WO2016165659A1 - 人工光源微藻养殖装备 - Google Patents

人工光源微藻养殖装备 Download PDF

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Publication number
WO2016165659A1
WO2016165659A1 PCT/CN2016/079498 CN2016079498W WO2016165659A1 WO 2016165659 A1 WO2016165659 A1 WO 2016165659A1 CN 2016079498 W CN2016079498 W CN 2016079498W WO 2016165659 A1 WO2016165659 A1 WO 2016165659A1
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light
light source
microalgae
culture
artificial light
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PCT/CN2016/079498
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English (en)
French (fr)
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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Publication of WO2016165659A1 publication Critical patent/WO2016165659A1/zh

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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

Definitions

  • the invention relates to the field of biotechnology, in particular to an artificial light source microalgae breeding equipment.
  • 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 an artificial light source microalgae breeding equipment capable of optimizing the light source, carbon dioxide, nutrition, temperature, flow rate and the like according to different breeding conditions of different kinds of algae, thereby realizing ultra-high density microalgae cultivation. It is stable and controllable and meets the needs of industrialization.
  • the present invention provides an artificial light source microalgae breeding equipment
  • the artificial light source Microalgae farming equipment includes:
  • reaction vessel having a cover plate disposed thereon and the reaction vessel containing a culture solution for growing microalgae
  • the inlet and outlet device is sealingly connected to the reaction vessel;
  • the light-emitting device 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 liquid, thereby providing light required for growth of the microalgae in the reaction container
  • 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;
  • a nutrient cloth device capable of supplying manganese ions to the culture solution
  • a gas supply device for supplying a gas required for the growth of microalgae into the reaction vessel.
  • the manganese ion source in the culture solution is derived from MnCl 2 ⁇ 4H 2 O, and the mass of MnCl 2 ⁇ 4H 2 O added per liter of the culture solution is 5-50 mg.
  • the illuminating device emits 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; /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 gas supply device may introduce 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; /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 comprises a gas refining distributor. During the process of introducing gas into the reaction vessel, a gas with a certain pressure uniformly escapes through the pores of the refiner, thereby promoting gas and nutrition. The substance is dispersed in a liquid culture system.
  • the artificial light source microalgae breeding equipment is further provided with a water quality monitor for monitoring the concentration of manganese ions or other ions in the culture liquid, and the water intake of the water quality monitor is located below the liquid surface. among them
  • the water quality monitor When the water quality monitor detects that the concentration of manganese ions or other ions in the culture solution is lower than a preset value, the water quality monitor can issue a reminder to make the nutritive cloth device perform manganese salt or other nutrient salt Added.
  • the artificial light source microalgae breeding equipment 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 equipment is further provided with a monitoring system for monitoring the liquid environment.
  • the parameters are selected from: 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 in accordance with one embodiment of the present invention
  • FIG. 2 is a perspective cross-sectional view showing an artificial light source microalgae breeding apparatus according to an embodiment of the present invention
  • FIG. 3 is a front cross-sectional view showing an artificial light source microalgae breeding apparatus according to an embodiment of the present invention
  • FIG. 4 is a top cross-sectional view showing an artificial light source microalgae breeding apparatus according to an embodiment of the present invention
  • FIG. 5 is a perspective view of a light-emitting device of an artificial light source microalgae breeding apparatus according to an embodiment of the present invention.
  • Figure 6a is a front elevational view of a light-emitting device of an artificial light source microalgae farming apparatus in accordance with one embodiment of the present invention
  • Figure 6b is a side elevational view of a light-emitting device of an artificial light source microalgae farming apparatus in accordance with one embodiment of the present invention
  • Figure 6c is a plan view of a light-emitting device of an artificial light source microalgae breeding apparatus according to an embodiment of the present invention.
  • Figure 6d is an enlarged view of the portion B according to Figure 6b;
  • FIGS. 7a-7c are simplified top plan views of an artificial microalgae culture apparatus in accordance with other embodiments of the present invention.
  • Figure 8 is a side cross-sectional view of an artificial microalgae culture apparatus in accordance with other embodiments of the present invention.
  • the inventors have developed an artificial light source microalgae breeding equipment for the first time through extensive and in-depth research.
  • the reactor of the present invention not only shortens the culture time but also significantly increases the biomass of the microalgae through a specially designed light-emitting device and the like. Density and total active content, which can produce better quality biomass raw materials and products.
  • the present invention has been completed on this basis.
  • the built-in light source bioreactor 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 vessel 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 breeding equipment, and the gas distributor 6 can be a rotary type, a trough type, a tubular type 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 increase the amount of manganese ions provided in the culture solution. Breeding 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 5 to facilitate 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, thus Does not affect the rotation of the gas distributor at the bottom of the vessel. It should be understood that the bracket of the lighting device can also be secured to the reaction vessel in any suitable location in any suitable manner.
