CN111100885B - Method for improving oil production of microalgae - Google Patents
Method for improving oil production of microalgae Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
-
- 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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Abstract
The invention relates to a method for improving oil production of microalgae, which comprises the steps of firstly adding a microalgae culture medium and a Chlorella Kelvinea FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 seed solution into a photoreactor, adjusting the pH to 10-12, keeping the illumination intensity to 8000-2Culturing for a certain time by gas with the content of 1-5 v%; then inoculating seed liquid of Monochoria SHJ-02 or/and Scenedesmus HCS-02, and introducing CO2Controlling the pH value of 5-45 v% of gas, reducing the illumination intensity to 1000-5000 Lux, changing the light-dark alternating reaction into light-dark alternating reaction, culturing to a stable stage, and harvesting microalgae cells, wherein the light-dark period is 0.5-5 s, and the light-dark time ratio is 1: 1-5. The invention can improve the microalgae culture system to high-concentration CO2The tolerance and the solubility of the compound improve the biomass and the grease content.
Description
Technical Field
The invention belongs to the field of biotechnology and biological energy, and particularly relates to a method for improving oil production of microalgae.
Background
As fossil energy is decreasing 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 the microalgae can reach 3.5 percent, the microalgae is a potential resource for producing medicines, fine chemicals and novel fuels, and fatty acid obtained from the 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 CO2The greenhouse effect is getting more and more serious due to the sharp increase of the amount of the organic compound. Short growth cycle of microalgae, high photosynthetic efficiency, CO2High fixing efficiency which is more than 10 times of that of terrestrial plants under certain conditions, and can reduce CO2The discharge and the culture cost are reduced; CO removal2Besides, some SOx, NOx and other components in the exhaust gas are purified along with the metabolism of the microalgae, so that the emission of harmful gases is effectively reduced. Therefore, biodiesel produced using microalgal oil as a feedstock is currently the most likely renewable energy source for fuels needed for world transportation.
At present, more researches are carried out on oil-producing microalgae such as chlorella and scenedesmus. CN102229889A discloses a Chlorella strain Chlorella sp, MRA-1, the growth of which can adapt to various culture media, temperature, nitrogen source concentration and CO2The concentration condition, the oil content and the yield under the low nitrogen condition are high, and the application field comprises CO2The 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 used2Above 5%, the growth of most microalgae will be inhibited. Moreover, the solubility of carbon dioxide has a certain relationship with the acid-base property of the solution, the solubility of carbon dioxide in the alkaline solution is obviously improved, and most microalgae do not have the capability of tolerating high pH value, so that the solubility of carbon dioxide is limited, and the fixation efficiency of the algal strains on carbon dioxide is influenced.
The illumination intensity is one of important elements influencing the growth and the change of biochemical components in the microalgae culture. The relationship between light intensity and fatty acid content of marine phytoplankton varies from species to species, but most species have the highest level of EPA at low light intensities, whereas DHA generally decreases with decreasing light intensity. Research on dinoflagellates such as nitzschia closterium and cocci finds that the fat content of the two microalgae is high under low illumination conditions, but the fat content is opposite under high illumination conditions, and similar results are obtained in research on other bait microalgae. Experiments also show that increasing the light intensity can promote the increase of the fatty acid content, for example, the polyunsaturated fatty acid content in chlorella and euglena can be increased along with the increase of the light intensity. However, more studies have shown that microalgae grown under high light intensity have a reduced proportion of unsaturated fatty acids. PUFAs are known to be mainly present in polar lipids (phospholipids and glycerolipids), the content of phospholipids is relatively stable, and the relative amount of glycerolipids is closely related to the photosynthetic activity of cells. At low light intensity, the synthesis rate of membrane lipids and light and pigments is maintained at a high level in order to increase light absorption and light utilization efficiency. When the illumination intensity exceeds the saturation illumination intensity, the content of organelles participating in light and action in the cells is reduced, the absorption capacity and the utilization efficiency of the light are reduced, the synthesis rate of membrane lipid is lower, and therefore the content of PUFAs is reduced.
