CN113604361A - Method for culturing modified living microalgae by using high-salinity wastewater and application thereof - Google Patents
Method for culturing modified living microalgae by using high-salinity wastewater and application thereof Download PDFInfo
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- CN113604361A CN113604361A CN202110809046.8A CN202110809046A CN113604361A CN 113604361 A CN113604361 A CN 113604361A CN 202110809046 A CN202110809046 A CN 202110809046A CN 113604361 A CN113604361 A CN 113604361A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F2101/00—Nature of the contaminant
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Abstract
The invention discloses a method for culturing modified living microalgae by using high-salinity wastewater, which comprises the following steps: placing the microalgae in high-salinity wastewater with different sulfate concentrations, and culturing the microalgae in 1000-1-100 volume ratio for 10-20 days at 20-25 ℃ in an alternating manner of illumination and no illumination to obtain the microalgae with high sulfur-loading surface. The invention also provides an application of the high-sulfur-content microalgae in treatment of heavy metal-containing wastewater, which comprises the step of directly adding the obtained high-sulfur-content microalgae into the heavy metal-containing wastewater for adsorption reaction. The high-salt wastewater threatens the growth of microorganisms, and the growth of microalgae is not inhibited by the high-salt wastewater. Meanwhile, the cultivated microalgae has the capability of effectively removing heavy metal ions in the wastewater. The invention realizes the purpose of treating high-salinity wastewater and heavy metal wastewater simultaneously.
Description
Technical Field
The invention belongs to the technical field of environmental engineering, and particularly relates to a method for culturing modified living microalgae by using high-salinity wastewater and application thereof.
Background
The high-salinity wastewater refers to wastewater with the total salt mass fraction of at least 1%. It is mainly from chemical plants and the collection and processing of petroleum and natural gas. This waste water contains a variety of substances (including salts, oils, heavy metals, and radioactive substances). The production route of the salt-containing wastewater is wide, and the water quantity is increased year by year. Most of salt substances in the high-salinity wastewater are Cl-、SO4 2-、Na+、Ca2+And the like. Although these ions are essential nutrients for the growth of microorganisms, if the concentration of these ions is too high, they may have inhibitory and toxic effects on microorganisms. Therefore, the problem of treating the salt substances in the high-salinity wastewater needs to be solved urgently.
Heavy metal ions in water can produce extremely harmful effects even at trace concentrations. The increased human activities of mining, smelting, fossil fuel combustion and battery production lead to the discharge of large amounts of heavy metal wastewater. Therefore, the development of an efficient adsorbent for fixing heavy metals is urgently required. The traditional methods for removing heavy metals in industrial wastewater comprise a chemical precipitation method, an oxidation-reduction method, a membrane separation method, an adsorption method and the like. However, such physical and chemical treatments have problems of generation of toxic sludge or low efficiency at low concentrations.
Biological adsorption is widely concerned as an environment-friendly, efficient and low-cost method, and the purpose of removing free heavy metal ions in sewage is achieved by utilizing the interaction of surface active functional groups of bacteria, fungi and algae and heavy metals. The microalgae is widely concerned due to the advantages of low price, easy obtaining, simple culture and the like. The cell wall of the algae cell is composed of polysaccharide, protein and lipid, has rich functional groups including amino, hydroxyl, carboxyl, sulfydryl and the like, and can be used as a metal binding site. At present, most of adsorbents adopted by heavy metal treatment by a biological method are microalgae biomasses, the heavy metal biomasses are prepared by collecting and inactivating microalgae, and pollution such as culturing the microalgae by using high-salinity wastewater and directly treating the heavy metal by using living cells of the microalgae is avoided.
Disclosure of Invention
The invention mainly solves the technical problems that:
one of the objectives of the present invention is to provide a method for culturing modified living microalgae using high-salinity wastewater, which is simple in operation, low in cost, and free from secondary pollution, and can degrade sulfate in high-salinity wastewater by using the growth of microalgae.