  • Figures 6a-6c are front, side and top views, respectively, of a light-emitting device of an artificial light source microalgae farming equipment;
  • 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 electricity generated by the solar panel absorbing solar energy. can.
  • 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 generated electricity is stably supplied to the light-emitting unit group of the bioreactor for continuous operation. Luminescence ensures 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 the nutrients (including nitrogen source, phosphorus source, inorganic salt (such as manganese, iron, zinc, magnesium, etc.) required for the survival of the microalgae to the reactor and Providing carbon dioxide or air to turn on the illuminating device to produce the light needed for the growth of the microalgae.
  • nutrients including nitrogen source, phosphorus source, inorganic salt (such as manganese, iron, zinc, magnesium, etc.) required for the survival of the microalgae to the reactor and Providing carbon dioxide or air to turn on the illuminating device to produce the light needed for the growth of the 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 diameter of 10nm
  • the carbon dioxide and air mix ratio is 2%-3%
  • 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

提供了一种人工光源微藻养殖装备，包括：反应容器、进出料装置、发光装置、营养布料装置和气体分布器。该装备中还设置红外光源，培养液中含有适量的锰离子，并通入适量的二氧化碳。

Description

人工光源微藻养殖装备 技术领域
本发明涉及生物技术领域，具体涉及人工光源微藻养殖装备。
背景技术
微藻生物质是生物质利用的未来发展方向，具有能量转化效率高的特点，与普通陆生农作物相比，单位面积的产率可高出数十倍，从而可实现工厂化养殖。微藻生长的基本原理是利用微藻光合作用，将二氧化碳转化为微藻自身的生物质从而固定了碳元素，并通过微藻自身的特性与机能合成油脂、蛋白质、多糖、纤维素等生物质。利用物理或化学方法把微藻细胞内的有用物质转化到细胞外，再进行提炼加工，从而生产出生物柴油、藻类多糖等产品。由此，在实际生产中，即可通过藻类的光合作用，将废水废气中的营养物质和二氧化碳转化为生物燃料、蛋白质等有高利用价值的产品。在石油价格大幅上升，粮食短缺问题日渐突出的今天，该产业有着广阔的发展前景。
为进一步提高微藻生物柴油的单位产量、实现立体化养殖，必须使用专用的微藻养殖光反应器。现有技术中的微藻养殖光反应器虽然相比较跑道池、多级池等露天直接养殖，养殖密度、单位面积产量都有明显提高，但是由于微藻生长与有效物定向诱导生产时所用的光源仍为自然光，微藻对太阳光的利用并不充分，具有不确定与不可控性，与工厂化生产稳定、持续的要求完全背道而驰，加之营养补充、二氧化碳充气等辅助养殖功能也未实现自动化，使其与实际产业化应用尚有较大距离。
因此，目前本领域迫切需要一种技术能够克服上述现有技术中的微藻养殖光反应器的可控性差、自动化程度低、效率低的弊端。
发明内容
本发明的目的是提供一种可根据不同种类藻类不同阶段的养殖需求配比最佳的光源、二氧化碳、营养、温度、流速等要素的人工光源微藻养殖装备，从而实现超高密度微藻养殖的稳定、可控，满足产业化需求。