In outdoor microalgae mass culture, two factors, illumination intensity and temperature, are closely related because they are both from solar radiation. Voltolina et al found that the oil content of Chaetoceros mulleri cultured outdoors is highest in winter, followed by spring and lowest in summer under different illumination intensities and temperatures. In addition, the open culture of microalgae is easily polluted by harmful organisms such as filamentous fungi, rotifers, protozoa, etc., and when the number of the harmful organisms reaches a certain density, the harmful organisms affect the growth and propagation of the cultured algae, and the polluted algae liquid is not favorable for re-expanding culture, and the serious organisms cause culture failure.
From the above, the production of oil by microalgae is closely related to the carbon dioxide content, the illumination intensity, the temperature and other factors, but different microalgae show different correlations, so the implementation mode of how specific types of microalgae can efficiently produce oil needs to be determined specifically.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for improving the production of oil by microalgae. The invention can not only improve the microalgae culture system to high-concentration CO2The tolerance and the solubility of the compound can improve the biomass and the grease content, and can inhibit the mixed bacteria pollution in the culture process.
The method for improving the oil production of microalgae comprises the following steps:
(1) mixing microalgae culture medium with Chlorella Kelvin: (Parachlorella kessleri) FSH-Y3 and/or Scenedesmus obliquus (Scenedesmus obliquus)Scenedesmus obliqnus) Adding the FSH-Y2 seed liquid into a photoreactor, adjusting the pH to 10-12, keeping the illumination intensity to 8000-20000Lux, and introducing CO2Culturing gas with the content of 1-5 v% for a certain time;
(2) inoculating the single needle algae (A) into the culture system in the step (1)Monoraphidium sp.) SHJ-02 and/or Scenedesmus(s) (SHJ-02 and/or Scenedesmus)Scenedesmus sp.) HCS-02 seed liquid, and introducing CO2Controlling the pH value of 5-45 v% of gas, reducing the illumination intensity to 1000-5000 Lux, changing the light-dark alternating reaction into light-dark alternating reaction, culturing for a stabilization period, wherein the light-dark period is 0.5-5 s and the light-dark time ratio is 1: 1-5, and harvesting microalgae cells.
In the present invention, the Chlorella Kelvin (A) is describedParachlorella kessleri) FSH-Y3, Scenedesmus obliquus (Scenedesmus obliquus), (A) and (B)Scenedesmus obliqnus) FSH-Y2, Monochoria algae (A. sp.)Monoraphidium sp.) SHJ-02, scenedesmus (Scenedesmus sp.) HCS-02 is respectively preserved in China general microbiological culture Collection center in 2014, 5-26 days, 2012, 9-11 days, 2015, 4-24 days and 2015, 4-24 days, wherein the preservation numbers are CGMCC No.9238, CGMCC No.6551, CGMCC No.10763 and CGMCC No.10766, the microalgae are respectively disclosed in CN106467896A, CN104611227A, CN106635807A and CN107177505A, 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, BBM, etc., which are well known to those skilled in the art, is used.
In the invention, the preparation method of the seed liquid of the Chlorella Kernesii 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 24h, light-dark time ratio of 14:10 and illumination intensity of 2000-20000 Lux. The volume ratio of the seed solution of the Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 to the microalgae culture medium added into the reactor is 1: 20-1: 10. When the 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 seed liquid of the single needle alga SHJ-02 and the scenedesmus HCS-02 comprises the following steps: adjusting the pH value of the culture medium to 7-9, carrying out shake culture at the temperature of 10-35 ℃, the illumination period of 24 hours, the light-dark time ratio of 14:10 and the illumination intensity of 1500-15000 Lux until the logarithmic growth phase. The volume ratio of the seed liquid of the single-needle alga SHJ-02 or/and the seed liquid of the scenedesmus HCS-02 to the microalgae culture medium added into the reactor is 1: 20-1: 10. When the microalgae contains two kinds of microalgae simultaneously, the volume ratio of the single-needle algae SHJ-02 seed liquid to the scenedesmus HCS-02 seed liquid is 1: 1-1: 5.
