WO2016000192A1 - Bioreactor with built-in light source and microalgae culture method - Google Patents

Abstract

A bioreactor with a built-in light source comprises a reaction vessel, a material feeding and discharging device, a light-emitting device, a nutrient distributing device and a gas distributor, wherein the reaction vessel is provided thereon with a cover plate and serves to accommodate a culture liquid for microalgae growth; the material feeding and discharging device is in sealing connection with the reaction vessel and provided thereon with a valve and a switch, and microalgae are pumped into or out of the reaction vessel through the material feeding and discharging device; the light-emitting device is used for generating a light source required for microalgae growth; the nutrient distributing device is used for supplying nutrients required for microalgae growth into the reaction vessel; and the gas distributor is used for supplying gases required for microalgae growth into the reaction vessel. The bioreactor with the built-in light source is not affected by weather changes, can realize stage-by-stage microalgae reproduction, has high controllability, and is beneficial to realization of stable and continuous industrialized production; and due to the serial and/or parallel connection system of the bioreactor, large-scale microalgae culture becomes more efficient.

Description

 Built-in light source bioreactor and microalgae breeding method

 The invention relates to the field of biotechnology, and particularly relates to a built-in light source bioreactor and a microalgae cultivation method. Background technique

 Microalgae biodiesel is the future development direction of liquid biofuels. It has high energy conversion efficiency, and the yield per unit area can be several tens of times higher than that of ordinary terrestrial crops, so that factory farming can be realized. The principle of microalgae oil production is to use microalgae photosynthesis to convert the carbon dioxide produced in the chemical production process into the microalgae's own biomass to fix the carbon element, and then induce the reaction to convert the microalgae's own carbon material into oil. Then, the oil or fat in the microalgae cells is transformed into the extracellular cells by physical or chemical methods, and then refined to produce biodiesel, which can convert the nutrients and carbon dioxide in the waste gas into biofuels through photosynthesis of algae. , protein. Today, with the sharp rise in oil prices and the growing shortage of food, the industry has broad prospects for development.

The cultivation of microalgae for the production of biodiesel is mainly divided into two stages, namely the breeding stage and the oil production stage. In the breeding stage of microalgae cultivation, microalgae mainly absorb red-yellow light and blue-violet light (wavelength range 620-700nm and 410-470nm), while absorbing carbon, nitrogen, phosphorus and other elements to reproduce without producing oil; During the oil production stage of algae culture, nitrogen source is no longer used to maintain a small amount of phosphorus and continuously enters C0 ₂ to maintain light. The microalgae is significantly rich in oil production due to carbon and nitrogen starvation, and the reproduction rate is significantly reduced.

 In order to further increase the unit yield of microalgae biodiesel and achieve three-dimensional culture, a dedicated microalgae culture photoreactor must be used. Although 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 since the light source of the microalgae growth and oil production is still natural light, The use of sunlight by microalgae is not sufficient, it is uncertain and uncontrollable, and it runs counter to the stable and continuous requirements of factory production. In addition, the auxiliary farming functions such as nutritional supplement and carbon dioxide aeration are not automated, making it practical. Industrialization applications still have a long way to go.

Therefore, there is an urgent need in the art for a technology to overcome the disadvantages of poor controllability, low automation, and low efficiency of the microalgae aquaculture photoreactor of the prior art described above. Summary of the invention The object of the present invention is to provide a built-in light source bioreactor which can optimize the source of light source, carbon dioxide, nutrition, temperature and flow rate according to different breeding requirements of different kinds of algae, thereby realizing the stability of ultra-high density seaweed culture. , controllable, to meet the needs of industrialization.

