CN115029248A - Method for improving microalgae lipid yield by utilizing recycled wastewater - Google Patents
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Abstract
The invention belongs to the technical field of bioengineering, and discloses a method for improving microalgae lipid yield by using recycled wastewater, which adopts a two-stage culture mode, carries out algae species mixotrophic culture in the first stage to enable algae cells to grow rapidly in a short period, then collects the wastewater after microalgae culture, mixes BG-11 after simple filtration to form a mixed culture medium, and combines a photobioreactor to culture microalgae by using the mixed culture medium. The method combines a mixotrophic-photoautotrophic two-stage culture mode, and in the photoautotrophic culture stage, when the concentration of the recycled wastewater is 40%, the lipid content and lipid yield of the microalgae are respectively improved by 14.95% and 14.00% compared with those of a single BG-11 group; the culture method can obviously improve the lipid content and the lipid yield of the microalgae, realizes the rapid synthesis of the lipid in the algae cells, simultaneously reduces the use of fresh water and the culture cost of the microalgae, and provides a new research direction for the culture of the microalgae by using waste water and the treatment of the recycling cost.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to a method for improving microalgae lipid yield by utilizing recycled wastewater.
Background
At present, with the exploitation of fossil fuels, energy crisis and environmental pollution bring serious problems to human society, and the development of novel sustainable resources to meet the increasing demand in the future is urgent. As an emerging raw material, microalgae has several advantages over biomass of traditional cash crops: 1) no competitive tillage; 2) the photosynthetic efficiency is high, and the growth period is relatively short; 3) wide adaptability to severe conditions. Therefore, third generation biofuels based on microalgal lipids are of great interest. Although microalgae can accumulate a large amount of lipid, high cost and low yield are main factors limiting the industrialization of microalgae, and how to reduce the culture cost of microalgae and improve the lipid content and yield of microalgae is a problem to be solved urgently in recent years. The two-stage culture mode is a strategy for microalgae to rapidly accumulate biomass and lipids.
In order to further reduce the cost of microalgae culture, the wastewater is used for culturing microalgae, so that organic matters such as nitrogen, phosphorus and the like in the wastewater can be absorbed and utilized, and the traditional culture medium can be replaced. In the process of microalgae culture, a large amount of wastewater can be generated and environmental pollution can also be caused, and the reports of re-culturing microalgae by using the wastewater generated by microalgae culture are less. On the other hand, when the pure wastewater is used for culturing the microalgae, the growth rate of the microalgae is slow due to toxic substances in the wastewater, so that the biomass yield of the microalgae is reduced, and the integral oil yield is influenced. Therefore, the biomass and lipid yield of the whole microalgae can be improved by proportioning certain nutrients, the strategy can be beneficial to increasing the lipid yield of the microalgae, reducing the culture cost of the microalgae and reducing the use of fresh water, and a new technical idea is provided for recycling the wastewater.
Through the above analysis, the problems and defects of the prior art are as follows: the existing microalgae has high cost and low yield; a large amount of wastewater is generated in the process of microalgae culture, and the reports of re-culturing microalgae by using the wastewater generated by microalgae culture are less; the growth rate of microalgae is slow due to toxic substances in the wastewater, and the lipid yield of microalgae cultured by using pure wastewater is low.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for improving the yield of microalgae lipid by utilizing recycled wastewater.
The invention is realized by a method for improving microalgae lipid yield by using recycled wastewater, which comprises the following steps:
the method comprises the steps of adopting a two-stage culture mode, carrying out algae species mixotrophic culture in the first stage to enable algae cells to quickly grow in a short period, then collecting wastewater after microalgae culture, simply filtering, proportioning BG-11 to form a mixed culture medium, and combining a photobioreactor to culture microalgae by using the mixed culture medium.
Further, the method for improving the yield of microalgae lipid by using the recycled wastewater further comprises the following steps:
taking glucose as a carbon source to culture fibrophyta mixotrophically, pouring out supernatant after microalgae grow to the late stage of logarithmic phase, collecting residual algae cells as seed liquid, and performing suction filtration on the supernatant to obtain algae culture wastewater; proportionally mixing the algae culture wastewater with BG-11 to serve as a mixed culture medium, re-suspending the microalgae in a column type photobioreactor by using the mixed culture medium with different proportions, and continuously introducing 10% of CO 2 Mixing with air, culturing at certain temperature and under certain illumination intensity, and detecting and analyzing microalgae biomass, lipid content and yield.