为达到上述目的，本发明提供了一种人工光源微藻养殖装备，所述人工光源 微藻养殖装备包括：
反应容器，所述反应容器上设有盖板且所述反应容器容纳供微藻生长的培养液；
进出料装置，所述进出料装置与所述反应容器密封连接；
发光装置，所述发光装置设在反应容器内部且当所述反应器工作时，所述发光装置至少部分或全部浸没于所述的培养液，从而在反应容器内提供微藻生长所需的光，其中所述发光装置所发出的光的光强在反应容器的深度方向(Z轴方向)是均匀的或大致均匀的；
营养布料装置，所述营养布料装置可向所述培养液中提供锰离子；和
供气装置，所述供气装置用于向所述反应容器内提供微藻生长所需的气体。
在一优选实施例中，所述培养液中的锰离子源来自MnCl2·4H2O，且每升所述培养液中添加的MnCl2·4H2O的质量为5-50mg。
在一优选实施例中，所述发光装置工作时发射至少三个不同波长的光，其中包括波长为600-680nm的光、波长为420-500nm的光，以及波长为700-780nm的光；和/或
所述发光单元发出的复合光的色温为1000-20000K，较佳地1500-6000K。
在一优选实施例中，所述三个不同波长的光包括波长为650-670nm的光、波长为440-460nm的光，以及波长为710-730nm的光。
在一优选实施例中，所述发光装置包括导光板和发光单元，所述发光单元嵌于所述导光板，所述发光单元产生的光透过导光板传导使导光板整体发光，所述发光装置还包括支架，所述支架用于固定支撑所述导光板，所述支架与所述反应容器和/或所述盖板可拆卸地连接；和/或
所述发光装置中，发光单元为LED，且LED的数量为1-10000/导光板；较佳地为10-1000/导光板。
在一优选实施例中，所述供气装置可向所述培养液中通入二氧化碳和空气的混合物，其中二氧化碳占混合气体总体积的0.1-10％，较佳地占0.5-5％；和/或
二氧化碳通入压力为0.1-1MPa；和/或
二氧化碳释出孔径为0.1-50nm。
在一优选实施例中，所述养殖设备中还设有用于检测和控制所述培养液中二氧化碳通入量的监测及自动化控制系统，其中
当所述监测及自动化控制系统检测到所述培养液中的pH≥5.5时，含二氧化碳的混合气体被持续通入所述培养液中；而当所述培养液中的PH<5.5时，所述培养液中暂停通入二氧化碳。
在一优选实施例中，所述反应容器的周围设有温控装置，所述温控装置用于维持反应容器内的液体环境温度处于适合微藻生长的范围内；和/或
所述供气装置包括气体细化分布器，在向反应容器中通入气体的过程中，带有一定压力的气体通过该细化器的小孔均匀细散地逸出，从而促使气体与营养物质分散于液态培养体系中。
在一优选实施例中，所述人工光源微藻养殖装备中还设有用于监测所述培养液中锰离子或其他离子浓度的水质监测仪，所述水质监测仪的取水口位于液面以下，其中
当所述水质监测仪监测到所述培养液中的锰离子或其他离子的浓度低于预设数值时，所述水质监测仪能够发出提醒，使得所述营养布料装置进行锰盐或其他营养盐的添加。
在一优选实施例中，所述人工光源微藻养殖装备适用于小球藻、栅藻、雨生红球藻等淡水真核藻的养殖。
在一优选实施例中，所养殖的藻类的接种浓度为0.1g/L-2g/L(藻类干重/培养液体积)，更佳地0.2g/L-0.6g/L。
在一优选例中，发光装置所发出的光的光强在反应容器的水平方向(包括X轴和Y轴方向)基本均匀。
在一优选例中，所述“均匀的或大致均匀的”指任一深度处的光强D1与在整个深度范围的平均光强Dm满足下式：
1.5≥D1/Dm≥0.7。
较佳地，1.2≥D1/Dm≥0.8；更佳地1.1≥D1/Dm≥0.9。
在一优选例中，反应容器的周围设有温控装置，该温控装置用于维持反应容器内的液体环境温度处于适合微藻生长的范围内。
在一优选例中，该养殖设备中的养殖温度为10-30℃，更佳地为20-30℃。
在一优选例中，该温控装置可包括电加热或蒸汽加热系统、冷水降温系统等，例如控温水管。
在一优选例中，人工光源微藻养殖装备还设有监测系统，用于监测液体环境 的参数，参数选自：PH值、温度和/或养料浓度。
在一优选例中，在盖板上设有出气孔。
在一优选例中，该人工光源微藻养殖装备中设有多块导光板，较佳地30-1000块，更佳地40-800块，最佳地50-500块。
在一优选例中，导光板由具有防弱酸性能的透明有机材料制成。
附图说明
图1是根据本发明一个实施例的人工光源微藻养殖装备的立体剖视图；
图2是根据本发明一个实施例的人工光源微藻养殖装备的立体剖视示意图；
图3是根据本发明一个实施例的人工光源微藻养殖装备的主视剖视示意图；
图4是根据本发明一个实施例的人工光源微藻养殖装备的俯视剖视示意图；
图5是根据本发明一个实施例的人工光源微藻养殖装备的发光装置的立体示意图；
图6a是根据本发明一个实施例的人工光源微藻养殖装备的发光装置的主视图；
图6b是根据本发明一个实施例的人工光源微藻养殖装备的发光装置的侧视图；
图6c是根据本发明一个实施例的人工光源微藻养殖装备的发光装置的俯视图；
图6d是根据图6b中B部分的放大图；
图7a-7c是根据本发明其他实施例的人工微藻养殖设备的简化俯视示意图；以及
