CN109880856B - Open type microalgae grease production method - Google Patents
Open type microalgae grease production method Download PDFInfo
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- CN109880856B CN109880856B CN201711279587.4A CN201711279587A CN109880856B CN 109880856 B CN109880856 B CN 109880856B CN 201711279587 A CN201711279587 A CN 201711279587A CN 109880856 B CN109880856 B CN 109880856B
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to an open production method of microalgae grease, which comprises the steps of firstly adding a microalgae culture medium and a Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 seed liquid into an open reaction tank, adjusting the pH to 10-12 and the temperature to 20-30 ℃, and introducing CO 2 Culturing for a certain time with gas with the volume content of 1-5 v%; (2) adjusting the pH value of a culture system to 8-10 and the temperature to 0-20 ℃, inoculating chlorella SF-B1 seed liquid and monoraphidium SHJ-02 seed liquid, and introducing CO 2 And (3) culturing the gas with the volume content of 5-45 v% under the illumination condition, and harvesting the microalgae cells. The method improves the microalgae culture system to high-concentration CO 2 The tolerance and the solubility of the composite are improved, and the carbon fixation efficiency and the grease yield are improved; meanwhile, the method can inhibit the mixed bacteria pollution in the culture process.
Description
Technical Field
The invention belongs to the field of biotechnology and biological energy, and particularly relates to an open method for producing microalgae grease.
Background
As fossil energy is decreasing day by day and the greenhouse effect is increasing due to the use of fossil energy, more and more researchers are focusing on the development and utilization of renewable energy. Biomass can be the most important renewable energy on earth, and comprises forestry biomass, crops, aquatic plants, agricultural wastes and the like. Among the many biomass energy sources, microalgae are important renewable resources. They have the characteristics of wide distribution, large biomass, high photosynthesis efficiency, strong environment adaptability, short growth period, high biomass yield and the like. The cells contain unique primary or secondary metabolites and are chemically complex. The solar energy conversion efficiency of microalgae can reach 3.5%, and the microalgae is a potential resource for producing medicines, fine chemicals and novel fuels, and fatty acid obtained from microalgae can be converted into fatty acid methyl ester, namely biodiesel.
With the development of the world economy, the use and consumption of a large amount of fossil energy, resulting in the shortage of energy and the increasing deterioration of the environment, particularly CO 2 The greenhouse effect is getting more and more serious due to the sharp increase of the amount of the organic compound. Short growth period of microalgae, high photosynthetic efficiency, and CO 2 High fixing efficiency which is more than 10 times of that of terrestrial plants under certain conditions, and can reduce CO 2 The discharge and the culture cost are reduced, so the biodiesel produced by using the microalgae grease as a raw material is a renewable energy source which most probably meets the fuel required by world transportation at present.
At present, more researches are carried out on oil-producing microalgae such as chlorella and scenedesmus. CN20110144545.6 discloses a Scenedesmus algae strain, which can grow by using artificial culture medium or properly treated waste water, and is characterized by that its oil yield is higher than that of most of present algae-separating strains, and its application field includes CO 2 The fixation, the purification of waste water, and the production of grease, protein, pigment, starch, polysaccharide and nucleic acid. CN20111019480.X discloses a microalgae strainMychonases sp.) And the application of the strain in producing biodiesel, and polyunsaturated fatty acids with high added values can be produced by using the strain, wherein the polyunsaturated fatty acids comprise linolenic acid C18:3 and nervonic acid C24:1, and the strain can obtain the biodiesel and a byproduct with high added value. CN102703326A discloses a high CO 2 The microalgae with tolerance and fixed rate and the breeding method thereof, but the algae strain provided by the patent does not relate to the oil content of the algae strainAmount (v). CN102229889A discloses a Chlorella strain Chlorella sp, MRA-1, the growth of which can adapt to various culture media, temperature, nitrogen source concentration and CO 2 The concentration condition, the oil content and the yield under the low nitrogen condition are high, and the application field comprises CO 2 The fixation, the purification of waste water, and the production of biomass such as grease, protein, pigment, starch, polysaccharide, nucleic acid, etc.
But in practical application, when CO is used 2 Above 5%, the growth of most microalgae will be inhibited. Moreover, the solubility of carbon dioxide has a certain relation with the acidity and alkalinity of the solution, the solubility of carbon dioxide in an alkaline solution is obviously improved, and most microalgae do not have the capability of tolerating high pH values, so that the solubility of carbon dioxide is limited, and the fixation efficiency of the microalgae strains on carbon dioxide is influenced.
