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WO2016165660A1 - Procédé pour l'élevage de micro-algues eucaryotes d'eau douce - Google Patents

Procédé pour l'élevage de micro-algues eucaryotes d'eau douce Download PDF

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Publication number
WO2016165660A1
WO2016165660A1 PCT/CN2016/079499 CN2016079499W WO2016165660A1 WO 2016165660 A1 WO2016165660 A1 WO 2016165660A1 CN 2016079499 W CN2016079499 W CN 2016079499W WO 2016165660 A1 WO2016165660 A1 WO 2016165660A1
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Prior art keywords
microalgae
light
culture
breeding
culture solution
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PCT/CN2016/079499
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English (en)
Chinese (zh)
Inventor
张玟籍
陈辉
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上海希明生物科技有限公司
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/04—Apparatus for enzymology or microbiology with gas introduction means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor

Definitions

  • the invention relates to the field of biotechnology, and particularly relates to a method for breeding freshwater eukaryotic microalgae.
  • Microalgae biomass is the future development direction of biomass utilization, and has the characteristics of high energy conversion efficiency. Compared with ordinary terrestrial crops, the yield per unit area can be several tens of times higher, so that factory farming can be realized.
  • the basic principle of microalgae growth is to use microalgae photosynthesis to convert carbon dioxide into microalgae's own biomass to fix carbon, and to synthesize oil, protein, polysaccharide, cellulose and other biomass through microalgae's own characteristics and functions. .
  • the useful substances in the microalgae cells are transformed into extracellular cells by physical or chemical methods, and then refined and processed to produce products such as biodiesel and algae polysaccharides. Therefore, in actual production, nutrients and carbon dioxide in waste water and waste gas can be converted into products having high utilization value such as biofuels and proteins by photosynthesis of algae.
  • microalgae cultivation photoreactor In order to further increase the unit yield of microalgae biodiesel and achieve three-dimensional culture, a dedicated microalgae cultivation photoreactor must be used.
  • the microalgae aquaculture photoreactor in the prior art is directly cultured in the open air compared with the runway pool and the multi-stage pool, the production density and the yield per unit area are obviously improved, but the microalgae growth and the effective substance are used to induce the production.
  • the light source is still natural light, the utilization of sunlight by microalgae is not sufficient, it is uncertain and uncontrollable, it runs counter to the stable and continuous requirements of factory production, and the auxiliary farming function such as nutritional supplement and carbon dioxide inflation is not automated. So that it has a long way to go with the actual industrial application.
  • the object of the present invention is to provide a breeding method according to the optimal ratio of light source, carbon dioxide, nutrition, temperature, flow rate and the like according to the breeding demand of microalgae, thereby realizing stable, controllable and high-density microalgae cultivation. Demand.
  • the present invention provides a freshwater eukaryotic microalgae breeding method, and the freshwater eukaryotic microalgae breeding method comprises the steps of:
  • an artificial light source microalgae breeding apparatus for breeding freshwater eukaryotic microalgae
  • the artificial light source microalgae breeding apparatus comprising a reaction container, a light emitting device, a nutrient distributing device and a gas supply device
  • the reaction container is for accommodating a culture solution for growing microalgae
  • the light-emitting device being disposed inside the reaction vessel, and when the reactor is operated, the light-emitting device is at least partially or completely immersed in the culture solution, thereby providing microalgae in the reaction vessel Light required for growth, wherein the light intensity of the light emitted by the light-emitting device is uniform or substantially uniform in the depth direction (Z-axis direction) of the reaction container, and the nutrient cloth device is used for the culture liquid Providing nutrients required for growth and reproduction of microalgae and the nutrient distribution device capable of supplying manganese ions to the culture solution, the gas supply device for supplying a gas required for growth of microalgae
  • the concentration of carbon dioxide in the culture solution is monitored and adjusted.
