CN104109631A - A microalgae cultivation reactor - Google Patents

Abstract

The invention relates to the field of biotechnology, in particular to a microalgae culture reactor capable of setting different concentrations of _CO2 culture environments, including an incubator, an ultraviolet lamp, an air inlet pipeline and an air outlet pipeline, wherein the box body of the incubator It is a transparent box, with an air inlet on the lower side of the front end of the incubator, and an air outlet on the upper side of the rear end of the incubator. Gas flowmeters and valves are arranged on the road, and a _CO2 gas analyzer is installed outside the incubator to detect the carbon dioxide concentration at the inlet and outlet of the incubator. The gas filter device is also equipped with a humidification system in the incubator, and multiple incubators can be set in parallel. The invention screens wild-type microalgae or corresponding mutants by setting culture environments with different concentrations of CO ₂ , and the whole device has a simple structure and is easy to operate.

Description

A microalgae culture reactor

technical field

The invention relates to the field of biotechnology, in particular to a microalgae culture reactor capable of setting different concentrations of _CO2 culture environments.

Background technique

Microalgae is a kind of low-level phytoplankton all over the world. It is widely distributed in oceans, freshwater lakes and brackish water environments. It is an important part of the primary productivity in global aquatic ecosystems. The CO ₂ fixed by the photosynthesis of microalgae accounts for more than 40% of the global CO ₂ fixation every year, and plays a pivotal role in energy conversion and carbon cycle. The photosynthetic productivity of microalgae can reach up to 50g/m ² /d, It is equivalent to 10-50 times the carbon sequestration capacity of the forest. In theory, cultivating 1 ton of microalgae can reduce carbon _dioxide emission by about 1.83 tons. A sustainable and green-friendly approach. When cultivating microalgae, the residual sulfur and residual nitrogen in the gas can become the sulfur source and nitrogen source for the growth of microalgae, and other nutrients required for the growth of microalgae can also be provided by nutrient wastewater to achieve the purpose of purifying wastewater. Therefore, the development of microalgae emission reduction _CO2 technology in China can simultaneously achieve the three goals of "reducing _CO2 emissions, replacing fossil energy, and purifying waste gas and sewage", and has broad industrial and environmental application prospects.

However, microalgae in different environments have different abilities to fix CO ₂ , mainly because microalgae can grow in different CO ₂ concentration environments, and in practical applications we need to screen microalgae that can tolerate high CO ₂ concentrations , usually 15% to 20% CO ₂ concentration, because the CO ₂ concentration in the flue gas can reach up to 20%. In addition, most microalgae have formed a CO ₂ concentration mechanism to assist in improving carbon sequestration capacity during the long-term evolution process, but this mechanism is only activated when the CO ₂ concentration is at or below the air level, so in order to study CO ₂ concentration mechanism, we need to cultivate microalgae under low CO ₂ concentration. Under laboratory conditions, in order to simulate the cultivation and screening of microalgae under high or low _CO2 concentration, a special _CO2 culture reactor is required to _make the microalgae grow under a specific _CO2 concentration.

At present, CO ₂ photobioreactors that are used more are mainly CO ₂ incubators for plant or animal cell culture research, among which CO ₂ incubators for animal cell culture usually do not have light sources, and CO 2 incubators for plant light ₂ It is generally difficult to ensure the sterility of the culture room in the incubator. In addition, whether it is a CO ₂ incubator for plant or animal cultivation, they have in common that they are relatively expensive, occupy a large space, and have high operating costs. In addition, it is easy to cause damage during actual operation. Pollution by bacteria, mold, etc., so it is rarely used in the related research of microalgae, and is also limited by the above factors.

Contents of the invention

The purpose of the present invention is to provide a microalgae culture reactor, which can effectively control the gas flow rate and carbon dioxide concentration, so as to achieve the purpose of screening wild-type microalgae or corresponding mutants in a culture environment with different concentrations of _CO2 . The whole device has a simple structure and is convenient. operate.

The purpose of the present invention is achieved by the following technical solutions:

A microalgae culture reactor, including an incubator, an ultraviolet lamp, an air inlet pipeline and an air outlet pipeline, wherein the air inlet pipeline connected to the gas source is connected to the air inlet on the incubator, and the air outlet pipeline is connected to the air inlet on the incubator. connected to the gas outlet of the incubator, and the ultraviolet lamp tube used to sterilize the incubator before use is set in the incubator, and the gas inlet line and the gas outlet line are provided to maintain the balance of the gas concentration in the incubator. Flow meters and valves. the

The outside of the incubator is provided with a _CO2 gas analyzer that adjusts or closes the gas flow meter by detecting the carbon dioxide concentration at the gas inlet and gas outlet.