In addition, the open culture of microalgae is performed by using an open pond culture apparatus, such as a raceway pond or a round shallow pond, which has the advantages of simple technology and low investment, and thus has recently received attention from researchers. However, the open culture is easily contaminated by harmful organisms such as filamentous fungi, rotifers, and protozoa, which grow and propagate in the algae culture solution. When the number of the harmful organisms reaches a certain density, the growth and the propagation of the cultured algae are influenced, and the polluted algae liquid is not favorable for the expanded culture, and the serious algae liquid can cause the culture failure. Therefore, how to effectively prevent pollution of harmful organisms is a key problem of microalgae large-scale culture.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides an open method for producing microalgae grease. The method improves the microalgae culture system to high-concentration CO 2 The tolerance and the solubility of the composite are improved, and the carbon fixation efficiency and the grease yield are improved; meanwhile, the method can inhibit the mixed bacteria pollution in the culture process.
The method for open production of microalgae grease comprises the following steps:
(1) mixing microalgae culture medium with Chlorella Kelvin: (Parachlorella kessleri) FSH-Y3 or/and Scenedesmus obliquus (Scenedesmus obliquus)Scenedesmus obliqnus) Adding the FSH-Y2 seed liquid into an open reaction tank, adjusting the pH to 10-12 and the temperature to 20-30 ℃, and introducing CO 2 Culturing for a period of time with a gas having a volume content of 1-5 v%;
(2) adjusting the pH value of the culture system to 8-10 and the temperature to 0-20 ℃, and inoculating chlorella (chlorella)Chlorella sp.) SF-B1 seed liquid and Monochoria sinensis SHJ-02 (Monoraphidium sp.) Seed liquid is introduced with CO 2 Gas with the volume content of 5-45 v% is cultured under the illumination condition to harvest the microalgae cells.
Wherein the chlorella (C. vulgaris: (C. vulgaris))Chlorella sp.) SF-B1 has been deposited in China general microbiological culture Collection center (CGMCC) at 7.6.2015 with a collection number of CGMCC 11005 and a collection address of China academy of sciences institute of microbiology 3, Beijing, Naja-Nah-1-Beichen-Xilu-Ministry of China. Under microscope, the chlorella SF-B1 shows green algae cell, single cell algae, single growth, spherical and oval cell shape, and has pigment body inside and diameter of 5-6 μm. Chlorella SF-B1 tolerant CO 2 The concentration can reach 40v%, and the concentration of the tolerant NOx can reach 700 multiplied by 10 -6 (v/v), CO-containing may be used 2 And the waste gas or the smoke of NOx is subjected to illumination autotrophic growth to obtain the biomass rich in grease, so that the carbon fixation efficiency is high, and the tolerance capability is strong.
Wherein the C.trivialis is (C.trivialis) ((C.kiwii))Parachlorella kessleri) FSH-Y3, Scenedesmus obliquus (Scenedesmus obliquus), (A) and (B)Scenedesmus obliqnus) FSH-Y2, Monochoria algae (A. sp.)Monoraphidium sp.) SHJ-02 is preserved in China general microbiological culture Collection center (CGMCC) at 26 days 5 and 24 days 4 and 2015 respectively, the preservation numbers are CGMCC No.9238, CGMCC No.6551 and CGMCC No.10763, the microalgae are disclosed in CN106467896A, CN104611227A and CN106635807A respectively, and preservation and survival certificates are submitted.
In the present invention, as the microalgae culture medium, a liquid culture medium for culturing microalgae, such as BG11, SE, or BBM, which is well known to those skilled in the art, is used.
In the invention, the preparation method of the seed liquid of the Chlorella Kelvin FSH-Y3 and Scenedesmus obliquus FSH-Y2 comprises the following steps: the culture medium isAdjusting the pH value to 10-12, and carrying out shake culture to logarithmic growth phase under the conditions that the temperature is 20-30 ℃, the illumination period is 24 hours, the light-dark time ratio is 14: 10-10: 14, and the illumination intensity is 2000-20000 Lux. The Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus added into the reaction pool (Scenedesmus obliqnus) The volume ratio of the FSH-Y2 seed liquid to the microalgae culture medium is 1: 20-1: 5. When two kinds of microalgae are contained simultaneously, the volume ratio of the seed liquid of the Chlorella Kelvin FSH-Y3 to the Scenedesmus obliquus FSH-Y2 is 1:1-1: 5.