图8是根据本发明其他实施例的人工微藻养殖设备的侧截面示意图。
具体实施方式
本发明人经过广泛而深入的研究，首次开发了一种人工光源微藻养殖装备，通过特殊设计的发光装置等结构，本发明的反应器不仅缩短培养时间，而且能够显著提高微藻的生物量密度和总有效物含量，从而可制备品质更好的生物质原料及产品。在此基础上完成了本发明。
以下将结合附图对本发明的较佳实施例进行详细说明，以便更清楚理解本发明的目的、特点和优点。应理解的是，附图所示的实施例并不是对本发明范围的限制，而只是为了说明本发明技术方案的实质精神。
生物反应器
本发明的内置光源生物反应器人工光源微藻养殖装备适用于小球藻、栅藻、雨生红球藻等淡水真核藻的高密度产业化养殖。
如图1和图2所示，所述人工光源微藻养殖装备包括反应容器1、发光装置2、进出料装置3、盖板5、气体分布器6、营养布料装置、温控装置7、以及监测和控制系统等。反应容器1上设有盖板5，反应容器1与进出料装置3密封连接，且进出料装置上设有阀门和开关，用于控制进出料的进行或停止，反应容器1内设有发光装置2用于产生微藻生长各阶段所需的稳定光源，反应容器1的底部设有气体分布器6用于通入微藻生长各阶段所需的气体，营养布料装置用于向反应容器中提供养料，反应容器的周围设有温控装置7(图示为控温水管)用于保持反应容器内溶液的温度在适合微藻数量繁殖与个体生长的温度范围内。
该生物反应器的下部(如约30％～90％，或50％～70％的高度)可埋于地下。
该生物反应器可设置在发电厂周围，利用发电产生的废水废气作为微藻养殖的物质和能量来源，具有环保减排的作用。该反应容器1可以为不透光的密闭容器，在阴雨天与夜间也可以通过自身的发光装置进行发光，供微藻生长所用。在该养殖装备中设有监测及自动化控制系统，用于监测水温、PH值、CO2浓度、TDS、吸光度、光强度等，并能够根据需要对上述条件进行自动化控制。
图3是该人工光源微藻养殖装备的主视剖视示意图；如图3所示，在反应容器的壁上设有温控装置7，温控装置7可以为任何合适的形式，包括电加热或蒸汽加热系统、冷水降温系统，例如控温隔套等。较佳地，温控装置7为控温水管，发电厂排出的具有一定温度的废水可用于在控温水管中流通以保持生物反应器内的温度在20℃～30℃之间。
在生物反应器的盖板5的顶部设有出气孔，用于释放微藻光合作用产生的氧气。
图4是该人工光源微藻养殖装备的俯视剖视示意图。如图所示，在人工光源微藻养殖装备的底部设有气体分布器6，气体分布器6可为旋转式、槽式、管式 或其他任何合适的形式。在微藻养殖过程中，通过气体分布器向反应容器中通入二氧化碳、空气或其他气体。较佳地，本实例中采用旋转式气体分布器，在通入气体的过程中，气体分布器6旋转带动通入的气体与底部容器内液体的扰动，从而有利于气体与营养物质在溶液中均匀分布。
在反应容器的一个或多个位置(例如容器壁上、容器底部、容器顶部和盖板内侧等)安装营养布料装置。营养布料装置在微藻生长的不同阶段提供不同的养料。
反应容器中其他营养物质的通入速率与CO2的通入速率可成一定的比例，通过CO2在线监测系统可以确定溶液中CO2的溶解量，同时通过监测系统监测溶液的PH值获知溶液的酸碱度。控制系统能够控制的CO2和营养物质的通入速率，保证溶液中营养物质的溶解量及PH值在适合微藻生长繁殖或产油的范围内。
本发明的反应容器可采用玻璃钢制作而成，且反应容器可具有任何合适的外形和尺寸。容器表面较光滑，且较佳地，容器表面有可移动的封闭式盖板。
如图7a-7c所示，该反应容器可以为长环形立体容器。该容器的总长度为2m-500m，较佳地为10m-200m；容器截面高宽比为0.5:1-1:0.5；高度为20cm-400m，较佳地为50cm-200cm。较佳地，如图8所示，容器的底部截面中间低两边高，最低点位于中心，两边底部斜度为5°-60°，更佳地为10°-30°。
容器在中间部分的截面可变化为封闭式圆筒，圆筒直径为20cm-400m，更佳地为50cm-200cm，该部分长度可为20cm-100cm，更佳地可为20cm-50cm。容器的不同截面形状的转换部分呈光滑连接。
本发明的养殖设备还包括其他供气装置，例如图中未示出的气源、混合器、输气管等。本发明中气源主要为空气与二氧化碳。混合器主要用于混合空气与二氧化碳，混合器可采用文丘里管或静态混合器。输气口材质可为陶瓷微孔或树脂微孔结构，微孔直径为0.1-10微米，更佳地为0.5-5微米。气体分布器设置于养殖容器底部，气体分布器的下端距离容器内底0-5cm。如图所示，反应容器底部可设有多个并排等间隔布置的气体分布器。使用时，气体分布器的供气压力为0.2-20Mpa，更佳地为0.5-5Mpa。
较佳地，本发明的养殖设备中的营养布料装置可为位于容器上部的带孔管道，营养液经管道流至容器上方后可经小孔均匀喷洒至容器内的培养液中。此外，发明人经过实验和探究，发现在培养液中提供一定量的锰离子，能够有效地提高 养殖效益。因此，本发明的养殖设备的营养布料装置会向培养液中提供锰离子。