Further, the method for improving the microalgae lipid yield by utilizing the recycled wastewater comprises the following steps:
step one, collecting recycled wastewater;
step two, culturing microalgae by using the recycled wastewater in a ratio BG-11;
and step three, extracting lipid in the algae cells in the culture solution obtained in the step two by using an organic solvent.
Further, the collecting of the recycled wastewater in the first step comprises:
the BG-11 culture medium with glucose as a carbon source is mixedly cultured in a 500mL triangular flask to culture fibroalgae, after the microalgae grow to the late stage of logarithmic phase, after the microalgae settle for two days, supernatant is poured out from a super clean bench, residual algae cells are mixed to be used as seed liquid, and collected supernatant is subjected to suction filtration to obtain the recycling wastewater.
Further, the microalgae is the fibrous algae Ankistrodesmus sp.
Further, the step two of culturing the microalgae by using the recycled wastewater to match BG-11 comprises the following steps:
fresh BG-11 in the ratio of the recycled wastewater is used as a mixed culture medium, and the seed solution in the first stage is resuspended in a 1L column type photobioreactor to ensure that the inoculation amount of microalgae is 0.9 g/L; continuously introducing 10% of CO 2 And mixing the air with the mixed gas, and culturing in an environment with certain temperature and illumination intensity to obtain the final culture solution.
Further, the concentration of the recycled wastewater in the mixed culture medium is 30-60%.
Further, the temperature is 25 ℃, and the illumination intensity is 7000-8000 lux.
Further, the lipid in the algal cells in the culture solution obtained in the second step of extracting with an organic solvent in the third step includes:
centrifuging the final culture solution obtained in the second step for 5min at 5000r/min, collecting the algae cells in a centrifuge tube, freezing overnight at-80 ℃, and weighing after freeze-drying for 24h by a freeze dryer; weighing 0.2g of dry algae powder, adding quartz sand, grinding and uniformly mixing, adding an organic solvent, repeatedly extracting until the algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by using a gravimetric method.
Further, the organic solvent is a chloroform-methanol solution, wherein the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; the mass of the quartz sand is 2 times of that of the dry algae powder.
In combination with the technical solutions and the technical problems to be solved, please analyze the advantages and positive effects of the technical solutions to be protected in the present invention from the following aspects:
first, aiming at the technical problems existing in the prior art and the difficulty in solving the problems, the technical problems to be solved by the technical scheme of the present invention are closely combined with results, data and the like in the research and development process, and some creative technical effects are brought after the problems are solved. The specific description is as follows:
the invention combines the mixotrophic photoautotrophic two stagesThe stage culture mode is that 40% of the recycled wastewater can obviously improve the lipid content and the lipid yield of the microalgae in the photoautotrophic culture stage, and by the culture method, the rapid synthesis of the lipid in the algae cells is realized, the use of fresh water and the culture cost of the microalgae are reduced, and a new research direction is provided for the treatment of the microalgae culture and recycling cost by utilizing the wastewater. The invention obviously improves the lipid content of the microalgae, and when the concentration of the recycled wastewater is 40 percent, the lipid content and the lipid yield of the microalgae are 52.01 percent (dry cell weight) and 250.72mg L -1 d -1 The improvement is 14.95 percent and 14.00 percent respectively compared with the BG-11 group alone.
Secondly, considering the technical scheme as a whole or from the perspective of products, the technical effect and advantages of the technical scheme to be protected by the invention are specifically described as follows:
the method has simple process, easy operation and environmental protection, can obviously improve the lipid content and the yield of the microalgae, reduces the production cost of the microalgae, and provides a new idea for recycling the algae cultivation wastewater. Reducing pollution caused by algae cultivation wastewater; the microalgae culture cost is reduced, organic matters in the recycled wastewater are absorbed and utilized, the use of fresh water is reduced, and the economic feasibility of microalgae culture is improved; obtaining high value-added metabolites for producing biodiesel.
