CN108546649B - Chlorella W2 capable of removing heavy metals in water with high heavy metal content and application thereof - Google Patents

Chlorella W2 capable of removing heavy metals in water with high heavy metal content and application thereof Download PDF

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CN108546649B
CN108546649B CN201810468898.3A CN201810468898A CN108546649B CN 108546649 B CN108546649 B CN 108546649B CN 201810468898 A CN201810468898 A CN 201810468898A CN 108546649 B CN108546649 B CN 108546649B
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chlorella
heavy metal
sewage
metal content
water
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CN108546649A (en
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王强
陈辉
何晨柳
汪杰
陈为先
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Institute of Hydrobiology of CAS
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    • 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
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    • 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
    • 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
    • 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
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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) in 11-month and 27-month in 2017, and the preservation number is CCTCC NO: M2017740. The chlorella W2 has high tolerance to heavy metal ions in a culture water body, has high removal rate to Cu, Zn, Ni, Co, Cr and Pb when growing in a water body with high heavy metal content, and is far higher than most of microalgae for removing heavy metal at present, so the chlorella W2 is particularly suitable for treating polluted water bodies with high heavy metal content. In addition, when heavy metals are removed, the chlorella W2 can also remove N, P and other nutrient substances in the water body, so that the risk of water body eutrophication is prevented.

Description

Chlorella W2 capable of removing heavy metals in water with high heavy metal content and application thereof
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 combined morphology shows that the protein belongs to the genus Chlorella of the phylum Chlorophyta, so the protein is named Chlorella W2 under the Latin chemical name Chlorella sp.W2.
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 (CCTCC) in 11-27 months in 2017 with the preservation number of M2017740.
The invention also provides application of the chlorella in removing heavy metals in water with high heavy metal content.
In one embodiment, the heavy metal is one or more combinations of Cu, Zn, Ni, Co, Cr, Pb.
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 to 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 in S2 is inoculated at an OD concentration700The value is 0.1-0.5.
In a preferred embodiment, the culturing conditions in S3 are 30 ℃, 80. + -. 5. mu. mol m-2s-1The intensity of the light.
The chlorella W2 has high tolerance to heavy metal ions in a culture water body, has high removal rate to Cu, Zn, Ni, Co, Cr and Pb when growing in a water body with high heavy metal content, and is far higher than most of microalgae for removing heavy metal at present, so the chlorella W2 is particularly suitable for treating polluted water bodies with high heavy metal content. In addition, when heavy metals are removed, the chlorella W2 can also remove N, P and other nutrient substances in the water body, so that the risk of water body eutrophication is prevented.
Preservation of microorganisms
The microalgae related to the invention is obtained by separating a water sample collected in the field, and belongs to the genus Chlorella of the phylum Chlorella of the green alga through 18S rDNA sequencing and morphological identification. The microalgae is preserved in 27 months 11 in 2017 in China Center for Type Culture Collection (CCTCC) delivered to Wuhan university, No. eight 299 in Wuchang district, Wuhan city, Hubei province, with the preservation number of CCTCC NO: M2017740, named Chlorella W2 and the Latin school name of Chlorella sp.W2.
Drawings
FIG. 1 is an optical micrograph of Chlorella vulgaris W2 according to the present invention;
FIG. 2 shows growth curves of Chlorella W2 cultured in BG11 medium and BG11 medium supplemented with heavy metals, respectively;
FIG. 3 shows the growth curves of a control strain Chlorella G32 cultured in BG11 medium and BG11 medium supplemented with heavy metals, respectively.
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 W2
The Chlorella W2(Chlorella sp. W2) of the present invention is obtained by separating from a water sample containing algae collected in the field, and the cell morphology thereof is as shown in fig. 1, and the cells are round or oval, green, without flagella and nucleated. By 18S rDNA binding morphology, the microalgae belong to the genus Chlorella, the phylum Chlorella. The microalgae has been preserved in China Center for Type Culture Collection (CCTCC) of Wuhan university in Wuhan City, Hubei province, China in 2017 at 11/27, the preservation number is CCTCC NO: M2017740, and the microalgae is named Chlorella W2 and Latin's name Chlorella sp.W2.
2. Tolerance of Chlorella W2 to heavy metals
Activating Chlorella W2 in small triangular flask, inoculating into 500ml conical flask containing 300ml BG11 culture medium with high heavy metal content, and inoculating to initial OD7000.1, the culture conditions are 30 ℃, 80 +/-5 mu mol m-2s-1Culturing in air-permeable mode under illumination.
BG11 medium with high heavy metal content was prepared by adding heavy metal mother liquor to BG11 medium. Wherein, the BG11 culture medium is prepared by the following method: 1.5g of NaNO was added to 1ml of each of Stock II, Stock III, Stock IV, Stock VI, Stock VII and Stock VIII3And adding water to a constant volume of 1000 ml. The components of the stock solutions are shown in Table 1, and the components of A5 are shown in Table 2As shown.
TABLE 1 composition of each stock solution in BG11 Medium preparation
Figure BDA0001662570250000041
TABLE 2A 5 composition
Figure BDA0001662570250000051
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 ×. Table 3 shows the heavy metal mother liquor concentration and working concentration.
TABLE 3 heavy metal mother liquor concentration and working concentration
Figure BDA0001662570250000052
After inoculation, the OD of the chlorella culture was measured daily700Values, a growth curve was prepared, and tolerance of chlorella W2 to heavy metals was observed with chlorella cultured in BG11 medium as a control.
As a result, as shown in FIG. 2, the growth curve of Chlorella W2 cultured in BG11 medium was not much different from that of BG11 medium supplemented with heavy metals, and it was found that Chlorella W2 had a high tolerance to heavy metals. FIG. 3 is a growth curve of another chlorella G32 isolated by the inventors during isolation, cultured in BG11 medium and BG11 medium with heavy metals, respectively, and the results show that the growth of the chlorella is greatly inhibited in BG11 medium with heavy metals, like most of the known chlorella.
3. Chlorella W2 removal efficiency of heavy metals in culture medium
After the chlorella W2 had grown 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 W2 has high removal rate of Cu, Zn, Ni, Co, Cr and Pb, which is much higher than that of most of the microalgae for removing heavy metals at present, and is particularly suitable for treating polluted water with high heavy metal content.
TABLE 4 Chlorella W2 pairs of heavy metals in the culture media
Figure BDA0001662570250000061
4. Application of chlorella W2 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 W2 into the sewage with the inoculation concentration of OD700To 0.1-0.5. 80. + -. 5. mu. mol m at 30 ℃-2s-1Culturing 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> institute of aquatic organisms of Chinese academy of sciences
<120> chlorella W2 capable of removing heavy metals in water with high heavy metal content and application thereof
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1831
<212>DNA
<213>Chlorella sp. W2
<400>1
tagcgttacg acggcagtgc 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 agccaaccgt 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 ccaggattgg 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 ctttgagaat ctctagagga tccccgggta 1800
ccgagctcga atcgtaatca tgtcaatgtt c 1831