In the invention, the preparation method of the chlorella SF-B1 seed solution comprises the following steps: adjusting the pH value of the culture medium to 7-9, and carrying out shaking culture to logarithmic growth phase at the temperature of 10-30 ℃, in an illumination period of 24 hours, in a light-dark time ratio of 14: 10-10: 14 and in an illumination intensity of 2000-20000 Lux. The volume ratio of the chlorella SF-B1 seed liquid to the microalgae culture medium added into the reaction tank is 1: 20-1: 5.
In the invention, the preparation method of the single-needle algae SHJ-02 seed liquid comprises the following steps: adjusting the pH value of the culture medium to 7-9, performing shaking culture at the temperature of 10-25 ℃, the illumination period of 24 hours, the light-dark time ratio of 14: 10-10: 14 and the illumination intensity of 2000-10000 Lux until the logarithmic phase. The volume ratio of the chlorella SF-B1 seed liquid to the microalgae culture medium added into the reaction tank is 1: 20-1: 5.
The total inoculation amount of the microalgae seed liquid is controlled to be 10% -30% of the total volume of the culture medium, wherein the volume ratio of the Kjeldahl quasi-Chlorella FSH-Y3 or/and the Scenedesmus obliquus FSH-Y2 seed liquid to the Chlorella SF-B1 seed liquid to the Monoraphidium SHJ-02 seed liquid is 1:6: 6-4: 1: 1.
In the present invention, the CO is 2 The gas with the volume content of 1-45 v% can be prepared by self or by adopting flue gas, the flue gas is from incineration tail gas of a sulfur recovery device, catalytic cracking regeneration tail gas or S-zorb regeneration tail gas, wherein CO 2 The content is 1v% -45 v%, and the NOx content is not more than 800 multiplied by 10 -6 （v/v）。
In the invention, the temperature of mixed culture is 5-10 ℃, the illumination intensity is 2000-10000 Lux, and the culture is finished until the growth stabilization period. Microalgae cells are harvested by centrifuging, settling and other modes, the dry weight of the cells and the oil content are measured, the dry weight of the cells can reach more than 13g/L, and the oil content can reach more than 48% of the dry weight of the cells.
The invention improves the carbon fixation efficiency and the oil content by the synergistic effect of the microalgae in high pH and low temperature environments, and can inhibit the mixed bacteria pollution in the culture process. Culturing by adopting the Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 which can tolerate the high pH environment, wherein the initial high pH and the intermittent illumination can inhibit the growth of mixed bacteria and plant diseases and insect pests at the initial stage of culturing the microalgae, and are beneficial to the growth advantage of the microalgae; and high pH favors CO 2 Dissolving to make CO 2 Is easier to be absorbed and utilized by microalgae, and is helpful for improving CO 2 The efficiency of the fixation of (c). And after the biomass is cultured for a certain time in a high pH environment, the reaction temperature is reduced to be below 20 ℃, so that the growth of mixed bacteria can be inhibited, the accumulation of cell grease is facilitated, and the obtained biomass contains more grease. When the flue gas is used as the culture gas, NOx in the flue gas can be removed, and the method is environment-friendly and economical.
Detailed Description
The present invention will be described in further detail by way of examples. The embodiments are implemented on the premise of the technical scheme of the invention, and detailed implementation modes and specific operation processes are given, but the protection scope of the invention is not limited by the following embodiments. In the present invention, wt% is a mass fraction, v% is a volume fraction, and v/v is a volume ratio.
The experimental procedures in the following examples are, unless otherwise specified, those conventional in the art. The experimental materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
The biomass yield of the invention is the mass g/(volume L of algae solution multiplied by culture time d) of the harvested algae powder, and the removal rate is (gas content-exhaust gas content)/gas content.
The culture of the microalgae adopts BG11 culture medium, and the formula is shown in tables 1 and 2.