Third, as an inventive supplementary proof of the claims of the present invention, there are also presented several important aspects:
the expected income and commercial value after the technical scheme of the invention is converted are as follows: provides a new technical approach for the reuse of wastewater in microalgae industrialization, reduces the culture cost of microalgae, and reduces the use of fresh water.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments of the present invention will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a flow chart of a method for improving microalgae lipid production by using recycled wastewater according to an embodiment of the present invention;
FIG. 2A is a graph showing the effect of different concentrations of recycled media on microalgae biomass as provided by an embodiment of the present invention;
FIG. 2B is a schematic diagram showing the effect of different concentrations of recycled media on oil content and high quality yield.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In view of the problems of the prior art, the present invention provides a method for improving microalgae lipid yield by using recycled wastewater, and the present invention is described in detail below with reference to the accompanying drawings.
First, an embodiment is explained. This section is an explanatory embodiment expanding on the claims so as to fully understand how the present invention is embodied by those skilled in the art.
As shown in fig. 1, the method for improving microalgae lipid yield by using recycled wastewater provided by the embodiment of the invention comprises the following steps:
s101, collecting the recycled wastewater;
s102, culturing microalgae by using the recycled wastewater in a ratio BG-11;
s103, extracting lipid in the algal cells in the culture solution obtained in S102 by using an organic solvent.
As a preferred embodiment, the method for improving microalgae lipid yield by using recycled wastewater provided by the embodiment of the invention specifically comprises the following steps:
(1) collecting the recycled wastewater: using BG-11 culture medium with glucose as carbon source to mixedly culture fibroalgae in a 500mL triangular flask, pouring out supernatant in a super clean bench after the microalgae grow to the late stage of logarithmic phase and settle for two days, mixing the residual algae cells to be used as seed liquid, and performing suction filtration on the collected supernatant to obtain the recycling wastewater; wherein the microalgae is fiber algae Ankistrodesmus sp.
(2) Proportioning BG-11 for culturing microalgae by using the recycled wastewater: proportioning fresh BG-11 of the recycled wastewater to serve as a mixed culture medium, wherein the concentration of the recycled wastewater in the mixed culture medium is 30-60%; resuspending the seed solution of the first stage into a 1L column type photobioreactor to ensure that the inoculation amount of microalgae is about 0.9g/L, and continuously introducing 10% of CO 2 And mixing the air and the mixed gas, and culturing at the temperature of 25 ℃ and the illumination intensity of 7000-8000 lux to obtain the final culture solution.
(3) Extracting lipid in algal cells in the culture solution obtained in the step (2) by using an organic solvent: centrifuging the final culture solution obtained in the step b for 5min at 5000r/min, collecting algae cells in a centrifuge tube, freezing at-80 ℃ for overnight, freeze-drying for 24h by using a freeze dryer, weighing, adding 0.2g of dry algae powder, adding quartz sand, grinding, uniformly mixing, adding an organic solvent, repeatedly extracting until the algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae algae powder by using a gravimetric method; the organic solvent is chloroform-methanol solution, and the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; the mass of the quartz sand is 2 times of that of the dry algae powder.
Second, the application embodiment. In order to prove the creativity and the technical value of the technical scheme of the invention, the part is the application example of the technical scheme of the claims on specific products or related technologies.
Example 1:
the method for detecting and analyzing the microalgae biomass and the lipid content under the condition that the concentration of the recycled wastewater is 30 percent comprises the following specific steps:
(1) collecting the recycled wastewater: the BG-11 culture medium with glucose as a carbon source mixedly cultures fibrophyta in a 500mL triangular flask, when the microalgae grow to the late stage of logarithmic phase, after settling for two days, supernatant is poured out from a super clean bench, residual algae cells are mixed to be used as seed liquid, and the collected supernatant is filtered to obtain the recycling wastewater;
(2) culturing microalgae with 30% of recycled wastewater: using 30% of recycled wastewater to match with fresh BG-11 as mixed culture medium to resuspendThe seed liquid at one stage is put into a 1L column type photobioreactor to ensure that the inoculation amount of the microalgae is about 0.9g/L, and 10 percent of CO is continuously introduced 2 And mixing the air and the mixed gas, and culturing at the temperature of 25 ℃ and the illumination intensity of 7000-8000 lux to obtain the final culture solution.