Further, the chlorella is inoculated in the step (2)Chlorella sp.) SF-B1 seed liquid, which has been preserved in China general microbiological culture Collection center (CGMCC) on 6.7.2015 with the preservation number of CGMCC 11005 and the preservation address of China academy of sciences microbial research institute No. 3 of Western Lu 1 on North Chen of the Korean district, Beijing. Chlorella SF-B1 tolerant CO2The concentration can reach 40v%, and the concentration of the tolerant NOx can reach 700 multiplied by 10-6(v/v) can utilize a CO-containing compound2And 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. By adding the chlorella SF-B1, the carbon fixation efficiency of a microalgae culture system is improved, and the microalgae culture system is more suitable for extreme environments; and the low temperature resistance of the system is improved, and the grease accumulation is facilitated. 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 and the illumination intensity of 2000-20000 Lux until the logarithmic growth phase. The volume ratio of the chlorella SF-B1 seed liquid to the microalgae culture medium added into the reactor is 1: 20-1: 100.
Further, on the basis of the reaction temperature in the step (1), the step (2) adopts a gradual cooling mode to culture, the temperature is reduced to 0-20 ℃, and the temperature is reduced by 0.1-5 ℃ every 0.5-2 hours.
In the present invention, the CO is2The gas with the volume content of 1-45 v% can be prepared by self or by adopting a chemical engineeringThe process flue gas is derived from incineration tail gas of a sulfur recovery device, catalytic cracking regeneration tail gas or S-zorb regeneration tail gas, wherein CO is2The content is 1v% -45 v%, and the NOx content is not more than 800 multiplied by 10-6（v/v）。
In the invention, the mixed culture in the step (2) is carried out until the growth stabilization phase is finished, microalgae cells are harvested by centrifugation, sedimentation 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 12g/L, and the oil content can reach more than 50 percent of the dry weight of the cells.
Compared with the prior art, the invention can bring the following beneficial effects:
(1) the microalgae grease is cultured and produced by adopting a microalgae two-stage method, and high-pH and high-continuous illumination is adopted in the first stage, so that the rapid growth of microalgae is facilitated, and the biomass is increased; and in the second stage, a conventional pH, low illumination intensity and light-dark alternative reaction mode is adopted, so that the respiration of the microalgae is enhanced, and the oil content is improved. Meanwhile, the culture mode can inhibit the growth of mixed bacteria pollution and plant diseases and insect pests of a microalgae culture system, and is suitable for open culture of microalgae.
(2) The microalgae grease is cultured and produced by adopting a two-section temperature-changing method, and the microalgae grease content is further improved and the pollution of mixed bacteria is avoided by adopting a gradual cooling mode for culture in the second section on the basis of the first section culture temperature.
(3) In the process of producing oil by culturing microalgae by adopting a two-stage method, the culture system is always in a proper state of accumulating oil and inhibiting the growth of other microorganisms by the microalgae through the conversion of high and low pH, high and low temperature environment and illumination intensity, so that the carbon fixation efficiency and the oil content can be improved, and the stability of the microalgae culture process is improved.
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, conventional in the art. The experimental materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
After the culture is finished, microalgae cells are harvested by centrifugation, the dry weight of algae powder is measured after vacuum freeze drying is carried out at the temperature of minus 60 ℃ to constant weight, the biomass yield is calculated, and the total lipid content is measured by adopting a normal hexane-ethyl acetate method. The biomass yield is the mass g/(volume L 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
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 Scenedesmus minutissima SHJ-02, Scenedesmus obliquus HCS-02 and Chlorella vulgaris SF-B1 was adjusted to 8.0, and then the Chlorella Kelvin FSH-Y3, Scenedesmus obliquus FSH-Y2, Scenedesmus obliquus SHJ-02, Scenedesmus obliquus HCS-02 and Chlorella SF-B1 were inoculated into the above medium, respectively. Culturing in constant temperature light shaking table at 25 deg.C for 24h with light-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, Monochorus hirsutus SHJ-02 seed solution, Scenedesmus obliquus HCS-02 seed solution and Chlorella vulgaris SF-B1 seed solution, and storing the seed solutions at 15 deg.C under weak light for use.