 According to one aspect of the invention, a built-in light source bioreactor is provided, the built-in light source bioreactor comprising:

 a reaction vessel, a reaction vessel is provided with a cover plate and the reaction vessel contains a culture liquid for the growth of the microalgae; an inlet and outlet device, the inlet and outlet device is sealingly connected with the reaction vessel, and a valve and a switch are arranged on the inlet and outlet device;

 a light-emitting device, wherein the light-emitting device is disposed inside the reaction vessel and when the reactor is in operation, the light-emitting device is at least partially or completely immersed in the culture liquid, thereby providing light required for the growth of the microalgae in the reaction container, wherein the light-emitting device emits The light intensity of the light is uniform or substantially uniform in the depth direction (Z-axis direction) of the reaction vessel;

 A gas distributor for supplying a gas required for the growth of microalgae into the reaction vessel. In another preferred embodiment, 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).

 In another preferred embodiment, the "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:

 1. 5 ^ D1 /Dm ^ 0. 7.

 Preferably, 1. 2 ^D 1/Dm^ 0. 8; more preferably 1 · 1 D1 /Dm 0. 9.

 In another preferred embodiment, 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.

 In another preferred embodiment, the reactor is further provided with a nutrient distribution device for providing the nutrients required for the growth of the microalgae in the reaction vessel.

 In another preferred embodiment, the gas distributor is a rotary gas distributor, and during the passage of the gas into the reaction vessel, the rotary gas distributor rotates to promote the dispersion of the gas and nutrients in the liquid culture system.

 In another preferred embodiment, the temperature control device is a temperature controlled water pipe.

In another preferred embodiment, the range suitable for the growth of the microalgae means 15-45 ° C, preferably 20-40 ° C. In another preferred embodiment, the built-in light source bioreactor is further provided with a monitoring system for monitoring parameters of the liquid environment, the parameters being selected from the group consisting of: ra value, temperature and/or nutrient concentration. In another preferred embodiment, the light emitting device includes a light guide plate and a light emitting unit, and the 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.

 In another preferred embodiment, the light-emitting device further includes a bracket for fixing and supporting the light guide plate, the bracket being detachably coupled to the reaction container and/or the cover plate, and the light-emitting unit being embedded in the light guide plate.

 In another preferred embodiment, an air vent is provided in the cover.

 In another preferred embodiment, the lighting unit is an LED lighting unit.

 In another preferred embodiment, the built-in light source bioreactor is provided with a plurality of light guide plates, preferably 3-100 pieces, more preferably 4-80 pieces, and most preferably 5-50 pieces.

 In another preferred embodiment, the light guide plate is made of a transparent organic material having weak acid resistance.

 In another preferred embodiment, the illumination device emits at least 2 different wavelengths of light when in operation.

 In another preferred embodiment, the light of the different wavelengths includes: light having a wavelength of 600-800 (preferably 650-750) nm and light having a wavelength of 400-480 (preferably 430-470) nm.

In another preferred embodiment, the color temperature of the light emitted by the light emitting unit is 1000-20000K, preferably

 In another preferred embodiment, in the light-emitting device, the light-emitting unit is an LED, and the number of the LEDs is 1-10000 / light guide plate; preferably 10-1000 / light guide plate.

 According to a second aspect of the invention, there is provided a production aquaculture apparatus comprising the built-in light source bioreactor of the first aspect of the invention.

 In another preferred embodiment, at least two of the built-in light source bioreactors are connected in series and/or in parallel.

 In another preferred embodiment, the interconnected reactors are connected by an inlet and outlet device.

 In another preferred embodiment, the production aquaculture equipment is used as a production and breeding system for Chlorella, Chlorella, Cyanophyta and Red algae microalgae.

 According to a third aspect of the invention, there is provided a method of cultivating, the method comprising the steps of:

 (a) providing the aforementioned built-in light source bioreactor;

 (b) Culture microalgae in a built-in light source bioreactor.