TABLE 1 BG11 culture Medium
Table 2 composition of a5+ Co solution in table 1
First, BG11 liquid medium was prepared according to tables 1 and 2, the pH of the medium for culturing the Chlorella Kelvin FSH-Y3 and Scenedesmus obliquus FSH-Y2 was adjusted to 10, the pH of the medium for culturing the Chlorella Kelvin SF-B1 and Scenedesmus obliquus SHJ-02 was adjusted to 8.0, and then the Chlorella Kelvin FSH-Y3, Scenedesmus obliquus FSH-Y2, Chlorella Kelvin SF-B1 and Scenedesmus obliquus SHJ-02 were inoculated into the above medium, respectively. Culturing in constant temperature light shaking table at 25 deg.C with light cycle of 24h and dark time ratio of 14:10 and light intensity of 5000Lux at 120rpm until logarithmic phase to obtain Chlorella Kelly FSH-Y3 seed solution, Scenedesmus obliquus FSH-Y2 seed solution, Chlorella vulgaris SF-B1 seed solution and Monoraphidium SHJ-02 seed solution, and storing the seed solutions at 15 deg.C under weak light.
Example 1
(1) Adding 400mL of quasi-chlorella kellogia FSH-Y3 seed liquid and 8L of microalgae culture medium into a 12L open photobioreactor, adjusting the pH value of the microalgae culture medium to 10, culturing at 25 deg.C under illumination intensity of 5000Lux for 24h in a light period at a light-dark time ratio of 14:10, introducing CO in flue gas 2 Has a content of 5v% and a NO content of 50X 10 -6 （v/v）。
(2) After culturing for 4 days, inoculating 400mL of a single-needle algae SHJ-02 seed solution and 400mL of a chlorella SF-B1 seed solution, adjusting the pH of a microalgae culture system to 8, controlling the temperature to be 10 ℃, and continuously culturing by illumination with the illumination intensity of 5000 Lux; introducing CO into the flue gas 2 Has a content of 40v% and a NO content of 800X 10 -6 （v/v）。
(3) And (4) after culturing for 7 days, entering a growth stabilization phase, finishing culturing, centrifugally harvesting microalgae cells, and measuring the dry weight and the oil content of the cells. And (3) carrying out vacuum freeze drying at the temperature of-60 ℃ to constant weight, measuring the dry weight of the algae powder, calculating the biomass yield, and measuring the total lipid content by adopting a normal hexane-ethyl acetate method. The detected dry cell weight can reach 14.1g/L, and the oil contentThe amount of CO in the culture medium is 48.5% of the dry weight of the cells 2 The removal rate was 51% and the NO removal rate was 80.9%.
Example 2
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 12L open photobioreactor, wherein the seed liquid is added in an amount of 800mL, the pH value of the microalgae culture medium is adjusted to 12, the addition amount is 8L, the illumination intensity of the culture is 5000Lux, the culture temperature is 20 ℃, the illumination period is 24h, the light-dark time ratio is 14:10, introducing CO in flue gas 2 In an amount of 5v%, NO and NO 2 The content is 80X 10 -6 （v/v）。
(2) After culturing for 2 days, inoculating 560mL of a single-needle algae SHJ-02 seed solution and 560mL of a chlorella SF-B1 seed solution, adjusting the pH of a microalgae culture system to 10, and continuously culturing under illumination at 5 ℃ with the illumination intensity of 5000 Lux; introducing CO into the flue gas 2 Has a content of 40v% and a NO content of 800X 10 -6 （v/v）。
(3) After culturing for 8 days, entering a stable period, finishing culturing, centrifugally harvesting microalgae cells, and measuring the dry weight and the oil content of the cells. And (3) carrying out vacuum freeze drying at the temperature of-60 ℃ to constant weight, measuring the dry weight of the algae powder, calculating the biomass yield, and measuring the total lipid content by adopting a normal hexane-ethyl acetate method. The dry weight of the cells can reach 13.6g/L, the oil content is 50.3 percent of the dry weight of the cells, and CO is generated in the culture process 2 The removal rate was 51.2%, and the NO removal rate was 81.1%.