  • the culture device is further provided with a water quality monitor for monitoring the concentration of manganese ions or other ions in the culture solution, wherein the water intake of the water quality monitor is below the liquid level, when the water quality
  • the water quality monitor can issue a reminder that the nutrient distribution device performs the addition of manganese salts or other salts.
  • the culture method comprises introducing a mixture of carbon dioxide and air into the culture solution, wherein the carbon dioxide accounts for 0.1-10%, preferably 0.5-5%, of the total volume of the mixed gas; and / or
  • the carbon dioxide pressure is 0.1-1 MPa; and/or
  • the carbon dioxide release pore size is from 0.1 to 50 nm.
  • the breeding apparatus is further provided with a monitoring and automatic control system for detecting and controlling the amount of carbon dioxide in the culture solution, wherein
  • the monitoring and automatic control system detects that the pH in the culture solution is ⁇ 5.5, the mixed gas containing carbon dioxide is continuously introduced into the culture solution; and when the pH in the culture solution is less than 5.5, The introduction of carbon dioxide into the culture solution is suspended.
  • a temperature control device is disposed around the reaction vessel, and the temperature control device is configured to maintain a liquid ambient temperature in the reaction vessel within a range suitable for the growth of the microalgae;
  • the gas supply device includes a gas refining distributor, in the process of introducing gas into the reaction vessel, the belt
  • the gas with a certain pressure escapes evenly and finely through the pores of the refiner, thereby promoting the dispersion of gas and nutrients in the liquid culture system.
  • the nutrient cloth device adds MnCl 2 ⁇ 4H 2 O to the culture solution, and the mass of MnCl 2 ⁇ 4H 2 O added per liter of the culture solution is 5-50 mg.
  • the illuminating device is capable of emitting at least three different wavelengths of light when operating, including light having a wavelength of 600-680 nm, light having a wavelength of 420-500 nm, and light having a wavelength of 700-780 nm; and / or
  • the color temperature of the composite light emitted by the light emitting unit is 1000-20000K, preferably 1500-6000K.
  • the three different wavelengths of light comprise light having a wavelength of 650-670 nm, light having a wavelength of 440-460 nm, and light having a wavelength of 710-730 nm.
  • the light-emitting device includes a light guide plate and a light-emitting unit, the light-emitting unit is embedded in the light guide plate, and light generated by the light-emitting unit is transmitted through the light guide plate to cause the light guide plate to emit light as a whole.
  • the device further includes a bracket for fixedly supporting the light guide plate, the bracket being detachably coupled to the reaction container and/or the cover plate; and/or
  • the light-emitting unit is an LED, and the number of the LEDs is 1-1000/light guide plate; preferably 10-1000/light guide plate.
  • the artificial light source microalgae culture method is suitable for the cultivation of freshwater eukaryotic algae such as chlorella, Scenedesmus, and Haematococcus pluvialis.
  • the cultured algae is inoculated at a concentration of from 0.1 g/L to 2 g/L (dry weight of algae/volume of culture medium), more preferably from 0.2 g/L to 0.6 g/L.
  • the light intensity of the light emitted by the light-emitting device is substantially uniform in the horizontal direction of the reaction vessel (including the X-axis and Y-axis directions).
  • the "uniform or substantially uniform" means that the light intensity D1 at any depth and the average light intensity Dm over the entire depth range satisfy the following formula:
  • a temperature control device is provided around the reaction vessel for maintaining the liquid ambient temperature within the reaction vessel within a range suitable for the growth of the microalgae.
  • the culture temperature in the culture apparatus is 10-30 ° C, more preferably 20-30 ° C.
  • the temperature control device may include an electric heating or steam heating system, a cold water cooling system, etc., such as a temperature controlled water pipe.
  • the artificial light source microalgae breeding apparatus is further provided with a monitoring system for monitoring parameters of the liquid environment, the parameters being selected from the group consisting of: pH, temperature and/or nutrient concentration.
  • an air vent is provided in the cover.