Gas filter devices are respectively installed at the gas inlet and the gas outlet of the incubator. the

The front end of the incubator is provided with openings for picking and placing culture devices such as solid plates, multi-well culture plates or Erlenmeyer flasks, and the openings are sealed and closed by flange covers. the

The ultraviolet lamp tube is installed on the flange cover. the

The incubator is provided with a humidification system. the

The humidification system is a water tray filled with sterile pure water. the

The box body of the incubator is a transparent box body, the air inlet is arranged on the lower side of the front end of the incubator, and the air outlet is arranged on the upper side of the rear end of the incubator. the

Multiple incubators are set up in parallel by installing a tee on the intake line. the

Advantage of the present invention and positive effect are:

1. The present invention is respectively equipped with gas flow meters on the inlet pipeline and the outlet pipeline, which can control and adjust the carbon dioxide concentration as required, and then screen wild-type microalgae or corresponding mutants by setting different concentrations of _CO2 in the culture environment. The whole device structure It is simple and easy to operate, and the invention adopts a closed incubator to cultivate and screen microalgae, avoiding the pollution of miscellaneous bacteria in the air and the like.

2. The incubator of the present invention adopts resin material, which can ensure the light transmittance during illumination, and at the same time, it is convenient to observe the growth of algae in the whole cultivation process, and an ultraviolet lamp is provided in the incubator, which is beneficial to Use ultraviolet disinfection to avoid contamination by bacteria and mold during the cultivation process. the

3. The present invention is equipped with a CO ₂ gas concentration analyzer outside the incubator to measure the carbon dioxide concentration at the inlet and outlet of the incubator, which can effectively control and adjust the carbon dioxide concentration in the incubator. The CO ₂ concentration at the gas outlet of the incubator is measured by a CO ₂ gas concentration analyzer. When the required CO ₂ concentration is reached, the gas flow meters at the inlet and gas outlet can also be closed at the same time to maintain the CO ₂ gas concentration inside the box, according to In the growth cycle (or growth generation) of different microalgae, the gas replacement is performed 2-3 times a day, which can greatly save the usage of CO ₂ gas.

4. The present invention is equipped with a gas filtration and sterilization device at the air inlet and outlet of the incubator, which is used to filter the miscellaneous bacteria in the gas, so as to avoid the contamination of the microalgae by the miscellaneous bacteria and the like during the cultivation process, and is arranged in the incubator The gas filtration and sterilization device at the gas outlet can also prevent gas from being sucked back. the

5. The present invention is provided with an air inlet and an air outlet along the diagonal direction of the incubator. This arrangement is conducive to the gas being fully diffused inside the incubator before being discharged, so that the _CO2 concentration in the incubator is as consistent as possible, avoiding the reaction Inhomogeneous gas concentration in the system.

6. The present invention has a humidification system in the incubator. The present invention adopts a passive humidity control method, that is, a water tray is placed in the incubator to contain sterile pure water, and rely on the natural evaporation of water to make the relative humidity in the box reach 85%-95%. This also prevents premature drying of the solid agar on the plate when used for solid plate screening. the

7. In the present invention, multiple incubators are installed in parallel by installing a tee on the air intake pipeline for large-scale screening. the

Description of drawings

Fig. 1 is a structural schematic diagram of the present invention. the

Among them, 1 is the gas flow meter; 2 is the valve; 3 is the flange cover; 4 is the UV light switch; 5 is the air inlet; 6 is the air outlet; 7 is the incubator; 8 is the ultraviolet lamp; 9 is the air inlet Road; 10 is the air outlet pipeline. the

Detailed ways

The present invention will be described in further detail below in conjunction with the accompanying drawings. the