Extracting lipid in algae cells in the culture solution obtained in the step (2) by using an organic solvent: extracting the grease in the algae cells in the step (2) by using an organic solvent: wherein the organic solvent is a chloroform-methanol solution, and the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; centrifuging the final culture solution obtained in the step (2) for 5min at 5000r/min, collecting algae cells in a centrifuge tube, freezing overnight at-80 ℃, freeze-drying for 24h by using a freeze dryer, weighing, then weighing 0.2g of dry algae powder, adding 0.4g of quartz sand, grinding, uniformly mixing, adding an organic solvent, repeatedly extracting until an algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by using a gravimetric method. The biomass was 4.44g L -1 The amplitude is slightly improved compared with that of a control group, but no significant difference exists; the oil content is 49.28 percent, and the oil yield is 243.38mg L -1 d -1 (see FIGS. 2A-2B), an increase of 8.93% and 10.66% over BG-11 group alone, respectively.
Example 2:
the method for detecting and analyzing the microalgae biomass and the lipid content under the condition that the concentration of the recycled wastewater is 40% comprises the following specific steps:
(1) collecting the recycled wastewater: using BG-11 culture medium with glucose as carbon source to mixedly culture fibroalgae in a 500mL triangular flask, pouring out supernatant in a super clean bench after the microalgae grow to the late stage of logarithmic phase and settle for two days, mixing the residual algae cells to be used as seed liquid, and performing suction filtration on the collected supernatant to obtain the recycling wastewater;
(2) culturing microalgae with 40% of recycled wastewater: using 40% of recycled wastewater and fresh BG-11 as a mixed culture medium, resuspending the seed solution in the first stage into a 1L column type photobioreactor, ensuring that the inoculation amount of microalgae is about 0.9g/L, and continuously introducing 10% of CO 2 Mixing with air, and placing at 25 deg.C with illumination intensity of 7000-8000 luxAnd (5) culturing to obtain a final culture solution.
Extracting lipid in algal cells in the culture solution obtained in the step (2) by using an organic solvent: extracting the grease in the algae cells in the step (2) by using an organic solvent: wherein the organic solvent is a chloroform-methanol solution, and the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; centrifuging the final culture solution obtained in the step (2) for 5min at 5000r/min, collecting algae cells in a centrifuge tube, freezing overnight at-80 ℃, freeze-drying for 24h by using a freeze dryer, weighing, then weighing 0.2g of dry algae powder, adding 0.4g of quartz sand, grinding, uniformly mixing, adding an organic solvent, repeatedly extracting until an algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by using a gravimetric method. The biomass was 4.33g L -1 No significant difference from the control group; the oil content is 52.01%, and the oil yield is 250.72mg L -1 d -1 (see FIGS. 2A-2B), which are 14.95% and 14.00% higher than BG-11 alone, respectively.
Example 3:
the embodiment of the invention provides a method for detecting and analyzing microalgae biomass and lipid content under the condition that the concentration of recycled wastewater is 50%, which comprises the following steps:
(1) collecting the recycled wastewater: using BG-11 culture medium with glucose as carbon source to mixedly culture fibroalgae in a 500mL triangular flask, pouring out supernatant in a super clean bench after the microalgae grow to the late stage of logarithmic phase and settle for two days, mixing the residual algae cells to be used as seed liquid, and performing suction filtration on the collected supernatant to obtain the recycling wastewater;
(2) culturing microalgae with 50% of recycled wastewater: using 50% of recycled wastewater and fresh BG-11 as a mixed culture medium, resuspending the seed solution in the first stage into a 1L column type photobioreactor, ensuring that the inoculation amount of microalgae is about 0.9g/L, and continuously introducing 10% of CO 2 And mixing the air and the mixed gas, and culturing at the temperature of 25 ℃ and the illumination intensity of 7000-8000 lux to obtain the final culture solution.
Extracting lipid in algal cells in the culture solution obtained in the step (2) by using an organic solvent: extracting the grease in the algae cells in the step (2) by using an organic solvent: wherein the organic solvent isChloroform-methanol solution, wherein the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; centrifuging the final culture solution obtained in the step (2) for 5min at 5000r/min, collecting algae cells in a centrifuge tube, freezing overnight at-80 ℃, freeze-drying for 24h by using a freeze dryer, weighing, then weighing 0.2g of dry algae powder, adding 0.4g of quartz sand, grinding and uniformly mixing, adding an organic solvent, repeatedly extracting until an algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae algae powder by using a gravimetric method. The biomass was 3.69g L -1 84.67% of the control group only; the oil content is 50.64%, which is 11.94% higher than that of the control group, but the oil yield is only 197.70mg L because the growth is inhibited - 1 d -1 (see FIGS. 2A-2B).