  • the artificial light source microalgae breeding apparatus is provided with a plurality of light guide plates, preferably 30-1000 pieces, more preferably 40-800 pieces, and most preferably 50-500 pieces.
  • the light guide plate is made of a transparent organic material having weak acid resistance.
  • FIG. 1 is a perspective cross-sectional view of an artificial light source microalgae breeding apparatus used in a method for breeding freshwater eukaryotic microalgae according to the present invention
  • FIG. 2 is a schematic cross-sectional view showing an artificial light source microalgae breeding apparatus used in a method for breeding freshwater eukaryotic microalgae according to the present invention
  • FIG. 3 is a front cross-sectional view showing the artificial light source microalgae breeding apparatus used in the freshwater eukaryotic microalgae culture method according to the present invention
  • Figure 4 is a top cross-sectional view showing the artificial light source microalgae breeding apparatus used in the freshwater eukaryotic microalgae culture method according to the present invention
  • FIG. 5 is a schematic perspective view of a light-emitting device of an artificial light source microalgae breeding apparatus used in a method for breeding freshwater eukaryotic microalgae according to the present invention
  • Figure 6a is a front elevational view of a light-emitting device of an artificial light source microalgae breeding apparatus used in a method for breeding freshwater eukaryotic microalgae according to the present invention
  • Figure 6b is a side view of a light-emitting device of an artificial light source microalgae breeding apparatus used in a freshwater eukaryotic microalgae culture method according to the present invention
  • Figure 6c is a plan view of a light-emitting device of an artificial light source microalgae breeding apparatus used in a method for breeding freshwater eukaryotic microalgae according to the present invention
  • Figure 6d is an enlarged view of the portion B according to Figure 6b;
  • FIG. 7a-7c are simplified top plan views of an artificial microalgae culture apparatus according to other embodiments of the freshwater eukaryotic microalgae culture method of the present invention.
  • Figure 8 is a side cross-sectional view showing an artificial microalgae culture apparatus of another embodiment employed in the freshwater eukaryotic microalgae culture method according to the present invention.
  • the present inventors have developed a method for breeding freshwater eukaryotic microalgae.
  • the cultivation time of microalgae can be shortened, and the breeding efficiency of microalgae can be greatly improved.
  • the artificial light source microalgae breeding equipment of the invention is suitable for high-density industrialized breeding of freshwater eukaryotic algae such as chlorella, Scenedesmus, and Haematococcus pluvialis.
  • the artificial light source microalgae breeding equipment comprises a reaction container 1, a light-emitting device 2, an inlet and outlet device 3, a cover plate 5, a gas distributor 6, a nutrient distribution device, a temperature control device 7, and Monitoring and control systems, etc.
  • the reaction vessel 1 is provided with a cover plate 5, and the reaction vessel 1 is sealingly connected with the inlet and outlet device 3, and a valve and a switch are arranged on the inlet and outlet device for controlling the start or stop of the inlet and outlet, and the reaction vessel 1 is provided with a light-emitting device.
  • a temperature control device 7 (shown as a temperature-controlled water pipe) is disposed around the reaction vessel for maintaining the temperature of the solution in the reaction vessel within a temperature range suitable for the number of microalgae to multiply and grow.
  • the lower portion of the bioreactor (e.g., about 30% to 90%, or 50% to 70% of the height) can be buried underground.
  • the bioreactor can be arranged around the power plant, and the waste water generated by the power generation is used as a material and energy source for microalgae cultivation, and has the function of environmental protection and emission reduction.
  • the reaction vessel 1 may be a closed container that is opaque to light, and may be illuminated by its own light-emitting device during rainy days and nights for the growth of microalgae.
  • a monitoring and automation control system is provided in the breeding equipment for monitoring water temperature, pH value, CO 2 concentration, TDS, absorbance, light intensity, etc., and can automatically control the above conditions as needed.
