CN210340879U - Automatic dissolved oxygen periodic fluctuation device - Google Patents
Automatic dissolved oxygen periodic fluctuation device Download PDFInfo
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- CN210340879U CN210340879U CN201920748506.9U CN201920748506U CN210340879U CN 210340879 U CN210340879 U CN 210340879U CN 201920748506 U CN201920748506 U CN 201920748506U CN 210340879 U CN210340879 U CN 210340879U
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Abstract
The utility model discloses an automatic dissolved oxygen periodic fluctuation device, which comprises a first gas source, a second gas source and a gas outlet, wherein the first gas source and the second gas source are respectively in fluid connection with the gas outlet through pipelines; wherein, a valve is respectively arranged on the pipeline between the first gas source and the gas outlet and the pipeline between the second gas source and the gas outlet and used for switching a ventilation pipeline. The utility model discloses an automatic dissolved oxygen periodic fluctuation device, the condition of accessible editorial outage timer is controlled opening and close of electric ball valve, and then realizes automatic dissolved oxygen periodic fluctuation. The utility model discloses can be applicable to the dissolved oxygen periodic pulse of various microbial reactor cultivates the in-process, provide the automatic periodically undulant environment of dissolved oxygen, provide good basis for the live body dynamics research of microorganism.
Description
Technical Field
The utility model relates to the technical field of biological cultivation, biological fermentation, etc., in particular to an automatic dissolved oxygen periodic fluctuation device.
Background
Dissolved oxygen is an important factor affecting the growth of microorganisms during the biological cultivation process. At present, few studies report the influence of fermentation yield and incomplete mixing during fermentation, and most of the reported studies focus only on dissolved oxygen concentration. The Vardar and Lilly studies found that penicillin production decreased dramatically below a critical oxygen concentration (around 30% of air saturation), and also found that penicillin production decreased as dissolved oxygen concentration fluctuated above and below this critical level (Vardar, F. and Lilly, M.D., Effect of closed isolated oxygen concentrations on product formation in cellulose transfer. European journal of Applied Microbiology & Biotechnology, 1982.14(4): p.203-. Larsson and Enfors utilized a dual chamber anaerobic plug flow reactor to find that prolonged (5-10 min) exposure of P. chrysogenum to an anaerobic environment reduces the respiratory capacity of microorganisms without any irreversible effect (Larsson, G. and Enforns, S.O., studios of inert in biologicals: Effects of inert microorganism restriction on microbial inoculum Engineering,1988.3(3): p. 123. 127.). However, the above experiments did not investigate the effect of dissolved oxygen fluctuations on penicillin production. Diano et al investigated the formation of polyols as by-products during continuous fermentation of Aspergillus niger under oxygen-limited conditions and showed that the biomass-synthesized polyols accounted for up to 22% of the carbon source under oxygen-limited conditions (Diano, A., et al., polyol synthesis in Aspergillus niger: fluorine of oxygen availability, carbon and nitrogen sources on the metabolism, Biotechnology and Bioengineering, 2010.94(5): p. 899-908.). Pedersen et al, in exploring the effects of oxygen and osmotic pressure on the production of saccharifying enzymes in submerged fermentation processes, found that appropriate oxygen limitation and high osmotic pressure can increase the production of saccharifying enzymes (Lasse, P., et al, Industrial glucoamylase fed-batch resins from oxidation and high-throughput experimentation, 2011.109 (1): p. 116-124.).