Example 4:
the embodiment of the invention provides a method for detecting and analyzing microalgae biomass and lipid content under the condition that the concentration of recycled wastewater is 50%, which comprises the following steps:
(1) collecting the recycled wastewater: using BG-11 culture medium with glucose as carbon source to mixedly culture fibroalgae in a 500mL triangular flask, pouring out supernatant in a super clean bench after the microalgae grow to the late stage of logarithmic phase and settle for two days, mixing the residual algae cells to be used as seed liquid, and performing suction filtration on the collected supernatant to obtain the recycling wastewater;
(2) culturing microalgae with 60% of recycled wastewater: 60% of recycled wastewater is used as a mixed culture medium in proportion to fresh BG-11, the seed solution in the first stage is resuspended in a 1L column type photobioreactor, the inoculation amount of microalgae is ensured to be about 0.9g/L, and 10% of CO is continuously introduced 2 And mixing the air and the mixed gas, and culturing at the temperature of 25 ℃ and the illumination intensity of 7000-8000 lux to obtain the final culture solution.
Extracting lipid in algal cells in the culture solution obtained in the step (2) by using an organic solvent: extracting the grease in the algae cells in the step (2) by using an organic solvent: wherein the organic solvent is a chloroform-methanol solution, and the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; centrifuging the final culture solution obtained in step (2) at 5000r/min for 5min, collecting algae cells in a centrifuge tube, freezing at-80 deg.C overnight, and usingFreeze-drying for 24h by a freeze dryer, weighing 0.2g of dry algae powder, adding 0.4g of quartz sand, grinding and uniformly mixing, adding an organic solvent, repeatedly extracting until the algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by a gravimetric method. The biomass was 3.04g L -1 Only 69.56% of the control group is reached; the oil content is 48.64%, the increase is 11.94% compared with the control group, the growth of the oil is obviously inhibited, and the oil yield is only 143.24mg L -1 d -1 Significantly lower than the control group (see fig. 2A to 2B).
And thirdly, evidence of relevant effects of the embodiment. The embodiment of the invention achieves some positive effects in the process of research and development or use, and has great advantages compared with the prior art, and the following contents are described by combining data, diagrams and the like in the test process.
Comparative example:
the invention provides a method for detecting and analyzing microalgae biomass and lipid content under a single BG-11 culture condition, which comprises the following steps:
(1) mixedly culturing fibrophyceae in a BG-11 culture medium taking glucose as a carbon source in a 500mL triangular flask, pouring out supernatant in a super clean bench after the microalgae grow to the late stage of logarithmic phase and settle for two days, and mixing the rest algae cells to serve as seed liquid;
(2) culturing microalgae by using BG-11: resuspending the first stage seed solution into a 1L column photobioreactor with fresh BG-11 to ensure microalgae inoculation amount of about 0.9g/L, and continuously introducing 10% CO 2 And mixing the air and the mixed gas, and culturing at the temperature of 25 ℃ and the illumination intensity of 7000-8000 lux to obtain the final culture solution.
Extracting lipid in algal cells in the culture solution obtained in the step (2) by using an organic solvent: extracting the grease in the algae cells in the step (2) by using an organic solvent: wherein the organic solvent is a chloroform-methanol solution, and the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; centrifuging the final culture solution obtained in step (2) for 5min at 5000r/min, collecting algae cells, freezing at-80 deg.C overnight, lyophilizing for 24 hr, weighing, adding 0.2g dry algae powder, and adding 0.4g dry algae powderAnd (3) grinding and uniformly mixing quartz sand, adding an organic solvent, repeatedly extracting until the algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by using a gravimetric method. The biomass was 4.37g L -1 The oil content is 45.24 percent, and the oil yield is 219.93mg L -1 d -1 (see FIGS. 2A-2B).