 According to a fourth aspect of the present invention, there is provided a method of preparing microalgae for producing a material diesel, the method comprising the steps of:

 (a) providing the aforementioned built-in light source bioreactor;

 (b) cultivating microalgae for the production of material diesel in a built-in light source bioreactor,

Wherein the culture comprises a first culture stage and a second culture stage, in the first stage of the growth of the microalgae, the illumination The light emitted by the unit has a wavelength of 350-900 nm, preferably 570-800 and 400-500 nm; in the second stage of microalgae growth, the light emitted by the light-emitting unit has a wavelength of 350-900 nm, preferably 600-800 and 400-480nm;

 (c) recovering the cultured microalgae from the built-in light source bioreactor.

 In another preferred embodiment, in the first stage of microalgae growth, the illuminating unit emits a wavelength of 570-800 nm; in the second stage of microalgae growth, the illuminating unit emits at a wavelength of 400-480 nm.

 In another preferred embodiment, a nitrogen source is provided to the built-in light source bioreactor during the first stage of microalgae growth; in the second stage of microalgae growth, the supply of nitrogen source to the built-in light source bioreactor is stopped.

 In another preferred embodiment, the method further comprises: processing the recovered microalgae (eg, drying, breaking, extracting, transesterifying, etc.) to produce biomass diesel.

 According to a fifth aspect of the present invention, there is provided a microalgae which can be used for the production of a substance diesel which is prepared by the method described in the fourth aspect.

 In another preferred embodiment, the microalgae has the following characteristics:

 (i) the size of the microalgae is 5-500 microns;

 (ϋ) Oil content (dry weight) 10%-70%.

 According to a sixth aspect of the invention, there is provided a method of producing biomass diesel, characterized in that it comprises the steps of: using the microalgae according to the fifth aspect of the invention as a raw material, and processing, thereby producing biomass diesel oil.

 In another preferred embodiment, the processing of the microalgae comprises: drying, extraction, extraction, transesterification, and the like. DRAWINGS

 1 is a perspective cross-sectional view of a built-in light source bioreactor according to an embodiment of the present invention; FIG. 2 is a perspective cross-sectional view of a built-in light source bioreactor according to an embodiment of the present invention; FIG. 3 is a schematic view of a built-in light source bioreactor according to an embodiment of the present invention; 4 is a schematic cross-sectional view of a built-in light source bioreactor; FIG. 4 is a top cross-sectional view of a built-in light source bioreactor according to an embodiment of the present invention; FIG. 5 is a light-emitting of a built-in light source bioreactor according to an embodiment of the present invention. a schematic view of the device;

 Figure 6a is a front elevational view of a light emitting device incorporating a light source bioreactor in accordance with one embodiment of the present invention;

Figure 6b is a side view of a light emitting device with a built-in light source bioreactor, in accordance with one embodiment of the present invention; Figure 6c is a top plan view of a light emitting device with a built-in light source bioreactor according to one embodiment of the present invention;

 Figure 6d is an enlarged view of the portion B according to Figure 6b. detailed description

 The inventors have extensively and intensively researched and developed a built-in light source bioreactor for the first time. The reactor of the present invention not only shortens the culture time but also significantly increases the biomass density of the microalgae through a specially designed light-emitting device or the like. And the total effective fat content, so that better quality biodiesel can be prepared. The present invention has been completed on this basis. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The preferred embodiments of the present invention will be described in detail with reference to the appended claims The embodiment shown in the drawings is not intended to limit the scope of the invention, but only to illustrate the spirit of the invention. Bioreactor

 As shown in FIG. 1 and FIG. 2, the built-in light source bioreactor includes 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. 2 for generating a stable light source required for each stage of microalgae growth, a gas distributor 6 is provided at the bottom of the reaction vessel 1 for introducing gas required for each stage of microalgae growth, and a nutrient distribution device is used for providing nutrients to the reaction vessel A temperature control device 7 (shown as a temperature control 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 microalgae propagation and oil production.

 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 installed around the power plant, and the waste water generated by the power generation can be used as a source of material and energy 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 system is provided in the built-in light source bioreactor for monitoring the solution ra value and temperature in the reactor.

In addition, the built-in light source bioreactor of the present invention can also be used for silk algae, dinoflagellates or other aquatic organisms. High-density industrial farming.