With the increasing research on the influence of incomplete mixing in the fermentation process, the role of dissolved oxygen in the fermentation process is gradually uncovered. According to the Hanison and Topiwala studies, microorganisms are insensitive to oxygen concentration variations between 10% and 100% in air saturation (Harrison, D.E.F. and Topiwala, H.H., transformed and scientific stations of continuous in biochemical engineering, 1974.3: p.167-219.). Sweer studied the periodic pulsing of the dissolved oxygen in yeast cells by circulating nitrogen and air through a single reactor and found that fluctuations in dissolved oxygen had little effect on the composition of the cells (Sweere, A.P., et al., Experimental simulation of oxygen profiles and the oxygen on baker's product: I. One-sensor system, Biotechnology and Biotechnology, 2010.31(6): p.567-578). Oosterhas uses a two-reactor Dissolved oxygen limiting-pulsing device to find that exposure of cultures to fluctuations in oxygen concentration decreases the yield of Gluconobacter oxydans gluconic acid, which decreases in relation to the time of exposure of the cells to oxygen-limiting conditions, however, the potential ability of the cells to produce gluconic acid is not affected (Oosterhas, N.M. and Kossen, N.W., Dissolved oxygen control profiles in a process-scale biorator, Biotechnology and Biotechnology engineering, 1984.26(5): p.546), and thus, Gluconobacter oxydans are not sensitive to externally imposed environmental fluctuations. In the above studies Vardar and Lilly also found a decrease in penicillin production in the dissolved oxygen fluctuation experiment of Penicillium chrysogenum. Furthermore, Moes et al found that the oxygen-rich region in Bacillus subtilis produced ethyl acetate, while the oxygen-limited region produced butanediol as a byproduct (Moes, J., et al, organic culture with oxygen-sensitive product distribution as a product for use in a fermentation bioreactor transport. Biotechnology and Biotechnology, 2010.27(4): p. 482. sup. 489.), so that the oxygen-limited region in the fermentor could be detected by the discharged product.
At present, no report about an automatic dissolved oxygen periodic fluctuation device exists in China, and dissolved oxygen fluctuation is only limited to manual regulation of ventilation volume in a period of time. The size of the dissolved oxygen is related to the oxygen content in the aeration and the aeration, and the method of changing the aeration has an effect on the growth of the microorganisms and the distribution of the dissolved oxygen, so that the simulated dissolved oxygen periodic fluctuation cannot meet the real situation of the dissolved oxygen gradient in a large-scale reactor. The utility model discloses do not change the air output in the design of dissolved oxygen periodic fluctuation device, but start with from the oxygen content of ventilating, design the periodic fluctuation device of control dissolved oxygen on the basis that does not change the air output, can simulate the periodic fluctuation environment of automatic dissolved oxygen.
Disclosure of Invention
An object of the utility model is to solve above-mentioned problem, provide an automatic dissolved oxygen periodic fluctuation device, can simulate out automatic dissolved oxygen periodic fluctuation environment, apply to the microbial fermentation, provide good basis for the living body dynamics research of microorganism.
In order to achieve the above object, the present invention provides an automatic dissolved oxygen periodic fluctuation device, which comprises a first gas source, a second gas source and a gas outlet, wherein the first gas source and the second gas source are respectively in fluid connection with the gas outlet through a pipeline; wherein, a valve is respectively arranged on the pipeline between the first gas source and the gas outlet and the pipeline between the second gas source and the gas outlet and used for switching a ventilation pipeline.
Further, the automatic dissolved oxygen periodic fluctuation device comprises a first pipeline, a second pipeline and a third pipeline; wherein the first gas source is connected to the third conduit via the first conduit and is in fluid connection with the gas outlet, and the second gas source is connected to the third conduit via the second conduit and is in fluid connection with the gas outlet; the first pipeline is provided with a first valve, and the second pipeline is provided with a second valve.
Further, the first valve and the second valve are electric ball valves, and the electric ball valves are connected to the first pipeline and the second pipeline through ball valve connecting pipelines.
Further, the electric ball valve is a power-off electric ball valve.
Further, the first valve is a normally open type electric ball valve or a normally closed type electric ball valve, and the second valve is a normally open type electric ball valve or a normally closed type electric ball valve.
Further, the first valve is of a different type than the second valve.
Further, the first valve and the second valve are respectively in conductive connection with a power-off timer, and the power-off timer controls the opening and closing of the first valve and the second valve.
Further, a mass flow meter is arranged on the third pipeline and used for controlling the flow.
Further, the first gas source is an inert gas source; the second gas source is an air gas source.
Further, the inert gas is nitrogen.