Example 1
(1) Adding Chlorella Kelvin FSH-Y into 20L photobioreactor3 seed liquid and microalgae culture medium, wherein the addition of the seed liquid is 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and CO in the flue gas is introduced2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 8, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
After detection, the dry weight of the cells can reach 12.8g/L, the oil content is 51.1 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate is 51.3 percent, and the NO removal rate is 50.3 percent.
Example 2
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L 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 culture temperature is 35 ℃, the continuous illumination intensity is 10000Lux, and introducing CO in smoke2In an amount of 5v%, NO and NO2The content is 80X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800ml of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 9, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v), changing continuous illumination into light-dark alternating reaction, wherein the illumination intensity is 2000 Lux; the light-dark period is 3s, and the light-dark time ratio is 1: 1.
(3) And (4) after culturing for 7 days, entering a stable period, finishing culturing, centrifugally harvesting microalgae cells, and measuring the dry weight and the oil content of the cells.
After detection, the dry weight of the cells can reach 12.3g/L, the oil content is 51.7 percent of the dry weight of the cells, and CO is generated in the culture process2Removal rate51.9% and the NO removal rate was 50.6%.
Example 3
(1) Adding the seed liquid of the Chlorella Kelvin FSH-Y3 and the microalgae culture medium into a 20L photobioreactor, wherein the addition amount of the seed liquid is 800mL, the pH value of the microalgae culture medium is adjusted to 11, the addition amount is 8L, the culture temperature is 30 ℃, the continuous illumination intensity is 15000Lux, and introducing CO in smoke2In an amount of 5v%, NO and NO2The content is 80X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800ml of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 10, culturing at 30 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v), changing continuous illumination into light-dark alternating reaction, wherein the illumination intensity is 4000 Lux; the light-dark period is 4s, and the light-dark time ratio is 1: 1.
(3) And (4) after culturing for 7 days, entering a stable period, finishing culturing, centrifugally harvesting microalgae cells, and measuring the dry weight and the oil content of the cells.
After detection, the dry weight of the cells can reach 13.1g/L, the oil content is 50.1 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate is 50.9%, and the NO removal rate is 50.3%.
Example 4
(1) Adding seed liquid of Scenedesmus obliquus FSH-Y2 and microalgae culture medium into a 20L photobioreactor, wherein the addition amount of the seed liquid is 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in flue gas2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 8, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 12.9g/L, the oil content is 51.7 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 51.9%, and the NO removal rate was 51.1%.
Example 5
(1) 400mL of each of the Chlorella Kelvin FSH-Y3 seed liquid and Scenedesmus obliquus FSH-Y2 seed liquid and a microalgae culture medium are added into a 20L photobioreactor, the addition amount of the seed liquid is 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and CO in smoke is introduced2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 8, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 13.1g/L, the oil content is 51.9 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 52.1%, and the NO removal rate was 51.3%.
Example 6
(1) Adding a pseudo chlorella cathayensis FSH-Y3 seed solution and a microalgae culture medium into a 20L photobioreactor, wherein the addition amount of the seed solution is 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in smoke2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of scenedesmus HCS-02 seed solution, adjusting the pH of the microalgae culture system to 8, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); changing continuous illumination into light-dark alternating reflectionThe illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 13.1g/L, the oil content is 51.3 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 51.9%, and the NO removal rate was 52.1%.
Example 7
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L photobioreactor, wherein the seed liquid is added in an amount of 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in smoke2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 400mL of each of single needle algae SHJ-02 seed solution and Scenedesmus HCS-02 seed solution, adjusting the pH of microalgae culture system to 8, culturing at 25 deg.C, introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 13.2g/L, the oil content is 51.8 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 52.7%, and the NO removal rate was 52.3%.