 Figure 3 is a front cross-sectional view of the built-in light source bioreactor; as shown in Figure 3, 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, such as a temperature control compartment. set. Preferably, the temperature control device 7 is a temperature control water pipe, and the wastewater having 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 15 ° C and 45 ° C.

 An air vent is provided at the top of the cover 5 of the bioreactor for releasing oxygen generated by photosynthesis of the microalgae.

 Figure 4 is a top cross-sectional view of the built-in light source bioreactor. As shown, a gas distributor 6 is provided at the bottom of the built-in light source bioreactor. The gas distributor 6 can be in the form of a rotary, trough, tubular or any other suitable form. In the microalgae culture process, carbon dioxide, air or other gas is introduced into the reaction vessel through a gas distributor. Preferably, in this example, a rotary gas distributor is used. During the process of introducing the gas, 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 flow speed of C0 ₂ may be such proportion, the amount of dissolution solution is C0 ₂ by the C0 ₂ online monitoring system may determine that, while the ra value obtained by the monitoring system monitoring the solution known solution The pH. The control system can control the rate of introduction of co ₂ and nutrients to ensure that the amount of nutrients dissolved in the solution and the ra value are within the range suitable for the growth or production of microalgae. Illuminating device

 Fig. 5 is a perspective view of the light-emitting device 2. The arrangement of the illuminators allows the bioreactor to operate normally in the absence of sunlight or other external light sources. As shown in FIG. 5, 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. At the top of the light guide plate 11, the light guide plate 11 or all of them protrude below the liquid surface. In the bioreactor of the present invention, 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 transmit light. The plate conducts and causes the light guide plate to emit light as a whole.

 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 not affecting the gas at the bottom of the container The rotation of the spreader. It should be understood that the holder of the illumination device can also be secured to the reaction container at any suitable location in any suitable manner.

 Figures 6a-6c are front, side and top views, respectively, of a light-emitting device with a built-in light source bioreactor; 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 range of the monochrome LED is 350-900 nm, and the color temperature range of the composite light is 1500-20000K. The number of LED lighting units on a single light guide plate can be from 1 to 10,000.

 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 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 the intensity D1 at any depth and the average intensity over the entire depth range. Dm satisfies the following formula:

 1 · 5 D1 /Dm 0. 7.

 Preferably, 1. 2 ^D 1/Dm^ 0. 8; more preferably 1 · 1 D1 /Dm 0. 9.

 The illuminating device is capable of emitting at least 2 different wavelengths of light when operating. The different wavelengths of light include: light having a wavelength of 570-800 nm and light having a wavelength of 400-500 nm.

 In fact, the LED lighting unit group 10 may be located at the bottom of the light guide plate 1 1 or at other suitable positions of the illuminating panel, as long as the light emitted by the LED light guide plate can be conducted in the entire light guide plate so that the entire light guide plate emits light.

 Preferably, the energy source of the LED lighting unit group is the electric energy generated by the solar panel absorbing solar energy. Thus, although 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 sustainability of microalgae culture. Production culture system

 The propagation and oil production stages of microalgae 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.

In order to achieve efficient large-scale farming, multiple bioreactors can be used in series or in parallel to form large gauges. Mold production 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.

 Considering the quantitative changes in the microalgae cultivation process, the demand for light sources and nutrients is also constantly changing. A plurality of bioreactors can be used in series, for example, a system in which three bioreactors are connected in series to complete micro The entire growth process of algae reproduction and oil production:

 Put the appropriate amount of algae into the first-stage bioreactor, increase the reproduction to a certain amount, enter the second-stage bioreactor through the inlet and outlet for further growth and reproduction, and then enter the third-stage bioreactor, no longer breeding. , mainly for oil production, but less breeding.