Example 3
(1) Adding pseudo-globule Kane FSH-Y3 seed liquid and microalgae culture medium into an open 12 photobioreactor, wherein the seed liquid is added in an amount of 800mL, the microalgae culture medium has a pH value adjusted to 11 and an addition amount of 8L, the illumination intensity for culture is 5000Lux, the culture temperature is 30 ℃, the illumination period is 24h, the light-dark time ratio is 14:10, introducing CO in flue gas 2 In an amount of 5v%, NO and NO 2 The content is 80X 10 -6 （v/v）。
(2) After 2 days of culture, 280mL of single-needle algae SHJ-02 seed solution and 280mL of chlorella SF-B1 seed solution are added, the pH of the microalgae culture system is adjusted to 9, the culture temperature is 15 ℃, continuous illumination culture is carried out, and the illumination intensity isIs 5000 Lux; introducing CO into the flue gas 2 Has a content of 40v% and a NO content of 800X 10 -6 （v/v）。
(3) After culturing for 8 days, entering a stable period, finishing culturing, centrifugally harvesting microalgae cells, and measuring the dry weight and the oil content of the cells. And (3) carrying out vacuum freeze drying at the temperature of-60 ℃ to constant weight, measuring the dry weight of the algae powder, calculating the biomass yield, and measuring the total lipid content by adopting a normal hexane-ethyl acetate method. The dry weight of the cells can reach 13.3g/L, the oil content is 48.9 percent of the dry weight of the cells, and CO is generated in the culture process 2 The removal rate was 52.1%, and the NO removal rate was 81.8%.
Example 4
The same culture procedure and culture conditions as in example 1 were used, except that: 400mL of Scenedesmus obliquus FSH-Y2 seed solution is added in the step (1). The dry weight of the cells can reach 14.2g/L, the oil content is 48.7 percent of the dry weight of the cells, and CO is generated in the culture process 2 The removal rate was 51.3%, and the NOx removal rate was 81.4%.
Example 5
The same culture procedure and culture conditions as in example 1 were used except that: 200mL of Chlorella Kelvin FSH-Y3 seed solution and 200mL of Scenedesmus obliquus FSH-Y2 seed solution are added in the step (1). The dry weight of the cells can reach 14.7g/L, the oil content is 49.9 percent of the dry weight of the cells, and CO is generated in the culture process 2 The removal rate was 51.9%, and the NOx removal rate was 83.2%.
Example 6
The same culture procedure and culture conditions as in example 3 were used, except that: 400mL of the Chlorella Kelvin FSH-Y3 seed solution and 400mL of the Scenedesmus obliquus FSH-Y2 seed solution are added in the step (2). The dry weight of the cells can reach 14.1g/L, the oil content is 50.4 percent of the dry weight of the cells, and CO is generated in the culture process 2 The removal rate was 52.5%, and the NOx removal rate was 83.9%.
Comparative example 1
The same culture procedure and culture conditions as in example 1 were used, except that: adding the seed liquid of the Chlorella Kelvin FSH-Y3, the seed liquid of the Monoraphidium SHJ-02 and the seed liquid of the Chlorella SF-B1 into a reaction tank at the beginning of culture under the culture conditions of the step (1). After the culture is finishedCentrifuging to obtain microalgae cells, measuring cell dry weight and oil content, wherein the cell dry weight can be 12.1g/L, the oil content is 39.2% of the cell dry weight, and CO is 2 The removal rate was 41.9%, and the NOx removal rate was 65.8%.
Comparative example 2
The same culture procedure and culture conditions as in example 1 were used except that: adding the seed liquid of the Chlorella Kelvin FSH-Y3, the seed liquid of the Monoraphidium SHJ-02 and the seed liquid of the Chlorella SF-B1 into a reaction tank at the beginning of culture under the culture conditions of the step (2). Centrifuging after the culture is finished to obtain microalgae cells, measuring the dry weight of the cells and the oil content, wherein the dry weight of the cells can be 9.9g/L, the oil content is 42.8 percent of the dry weight of the cells, and CO 2 The removal rate was 42.3%, and the NOx removal rate was 70.2%.
Comparative example 3
The same culture procedure and culture conditions as in example 1 were used, except that: and (3) in the step (2), chlorella SF-B1 is not added, the culture is finished after 11 days of culture, microalgae cells are obtained through centrifugation, and the dry weight and the oil content of the cells are measured. The dry weight of the cells can reach 11.9g/L, the content of oil is 43.9 percent of the dry weight of the cells, and CO 2 The removal rate is 40.3%, and the NOx removal rate is 10.2%.
In conclusion, compared with a single algae species, the Chlorella Kelvin FSH-Y3, the Chlorella simplex SHJ-02 and the Chlorella vulgaris SF-B1 adopt two-step mixed culture, which is beneficial to improving the tolerance capability of a culture system and can obtain higher biomass and oil content. The invention utilizes the flue gas to prepare the microalgae grease, thereby realizing the production of the grease, purifying the waste gas and obviously improving the economic benefit and the environmental benefit.