The invention also aims to provide an application of living microalgae in treating heavy metal wastewater, so as to solve the problems of unsatisfactory treatment effect, high treatment cost, secondary pollution and the like in the conventional treatment of heavy metal wastewater.
In order to solve the technical problems, the invention is realized by the following technical scheme:
in a first aspect, the invention provides a method for culturing modified living microalgae by using high-salinity wastewater, which comprises the following steps:
placing the microalgae in high-salinity wastewater with different sulfate concentrations, and culturing the microalgae in 1000-1-100 volume ratio for 10-20 days at 20-25 ℃ in an alternating manner of illumination and no illumination to obtain the microalgae with high sulfur-loading surface.
Preferably, the high-salinity wastewater with different sulfate concentrations is prepared by changing the sulfate concentration in the microalgae culture medium, and the sulfate comprises one or more of magnesium sulfate, sodium sulfate and potassium sulfate.
Preferably, the sulfur concentration of the high-salt wastewater with different sulfate concentrations ranges from 10mg/L to 80 mg/L.
Preferably, the high-salinity wastewater source comprises industrial wastewater treated by a physicochemical method, and the industrial wastewater comprises one or more of acid mine wastewater, pharmaceutical wastewater and paper-making wastewater.
Preferably, the microalgae culture period is 14 hours of illumination, and 10 hours of no illumination is performed alternately.
In a second aspect, the invention provides application of microalgae with high sulfur content in treatment of wastewater containing heavy metals.
Preferably, the application method of the high-sulfur-content microalgae comprises the following steps: the high-sulfur-loading microalgae is directly added into the wastewater containing heavy metal for adsorption reaction, so that the concentration of the heavy metal ions in the wastewater is reduced.
Further preferably, the volume ratio of the high-sulfur-carrying microalgae to the heavy metal solution is 1:100-1000, the pH value of the wastewater in the adsorption process is 3-7, the adsorption time is 1-3h, and the adsorption temperature is 25-30 ℃.
Further preferably, the concentration range of the heavy metal ions in the heavy metal wastewater is 0-30 mg/L.
It is further preferred that the heavy metal wastewater comprises cationic heavy metal wastewater, and the cationic heavy metal wastewater comprises one or more of mercury wastewater, lead-containing wastewater and cadmium-containing wastewater.
Compared with the prior art, the method for culturing the modified living microalgae by using the high-salinity wastewater has the following advantages:
1) according to the invention, the high-salinity wastewater is used for carrying out modified culture on the surface of the microalgae, the growth of the microalgae is not inhibited by the high-salinity wastewater, the microalgae with high sulfur-carrying capacity on the surface is successfully obtained, the fixing capacity of the microalgae with high sulfur-carrying capacity on the surface is enhanced, and the heavy metal ions are mainly fixed on the surface of the microalgae in the form of sulfide precipitates. Compared with other methods for modifying the biological adsorbent, the method for modifying and culturing the biological adsorbent has the advantages of simple operation, low cost and no secondary pollution.
2) The microalgae adopted by the invention can normally grow in the high-salt wastewater, and the growth process of the microalgae achieves the purpose of degrading the sulfate content in the high-salt wastewater while treating heavy metal ions.
3) The invention utilizes the living microalgae for direct adsorption, saves the steps of collecting, centrifuging, drying and the like of microorganisms compared with a biomass adsorbent, has higher treatment efficiency and is more environment-friendly.
4) The invention successfully fixes the heavy metal on the cell surface, thereby reducing the toxic action of the heavy metal on organisms. Moreover, the adsorption capacity of the microalgae modified and cultured by the invention to heavy metals is much higher than that of other microalgae, for example, the maximum adsorption capacity of adsorbing heavy metal mercury by directly utilizing Chlorella (Chlorella sp DT) is 3.33mg/L (Appl Microbiol Biotechnol,2006,72: 197-.
Detailed Description
In order that those skilled in the art will better understand the technical solutions of the present invention, the following description of the preferred embodiments of the present invention is provided in connection with specific examples, which should not be construed as limiting the present patent.