 In the first-stage and second-stage bioreactors, a light-emitting device having an emission wavelength suitable only for facilitating the propagation of microalgae is provided, and the nutrient distribution device is supplied with ammonium phosphate, potassium dihydrogen phosphate or dipotassium hydrogen phosphate into the reaction container. Nitrogen oxides are used as nitrogen and phosphorus sources, and elements such as iron and zinc are added at the same time. The gas distributor introduces nitrogen oxides and CO 2 into the reaction vessel, and the luminous intensity and nutrient supply rate in the second-stage bioreactor are higher than those of the first-order organisms. The reactor is large to accommodate the growth requirements of the microalgae with increased reproduction; in the third-stage bioreactor, the illuminating wavelength is only suitable for the light-emitting device which is beneficial to the production of microalgae, and the PH and temperature are simultaneously adjusted to be suitable for microalgae production. Under the condition of the oil, and the nutrient distribution device does not provide a nitrogen source, the gas distributor introduces C02 into the reaction vessel.

 Different grades of bioreactors can be sized according to their needs. For example, the size of the first stage bioreactor is smaller than the latter two stages.

 After the microalgae in the first-stage bioreactor is discharged to the second-stage bioreactor, new algae species can be introduced into the first-stage bioreactor while a new round of culture is carried out. In order to increase the purity of the algae species, a portion of the algae species as the first stage bioreactor can be filtered from the microalgae discharged from the second stage bioreactor. Training method

 The present invention provides a method of culturing microalgae, the method comprising the steps of:

 1. Providing a built-in light source bioreactor of the invention;

 2. The algae species are placed in the built-in light source bioreactor to provide the nutrients (including nitrogen sources, phosphorus sources, inorganic salts (such as iron, zinc), etc.) required for the survival of the microalgae and provide carbon dioxide or air. Turn on the illuminating device to generate the light needed for the growth of the microalgae.

In particular, the present invention provides a method for culturing microalgae producing biomass diesel, the method comprising the steps of: 1. Providing a built-in light source bioreactor of the invention;

 2. The algae species are placed in the built-in light source bioreactor to provide the nutrients (including nitrogen sources, phosphorus sources, inorganic salts (such as iron, zinc), etc.) required for the survival of the microalgae and provide carbon dioxide or air. Opening the illuminating device to generate light required for the growth of the microalgae, wherein

 In the first stage of microalgae growth, the light emitted by the light emitting unit has a wavelength of 350-900 nm, preferably

570-800 and 400-500 nm; In the second stage of microalgae growth, the light emitted by the light-emitting unit has a wavelength of 350-900 nm, preferably 600-800 and 400-480 nm.

 Preferably, in the first stage and the second stage of the growth of the microalgae, different wavelengths of light are used as the light source, and the supply of the nutrient and the gas is used to make the microalgae mainly propagate in the first stage, and the second stage mainly produces oil. For example, in the first stage, red light is used as the light source and the nitrogen source is passed, and in the second stage, blue light is used as the light source and the nitrogen source is stopped.

 The 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): Chlorophyta, Cyanophyta, Chlorella, and Red algae microalgae. A preferred microorganism is a freshwater species of Nannochlorops i s l imne t i ca. Preparation

 The present invention provides a method for preparing biomass diesel using a microalgae prepared by the aforementioned method for producing biomass diesel as a raw material, thereby processing to produce biomass diesel, and a typical processing process includes Steps:

 The microalgae are collected by filtration, pressure filtration or bubble suspension, and then the algae oil and the algae are separated by cellulase hydrolysis and homogenization, and the pure algae oil is obtained by extraction, leaching or pressure filtration. Biodiesel that can be directly used in diesel engines is obtained after transesterification, solvent dilution or thermal decomposition. Advantages of the invention

 Compared to the prior art, the present invention has the following main advantages:

 1. The bioreactor in the prior art relies on external natural light, and in the case of weak weather such as rainy days, the microalgae cannot obtain sufficient light source to slow down growth, and the built-in light source bioreactor of the present invention is not The impact of weather changes;