The invention combines the culture mode of the mixotrophic-photoautotrophic two stages, 40 percent of recycled wastewater can obviously improve the lipid content and the lipid yield of the microalgae in the photoautotrophic culture stage, and by the culture method, the rapid synthesis of the lipid in the algae cells is realized, the use of fresh water and the culture cost of the microalgae are reduced, and a new research direction is provided for the treatment of the cost for culturing the microalgae by using the wastewater and recycling the microalgae.
The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A method for improving microalgae lipid yield by using recycled wastewater is characterized by comprising the following steps:
the method comprises the steps of adopting a two-stage culture mode, carrying out algae species mixotrophic culture in the first stage to enable algae cells to quickly grow in a short period, then collecting wastewater after microalgae culture, simply filtering, proportioning BG-11 to form a mixed culture medium, and combining a photobioreactor to culture microalgae by using the mixed culture medium.
2. The method for improving microalgae lipid production using recycled wastewater of claim 1, wherein the method for improving microalgae lipid production using recycled wastewater further comprises:
culturing fibrophyceae with glucose as carbon source, collecting supernatant after microalgae grows to late logarithmic phase, collecting residual algae cells as seedsFiltering the supernatant to obtain algae-raising wastewater; proportionally mixing the algae-culturing wastewater BG-11 as a mixed culture medium, re-suspending the microalgae in a column type photobioreactor by using the mixed culture medium with different proportions, and continuously introducing 10% of CO 2 Mixing with air, culturing at certain temperature and under certain illumination intensity, and detecting and analyzing microalgae biomass, lipid content and yield.
3. The method for improving lipid production of microalgae using the recycled wastewater as claimed in claim 1, wherein the method for improving lipid production of microalgae using the recycled wastewater comprises the following steps:
step one, collecting recycled wastewater;
step two, culturing microalgae by using the recycled wastewater in a ratio BG-11;
and step three, extracting lipid in the algae cells in the culture solution obtained in the step two by using an organic solvent.
4. The method for improving lipid production in microalgae according to claim 3, wherein the collecting of the recycled wastewater in the first step comprises:
the BG-11 culture medium with glucose as a carbon source is mixedly cultured in a 500mL triangular flask to culture fibroalgae, after the microalgae grow to the late stage of logarithmic phase, after the microalgae settle for two days, supernatant is poured out from a super clean bench, residual algae cells are mixed to be used as seed liquid, and collected supernatant is subjected to suction filtration to obtain the recycling wastewater.
5. The method for improving lipid production of microalgae using recycled wastewater as claimed in claim 4, wherein the microalgae is Fibrospira ansustrates sp.
6. The method for improving lipid production of microalgae according to claim 3, wherein the step two of culturing microalgae by using the recycled wastewater to match BG-11 comprises:
the recycled wastewater is proportioned with fresh BG-11 and used as a mixed culture medium to resuspend the seed liquid of the first stage to a 1L columnIn the formula photobioreactor, the inoculation amount of microalgae is ensured to be 0.9g/L, and 10 percent of CO is continuously introduced 2 And mixing the air with the mixed gas, and culturing in an environment with certain temperature and illumination intensity to obtain the final culture solution.
7. The method for improving lipid production of microalgae by using the recycled wastewater as claimed in claim 6, wherein the concentration of the recycled wastewater in the mixed culture medium is 30-60%.
8. The method for improving the lipid yield of microalgae by using the recycled wastewater as claimed in claim 6, wherein the temperature is 25 ℃ and the illumination intensity is 7000-8000 lux.
9. The method for increasing lipid yield of microalgae according to claim 3, wherein the lipid in algal cells in the culture solution obtained from the second step of organic solvent extraction in the third step comprises:
centrifuging the final culture solution obtained in the second step for 5min at 5000r/min, collecting the algae cells in a centrifuge tube, freezing overnight at-80 ℃, and weighing after freeze-drying for 24h by a freeze dryer; weighing 0.2g of dry algae powder, adding quartz sand, grinding and uniformly mixing, adding an organic solvent, repeatedly extracting until the algae body turns white, centrifuging and collecting an organic phase to obtain total lipid, and measuring the lipid content of the microalgae powder by using a gravimetric method.
10. The method for improving lipid production of microalgae using the recycled wastewater as claimed in claim 9, wherein the organic solvent is chloroform-methanol solution, wherein the volume ratio of chloroform to methanol in the chloroform-methanol solution is 2: 1; the mass of the quartz sand is 2 times of that of the dry algae powder.
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