As shown in Figure 1, the present invention comprises an incubator 7, an ultraviolet lamp tube 8, an air inlet pipeline 9 and an air outlet pipeline 10, wherein an air inlet 5 is arranged on the lower side of the front end of the incubator 7, and an air inlet 5 is arranged at the rear of the incubator 7. The upper side of the end is provided with an air outlet 6, and the air inlet pipeline 9 is connected to the air inlet 5 on the incubator 7, and the air outlet pipeline 10 is connected to the air outlet 6 on the incubator 7, and the air inlet 5 and the air outlet are connected to each other. The gas ports 6 are respectively arranged on the lower front side and the upper side of the rear end of the incubator 7 to facilitate the gas to fully diffuse in the incubator 7 before being discharged, so that the gas concentration in the incubator 7 is uniform. Ultraviolet lamp tube 8 is arranged in described incubator 7, is used for carrying out ultraviolet disinfection to incubator 7 before use, prevents pollution such as bacterium, mould, and described ultraviolet lamp tube 8 is installed in the ultraviolet lamp switch on the outside of incubator 7 4 controls on and off. The front end of the incubator 7 is provided with openings for taking and placing culture devices such as solid flat plates, porous plates or triangular flasks. The opening is sealed and closed by a flange cover 3, and the ultraviolet lamp tube 8 is installed on the flange cover 3. When the flange cover 3 is opened, the ultraviolet lamp 8 can move with the flange cover 3 . The incubator 7 is made of light-transmitting material. In this embodiment, the incubator 7 is made of transparent resin material, and the light transmittance can reach 90%. the

A gas flow meter 1 for controlling the gas flow rate and a valve 2 for controlling the opening and closing of the pipeline and adjusting the gas flow are respectively arranged on the inlet pipeline 9 and the gas outlet pipeline 10. In this embodiment, the model of the gas flowmeter 1 is Hongqi LZB-10, the manufacturer is Changzhou Shuangbo Instrument Co., Ltd., and a _CO2 gas analyzer for detecting the carbon dioxide concentration at the inlet 5 and the gas outlet 6 of the incubator 7 is arranged outside the incubator 7. When cultivating When the carbon dioxide concentration in the box 7 does not meet the requirements, the _CO2 gas analyzer can send a signal to adjust the gas flow meter 1, so as to prevent the difference between the carbon dioxide concentration in the incubator 7 and the carbon dioxide concentration of the gas fed into the incubator from being too large, and ensure that the microalgae are in the pre-prepared state. Cultivate under the set carbon dioxide concentration, when the _CO concentration at the gas outlet 6 of the incubator 7 reaches the required concentration, the _CO gas analyzer can also close the gas inlet line 9 and the gas outlet line 10 at the same time. Gas flow meter 1 to maintain the _CO2 gas concentration inside the incubator 7, and then perform gas replacement 2 to 3 times a day according to the growth cycle (or growth generation) of different microalgae, which can greatly save the use of _CO2 gas quantity. In this embodiment, the model of the CO ₂ gas analyzer is C150 CO ₂ concentration analyzer, and the manufacturer is British Geotech Company.

The air inlet 5 and the air outlet 6 places of the incubator 7 are respectively provided with a gas filter, wherein the gas filter installed at the air inlet 5 is used for filter sterilization, and the gas filter installed at the air outlet 6 is installed at the air outlet 6. Filtration is also used to prevent external air from being sucked back into the incubator 7. In the present embodiment, the gas filter installed at the air inlet 5 of the incubator 7 is a microporous membrane, and the microporous membrane is used to filter gas. Known technology in the field, the gas filtering device installed at the gas outlet 6 of the incubator 7 is an air filter, and the air filter model is Sartorius Midisart200017805, and the manufacturer is Sartorius/Sartorius, Germany. the

The gas source communicates with the inside of the incubator 7 through the air intake line 9, and the gas source is an air pump (for feeding compressed air into the incubator 7) or a gas cylinder (for feeding in the incubator 7 according to different conditions in advance). Proportionally configured gas), the flow rate and flow rate of the gas can be adjusted by the gas flow meter 1 and valve 2 on the pipeline. The present invention can also be provided with separate pure _CO gas cylinder pipelines and compressed air pipelines, and the above two gas pipelines are connected to a gas buffer bottle through a tee, and the gas buffer bottle is connected to the incubator 7 through the pipeline , Gas flowmeters are respectively arranged on the pure _CO2 gas cylinder pipeline and the compressed air pipeline to adjust the gas flow rate, and the gas buffer bottle is used for premixed gas.

A humidification system is provided in the incubator 7, because in addition to ensuring accurate _CO content in the incubator 7, stable and accurate humidity is also an important indicator of an ideal physiological environment. The present invention adopts a passive humidity control method, that is, in the cultivation A large water tray is placed in the box 7, which contains sterile pure water, and relies on the natural evaporation of water to make the relative humidity in the box reach 85%-95%. This also prevents premature drying of the solid agar on the plate when used for solid plate screening. In this embodiment, a 1L water tray with a capacity of 0.5L of sterile pure water is placed in the incubator 7 .