2. The prior art is not suitable for large-scale aquaculture, because only a microalgae close to the upper surface of the pool can obtain a sufficient light source, and the bioreactor of the present invention has a plurality of uniformly distributed light-emitting devices built therein, and the present invention The gas distributor and nutrient distribution device in the bioreactor of Mingming are beneficial to the uniform distribution of nutrients in the reaction vessel, so that large-scale aquaculture, three-dimensional aquaculture can be realized, and the breeding efficiency can be improved;

 3. The bioreactor of the invention can adopt different built-in light sources and nutrition, temperature, pH and the like according to different growth stages of the microalgae, so that the microalgae propagation is carried out in stages, and the controllability is strong, which is favorable for achieving stable and continuous Factory production;

 4. The series and/or parallel system of the bioreactor of the present invention makes the large-scale cultivation of microalgae more efficient.

 The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in which the specific conditions are not specified in the following examples are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are percentages by weight and parts by weight.

 In Examples 1-3, the built-in light source bioreactor of the present invention as shown in Fig. 1 was employed. Example 1

The conventional Nannochlorops isl imnetica freshwater species was selected, the initial culture density was 0. lg/L, and the reactor liquid depth was 80 cm (the cross-sectional area was lm X 1.5 m). In the first stage, a red LED with a wavelength of 706 nm is used as a light source, and the temperature is constant at 24 ° C. A trace amount of ammonium phosphate, zinc, iron and other trace element fertilizers are preliminarily added to the culture solution, and a certain amount of C0 _{2 is introduced} (about 90%). And nitrogen oxides such as NO or N0 ₂ as a nitrogen source are placed in the algae species at the initial culture density after one hour, and the culture is started. After 36 hours, the biomass density was increased to 6.86 g/L (dry weight/solution, the same below). After 72 hours, the biomass density was 9.71 g/L, and the microalgae in the reactor was moved to another reaction. In the device (liquid surface depth 80cm, cross-sectional area 2m X 3m), stop the supply of nitrogen oxides, keep C0 ₂ supply, use blue LED with wavelength of 450nm as the light source, the temperature is constant at 28 °C, move to another reaction After 24 hours, the water sample was taken, the nitrogen source was almost exhausted, and the biomass density was 10.64 g/L. After 72 hours of moving to another reactor, the biomass density increased to 12.88 g/L. A total of 144 hours.

The distribution of microalgae in the reactor was visually observed. 3%, The total effective fat content is 40. 39%, the total lipid content (dry weight) is 45.08%, the triglyceride fatty acid ester (triglyceride) accounts for 89.6%, the total effective fat content is 40.39%. , total effective lipid content density of 5. 20g / L. Example 2 The freshwater species of Nannochloropsis 1 imnetica was selected, the initial culture density was 0. lg/L, and the liquid level of the reactor was 80 cm (the cross-sectional area was ImX1.5 m). In the first stage, a blue LED with a wavelength of 450 nm is used as a light source, and the temperature is constant at 24 ° C. A trace amount of ammonium phosphate, zinc, iron and other trace element fertilizers are preliminarily added to the culture solution, and a certain amount of C0 ₂ (about 90%) is respectively introduced. And nitrogen oxides such as NO or N0 ₂ as a nitrogen source are placed in the algae species at the initial culture density after one hour, and the culture is started. The biomass density increased to 5.66 g/L after 36 hours, and the biomass density was 8.03 g/L after 72 hours. The microalgae in the reactor was moved to another reactor (the depth of the liquid surface was 80 cm, and the cross-sectional area was In 2mX3m), the supply of nitrogen oxides is stopped, the supply of C0 _{2 is} kept, and the red LED with a wavelength of 706 nm is used as the light source. The temperature is constant at 30 ° C. After moving to another reactor for 24 hours, the water sample is taken, and the nitrogen source is basically After depletion, the biomass density was 9.21 g/L. After 72 hours of moving to another reactor, the biomass density increased to 10.52 g/L, and the culture ended, for a total of 144 hours.