Furthermore, the first pipeline, the second pipeline and the third pipeline are made of thermoplastic polyurethane, and the ball valve connecting pipeline is made of fluororubber.
Further, the full-closing time of the electric ball valve is 3s, and the full-opening time is 12.5 s.
The utility model discloses in outage formula electric ball valve's inner core is ceramic ball core, and the undergauge ball valve is 14.5mm, and the valve body material is reinforcing polyphenyl ether (PPO), and the interface is the external 4 outer silk straight-through pipes of plastic-aluminum (POM) of branch of internal thread, and electric ball valve length is 14mm, highly is 144mm, and the straight pipe diameter of outer silk is 20 mm.
The utility model has the advantages that:
① the utility model discloses in adopt the outage timer to control the switching of two outage formula electric ball valves, the switching of air pipe can be controlled again to opening and close of two outage formula electric ball valves, and then realize normally ventilating and dissolve the conversion between the oxygen restriction mode, change the oxygen content who lets in gas, wherein, the condition of outage timer can be edited, controls through the editor condition the utility model discloses an opening and close of air pipe, automatic normally ventilate and dissolve and change between the oxygen restriction mode, provide the periodic fluctuation environment of automatic dissolved oxygen, the utility model discloses still set up a mass flow meter in the place ahead of gas outlet, can control gas flow, make gas flow more stable, thereby reduce because the undulant new error of introducing of flow.
② the utility model discloses a device has opened up the buildding of automatic dissolved oxygen periodic fluctuation device, has realized automatic dissolved oxygen periodic fluctuation simultaneously, has saved manpower and material resources in the microorganism live body dynamics research greatly, the utility model discloses the time of well switching step is very brief, can realize switching in the twinkling of an eye, reduces the interference of switching time to the intracellular metabolite change of microorganism, can simulate out the condition of microorganism cell is intracellular more really.
③ the utility model can realize normal ventilation in 3s and switch between the dissolved oxygen limiting modes, and can select single mode, the switching time and frequency can be regulated and controlled arbitrarily according to the experimental requirements, and the precedence order of different modes can be selected according to the experimental requirements, the utility model discloses a device is simple and convenient to construct and operate, and various reactors can also be connected in a flexible way.
④ adopts the utility model discloses an automatic dissolved oxygen periodic fluctuation environment can be provided to the device, is applicable to the dissolved oxygen periodic pulse among the various microbial reactor culture processes, provides good basis for the live body dynamics research of microorganism.
Drawings
Fig. 1 is a schematic diagram of a device for periodic dissolved oxygen limiting-pulse experiment in embodiment 1 of the present invention.
Fig. 2 is a diagram showing the fluctuation of dissolved oxygen in the cold mold experiment in embodiment 1 of the present invention.
Fig. 3 is a dissolved oxygen fluctuation diagram of the periodic dissolved oxygen limiting-pulse experiment in example 1 of the present invention.
The reference numerals in the figures are respectively:
1. a first source of gas;
2. a second gas source;
3. a power-off timer;
4. a mass flow meter;
5. a reactor;
6. a first valve;
7. a second valve;
8. a first pipeline;
9. a second pipeline;
10. a third pipeline;
11. and a gas outlet.
Detailed Description
The following embodiments of the present invention will be further explained with reference to the drawings, and it should be noted that the following embodiments should not be considered as limitations to the specific implementation of the present invention.
Example 1
In this embodiment, an automatic dissolved oxygen periodic fluctuation device is provided, referring to fig. 1, the automatic dissolved oxygen periodic fluctuation device includes: a first gas source 1, a second gas source 2 and a gas outlet 11. The first gas source 1 and the second gas source 2 are respectively in fluid connection with the gas outlet 11 through pipelines. As shown in fig. 1, a valve is disposed on each of the pipeline between the first gas source 1 and the gas outlet 11 and the pipeline between the second gas source 2 and the gas outlet 11 for switching the ventilation pipeline.