  • a temperature control device 7 is provided on the wall of the reaction vessel, and the temperature control device 7 can be in any suitable form, including electric heating. Or steam heating system, cold water cooling system, such as temperature control spacers.
  • the temperature control device 7 is a temperature-controlled water pipe, and the wastewater with a certain temperature discharged from the power plant can be used to circulate in the temperature control water pipe to keep the temperature in the bioreactor between 20 ° C and 30 ° C.
  • An air outlet is provided at the top of the cover plate 5 of the bioreactor for releasing oxygen generated by photosynthesis of the microalgae.
  • a gas distributor 6 is provided at the bottom of the artificial light source microalgae cultivation apparatus, which may be in the form of a rotary, trough, tubular or any other suitable form.
  • carbon dioxide, air or other gas is introduced into the reaction vessel through a gas distributor.
  • a rotary gas distributor is used.
  • the gas distributor 6 rotates to drive the gas in the bottom container and the liquid in the bottom container, thereby facilitating the gas and nutrient in the solution. Evenly distributed.
  • a nutritional drape device is installed at one or more locations of the reaction vessel (e.g., on the vessel wall, at the bottom of the vessel, at the top of the vessel, and inside the cover, etc.).
  • the nutrient cloth device provides different nutrients at different stages of microalgae growth.
  • the reaction vessel into the rate of other nutrients and the CO flow speed may be such proportion, dissolved amount of 2 was CO by CO 2 line monitoring system may determine that, while the PH value of the solution was monitored in known solutions by monitoring the system The pH.
  • the control system can control the rate of introduction of CO 2 and nutrients to ensure that the amount of dissolved nutrients in the solution and the pH are within the range suitable for the growth or production of microalgae.
  • the reaction vessel of the present invention can be fabricated from fiberglass and the reaction vessel can have any suitable shape and size.
  • the surface of the container is relatively smooth and, preferably, the surface of the container has a removable closure.
  • the reaction vessel can be a long annular solid vessel.
  • the container has a total length of from 2 m to 500 m, preferably from 10 m to 200 m; a cross-sectional aspect ratio of the container of from 0.5:1 to 1:0.5; and a height of from 20 cm to 400 m, preferably from 50 cm to 200 cm.
  • the bottom section of the container has a low height on both sides, a lowest point at the center, and a bottom slope of 5 to 60 degrees, more preferably 10 to 30 degrees.
  • the cross section of the container in the intermediate portion may be changed to a closed cylinder having a diameter of 20 cm to 400 m, more preferably 50 cm to 200 cm, and the length of the portion may be 20 cm to 100 cm, more preferably 20 cm to 50 cm.
  • the transition portions of the different cross-sectional shapes of the container are in smooth connection.
  • the culture apparatus of the present invention also includes other gas supply means such as a gas source, a mixer, a gas pipe, and the like not shown.
  • the gas source in the present invention is mainly air and carbon dioxide.
  • the mixer is mainly used for mixing air and carbon dioxide, and the mixer can be a venturi or a static mixer.
  • the material of the gas transmission port may be a ceramic microporous or resin microporous structure, and the micropore diameter is 0.1-10 micrometers, more preferably 0.5-5 micrometers.
  • the gas distributor is placed at the bottom of the culture vessel, and the lower end of the gas distributor is 0-5 cm from the inner bottom of the vessel. As shown, the bottom of the reaction vessel may be provided with a plurality of gas distributors arranged side by side at equal intervals. When used, the gas distributor has a supply pressure of 0.2-20 MPa, more preferably 0.5-5 MPa.
  • the nutrient distribution device in the culture equipment of the present invention may be a perforated pipe located in the upper portion of the container, and the nutrient solution may be evenly sprayed through the small holes to the culture liquid in the container after flowing through the pipe to the upper portion of the container.
  • the inventors discovered through experiments and exploration that it is possible to effectively raise a certain amount of manganese ions in the culture solution. Benefits. Therefore, the nutrient cloth device of the culture apparatus of the present invention supplies manganese ions to the culture liquid.