 The distribution of microalgae in the reactor was visually observed. After filtration, the microalgae were obtained. The total lipid content (dry weight) was 43.98%, the triacylglycerol fatty acid ester (triglyceride) accounted for 77.7% of the total lipid content, the total effective fat content was 34.17%, and the total effective fat content was The density is 3.59 g/L. Example 3

The fresh water species of Nannochloropsis 1 imnetica was selected, and the initial culture density was 0. lg/L, and the liquid level of the reactor was 80 cm (the cross-sectional area was 2 m×3 m). In the first stage, a red LED with a wavelength of 706 nm is used as a light source, and the temperature is constant at 24 ° C. A trace amount of ammonium phosphate, zinc, iron and other trace element fertilizers are preliminarily added to the culture solution, and a certain amount of C0 _{2 is introduced} (about 90%). And nitrogen oxides such as N0 or N0 ₂ as a nitrogen source are placed in the algae species at the initial culture density after one hour, and the culture is started. Samples were taken at regular intervals to determine biomass density. The breeding ended after 144 hours.

 Results: After 36 hours, the biomass density increased to 6.86 g/L. After 72 hours, the biomass density was 9.91 g/L. After 108 hours, the biomass density was 12.22 g/L. After 144 hours, the culture was carried out. The final biomass density was determined to be 14.34 g/L after the end. The distribution of microalgae in the reactor was visually observed. After filtration, the microalgae were obtained. The total lipid content (dry weight) was 24.52%, the triacylglycerol fatty acid ester (triglyceride) accounted for 81.6% of the total lipid content, the total effective fat content was 20.01%, and the total effective fat content was The density is 2.45 g/L. Comparative example 1

The fresh water species of Nannochloropsis 1 imnetica was selected and the initial culture density was 0. lg/L, the reactor liquid depth is 80cm (cross-sectional area is 2mX3m). Using natural light as the light source, laboratory culture, the culture medium is pre-added with trace amounts of ammonium phosphate, zinc, iron and other trace element fertilizers, and a certain amount of C0 ₂ (about 90%) and NO or N0 ₂ as nitrogen source are respectively introduced. Nitrogen oxides, cultured after 144 hours. Other culture conditions were the same as in Example 1 unless otherwise stated.

对比例 2 Results: The density of microalgae on the surface of the liquid surface was observed by the naked eye to be significantly larger, and the aggregate layer was formed. The final average biomass density was only 3.01 g/L, the total effective fat content was 19.99%, and the total effective fat content was only

Comparative example 2

 The fresh water species of Nannochloropsis 1 imnetica was selected and the initial culture density was

0. lg/L, the reactor liquid depth is 80cm (the cross-sectional area is 2mX3m). Using natural light as the light source, laboratory culture, the culture medium is pre-added with trace amounts of ammonium phosphate, zinc, iron and other trace element fertilizers, and a certain amount of air and NO or N0 ₂ nitrogen oxides as nitrogen sources, 144 After the hour, the breeding is over. Other culture conditions were the same as in Example 1 unless otherwise stated.

更清楚地， 上述实施例与对比例的部分实验结果对比如表 1所示： 表 1 Results: The density of microalgae on the surface of the liquid surface was observed by the naked eye to be significantly larger, and the aggregate layer was formed. The final average biomass density was only 1.29 g/L, the total effective fat content was 28.32%, and the total effective fat content was only

More clearly, some experimental results of the above examples and comparative examples are shown in Table 1: Table 1

结论

in conclusion

The experimental results of the above examples and comparative examples can be seen: 1. The embodiment of the present invention using the built-in light source is compared with the comparative example using natural light as the light source, and after the same time (144 hours), as shown in Table 1, the biomass density is increased by about 3-10 times, and the total effective content is The lipid density increased by about 4-14 times, and the breeding efficiency and oil production efficiency were significantly improved;

