CN109576159B - Chlorella W4 capable of removing heavy metals in water with high heavy metal content and application thereof - Google Patents
Chlorella W4 capable of removing heavy metals in water with high heavy metal content and application thereof Download PDFInfo
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- CN109576159B CN109576159B CN201710907107.8A CN201710907107A CN109576159B CN 109576159 B CN109576159 B CN 109576159B CN 201710907107 A CN201710907107 A CN 201710907107A CN 109576159 B CN109576159 B CN 109576159B
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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/125—Unicellular algae isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/89—Algae ; Processes using algae
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- 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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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
- C02F2101/22—Chromium or chromium compounds, e.g. chromates
Abstract
The invention relates to chlorella capable of removing heavy metals in a water body with high heavy metal content, which is preserved in China Center for Type Culture Collection (CCTCC) within 2 months and 22 days in 2017, and the preservation number is M2017059. The chlorella W4 has high tolerance to heavy metal ions in a culture water body, has high removal rate to Zn, Ni and Cr when growing in a water body with high heavy metal content, has the removal rate to Zn of more than 98 percent, and is far higher than the removal efficiency of most of microalgae Zn for removing heavy metals at present, thereby being particularly suitable for treating polluted water bodies with high Zn content. Besides, heavy metals are removed, and meanwhile, N, P and other nutrient substances in the water body can be removed by the chlorella W4, so that the risk of water body eutrophication is prevented.
Description
Technical Field
The invention relates to the field of microalgae culture and sewage treatment, in particular to chlorella capable of removing heavy metals in a water body with high heavy metal content and application thereof.
Background
The heavy metals discharged to the environment by human beings are increasing day by day, which not only seriously pollutes the soil and the water environment, but also causes great harm to the health of the human beings. The traditional method for treating the heavy metal polluted water mainly comprises physical, chemical and biological methods, including a precipitation method, a chelating resin method, a high-molecular trapping agent method, a natural zeolite adsorption method, a membrane technology, an activated carbon adsorption process, an ion exchange method and the like. These methods have good purification effect, but have complex process flow, complicated operation and high cost, and are not suitable for large-scale application.
Microalgae is a ubiquitous microorganism in various water bodies, is fast in propagation and strong in adaptability, and can reach the concentration required for sewage treatment only by inoculating a small amount of microalgae mother liquor in the water body and propagating in a proper environment for a period of time. The sewage is rich in nitrogen and phosphorus, and provides good inorganic nutrition for the propagation of microalgae. In addition, microalgae can obtain carbon source and energy by photosynthesis, and the growth of microalgae is ensured without additionally providing a carbon source, so that the cost is reduced. Therefore, the microalgae is used for treating the sewage, which is a sewage treatment mode with good effect, economy and practicability.
Research shows that some microalgae have an immobilization effect on heavy metal ions, and the process mainly comprises physical adsorption and chemical adsorption. The microalgae is applied to fixing heavy metals in sewage, and meanwhile, the microalgae can reduce the content of nitrogen, phosphorus and other nutrient substances in water and absorb other harmful substances in the water, so that the method has multiple purposes, and is an economical and effective sewage treatment method. However, although many microalgae have a certain heavy metal ion adsorption capacity, the current microalgae have a limited heavy metal ion adsorption efficiency, and many microalgae have poor tolerance to heavy metal ions, which greatly limits the application of microalgae in sewage treatment.
Therefore, there is a need for microalgae that can tolerate high heavy metal content and efficiently adsorb heavy metal ions.
Disclosure of Invention
The inventor collects algae samples from the field, and obtains an algae strain capable of removing some heavy metal ions in the environment through culture and separation. The cells are round or oval, green, without flagella and with a nucleus. The 18S rDNA sequence obtained by sequencing is shown as SEQ ID NO 1, and the sequence is compared in the ncbi database, so that the similarity of the sequence and Chlorella sorokiniana of Chlorella is higher, and the sequence is named as Chlorella W4.
Based on the findings, the invention provides chlorella capable of removing heavy metals in water with high heavy metal content, which is preserved in China center for type culture Collection in 2017, 2 months and 22 days, and the preservation number is CCTCC NO: M2017059.
The invention also provides application of the chlorella in removing heavy metals in a water body with high heavy metal content.