  • the culture apparatus of the present invention is further provided with a circulation propulsion device which may have a propeller type or a screw type propulsion blade for the flow and circulation of the water body in the system.
  • the propulsion blade speed is from 10 to 1000 rpm, more preferably from 20 to 400 rpm.
  • the breeding apparatus of the present invention can also integrate an on-line water quality monitor for detecting the concentration of manganese ions in the culture solution, so as to control the concentration of manganese ions in the culture solution to the extent that is most favorable for microalgae cultivation.
  • the water sample water inlet of the water quality monitor is located below the liquid level and can be automatically sampled at regular intervals.
  • the water filtration system is passed through a microfiltration system before the water sample is automatically fed into the detector.
  • the filtration accuracy is T grade.
  • the pH of the culture solution in the culture apparatus of the present invention ranges from 5 to 10, more preferably from 6 to 8.
  • the breeding apparatus of the present invention can also integrate a pH online monitoring sensor for measuring the pH value in the culture solution to control the concentration of carbon dioxide introduced into the culture solution.
  • the system automatically opens the carbon dioxide valve and introduces carbon dioxide gas into the pool; when the pH is lower than 6.5 (or other set value), the system turns off the carbon dioxide.
  • the valve and the venting device only pass air into the pool or directly stop the gas, so as to effectively adjust the amount of carbon dioxide to obtain the optimal carbon dioxide condition for microalgae reproduction and growth.
  • FIG. 5 is a schematic perspective view of the light-emitting device 2.
  • the arrangement of the illuminating device allows the bioreactor to perform normal operations such as aquaculture without sunlight or other external light sources.
  • the light-emitting device is composed of a bracket 9, an LED light-emitting unit group 10 and a light guide plate 11.
  • the light guide plate 11 is fixed by a bracket 9, and the top end of the bracket 9 is fixed on the cover plate 5, and the LED light-emitting unit group 10 is located.
  • the light guide plate 11 or all protrudes below the liquid surface.
  • a plurality of light-emitting devices 2 are provided, and each of the light guide plates can be separately mounted or detached.
  • the light guide plate can be made of a transparent organic material such as acrylic and has weak acid resistance.
  • the LED light emitting unit group is in direct contact with the light guide plate at the top of the light guide plate, and the light emitted by the LED light emitting unit group can be transmitted through the light guide plate and the light guide plate is entirely illuminated.
  • the light guide plates are placed in parallel, and the distance between the two plates may be from 5 cm to 50 cm, more preferably from 10 cm to 20 cm.
  • the illuminance of the illuminating panel can be adjusted as needed.
  • the top end of the bracket 9 of the illuminating device 2 is detachably connected to the cover plate 5, thereby facilitating the removal, replacement or reinstallation of the illuminating device; the illuminating device 2 is suspended in the reaction container without contacting the bottom of the container, thereby not affecting the gas at the bottom of the container The rotation of the spreader.
  • the bracket of the illuminating device can also be any other A suitable means is fixed to the reaction vessel at any suitable location.
  • Figures 6a-6c are front, side and top views, respectively, of a light-emitting device of an artificial light source microalgae culture apparatus;
  • Figure 6d is an enlarged view of a portion B of Figure 6b.
  • the LED lighting unit group 10 includes a small LED lighting unit.
  • the light generated by the light unit can be composite color or monochromatic light.
  • the color LED has a wavelength range of 350-900 nm and a composite color temperature range of 1500-20000K.
  • the number of LED lighting units on a single light guide plate can be 1-1000.
  • the light source is an LED light source
  • the illuminating device is capable of emitting light of different wavelengths when it is in operation.
  • the light of different wavelengths includes: light A having a wavelength of 600-680 nm, more preferably 650-670 nm; light B having a wavelength of 420-500 nm, more preferably 440-460 nm; 700-780 nm, more preferably 710-730 nm.