Specifically, as shown in fig. 1, the automatic dissolved oxygen periodic fluctuation device includes a first pipeline 8, a second pipeline 9 and a third pipeline 10, and the first pipeline 8, the second pipeline 9 and the third pipeline are connected by a three-way connector; wherein the first gas source 1 is connected to the third pipeline 10 via the first pipeline 8 and further fluidly connected to the gas outlet 11, and the second gas source 2 is connected to the third pipeline 10 via the second pipeline 9 and further fluidly connected to the gas outlet 11. A first valve 6 is disposed on the first pipeline 8, and a second valve 7 is disposed on the second pipeline 9, so that the first pipeline 8 and the second pipeline 9 can be controlled by the first valve 6 and the second valve 7, respectively. In this embodiment, the material of the first pipeline 8, the second pipeline 9 and the third pipeline 10 for gas circulation is thermoplastic polyurethane.
In this embodiment, the first gas source 1 is a nitrogen gas source; the second gas source 2 is an air gas source. The first valve 6 and the second valve 7 are electrically operated ball valves, more specifically, power-off electrically operated ball valves. And, the first valve 6 is different from the second valve 7 in the type of electric ball valve, so that the vent line can be controlled by controlling the opening and closing of the valves, and the oxygen content of the gas can be adjusted. For example, the first valve 6 is a normally open type electric ball valve, i.e., closed and opened by power supply, and the second valve 7 is a normally closed type electric ball valve, i.e., closed and opened by power supply. The electric ball valve is connected with the first pipeline 8 or the second pipeline 9 through a ball valve connecting pipeline. In this embodiment, the material of the ball valve connecting line for connecting the power-off electric ball valve to the first and second lines 8 and 9 is fluororubber.
In this embodiment, the electric ball valves as the first valve 6 and the second valve 7 may be a power-off reset/two-line/DC 9-24V, a full-close time of 3s, and a full-open time of 12.5 s.
As shown in fig. 1, the first valve 6 and the second valve 7 are respectively in conductive connection with a power-off timer 3, the power-off timer 3 controls the opening and closing of the first valve 6 and the second valve 7, the opening and closing of the electric ball valve is controlled by editing the conditions of the power-off timer, the type of the gas flowing in is changed, and therefore the oxygen content of the gas is automatically adjusted.
In this embodiment, the power-off timer 3 may be a timer B14M [2200 ].
In addition, as shown in fig. 1, a mass flow meter 4 is further disposed on the third pipeline 10 for controlling the flow rate, and the ventilation amount can be set, so that the flow rate can be controlled more stably, and new errors caused by flow rate fluctuation can be reduced.
In this embodiment, the mass flow meter 4 may be a seven-star Huachun gas mass flow meter CS 200.
In a specific embodiment, the operation of the automatic dissolved oxygen periodic oscillation device of the present invention is described in detail below.
In order to simulate the real situation of dissolved oxygen fluctuation in a large-scale production fermentation tank and clarify the actual dynamic response in an aspergillus niger cell, the automatic dissolved oxygen periodic fluctuation device carries out periodic dissolved oxygen limitation-pulse experiment and records the fluctuation change of the dissolved oxygen in the experiment. In the experiment, the automatic dissolved oxygen periodic fluctuation device of the utility model is utilized to simulate the fluctuation environment and record the dissolved oxygen change of the aspergillus niger fermentation process in the experiment; the blank control group was a periodic dissolved oxygen limitation-pulse experiment (cold mold experiment) performed under aseptic conditions.
1) Experimental apparatus: adopt the utility model discloses in the device of embodiment carry out periodic dissolved oxygen restriction-pulse experiment, gaseous from gas outlet 11 flow back through an air cleaner enter into reactor 5, simulate out the undulant environment of dissolved oxygen.
The power-off timer 3 can set the running time length, the time intervals and the sequence of power-off and power-on, the timing can be set to be eight groups at most, and the time can be accurate to the second. Rated voltage 220V, working frequency 50Hz, maximum current 10A, and maximum power (resistive load 2200W, inductive load 350W, capacitive load 200W).