In one embodiment, the heavy metal is one or more combinations of Zn, Ni, Cr.
In a preferred embodiment, the heavy metal is Zn.
The invention also provides a method for treating sewage with high heavy metal content, which comprises the step of treating the sewage by using the chlorella.
In one embodiment, the method comprises the steps of:
s1: filtering the sewage to remove large-particle substances;
s2: inoculating the chlorella into the filtered sewage to obtain a chlorella-sewage culture;
s3: culturing the chlorella-sewage culture by introducing air for illumination;
s3: removing chlorella in the chlorella-sewage culture after culturing for 8-15 days.
In a preferred embodiment, the chlorella is inoculated at OD concentration in S2 700 The value is 0.1-0.5.
In a preferred embodiment, the culturing conditions in S3 are 30 ℃, 80. + -. 5. mu. mol m -2 s -1 The intensity of the light.
The chlorella W4 has extremely high tolerance to heavy metal ions in a culture water body, has high removal rate to Zn, Ni and Cr when growing in a water body with high heavy metal content, has the removal rate to Zn as high as about 98 percent, and is far higher than the removal efficiency of most of microalgae Zn for removing heavy metals at present, thereby being particularly suitable for treating polluted water bodies with high Zn content. Besides, heavy metals are removed, and meanwhile, N, P and other nutrient substances in the water body can be removed by the chlorella W4, so that the risk of water body eutrophication is prevented. The strain can be used for treating industrial sewage, agricultural sewage and other water bodies with high heavy metal content.
Preservation of microorganisms
The microalgae related to the invention is obtained by separating a water sample collected in the field, and the microalgae belongs to the genus Chlorella through 18S rDNA sequencing and morphological identification, and the 18S rDNA sequence has higher similarity with Chlorella sorokiniana. The microalgae has been preserved in the China Center for Type Culture Collection (CCTCC) of Wuhan university in Wuhan City, Hubei province in 2017 in 22 months, the preservation number is CCTCC NO: M2017059, which is named Chlorella W4, and the Latin chemical name is Chlorella sorokiniana W4.
Drawings
FIG. 1 is an optical micrograph of Chlorella vulgaris W4 according to the present invention;
FIG. 2 shows the growth curves of Chlorella W4 cultured in BG11 medium and BG11 medium supplemented with heavy metals;
FIG. 3 shows the growth curves of a control strain Chlorella vulgaris G32 cultured in BG11 medium and BG11 medium supplemented with heavy metals.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
1. Obtaining Chlorella W4
The Chlorella W4(Chlorella sorokiniana W4) is obtained by separating water containing algae collected from the field, and the cell morphology of the Chlorella W4 is as shown in figure 1, and the cells are round or oval, green, without flagella and with a nucleus. The microalgae belongs to the genus Chlorella through 18S rDNA sequencing identification. The microalgae is preserved in China Center for Type Culture Collection (CCTCC) of Wuhan university in Wuhan City, Hubei province, China for 2 months and 22 days in 2017, the preservation number is CCTCC NO: M2017059, the Latin school name is Chlorella sorokiniana W4, the Latin school name is Chlorella W4, and the Latin school name is Chlorella sorokiniana W4.
2. Tolerance of Chlorella W4 to heavy metals
Activating Chlorella W4 in small triangular flask, inoculating into 500ml conical flask containing 300ml BG11 culture medium with high heavy metal content, and inoculating to initial OD 700 0.1, the culture conditions are 30 ℃, 80 +/-5 mu mol m -2 s -1 Culturing with aeration under illumination.
BG11 culture medium with high heavy metal content is prepared by adding BG11 culture mediumPreparing heavy metal mother liquor. Wherein, the BG11 culture medium is prepared by the following method: 1ml of Stock II, Stock II I, Stock IV, Stock VI, Stock VII and Stock VIII are taken, and 1.5g of NaNO is added 3 And adding water to a constant volume of 1000 ml. The respective stock solutions are shown in Table 1, and the composition of A5 is shown in Table 2.
TABLE 1 composition of each stock solution in BG11 Medium preparation
TABLE 2A5 composition
The addition of heavy metals is set according to the water quality standard of class V (heavy pollution) in the environmental quality standard of surface water (GB3838-2002) and the pollutant discharge standard of urban sewage treatment plants. The heavy metal mother liquor was set to 2000 x. Table 3 shows the heavy metal mother liquor concentration and working concentration.