  • Light C infrared light
  • the inventors found that the ratio of light of different wavelengths emitted by the illuminating device is in the following range, and the microalgae breeding efficiency in the breeding equipment can reach a higher level.
  • the inventors found through experiments that, with the same illumination power, after supplementing the near-infrared light of a specific wavelength, the yield of microalgae can be increased by 10-25%:
  • A: B 10: 1 ⁇ 2: 1;
  • the illumination intensity of the illuminating device can be adjusted, and the light and dark are alternated during illumination (change in illuminating power).
  • 0%-80% of the luminous power when the luminous power is bright light is 0%-60%, more preferably 0%-60%.
  • the dark light irradiation time is 10%-300% of the bright light irradiation time, more preferably 20%-100%.
  • the light intensity of the light emitted by the light-emitting device is uniform or substantially uniform in the depth direction (Z-axis direction) of the reaction vessel; the light intensity of the light emitted by the light-emitting device is in the horizontal direction of the reaction vessel (including the X-axis and the Y-axis)
  • the axis direction is basically uniform.
  • "Uniform or substantially uniform" means that the intensity D1 at any depth and the average intensity Dm over the entire depth range satisfy the following formula:
  • the LED light emitting unit group 10 may be located at the bottom of the light guide plate 11 or at other suitable positions of the light emitting plate as long as the light emitted by the light guide plate 11 can be conducted in the entire light guide plate so that the entire light guide plate emits light.
  • the energy source of the LED lighting unit group is the electric energy generated by the solar panel absorbing solar energy.
  • the bioreactor of the present invention does not directly utilize solar energy, the unstable solar energy is collected by the solar panel for power generation, and the electricity generated therefrom is stably supplied to the luminescence of the bioreactor.
  • the unit group keeps it glowing continuously, ensuring the stability and persistence of microalgae culture.
  • the number of microalgae reproduction and individual weight gain stages can be carried out in the same bioreactor, but it is necessary to change the type and rate of nutrient access as the growth progresses, and the process is complicated.
  • a plurality of bioreactors can be used in series or in parallel to form a large-scale production and breeding system.
  • the series or parallel connection between the bioreactors is connected through the inlet and outlet ports, and the feed and discharge between the bioreactors can be completed by the pump system.
  • the first-stage and second-stage bioreactors are provided with a light-emitting device whose light-emitting wavelength is only suitable for the propagation of microalgae, and the nutrient-distributing device introduces ammonium phosphate, potassium dihydrogen phosphate or dipotassium hydrogen phosphate into the reaction vessel.
  • Nitrogen oxides are used as nitrogen and phosphorus sources, and nutrients necessary for the growth of other microalgae such as iron and zinc are added.
  • the gas distributor introduces nitrogen oxides and CO 2 into the reaction vessel, and the luminous intensity in the second-stage bioreactor is
  • the nutrient supply rate is larger than that of the first-stage bioreactor to accommodate the growth requirements of the increased microalgae;
  • the third-stage bioreactor is provided with a luminescent device that is only suitable for the growth of microalgae individuals, PH and
  • the temperature and the like are simultaneously adjusted to conditions suitable for the growth of the microalgae individual, and the gas distributor introduces CO 2 into the reaction vessel.
  • Different grades of bioreactors can be selected according to different sizes, for example, the size of the first stage bioreactor is smaller than the latter two stages.
  • new algae species can be introduced into the first-stage bioreactor while a new round of culture is carried out.
  • a portion of the algae species as the first stage bioreactor can be filtered from the microalgae discharged from the second stage bioreactor.
  • the present invention provides a method of culturing microalgae, the method comprising the steps of:
  • the algae species are placed in the artificial light source microalgae breeding equipment to provide microalgae for survival in the reactor.
  • the nutrients including nitrogen sources, phosphorus sources, inorganic salts (such as manganese, iron, zinc, magnesium, etc.) and provide carbon dioxide or air, turn on the light-emitting device to produce the light required for the growth of microalgae.