Claims (9)
1. The method for producing the microalgae grease in an open mode is characterized by comprising the following steps:
(1) mixing microalgae culture medium with Chlorella Kelvin: (Parachlorella kessleri) FSH-Y3 or/and Scenedesmus obliquus (Scenedesmus obliquus)Scenedesmus obliqnus) Adding the FSH-Y2 seed liquid into an open reactor, adjusting the pH to 10-12 and the temperature to 20-30 ℃, and introducing CO 2 1-5 v% of gas, culturing for one sectionTime;
(2) adjusting the pH value of the culture system to 8-10 and the temperature to 5-20 ℃, and inoculating chlorella (chlorella)Chlorella sp.) SF-B1 seed liquid and Monochoria algae (Michthyophthiria)Monoraphidium sp.) SHJ-02 seed liquid, and introducing CO 2 Culturing gas with volume content of 5-45 v% under illumination condition to obtain microalgae cells;
the chlorella (A) is preparedChlorella sp.) SF-B1 has been deposited in China general microbiological culture Collection center (CGMCC) on 7/6/2015 with the collection number of CGMCC 11005;
the C.kei (C.Kelvin) quasi-chlorellaParachlorella kessleri) FSH-Y3, Scenedesmus obliquus (Scenedesmus obliquus)Scenedesmus obliqnus) FSH-Y2, Aphanizomenon mono-dentale: (A)Monoraphidium sp.) SHJ-02, respectively stored in China general microbiological culture Collection center in 2014, 26 months and 24 months and 2015, 4 months and 24 months, with the storage numbers of CGMCC No.9238, CGMCC No.6551 and CGMCC No. 10763;
the gas in the steps (1) and (2) is flue gas, and the content of NOx in the flue gas does not exceed 800 multiplied by 10 -6 （v/v）。
2. The method of claim 1, wherein: the microalgae culture medium is a liquid culture medium for culturing microalgae by adopting BG11, SE and BBM.
3. The method of claim 1, wherein: the preparation method of seed liquid of the Chlorella Kelvin FSH-Y3 and Scenedesmus obliquus FSH-Y2 comprises the following steps: adjusting the pH value of the culture medium to 10-12, and carrying out shake culture to logarithmic phase under the conditions of temperature of 20-30 ℃, illumination period of 24 hours, light-dark time ratio of 14: 10-10: 14 and illumination intensity of 2000-20000 Lux.
4. A method according to claim 1 or 3, characterized in that: the pseudo-chlorella kelloggi FSH-Y3 or/and Scenedesmus obliquus are added into the reactor (Scenedesmus obliqnus) The volume ratio of the FSH-Y2 seed liquid to the microalgae culture medium is 1: 20-1: 5; when containing two kinds of microalgae simultaneously, the Chlorella KelvinThe volume ratio of the FSH-Y3 to the Scenedesmus obliquus FSH-Y2 seed liquid is 1:1-1: 5.
5. The method of claim 1, wherein: the preparation method of the chlorella SF-B1 seed liquid comprises the following steps: adjusting the pH value of the culture medium to 7-9, performing shaking culture at the temperature of 10-30 ℃, the illumination period of 24 hours, the light-dark time ratio of 14: 10-10: 14 and the illumination intensity of 2000-20000 Lux until the logarithmic growth phase.
6. The method according to claim 1 or 5, characterized in that: the volume ratio of the chlorella SF-B1 seed liquid to the microalgae culture medium added into the reactor is 1: 20-1: 5.
7. The method of claim 1, wherein: the preparation method of the single-needle algae SHJ-02 seed liquid comprises the following steps: adjusting the pH value of the culture medium to 7-9, and carrying out shaking culture to logarithmic growth phase at the temperature of 10-25 ℃, the illumination period of 24 hours, the light-dark time ratio of 14: 10-10: 14 and the illumination intensity of 2000-10000 Lux.
8. The method of claim 1, wherein: controlling the total inoculation amount of the microalgae seed liquid to be 10-30% of the total volume of the culture medium, wherein the volume ratio of the Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 seed liquid, the Chlorella vulgaris SF-B1 seed liquid and the Monoraphidium shii SHJ-02 seed liquid is 1:6: 6-4: 1: 1.
9. The method of claim 1, wherein: and (3) performing mixed culture in the step (2) at the temperature of 5-10 ℃ and the illumination intensity of 2000-10000 Lux, and finishing the culture until the growth stabilization period is finished.
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