较佳地，本发明的养殖设备还设有循环推进装置，循环推进装置可具有螺旋桨式或螺杆式的推进桨叶，用于系统内水体的流动与循环。推进桨叶转速为10-1000rpm，更佳地为20-400rpm。
此外，本发明的养殖设备还可整合用于检测培养液中锰离子浓度的在线式水质监测仪，以便控制培养液中的锰离子浓度达到最有利于微藻养殖的范围。该水质监测仪的水样取水口位于液面以下，可定时自动采样。在水样自动送入检测仪之前经过微过滤系统，过滤精度为T级。当监测到锰离子含量低于5mg/L(或其他设定的数值)时，系统自动开启营养盐添加或者提醒管理人员进行营养盐补料。
本发明的养殖设备内的培养液的pH范围为5-10,更佳地为6-8。本发明的养殖设备还可整合PH在线监测传感器，用于测量培养液中的pH值，以便控制培养液中通入二氧化碳的浓度。当培养液中pH高于6.5(或其他设定的数值)时，系统自动开启二氧化碳阀门，向池中通入二氧化碳气体；当ph低于6.5(或其他设定的数值)时，系统关闭二氧化碳阀门，通气装置只向池中通入空气或者直接停止通入气体，从而有效调节二氧化碳的通入量，以获得微藻繁殖与生长的最佳二氧化碳条件。
发光装置
图5为发光装置2的立体示意图。发光装置的设置使得生物反应器可在无阳光或其他外部光源的情况下进行养殖生产等正常运作。如图5所述，所述发光装置由支架9、LED发光单元组10和导光板11构成，导光板11由支架9固定，支架9的顶端固定在盖板5上，LED发光单元组10位于导光板11的顶部，导光板11或全部伸入液面之下。在本发明的生物反应器中，设有多个发光装置2，各个导光板均可以单独地安装或拆卸。
导光板可用透明有机材料(例如亚克力)制成，且具有防弱酸性能。LED发光单元组在导光板的顶部与导光板直接接触，LED发光单元组发出的光可透过导光板传导并使导光板整体发光。较佳地，导光板平行放置，两块板之间的距离为可为5cm-50cm，更佳地，为10cm-20cm。发光板照度可以根据需要进行调整。
发光装置2的支架9的顶端与盖板5可拆卸地连接，从而便于发光装置的拆除、更换或重新安装；发光装置2悬挂在反应容器中而不接触容器的底部，因而 不影响容器底部气体分布器的旋转。应理解，发光装置的支架也可以以其他任何合适的方式在任何合适的位置与反应容器固定。
图6a-6c分别是人工光源微藻养殖装备的发光装置的主视图、侧视图和俯视图；图6d是图6b中B部分的放大图。
LED发光单元组10包括一个个小的LED发光单元，发光单元所产生的光可以是复合色或者单色光，单色LED发光波长范围为350-900nm，复合光色温范围为1500-20000K。单个导光板上的LED发光单元的数量可以是1-10000个。
光源为LED光源
发光装置工作时能够发射不同波长的光。该不同波长的光包括：波长为:600-680nm，更佳地650-670nm的光A；波长为420-500nm，更佳地440-460nm的光B；700-780nm，更佳地710-730nm的光C(红外光)。
发明人经过长期深入的实验探究，发现发光装置所发出不同波长的光的配比在以下范围时，养殖设备内的微藻养殖效益可达到较高的水平。发明人通过实验发现，相同照射功率的情况下，补充特定波长的近红外光后，可使微藻产量提高10-25％：
A:B＝10:1～2:1；
A:C＝20:1～3:1。
此外，发光装置的光照强度可以调整，光照时亮暗交替(发光功率变化)。较佳地，暗光时发光功率为亮光时发光功率的0％-80％更佳地为0％-60％。较佳地，暗光照射时间为亮光照射时间的10％-300％更佳地20％-100％
其中发光装置所发出的光的光强在反应容器的深度方向(Z轴方向)是均匀的或大致均匀的；发光装置所发出的光的光强在反应容器的水平方向(包括X轴和Y轴方向)基本均匀。“均匀的或大致均匀的”指任一深度处的光强D1与在整个深度范围的平均光强Dm满足下式：
1.5≥D1/Dm≥0.7。
较佳地，1.2≥D1/Dm≥0.8；更佳地1.1≥D1/Dm≥0.9。
实际上，LED发光单元组10可以位于导光板11的底部或者嵌于发光板的其他合适的位置，只要保证其发出的光可在整块导光板中传导使得整块导光板发光即可。
较佳地，LED发光单元组的能量来源为太阳能光板吸收太阳能而产生的电 能。如此以来，本发明的生物反应器虽没有直接利用太阳能，但通过太阳能光板将不稳定的太阳能收集起来用于发电，将其产生的电稳定地供应给生物反应器的发光单元组使其持续地发光，保证微藻养殖的稳定、持续性。
生产养殖系统
微藻的数量繁殖和个体增重阶段可在同一个生物反应器中进行，但需要随着生长的进行，更换发光装置，改变营养物质通入的种类与速率，过程较为复杂。
为了实现高效的大规模养殖，可采用多个生物反应器串联或并联组成大规模的生产养殖系统。生物反应器之间的串联或并联通过进出料口连接，通过泵系统可以完成生物反应器之间的进料和出料。
考虑到微藻养殖过程中的数量变化，对光源和营养物质的需求量也在不断发生变化，可采用多个生物反应器串联的养殖系统，例如，采用三个生物反应器串联的系统完成微藻生产的全部生长过程：