The working principle of the present invention is:

Before the present invention is used, it can be sterilized by the built-in ultraviolet lamp tube 8. After the disinfection is completed for 3 hours, the ultraviolet lamp tube 8 is turned off, and then the flange cover 3 is opened, and the pre-coated solid plate or liquid culture porous plate (24-well or 96-well), and at the same time put it into a water tray filled with sterilized pure water, then close the flange cover 3 and tighten the screws. The gas source enters air and carbon dioxide mixture through the intake pipeline 9, the ventilation rate is measured by the gas flow meter 1 on the intake pipeline 9, and the flow rate is adjusted through the valve 2 on the intake pipeline 9, and the gas passes through the gas flow at the inlet 5. The gas filtering device enters the incubator 7 after being filtered, and the valve 2 and the gas flow meter 1 on the gas outlet line 10 are also adjusted accordingly to ensure that the carbon dioxide concentration in the incubator 7 is constant and the gas flow is stable. the

There is a _CO2 concentration analyzer outside the incubator 7, which is used to measure the gas concentration of carbon dioxide entering and exiting the incubator 7, and can also measure the _O2 concentration and relative humidity in the incubator, and measure the concentration of carbon dioxide at the gas outlet 6 , adjust the gas flow meter 1 on the inlet pipeline 9 and the outlet pipeline 10 to ensure that the microalgae are cultivated at a preset carbon dioxide concentration and keep the relative humidity constant.

When the microalgae solid plate grows large enough to be visible to the naked eye, or when the green color can be clearly seen by the naked eye on the liquid culture 24-well plate (or in the Erlenmeyer flask), first close the air source valve, and then close the air inlet line 9 Open the valve 2 on the top, then open the flange cover 3, take out the solid plate sample or the multi-well plate liquid culture sample, and select the algae colonies of interest to expand the culture and conduct other biological research. the

In the present invention, a plurality of incubators 7 can be arranged in parallel by installing a tee on the intake pipeline 9 for large-scale screening. the

Claims (9)

1. a micro-algae is cultivated reactor, it is characterized in that: comprise incubator (7), ultraviolet lamp tube (8), admission passage (9) and outlet pipe (10), the admission passage (9) being wherein connected with source of the gas is connected with the inlet mouth (5) on incubator (7), outlet pipe (10) is connected with the air outlet (6) on incubator (7), be arranged at described incubator (7) for the ultraviolet lamp tube (8) before use incubator (7) being carried out disinfection, on described admission passage (9) and outlet pipe (10), be equipped with gas meter (1) and valve (2) for keeping gas concentration equilibrium in incubator (7).

2. micro-algae according to claim 1 is cultivated reactor, it is characterized in that: described incubator (7) outside is provided with the CO that locates gas concentration lwevel and regulate or close gas meter (1) by detecting described inlet mouth (5) and air outlet (6) ₂gas analyzer.

3. micro-algae according to claim 1 and 2 is cultivated reactor, it is characterized in that: gas-filtering device is located to be separately installed with in inlet mouth (5) and air outlet (6) of described incubator (7).

4. micro-algae according to claim 1 is cultivated reactor, it is characterized in that: described incubator (7) front end is provided with the opening for picking and placeing the culture apparatuses such as solid plate, porous culture plate or triangular flask, described opening is by blind flange (3) hermetically closing.

5. micro-algae according to claim 4 is cultivated reactor, it is characterized in that: described ultraviolet lamp tube (8) is arranged on described blind flange (3).

6. micro-algae according to claim 1 is cultivated reactor, it is characterized in that: described incubator is provided with humidification system in (7).

7. micro-algae according to claim 7 is cultivated reactor, it is characterized in that: described humidification system is the water pond that fills sterile pure water in.

8. micro-algae according to claim 1 is cultivated reactor, it is characterized in that: the casing of described incubator (7) is transparent casing, described inlet mouth (5) is arranged at the front end downside of described incubator (7), and described air outlet (6) are arranged at the rear end upside of described incubator (7).

9. micro-algae according to claim 1 is cultivated reactor, it is characterized in that: multiple incubators (7) are by being arranged in parallel in the upper installation of admission passage (9) threeway.