 2. In Example 1 and Example 2, the microalgae culture was carried out in different bioreactors in different stages and different stages were used in different stages. In Example 3, the microalgae were always cultured under the same conditions. The results of the three examples show that the total lipid content and total effective fat content of the microalgae cultured in stages are high, and the oil production efficiency is high;

 3. The difference between Embodiment 1 and Embodiment 2 is that the first stage in Embodiment 1 uses red light as the light source, the second stage uses blue light, the second stage in Embodiment 2 uses blue light, and the second stage uses red light. The experimental results show that the breeding effect of Example 1 is better than that of Example 2. It can be considered that the use of light sources of different wavelengths in the first and second stages of microalgae cultivation for different algae species will result in a result of the region|J According to the characteristics of the algae species, suitable and different light sources can be selected in the above two stages to improve the overall production efficiency. All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the present invention.

Claims

m 3⁄4  A built-in light source bioreactor, characterized in that: the built-in light source bioreactor comprises: a reaction vessel, the reaction vessel is provided with a cover plate and the reaction vessel contains a culture liquid for growing microalgae;  An inlet and outlet device, the inlet and outlet device is sealingly connected to the reaction vessel;  a light-emitting device, the light-emitting device is 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 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 vessel;  A gas distributor for supplying a gas required for the growth of microalgae into the reaction vessel.  2. The built-in light source bioreactor according to claim 1, wherein a temperature control device is disposed around the reaction vessel, and the temperature control device is configured to maintain a liquid environment temperature in the reaction vessel at a suitable microalgae. Within the scope of growth; and / or  The reactor is further provided with a nutrient distribution device for providing nutrients required for the growth of microalgae into the reaction vessel; and/or  The gas distributor is a rotary gas distributor that rotates in a process of introducing a gas into the reaction vessel to promote dispersion of gas and nutrients in the liquid culture system.  3. The built-in light source bioreactor according to claim 1, wherein the built-in light source bioreactor further comprises a monitoring system for monitoring parameters of the liquid environment, the parameters being selected from the group consisting of: ra value, temperature And / or nutrient concentration.  The built-in light source bioreactor according to claim 1, wherein the light-emitting device comprises a light guide plate and a light-emitting unit, and the 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.  5. The built-in light source bioreactor according to claim 4, wherein the light emitting device further comprises a bracket for fixing and supporting the light guide plate, the bracket and the reaction container and/or The cover plate is detachably connected, the light emitting unit is embedded in the light guide plate; and/or  The illuminating device emits at least 2 different wavelengths of light when operating; and/or  The color temperature of the composite light emitted by the light emitting unit is 1000-20000K, preferably 1500-6000K; and / or In the light-emitting device, the light-emitting unit is an LED, and the number of LEDs is 1-1000/light guide plate; The good ground is 10-1000/light guide.  A production aquaculture apparatus, characterized in that it comprises two or more internal light source bioreactors according to claim 1.  7. A method of cultivating, comprising the steps of:  (a) providing the built-in light source bioreactor of claim 1;  (b) cultivating the microalgae in the built-in light source bioreactor.  A method for preparing microalgae for producing biomass diesel, characterized in that it comprises the steps of: (a) providing the built-in light source bioreactor of claim 1; (b) cultivating microalgae for producing biomass diesel in the built-in light source bioreactor, wherein the culturing comprises a first culture stage and a second culture stage, in the first stage of microalgae growth, The light emitted by the light emitting unit has a wavelength of 350-900 nm, preferably 570-800 and 400-500 nm; in the second stage of microalgae growth, the light emitted by the light-emitting unit has a wavelength of 350-900 nm, preferably 600. -800 and 400-480nm _;  (c) recovering the microalgae from the built-in light source bioreactor.  A microalgae which can be used for the production of a substance diesel, characterized in that the microalgae are prepared by the method of claim 8.  A method of producing biomass diesel fuel, comprising the steps of: processing microalgae according to claim 9 for use as a raw material to produce biomass diesel oil.