将适量的藻种放入第一级生物反应器中，繁殖增加至一定数量，通过进出料口进入第二级生物反应器进行进一步生长繁殖，而后再进入第三级生物反应器，不再繁殖，只主要进行个体生长，而较少繁殖。
在第一级和第二级生物反应器中设置有发光波长仅适合有利于微藻繁殖的发光装置，且营养布料装置向反应容器中通入磷酸铵、磷酸二氢钾或磷酸氢二钾等氮氧化物作为氮磷源，同时添加铁、锌等其他微藻生长所必须的营养盐，气体分布器向反应容器中通入氮氧化物和CO2，第二级生物反应器中发光强度和营养物质供应速率较第一级生物反应器大，以适应繁殖增多的微藻的生长需求；在第三级生物反应器中设置有发光波长仅适合有利于微藻个体生长的发光装置，PH和温度等同时调节至适合微藻个体生长的条件，气体分布器向反应容器中通入CO2。
不同级的生物反应器可根据需求选择不同的尺寸，例如第一级生物反应器的尺寸较后两级较小。
第一级生物反应器中的微藻排出至第二级生物反应器后，可向第一级生物反应器通入新的藻种，同时进行新一轮养殖。为了提高藻种的纯度，可从第二级生物反应器排出的微藻中过滤出部分作为第一级生物反应器的藻种。
培养方法
本发明提供了一种培养微藻的方法，该方法包括步骤：
1.提供本发明的人工光源微藻养殖装备；
2.将藻种投放于该人工光源微藻养殖装备中，向反应器中提供微藻生存所需的养料(包括氮源、磷源、无机盐(例如锰、铁、锌、镁等)并提供二氧化碳或空气，开启发光装置产生微藻生长所需的光。
适用于本发明的微生物没有特别限制，只要其可利用光源进行生长。代表性的微生物包括(但并不限于)：绿藻门、蓝藻门、金藻门和红藻门微藻。一种优选的微生物是小球藻(Chlorellales)淡水种。本发明的养殖设备和养殖工艺尤其适用于小球藻、栅藻、雨生红球藻等淡水真核藻的养殖。
营养成分
本发明的养殖设备的反应容器中所养殖微藻的接种浓度为0.1g/L-2g/L(微藻干重/培养液体积)，更佳地0.2g/L-0.6g/L。营养布料装置以及供气装置所提供的营养成分的供应量应根据接种和饲养浓度来确定，当接种和饲养浓度提高时应适当提高营养盐浓度，并在养殖过程中保持和补充营养盐的消耗。
本发明的养殖设备可通入二氧化碳作为碳源。较佳地，二氧化碳与空气混合通入，其中二氧化碳占混合气体总体积的0.1-10％，较佳地为0.5-5％；二氧化碳通入压力为0.1-1MPa；二氧化碳释出孔径为0.1-50nm更佳地1-20nm。
本发明的养殖设备内的培养液的pH范围为5-10,更佳地为6-8。为了更好地控制培养液中二氧化碳的通入两，在一优选实施例中，当培养液中的pH≥5.5时，可持续通入含二氧化碳的混合气体；而当PH<5.5时，可暂停通气。
培养液中的氮源可选择以下含氮化合物的一种或几种，且各种每升培养液中含氮化合物的通入量大致如下：
CO(NH2)2，500-1500mg/L；
NaNO3，1000-3000mg/L；
NH4HCO3，1000-3000mg/L；
NH4NO3，500-1500mg/L。
培养液中的磷源可选择以下含磷化合物的一种或几种，且各种每升培养液中含磷化合物的通入量大致如下：
K2HPO4，20-80mg/L；
KH2PO4，20-80mg/L。
发明人经过长期而深入的研究，发现在培养液中加入适量的锰离子时，养殖效率大大提升。例如，可在培养液中添加浓度为5-50mg/L(添加剂重量/培养液体积)的MnCl2·4H2O。较佳地，通常藻种使用的培养基BG11培养基配比成的养殖系统中MnCl2·4H2O浓度为1.86mg/L。
除了上述碳源、氮源、磷源、锰离子以外，培养液中常常加入以下其他营养盐，且其他营养盐的浓度范围如下：
MgSO4·7H2O，70-150mg/L；
CaCl2·2H2O，30-80mg/L；
柠檬，5-10mg/L；
柠檬酸铁，5-50mg/L；
H3BO3，1.5-5mg/L；
ZnSO4·7H2O，0.2-0.5mg/L；
Na2MoO4·2H2O，0.2-0.8mg/L；
CuSO4·5H2O，0.05-0.3mg/L；
Co(NO3)2·6H2O 0.03-0.2mg/L。
制备方法
本发明提供了一种制备微藻生物质的方法，该方法采用通过前述方法制备的微藻藻液作为原料，进行加工，从而制得微藻成品，一种典型的加工过程包括步骤：
利用过滤、压滤或者气泡悬浮的方法收集微藻，低温烘干然后粉碎后即可得到微藻成品。
实施例1
在小型人工光源实验箱内进行微藻繁殖，所选藻种为蛋白核小球藻，接种浓度为0.3g/L(干重)。人工光源为三色LED光源，波长分别为650±5nm，440±5nm，720±10nm，三色光源配比为10:3:2，光线最暗处光强约为3000lx，亮暗交替时间为10小时(亮)：2小时(暗)，暗光功率约为亮光功率的40％。通入二氧化碳， 压力为0.4Mpa，释出的陶瓷膜压力管的孔径为10nm，二氧化碳与空气混合占比为2％-3％，持续通气至PH＝6.5左右停止，降低二氧化碳与空气混合占比，直至PH区间保持为6.5-6.8之间。养殖温度保持为27度±1度。
本实施例中培养液的营养选择为：
CO(NH2)2，800mg/L；
NaNO3，1200mg/L；
KH2PO4，50mg/L；