  • microorganism suitable for use in the present invention is not particularly limited as long as it can be grown using a light source.
  • Representative microorganisms include, but are not limited to, Chlorella, Chlorella, Cyanophyta, and Red algae microalgae.
  • a preferred microorganism is the Chlorellales freshwater species.
  • the breeding equipment and breeding process of the invention are particularly suitable for the cultivation of freshwater eukaryotic algae such as chlorella, Scenedesmus, and Haematococcus pluvialis.
  • the inoculating concentration of the cultured microalgae in the reaction vessel of the culture apparatus of the present invention is 0.1 g/L to 2 g/L (microalgae dry weight/culture liquid volume), more preferably 0.2 g/L to 0.6 g/L.
  • the supply of nutrients provided by the nutrient distribution device and the gas supply device should be determined according to the inoculation and feeding concentration. When the inoculation and feeding concentration are increased, the nutrient salt concentration should be appropriately increased, and the nutrient salt consumption should be maintained and supplemented during the breeding process. .
  • the culture apparatus of the present invention can pass carbon dioxide as a carbon source.
  • the carbon dioxide is mixed with air, wherein the carbon dioxide accounts for 0.1-10%, preferably 0.5-5% of the total volume of the mixed gas; the carbon dioxide pressure is 0.1-1 MPa; and the carbon dioxide release pore size is 0.1-50 nm. More preferably 1-20 nm.
  • the pH of the culture solution in the culture apparatus of the present invention ranges from 5 to 10, more preferably from 6 to 8.
  • the mixed gas containing carbon dioxide can be continuously introduced; and when the pH is less than 5.5, the suspension can be suspended. Ventilation.
  • the nitrogen source in the culture solution may be selected from one or more of the following nitrogen-containing compounds, and the amount of nitrogen-containing compounds per liter of the culture solution is as follows:
  • the phosphorus source in the culture solution may be selected from one or more of the following phosphorus-containing compounds, and the amount of the phosphorus-containing compound per liter of the culture solution is as follows:
  • MnCl 2 ⁇ 4H 2 O may be added to the culture solution at a concentration of 5 to 50 mg/L (additive weight/culture volume).
  • concentration of MnCl 2 ⁇ 4H 2 O in the culture system of the culture medium BG11 medium used in the usual algae species is 1.86 mg/L.
  • Iron citrate 5-50mg / L
  • the invention provides a method for preparing microalgae biomass, which adopts the microalgae liquid prepared by the foregoing method as a raw material, and processes the microalgae product, and a typical processing process comprises the following steps:
  • microalgae are collected by filtration, pressure filtration or bubble suspension, dried at a low temperature and then pulverized to obtain a microalgae product.
  • the artificial light source is a three-color LED light source with wavelengths of 650 ⁇ 5nm, 440 ⁇ 5nm, 720 ⁇ 10nm, and the ratio of three-color light source is 10:3:2.
  • the light intensity at the darkest point is about 3000lx, and the light and dark alternate time is 10 hours (bright): 2 hours (dark), the dark light power is about 40% of the brightness power.
  • the pressure is 0.4Mpa
  • the released ceramic membrane pressure tube has a pore size of 10nm
  • the ratio of carbon dioxide to air is 2%-3%
  • the continuous ventilation to PH 6.5 stops, reducing the ratio of carbon dioxide to air. Until the PH interval remains between 6.5-6.8.
  • the culture temperature was maintained at 27 degrees ⁇ 1 degree.
  • Citric acid 6mg/L
  • Ammonium ferric citrate 25mg/L
  • the artificial light source is a two-color LED light source with wavelengths of 650 ⁇ 5 nm and 440 ⁇ 5 nm respectively.
  • the ratio of the two-color light source is 10:3. After 7 days of cultivation, it is collected, filtered, dried and weighed. The biomass concentration was 0.82 g/L.