The test methods or test methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials, unless otherwise indicated, are conventionally obtained commercially or prepared by conventional methods.
The microalgae used in the embodiment of the invention is specifically Chlorella sorokiniana FK which is obtained by separating and purifying certain waste tailings in Guangdong province according to a conventional method, and has the biological characteristics of strong heavy metal ion resistance, rapid growth, rich surface functional groups and the like. The high-salt wastewater used in the embodiments of the invention is MgSO 24The mercury-containing wastewater is prepared by diluting mercury standard solution (1g/L) purchased from Aladdin company to a set concentration.
Example 1
The embodiment of the method for fixing the heavy metal mercury by using the microalgae modified by the high-salinity wastewater comprises the following steps:
(1) the method comprises the following steps of carrying out sulfur modification on chlorella by utilizing high-salinity wastewater without sulfate, measuring the biomass of the chlorella and the sulfate content in culture medium supernatant, and converting the sulfate content into the sulfur content, and specifically comprises the following steps:
(1a) preparing BG-11 culture medium for microalgae growth by adding 1.5g NaNO3，40mg K2HPO4，36mg CaCl2·2H2O, 6mg ammonium citrate-water, 6mg ferric ammonium citrate, 1mg EDTA, 2.86. mu. g H3BO3，1.81μg MnCl2·4H2O,0.222μg ZnSO4·7H2O，0.39μg NaMoO4·5H2O，0.079μg CuSO4·5H2O，0.050μg CoCl2·6H2O to 1L of sterile distilled water.
(1b) The samples were taken to determine the sulfate concentration in the culture supernatant (every 48h, measured by barium chromate spectrophotometry at 420nm wavelength) and at the same time to determine the Chlorella sorokiniana FK chlorophyll content (lower layer at 700nm wavelength by UV spectrophotometry). After 17 days of culture, centrifugation is carried out and deionized water is used for washing for 3 times, so that the influence of sulfate in the culture medium on the adsorption experiment is avoided. The fresh algal solution was stored at 4 ℃ (adsorption experiment was completed in less than 3 days to maintain freshness of the algal sample).
(2) Treating mercury-containing wastewater by using the microalgae obtained in the step (1), and specifically comprising the following steps:
(2a) respectively taking 5mg algae samples in 8mL mercury-containing solution with the concentration of 10mg/L (3 groups of parallel samples are set in an adsorption experiment), and adjusting the pH value to 3.0, 4.0, 5.0, 6.0 and 7.0; adsorbing for 3h on a vibration table with the temperature of 25 ℃ and the rotating speed of 150rad/min, separating supernatant after complete adsorption, and measuring the concentration of Hg (II) in the supernatant by using a flame atomic absorption method.
(2b) According to the experimental result in the step (2a), under the condition of pH 6, respectively taking 5mg algae samples in 8mL mercury-containing wastewater with the concentration of 0-30mg/L, adsorbing for 3h on a vibration table with the temperature of 25 ℃ and the rotating speed of 150rad/min, separating the supernatant after complete adsorption, and measuring the concentration of Hg (II) in the supernatant by using a flame atomic absorption method.
Example 2
This example refers to the method of example 1, with the difference that: prepared culture medium BG-11, by adding MgSO4(10mg/L in S) to simulate high salinity wastewater.
Example 3
This example refers to the method of example 1, with the difference that: prepared culture medium BG-11, by adding MgSO4(20mg/L in S) to simulate high salinity wastewater.
Example 4
This example refers to the method of example 1, with the difference that: prepared culture medium BG-11, by adding MgSO4(40mg/L in S) to simulate high salinity wastewater.
Example 5
This example refers to the method of example 1, with the difference that: in being equipped withCulture medium BG-11 by addition of MgSO4(80mg/L in S) to simulate high salinity wastewater.