MgSO4·7H2O，100mg/L；
CaCl2·2H2O，40mg/L；
柠檬酸，6mg/L；
柠檬酸铁铵，25mg/L；
H3BO3，3mg/L；
MnCl2·4H2O，12mg/L；
ZnSO4·7H2O，0.3mg/L；
Na2MoO4·2H2O，0.4mg/L；
CuSO4·5H2O，0.1mg/L；
Co(NO3)2·6H2O，0.06mg/L。
每日监测三次以上离子浓度，并适当补料保持营养。
养殖7天后，收集、过滤、烘干后称重，得最终生物质浓度为1.07g/L。
对比例1：
采用同样方法，但MnCl2·4H2O浓度变为1.8mg/L，养殖7天后，收集、过滤、烘干后称重，得最终生物质浓度为0.71g/L。
对比例2：
采用同样方法，但培养基中不添加MnCl2·4H2O，养殖7天后，收集、过滤、烘干后称重，得最终生物质浓度为0.42g/L，且颜色发黄，出现漂浮死藻。
对比例3：
采用同样方法，但MnCl2·4H2O浓度变为100mg/L，养殖7天后，收集、 过滤、烘干后称重，得最终生物质浓度为0.52g/L，出现漂浮死藻。
对比例4：
采用同样方法，但人工光源为两色LED光源，波长分别为650±5nm，440±5nm，两色光源配比为10:3，养殖7天后，收集、过滤、烘干后称重，得最终生物质浓度为0.82g/L。
对比例5：
采用同样方法，但不通入二氧化碳，只通入空气，养殖7天后，收集、过滤、烘干后称重，得最终生物质浓度为0.79g/L。
对比例6：
采用同样方法，但不通任何气体，藻体发生沉降，无法进行养殖。
结论
将上面的实施例和对比例的实验结果可以看出：
1.实施例1中MnCl2·4H2O的浓度为12mg/L，养殖7天后得到的最终生物质浓度为1.07g/L，而对比例1-3中或者减少或者增加MnCl2·4H2O的浓度，发现养殖7天后得到的最终生物质浓度都较实施例1有所减少，且一些对比例中出现了漂浮死藻，由此可以看出，在培养液中加入适量的锰离子的确有利于养殖设备中所养殖藻类的生长；
2.实施例1中采用配比为10:3:2的三色LED光源，而对比例4中采用了不发射红外光的两色LED光源，在其他培养条件相同的条件下，实施例1的养殖成果明显优于对比例4，由此可以看出，一定比例的红外光源有利于养殖设备中所养殖藻类的生长；
3.实施例1中通入一定混合占比的二氧化碳与空气的混合气体，而对比例5中不通入二氧化碳，对比例6中不通入任何气体，从养殖成果可以看出，通入适量混合比例的二氧化碳和空气，对藻类的生长和繁殖十分必要。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被 单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (20)

  1. 一种人工光源微藻养殖装备，其特征在于，所述人工光源微藻养殖装备包括：
    反应容器，所述反应容器上设有盖板且所述反应容器容纳供微藻生长的培养液；
    进出料装置，所述进出料装置与所述反应容器密封连接；
    发光装置，所述发光装置设在反应容器内部且当所述反应器工作时，所述发光装置至少部分或全部浸没于所述的培养液，从而在反应容器内提供微藻生长所需的光，其中所述发光装置所发出的光的光强在反应容器的深度方向(Z轴方向)是均匀的或大致均匀的；
    营养布料装置，所述营养布料装置可向所述培养液中提供锰离子；和
    供气装置，所述供气装置用于向所述反应容器内提供微藻生长所需的气体。
  2. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述培养液中的锰离子源来自MnCl2·4H2O，且每升所述培养液中添加的MnCl2·4H2O的质量为5-50mg。
  3. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述发光装置工作时发射至少三个不同波长的光，其中包括波长为600-680nm的光、波长为420-500nm的光，以及波长为700-780nm的光；和/或
    所述发光单元发出的复合光的色温为1000-20000K，较佳地1500-6000K。
  4. 如权利要求3所述的人工光源微藻养殖装备，其特征在于，所述三个不同波长的光包括波长为650-670nm的光、波长为440-460nm的光，以及波长为710-730nm的光。
  5. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述发光装置包括导光板和发光单元，所述发光单元嵌于所述导光板，所述发光单元产生的光透过导光板传导使导光板整体发光，所述发光装置还包括支架，所述支架用于固定支撑所述导光板，所述支架与所述反应容器和/或所述盖板可拆卸地连接；和/或
    所述发光装置中，发光单元为LED，且LED的数量为1-10000/导光板；较佳 地为10-1000/导光板。