Example 8
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L photobioreactor, wherein the seed liquid is added in an amount of 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in smoke2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After 4 days of culture, the monochoria SHJ-02 species were inoculated400mL of seed liquid and 400mL of chlorella SF-B1 seed liquid respectively, adjusting the pH of a microalgae culture system to 8, the culture temperature to 25 ℃, and introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(3) And (4) after 7 days of culture, entering a growth stabilization phase, finishing the culture, centrifuging to obtain microalgae cells, and measuring the dry weight and the oil content of the cells.
Detection proves that the dry weight of the cells can reach 13.5g/L, the oil content is 52.3 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 53.9%, and the NO removal rate was 78.9%.
Example 9
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L photobioreactor, wherein the seed liquid is added in an amount of 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in smoke2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 300mL of each of SHJ-02 seed solution of Monochoria, HCS-02 seed solution of Scenedesmus and 200mL of seed solution of Chlorella SF-B1, adjusting pH of microalgae culture system to 8, culturing at 25 deg.C, and introducing CO in flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 13.8g/L, the oil content is 52.9 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate is 54.6%, and the NO removal rate is 72.1%.
Example 10
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L photobioreactor, wherein the addition amount of the seed liquid is 800mL, and the pH value of the microalgae culture medium isAdjusting to 10, adding 8L, culturing at 25 deg.C, continuously illuminating at 13000Lux, introducing CO in flue gas2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 8, and culturing by gradually cooling to 17.8 deg.C, and cooling to 0.1 deg.C every 1 h. Introducing CO into the flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Detection shows that the dry cell weight can reach 12.3g/L, the oil content is 55.7% of the dry cell weight, and CO is generated in the culture process2The removal rate was 53.9%, and the NO removal rate was 51.4%.
Example 11
(1) Adding a Chlorella Kelvin FSH-Y3 seed liquid and a microalgae culture medium into a 20L photobioreactor, wherein the seed liquid is added in an amount of 800mL, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and introducing CO in smoke2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, inoculating 800mL of single needle algae SHJ-02 seed solution, adjusting the pH of the microalgae culture system to 8, culturing in a gradual cooling mode, cooling to 7 ℃, and reducing the temperature by 2 ℃ every 8 h. Introducing CO into the flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
The detection shows that the dry weight of the cells can reach 12.1g/L, the oil content is 56.7 percent of the dry weight of the cells, and the cells are culturedIn-process CO2The removal rate was 52.9%, and the NO removal rate was 51.6%.
Example 12
(1) 400mL of each of the Chlorella chevalieri FSH-Y3 seed liquid and Scenedesmus obliquus FSH-Y2 seed liquid and a microalgae culture medium are added into a 20L photobioreactor, the pH value of the microalgae culture medium is adjusted to 10, the addition amount is 8L, the culture temperature is 25 ℃, the continuous illumination intensity is 13000Lux, and CO in smoke is introduced2Has a content of 5v% and a NO content of 50X 10-6（v/v）。
(2) After culturing for 4 days, respectively inoculating 300mL of single needle algae SHJ-02 seed liquid and Scenedesmus HCS-02 seed liquid and 200mL of chlorella SF-B1 seed liquid, adjusting the pH of the microalgae culture system to 8, culturing by adopting a gradual cooling mode, cooling to 7 ℃, and reducing the temperature by 2 ℃ every 8 hours. Introducing CO into the flue gas2Has a content of 40v% and a NO content of 800X 10-6(v/v); the continuous illumination is changed into light-dark alternating reaction, the illumination intensity is 3000Lux, the light-dark period is 2s, and the light-dark time ratio is 1: 1.
(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.
Through detection, the dry weight of the cells can reach 12.2g/L, the oil content is 57.7 percent of the dry weight of the cells, and CO is generated in the culture process2The removal rate was 51.9%, and the NO removal rate was 75.1%.