In order to illustrate the effect of the high-salt wastewater modified microalgae and the heavy metal fixing effect thereof in comparison in each embodiment of the invention, the biomass of the microalgae and the removal rate of the microalgae to sulfur in high-salt wastewater with different sulfate concentrations are determined, the test results are shown in table 1, and the heavy metal fixing ability of different modified microalgae is shown in table 2. The sulfate content in the culture solution supernatant is obviously reduced along with the growth of the microalgae, which shows that the microalgae has the capability of degrading sulfate in high-salt wastewater and has good treatment effect. In addition, the biomass of the sulfur-modified microalgae is increased by a certain amount, which indicates that the growth of the microalgae is not influenced by high-salinity wastewater. The high-sulfur modified microalgae has obviously improved mercury adsorption effect. Wherein the adsorption effect of the B-40 is best, the maximum adsorption capacity reaches 21.58mg/g in the equilibrium, and the adsorption capacity is 2 times of that of the unmodified microalgae B-0.
TABLE 1 Biomass and sulfur uptake related parameters of microalgae cultured for 17 days
TABLE 2 maximum mercury adsorption by different high sulfur-bearing microalgae
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (10)
1. A method for culturing modified living microalgae by using high-salinity wastewater is characterized by comprising the following steps:
placing the microalgae in high-salinity wastewater with different sulfate concentrations, and culturing the microalgae in 1000-1-100 volume ratio for 10-20 days at 20-25 ℃ in an alternating manner of illumination and no illumination to obtain the microalgae with high sulfur-loading surface.
2. The method of claim 1, wherein the high-salinity wastewater with different sulfate concentrations is prepared by changing the sulfate concentration in the microalgae culture medium, and the sulfate comprises one or more of magnesium sulfate, sodium sulfate and potassium sulfate.
3. The method of claim 1, wherein the high-salinity wastewater with different sulfate concentrations has a sulfur concentration ranging from 10mg/L to 80 mg/L.
4. The method of claim 3, wherein the high salinity wastewater source comprises industrial wastewater treated by a physicochemical method, and the industrial wastewater comprises one or more of acid mine wastewater, pharmaceutical wastewater and paper-making wastewater.
5. The method for culturing modified living microalgae using high-salinity wastewater as claimed in claim 1, wherein the microalgae culturing period is 14h light and 10h no light are alternately performed.
6. Use of the high sulfur-bearing microalgae according to any one of claims 1 to 5 in treatment of heavy metal-containing wastewater.
7. The application of the microalgae with high sulfur content in the treatment of wastewater containing heavy metals according to claim 6, wherein the application method of the microalgae with high sulfur content is as follows: the high-sulfur-loading microalgae is directly added into the wastewater containing heavy metal for adsorption reaction, so that the concentration of the heavy metal ions in the wastewater is reduced.
8. The application of the microalgae with high sulfur content in treating heavy metal-containing wastewater as claimed in claim 7, wherein the volume ratio of the microalgae with high sulfur content to the heavy metal solution is 1:100-1000, the pH value of the wastewater in the adsorption process is 3-7, the adsorption time is 1-3h, and the adsorption temperature is 25-30 ℃.
9. The use of the sulfur-enriched microalgae according to claim 7 for treating heavy metal-containing wastewater, wherein the concentration of heavy metal ions in the heavy metal-containing wastewater is in the range of 0-30 mg/L.
10. The use of the high sulfur-bearing microalgae according to claim 7 for treating heavy metal-containing wastewater, wherein the heavy metal-containing wastewater comprises cationic heavy metal wastewater, and the cationic heavy metal wastewater comprises one or more of mercury wastewater, lead-containing wastewater and cadmium-containing wastewater.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115026128A (en) * | 2022-06-17 | 2022-09-09 | 中国科学院南京土壤研究所 | Rice field heavy metal inhibition control and absorption method based on periphyton |
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| CN115026128A (en) * | 2022-06-17 | 2022-09-09 | 中国科学院南京土壤研究所 | Rice field heavy metal inhibition control and absorption method based on periphyton |
| CN115026128B (en) * | 2022-06-17 | 2023-08-18 | 中国科学院南京土壤研究所 | Rice field heavy metal resistance control and digestion method based on periphyton |
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