TABLE 3 heavy metal mother liquor concentration and working concentration
After inoculation, the OD of the chlorella culture was measured daily 700 Values, a growth curve was prepared, and tolerance of chlorella W4 to heavy metals was observed with chlorella cultured in BG11 medium as a control.
The result is shown in fig. 2, the growth curve of chlorella W4 cultured in BG11 medium is not much different from the growth curve of chlorella W4 cultured in BG11 medium added with heavy metal in the early stage, and chlorella W4 in BG11 medium added with heavy metal grows better even in the later stage, and it can be seen that chlorella W4 has extremely high tolerance to heavy metal. FIG. 3 is a graph showing the growth curves of another chlorella G32 isolated by the inventors during the isolation process, cultured in BG11 medium and BG11 medium supplemented with heavy metals, respectively, and the results show that the growth of the chlorella G32 is greatly inhibited in BG11 medium supplemented with heavy metals, as is known for most chlorella.
3. Chlorella W4 removal efficiency of heavy metals in culture medium
After the chlorella W4 grew to a stable period, 6000g of algal cells were collected by centrifugation, and heavy metal ions remaining in the culture solution were detected by ICP-MS. As shown in Table 4, the chlorella W4 has high removal rate of Zn, Ni and Cr, and the removal rate of Zn is as high as about 98%, which is far higher than that of most of the microalgae for removing heavy metals at present, so the chlorella W4 is particularly suitable for treating polluted water with high Zn content. TABLE 4 Chlorella W4 pairs of heavy metals in the culture media
4. Application of chlorella W4 in treatment of industrial sewage with high heavy metal content
Filtering the industrial sewage with high heavy metal content to remove large granular substances in the sewage. Then inoculating chlorella W4 into the sewage with the inoculation concentration of OD 700 To 0.1-0.5. 80. + -. 5. mu. mol m at 30 ℃ -2 s -1 Culturing for 7-15 days under illumination intensity, separating algae cells from water by flocculation, and detecting metal ion content and nutrient element content such as nitrogen and phosphorus in water. The result shows that the treated water is clear, the content of metal ions and the content of nutrient elements are reduced to the III-class water standard, and the discharge requirement is met.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Sequence listing
<110> Wuhan algae Yoghurt Biotechnology GmbH
<120> chlorella W4 capable of removing heavy metals in water with high heavy metal content and application thereof
<130> 1
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1831
<212> DNA
<213> Chlorella W4()
<400> 1
atgggtacga cggtcagtgc cagcttgcat gcctgcaggt cgacgatttg ttacgacttc 60
accttcctct aggtgggagg gtttaatgaa cttctcggcg gccgagagcg gagaccgccc 120
ccggtcgcca atccgaacac ttcaccagca cacccaatcg gtaggagcga cgggcggtgt 180
gtacaaaggg cagggacgta atcaacgcaa gctgatgact tgcgcttact aggcattcct 240
cgttgaagat taataattgc aataatctat ccccatcacg atgcagtttc aaagattacc 300
cgggcctctc ggccaaggct aggctcgttg aatgcatcag tgtagcgcgc gtgcggccca 360
gaacatctaa gggcatcaca gacctgttat tgcctcatgc ttccattggc tagtcgccaa 420
tagtccctct aagaagtccg ccggctggcg aaccaaccgt gactatttag caggctgagg 480
tctcgttcgt taccggaatc aacctgacaa ggcaacccac caactaagaa cggccatgca 540
ccaccaccca tagaatcaag aaagagctct caatctgtca atcctcacta tgtctggacc 600
tggtaagttt tcccgtgttg agtcaaatta agccgcaggc tccacgcctg gtggtgccct 660
tccgtcaatt cctttaagtt tcagccttgc gaccatactc cccccggaac ccaaaaactt 720
tgatttctca taaggtgccg gcggagtcat cgaagaaaca tccgccgatc cctagtcggc 780