The inner core of the electric ball valve is a ceramic ball core, the diameter-reducing ball valve is 14.5mm, the valve body is made of reinforced PPO, the working pressure is 0-1MPa, the working temperature is 0-60 ℃, the connector is an internal thread external connection 4-branch external thread aluminum-plastic straight-through Pipe (POM), the length of the electric ball valve is 14mm, the height of the electric ball valve is 144mm, and the diameter of the external thread straight-through pipe is 20 mm. The input voltage is DC 12V, AC220V, the motor power is 2-3W, the actual power is 0.4-2.5W, the full-open time is 12.5 seconds, and the full-close time is 3 seconds.
The mass flow meter 4 is used for controlling ventilation volume, the ventilation volume can be set according to experiment requirements, the flow specification is less than or equal to 30SLM, the working pressure difference range is 0.1-0.35MPa, the response time is less than or equal to 1s, the leakage rate is 1 multiplied by 10-10 Pa.m 3/sec He, the sealing material is fluororubber, the working environment temperature is 5-45 ℃, and the power supply is +/-8 to +/-16 VDC.
The first pipeline 8 (nitrogen pipeline), the second pipeline 9 (air pipeline) and the third pipeline 10 adopt German BJARCRAG transparent thermoplastic polyurethane PU pipes, the working pressure is 10KGF/CM, the working temperature is-5-50 ℃, and the inner diameter and the outer diameter are 8 x 12 mm.
The ball valve connecting pipeline used for connecting the electric ball valve with the nitrogen pipeline and the air pipeline adopts an American MasterFlex fluororubber pipe, the temperature range is-20-260 ℃, and the inner diameter and the outer diameter are 10 × 17 mm.
2) The experimental method comprises the following steps: utilize the utility model discloses an automatic periodic dissolved oxygen wave motion device establishes the electric ball valve who connects on the nitrogen gas pipeline as: closed when electrified and opened when power is off; an electric ball valve connected to the air line is configured to: open when power is on and close when power is off. The parameters of the power down timer are set as: electrifying for 5min, and deenergizing for 5min, wherein 10min is a cycle of reciprocating circulation. Because air is introduced when the steady state is achieved, the first period is set to firstly cut off the power and introduce the nitrogen for 5min and then introduce the air for 5 min; the sampling period was similarly started by introducing nitrogen gas and ended by introducing nitrogen gas again.
This example selects a single reactor for periodic dissolved oxygen limiting-pulsing experiments and records the experimental data, as shown in fig. 2 and 3.
The experiment is designed to research the influence of cells in a large-scale fermentation tank on the physiological metabolic characteristics of thalli when the cells pass through an oxygen enrichment area and an oxygen deficiency area under extreme conditions, the fluctuation period is set to be 10min, and the normal ventilation and dissolved oxygen limiting modes are respectively 5 min. The purpose is to amplify the intracellular stress response that a cell makes when it passes through different regions. The simulation of the dissolved oxygen fluctuation environment in the large-scale reactor can be carried out in a single reactor or can be realized in a double reactor. The greatest advantage of the simulation in a single reactor is that all cells are in the same physiological state, while in a double reactor it is more consistent with the fact that in large scale fermentations the switch from one zone to another is made. However, the operation process of the double reactors is more complicated and the stability is not as good as that of the single reactor, so that the periodic dissolved oxygen limitation-pulse is selected in the single reactor in the experiment, the physiological metabolic characteristics of all cells can be consistent, and the change condition of the measured intracellular metabolites is more accurate.
