to be tested are inserted into the faucet ports;

the third air inlet pipeline and the third air outlet pipeline are arranged on the tank body or the tank cover, the third air inlet pipeline comprises a plurality of air inlet branches which are arranged in parallel, each air inlet branch is provided with a gas flowmeter and a third air inlet regulating valve, and the third air outlet pipeline is provided with a pressure detection device and a third air outlet regulating valve;

a temperature adjusting mechanism for heating and cooling the tank body;

6. A parallel reactor system according to any one of claims 1 to 4, characterized in that,

and/or;

And/or;

the buffer bottle is further provided with a drain pipeline, the drain pipeline is led out of the buffer bottle from the bottom of the buffer bottle, and the drain pipeline is provided with a drain valve.

7. The parallel reactor system according to any one of claims 1 to 4, wherein the sensing unit comprises one or more of a temperature sensor, a pH sensor, a dissolved oxygen sensor, a full tank weighing sensor, an exhaust gas analyzer, a velocimeter sensor, a pressure sensor, a defoaming sensor, a feed weighing sensor, a cell microscopic on-line observer, a viable bacteria amount sensor, an oxidation-reduction potential ORP sensor, a bacteria concentration OD sensor, and an air flow sensor.

8. The parallel reactor system according to claim 5, wherein,

further comprising a monitoring system and an analysis system; the monitoring system is communicated with the sensing unit, the parallel sampling device, the electrode calibration device, the main exhaust regulating valve and the pressure detection mechanism for data acquisition and control, and the analysis system analyzes the data acquired by the monitoring system.

9. The parallel reactor system according to any one of claims 1-4, wherein the first and second stirring mechanisms each comprise a stirring paddle and a stirring motor driving the stirring paddle.

10. A method of parallel control testing, characterized in that the parallel reactor system according to claim 8 is used, comprising the steps of:

s100, calibrating the sensing unit through the electrode calibration device;

s300, adding a culture medium into the reactor tank, and sterilizing;

s400, inoculating the seeds cultured in the threshing tanks into each reactor tank;