atcgtttatg gttgagacta ggacggtatc taatcgtctt cgagccccca actttcgttc 840
ttgattaatg aaaacatcct tggcaaatgc tttcgcagta gttcgtcttt cataaatcca 900
agaatttcac ctctgacaat gaaatacgaa tgcccccgac tgtccctctt aatcattact 960
ccggtcctac agaccaacag gataggccag agtcctatcg tgttattcca tgctaatgta 1020
ttcagagcgt aggcctgctt tgaacactct aatttactca aagtaacagc gccgactccg 1080
agtcccggac agtgaagccc aggagcccgt ccccggcaac aaggtgggcc ctgccagtgc 1140
acaccgaaac ggcggaccgg caggccccac ccgaaatcca actacgagct ttttaactgc 1200
agcaacttaa atatacgcta ttggagctgg aattaccgcg gctgctggca ccagacttgc 1260
cctccaattg atcctcgtta aggggtttag attgtactca ttccaattac cagacctgaa 1320
aaggcccagt attgttattt attgtcacta cctccctgtg tcaggattgg gtaatttgcg 1380
cgcctgctgc cttccttgga tgtggtagcc gtttctcagg ctccctctcc ggaatcgaac 1440
cctaatcctc cgtcacccgt taccaccatg gtaggcctct atcctaccat cgaaagttga 1500
tagggcagaa atttgaatga aacatcgccg gcgcaaggcc atgcgattcg tgaagttatc 1560
atgattcacc gcgagtcggg cagagcccgg tcggcctttt atctaataaa tacgtccctt 1620
ccagaagtcg ggatttacgc acgtattagc tctagattta ctacgggtat ccgagtagta 1680
ggtaccatca aataaactat aactgattta atgagccatt cgcagtttca cagtataaag 1740
cagtttatac ttagacatgc atggcttaat ctttgagaca atctctagag gatccccggg 1800
taccgagctc gaatcgtaat tcatgttcca a 1831
Claims (8)
1. The chlorella is preserved in a China center for type culture collection (CCTCC NO: M2017059) within 2 months and 22 days of 2017.
2. Use of the chlorella of claim 1 for removing heavy metals from a water body with a high heavy metal content.
3. The use according to claim 2, wherein the heavy metal is one or more of Zn, Ni, Cr in combination.
4. Use according to claim 3, wherein the heavy metal is Zn.
5. A method for treating sewage with high heavy metal content, which comprises the step of treating heavy metals in the sewage with the chlorella of claim 1.
6. The method of claim 5, comprising the steps of:
s1: filtering the sewage to remove large-particle substances;
s2: inoculating the chlorella into the filtered sewage to obtain a chlorella-sewage culture;
s3: culturing the chlorella-sewage culture by ventilation and illumination;
s4: removing chlorella from the chlorella-wastewater culture after culturing for 8-15 days.
7. The method of claim 6, wherein the chlorella in S2 is inoculated at an OD 700 The value is 0.1-0.5.
8. The method according to claim 6 or 7, wherein the culturing conditions in S3 are 30 ℃, 80 ± 5 μmol m -2 s -1 The intensity of the light.
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| CN111621423A (en) * | 2020-05-27 | 2020-09-04 | 武汉理工大学 | High-phosphorus-loading living microalgae and preparation method and application thereof |
| CN114260307B (en) * | 2021-12-21 | 2022-08-19 | 海南大学 | Application of tubulose algae and microbial agent and method for removing environmental heavy metals |
| 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 |
| CN116836808B (en) * | 2023-08-24 | 2023-12-12 | 广东省科学院生态环境与土壤研究所 | Acidophilic cadmium-resistant microalgae and application thereof in soil remediation |
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| CN104129860A (en) * | 2014-08-01 | 2014-11-05 | 浙江优创材料科技股份有限公司 | Method for purification of alkaline heavy metal wastewater by chlorella |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104129860A (en) * | 2014-08-01 | 2014-11-05 | 浙江优创材料科技股份有限公司 | Method for purification of alkaline heavy metal wastewater by chlorella |
Non-Patent Citations (2)
| Title |
|---|
| Microbial and plant derived biomass for removal of heavy metals from wastewater;Sarabjeet Singh Ahluwalia;《Bioresour Technol》;20060119;第98卷(第12期);第2243-2257页 * |
| 椭圆小球藻(Chlorella ellipsoidea)对4种重金属的耐受性及富集;浩云涛;《湖泊科学》;20010630;第13卷(第2期);第158-162页 * |
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