3) The experimental results are as follows: referring to fig. 2 and fig. 3, observing the macroscopic data of the whole fermentation process can find that the Dissolved Oxygen (DO) is obvious periodic fluctuation, and it can be seen from the dissolved oxygen data that the automatic dissolved oxygen periodic fluctuation device of the utility model is adopted, the stability and the repeatability of the dissolved oxygen fluctuation are very good. For example, in fig. 2, the periodic fluctuation condition of the dissolved oxygen in 1h of pulse and the change of the dissolved oxygen in one period in the cold die experiment can be seen, and 6 complete and parallel peak shapes can be clearly seen in 1h due to the limitation-the pulse period is 10 min; since there was no cell in the cold mould experiment, there was no consumption of oxygen and the dissolved oxygen rose to a maximum of 130% after aeration. In FIG. 3, the wave shape of the dissolved oxygen within 1h of the start-up dissolved oxygen limitation-pulse is "comb-shaped", which is similar to the wave shape of the dissolved oxygen in the cold mold experiment under aseptic conditions; in contrast, when the dissolved oxygen is consumed in 1 hour of the dissolved oxygen limiting pulse, the dissolved oxygen is only 110% by aeration. After the nitrogen is introduced, both dissolved oxygen can drop to zero.
Can know through the above-mentioned experimental data of analysis, the utility model discloses a design of device can realize automatic dissolved oxygen periodic fluctuation well, is applicable to the dissolved oxygen periodic pulse of various microbial reactor culture in-process, provides good basis for the living body dynamics research of microorganism.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of improvements and decorations can be made without departing from the principle of the present invention, and these improvements and decorations should also be regarded as the protection scope of the present invention.
Claims (12)
1. An automatic dissolved oxygen periodic fluctuation device is characterized by comprising a first gas source, a second gas source and a gas outlet, wherein the first gas source and the second gas source are respectively in fluid connection with the gas outlet through pipelines; wherein, a valve is respectively arranged on the pipeline between the first gas source and the gas outlet and the pipeline between the second gas source and the gas outlet and used for switching a ventilation pipeline.
2. The automatic dissolved oxygen periodic oscillation device of claim 1, wherein the automatic dissolved oxygen periodic oscillation device comprises a first pipeline, a second pipeline and a third pipeline; wherein the first gas source is connected to the third conduit via the first conduit and is in fluid connection with the gas outlet, and the second gas source is connected to the third conduit via the second conduit and is in fluid connection with the gas outlet; the first pipeline is provided with a first valve, and the second pipeline is provided with a second valve.
3. The automatic dissolved oxygen periodic oscillation device of claim 2, wherein the first valve and the second valve are an electric ball valve, and the electric ball valve is connected to the first pipeline and the second pipeline through a ball valve connecting pipeline.
4. The automatic dissolved oxygen periodic oscillation device of claim 3 wherein the motorized ball valve is a power-off motorized ball valve.
5. The automatic dissolved oxygen periodic oscillation device of claim 3 or 4, wherein the first valve is a normally open electric ball valve or a normally closed electric ball valve, and the second valve is a normally open electric ball valve or a normally closed electric ball valve.
6. The automatic dissolved oxygen periodic oscillation device of claim 5 wherein the first valve is of a different type than the second valve.
7. The automatic dissolved oxygen periodic oscillation device of claim 2, wherein the first valve and the second valve are respectively in conductive connection with a power-off timer, and the power-off timer controls the opening and closing of the first valve and the second valve.
8. The automatic dissolved oxygen periodic oscillation device of claim 2, wherein a mass flow meter is disposed on the third pipeline for controlling the flow rate.
9. The automated dissolved oxygen periodic oscillation device of claim 1, wherein the first gas source is an inert gas source; the second gas source is an air gas source.
10. The automatic dissolved oxygen periodic oscillation device of claim 9 wherein the inert gas is nitrogen.
11. The automatic dissolved oxygen periodic fluctuation device of claim 3, wherein the material of the first pipeline, the second pipeline and the third pipeline is thermoplastic polyurethane, and the material of the ball valve connecting pipeline is fluororubber.
12. The automatic dissolved oxygen periodic fluctuation device of claim 3 or 4, wherein the full-closing time of the electric ball valve is 3s, and the full-opening time is 12.5 s.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110093256A (en) * | 2019-05-23 | 2019-08-06 | 华东理工大学 | A kind of automatic dissolved oxygen cyclic fluctuation device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110093256A (en) * | 2019-05-23 | 2019-08-06 | 华东理工大学 | A kind of automatic dissolved oxygen cyclic fluctuation device |
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