Comparative example 1
The same as example 5, except that: adding the seed liquid of the Chlorella Kelvin FSH-Y3, the seed liquid of the Monochoria sinensis SHJ-02 and the seed liquid of the Scenedesmus HCS-02 into a reaction tank at the beginning of culture, and adopting the culture conditions in the step (1). The detection proves that the dry cell weight is 10.1g/L, the oil content is 42.1 percent of the dry cell weight, and the CO content is2The removal rate was 40.9%, and the NOx removal rate was 50.8%.
Comparative example 2
The difference from example 5 is that: adding the seed liquid of the Chlorella Kelvin FSH-Y3, the seed liquid of the Monochoria sinensis SHJ-02 and the seed liquid of the Scenedesmus HCS-02 into the reaction tank at the beginning of the culture, and adopting the culture conditions in the step (2). The detection proves that the dry cell weight can be 11.3g/L, and the oil content is the dry cell weight43.1% of (C), CO2The removal rate was 42.7%, and the NOx removal rate was 51.8%.
In conclusion, compared with single algae species or direct mixed culture, the two-step mixed culture is adopted, 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 (7)
1. The method for improving the oil production of microalgae 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 a photoreactor, adjusting the pH to 10-12, keeping the illumination intensity to 10000-2Gas with the content of 1-5 v% is cultured for 4 days;
(2) inoculating the single needle algae (A) into the culture system in the step (1)Monoraphidium sp.) SHJ-02 and Scenedesmus sp (S. Scenedesmus)Scenedesmus sp.) One or two kinds of HCS-02 seed liquid are simultaneously inoculated into chlorella SF-B1 seed liquid, the volume ratio of the chlorella SF-B1 seed liquid to the microalgae culture medium is 1: 20-1: 100, and CO is introduced into the chlorella SF-B1 seed liquid2Controlling the pH value of 40-45 v% of gas, reducing the illumination intensity to 2000-4000 Lux, changing the light-dark alternating reaction into light-dark alternating reaction, culturing for a stabilization period, wherein the light-dark period is 0.5-5 s and the light-dark time ratio is 1: 1-5, and harvesting microalgae cells;
the volume ratio of the Chlorella Kelvin FSH-Y3 or/and Scenedesmus obliquus FSH-Y2 seed liquid to the microalgae culture medium added into the reactor is 1: 20-1: 10; when the chlorella vulgaris FSH-Y3 and Scenedesmus obliquus FSH-Y2 are simultaneously contained, the volume ratio of the seed liquid of the chlorella vulgaris FSH-Y3 to the seed liquid of the Scenedesmus obliquus FSH-Y2 is 1: 1-1: 5;
the volume ratio of the single-needle alga SHJ-02 seed liquid or/and scenedesmus HCS-02 seed liquid added into the reactor to the microalgae culture medium is 1: 20-1: 10; when the single-needle alga SHJ-02 and the scenedesmus HCS-02 are contained simultaneously, the volume ratio of the single-needle alga SHJ-02 seed liquid to the scenedesmus HCS-02 seed liquid is 1: 1-1: 5.
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. The method of claim 1, wherein: the preparation method of seed liquid of the monochoria SHJ-02 and scenedesmus HCS-02 comprises the following steps: adjusting the pH value of the culture medium to 7-9, carrying out shake culture at the temperature of 10-35 ℃, the illumination period of 24 hours, the light-dark time ratio of 14:10 and the illumination intensity of 1500-15000 Lux until the logarithmic growth phase.
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, 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 and in an illumination intensity of 2000-20000 Lux.
6. The method of claim 1, wherein: on the basis of the reaction temperature in the step (1), the step (2) adopts a gradual cooling mode to culture, the temperature is reduced to 7-20 ℃, and the temperature is reduced by 0.1-5 ℃ every 0.5-2 hours.
7. The method of claim 1, wherein: CO described in step (1)21-5 v% of gas and CO in the step (2)2The gas with the content of 40-45 v% is self-made or process flue gas, and the process flue gas is from incineration tail gas, catalytic cracking regeneration tail gas or S-zorb regeneration tail gas of a sulfur recovery deviceGas in which the NOx content does not exceed 800X 10-6（v/v）。
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