  • Example 1 The concentration of MnCl 2 ⁇ 4H 2 O in Example 1 was 12 mg/L, and the final biomass concentration obtained after 7 days of culture was 1.07 g/L, while in Comparative Examples 1-3, the MnCl 2 ⁇ 4H 2 was decreased or increased.
  • the concentration of O was found to be lower than that of Example 1 after 7 days of culture, and some of the comparative examples showed floating dead algae. It can be seen that adding proper amount of manganese ions to the culture solution is indeed Conducive to the growth of algae cultured in breeding equipment;
  • Example 1 a three-color LED light source with a ratio of 10:3:2 was used, and in Comparative Example 4, a two-color LED light source that does not emit infrared light was used, and under the same conditions of other culture conditions, Example 1
  • the breeding results are obviously better than the comparative example 4, from which it can be seen that a certain proportion of infrared light source is beneficial to the growth of algae cultured in the culture equipment;
  • Example 3 a mixed gas of carbon dioxide and air with a certain mixing ratio is introduced, and in the comparative example 5, no carbon dioxide is introduced, and no gas is introduced into the comparative example 6. As can be seen from the breeding results, an appropriate mixing ratio is introduced. Carbon dioxide and air are necessary for the growth and reproduction of algae.

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Abstract

L'invention concerne un appareil d'élevage de micro-algues éclairé artificiellement destiné à être utilisé dans l'élevage de micro-algues eucaryotes d'eau douce, comprenant un réacteur, un appareil d'émission de lumière, un appareil de distribution de nutriments et un appareil d'alimentation en gaz. Le réacteur est utilisé pour recevoir une solution de culture pour la croissance de micro-algues. Le dispositif d'émission de lumière fournit dans le réacteur une lumière nécessaire à la croissance de micro-algues. L'appareil de distribution de nutriments est utilisé pour fournir dans la solution de culture des nutriments nécessaires à la croissance et à la reproduction et l'appareil de distribution de nutriments est capable de fournir des ions manganèse dans la solution de culture. L'appareil d'alimentation en gaz est utilisée pour fournir dans le réacteur un gaz nécessaire à la croissance de micro-algues. L'invention concerne également un procédé d'élevage de micro-algues eucaryotes d'eau douce dans l'appareil d'élevage de micro-algues artificiellement éclairé, comprenant les étapes consistant à : (1) fournir l'appareil d'élevage de micro-algues éclairé artificiellement destiné à être utilisé dans l'élevage de micro-algues eucaryotes d'eau douce, et (2) cultiver des micro-algues eucaryotes d'eau douce dans l'appareil d'élevage.
PCT/CN2016/079499 2015-04-17 2016-04-15 Procédé pour l'élevage de micro-algues eucaryotes d'eau douce WO2016165660A1 (fr)

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CN110777067A (zh) * 2019-11-08 2020-02-11 华南理工大学 一种光热质耦合的微藻养殖方法及其装置
CN114989949A (zh) * 2022-06-21 2022-09-02 中国科学院青岛生物能源与过程研究所 一种倒v形结构的微藻多表面贴壁培养反应器及培养方法
CN118308191A (zh) * 2024-06-06 2024-07-09 江西江投能源技术研究有限公司 一种基于燃煤烟气再利用的微藻养殖装置

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Cited By (4)

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Publication number Priority date Publication date Assignee Title
CN108865651A (zh) * 2018-06-05 2018-11-23 李盈贤 一种密封养殖藻培养装置及其组装方法
CN110777067A (zh) * 2019-11-08 2020-02-11 华南理工大学 一种光热质耦合的微藻养殖方法及其装置
CN114989949A (zh) * 2022-06-21 2022-09-02 中国科学院青岛生物能源与过程研究所 一种倒v形结构的微藻多表面贴壁培养反应器及培养方法
CN118308191A (zh) * 2024-06-06 2024-07-09 江西江投能源技术研究有限公司 一种基于燃煤烟气再利用的微藻养殖装置

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