Claims (7)

1. The chlorella is preserved in China center for type culture collection (CCTCC NO: M2017740) in 11-27 months in 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 combinations of Cu, Zn, Ni, Co, Cr, Pb.
4. 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.
5. The method according to claim 4, characterized in that it comprises the following steps:
s1: filtering the sewage to remove large-particle substances;
s2: inoculating the chlorella to the filtered sewage to obtain a chlorella-sewage culture;
s3: culturing the chlorella-sewage culture by introducing air for illumination;
s4: removing chlorella in the chlorella-sewage culture after culturing for 8-15 days.
6. The method of claim 5, wherein the chlorella in S2 is inoculated at an OD700The value is 0.1-0.5.
7. The method according to claim 5 or 6, wherein the culturing in S3 is carried out at 30 ℃ and 80. + -. 5. mu. mol m-2s-1The intensity of the light.
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CN109626582A (en) * 2018-12-25 2019-04-16 武汉理工大学 The method for removing lead in water removal using the modified microalgae of high phosphorus culture
CN114394670B (en) * 2021-12-15 2023-05-23 海南大学 Chlorella and application thereof in heavy metal wastewater treatment
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
CN114772742A (en) * 2022-05-19 2022-07-22 武汉轻工大学 Method for improving selenium-containing wastewater treatment capacity of chlorella
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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椭圆小球藻(Chlorella ellipsoidea)对4种重金属的耐受性及富集;浩云涛等;《湖泊科学》;20010630;第13卷(第2期);摘要，结果与讨论部分 *

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