  6. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述供气装置可向所述培养液中通入二氧化碳和空气的混合物，其中二氧化碳占混合气体总体积的0.1-10％，较佳地占0.5-5％；和/或
    二氧化碳通入压力为0.1-1MPa；和/或
    二氧化碳释出孔径为0.1-50nm。
  7. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述养殖装备中还设有用于检测和控制所述培养液中二氧化碳通入量的监测及自动化控制系统，其中
    当所述监测及自动化控制系统检测到所述培养液中的pH≥5.5时，含二氧化碳的混合气体被持续通入所述培养液中；而当所述培养液中的PH<5.5时，所述培养液中暂停通入二氧化碳。
  8. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述反应容器的周围设有温控装置，所述温控装置用于维持反应容器内的液体环境温度处于适合微藻生长的范围内；和/或
    所述供气装置包括气体细化分布器，在向反应容器中通入气体的过程中，带有一定压力的气体通过该细化器的小孔均匀细散地逸出，从而促使气体与营养物质分散于液态培养体系中。
  9. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述人工光源微藻养殖装备中还设有用于监测所述培养液中锰离子或其他离子浓度的水质监测仪，所述水质监测仪的取水口位于液面以下，其中
    当所述水质监测仪监测到所述培养液中的锰离子或其他离子的浓度低于预设数值时，所述水质监测仪能够发出提醒，使得所述营养布料装置进行锰盐或其他营养盐的添加。
  10. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述人工光源微藻养殖装备适用于小球藻、栅藻、雨生红球藻等淡水真核藻的养殖。
  11. 如权利要求10所述的人工光源微藻养殖装备，其特征在于，所养殖的藻类的接种浓度为0.1g/L-2g/L(藻类干重/培养液体积)，更佳地0.2g/L-0.6g/L。
  12. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述的发光 装置所发出的光的光强在反应容器的水平方向(包括X轴和Y轴方向)基本均匀。
  13. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述“均匀的或大致均匀的”指任一深度处的光强D1与在整个深度范围的平均光强Dm满足下式：
    1.5≥D1/Dm≥0.7；
    较佳地，1.2≥D1/Dm≥0.8；更佳地1.1≥D1/Dm≥0.9。
  14. 如权利要求8所述的人工光源微藻养殖装备，其特征在于，反应容器的周围设有温控装置，所述的温控装置用于维持反应容器内的液体环境温度处于适合微藻生长的范围内。
  15. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述养殖装备中的养殖温度为10-30℃，更佳地为20-30℃。
  16. 如权利要求8所述的人工光源微藻养殖装备，其特征在于，所述的温控装置可包括电加热或蒸汽加热系统、冷水降温系统。
  17. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述人工光源微藻养殖装备还设有监测系统，用于监测液体环境的参数，参数选自：PH值、温度和/或养料浓度。
  18. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述的盖板上设有出气孔。
  19. 如权利要求1所述的人工光源微藻养殖装备，其特征在于，所述人工光源微藻养殖装备中设有多块导光板，较佳地30-1000块，更佳地40-800块，最佳地50-500块。
  20. 如权利要求5所述的人工光源微藻养殖装备，其特征在于，导光板由具有防弱酸性能的透明有机材料制成。
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CN107227252A (zh) * 2017-07-03 2017-10-03 深圳市华明佰利科技有限公司 一种全密闭的垂直led光能生物反应釜
CN112739813A (zh) * 2018-05-03 2021-04-30 瓦克萨科技有限公司 用于储存活微藻的装置和方法
CN114438070A (zh) * 2022-02-15 2022-05-06 广东海纳川生物科技股份有限公司 一种悬挂式固体微生物及其制备方法、养殖池和应用
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CN114703051A (zh) * 2022-03-11 2022-07-05 湖南康大生物药品有限责任公司 一种硅藻生物养殖系统及养殖方法
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