CN114381377B - Aspergillus MF1 for removing heavy metal ions, microbial inoculum, preparation method and application thereof - Google Patents
Aspergillus MF1 for removing heavy metal ions, microbial inoculum, preparation method and application thereof Download PDFInfo
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- CN114381377B CN114381377B CN202111334940.0A CN202111334940A CN114381377B CN 114381377 B CN114381377 B CN 114381377B CN 202111334940 A CN202111334940 A CN 202111334940A CN 114381377 B CN114381377 B CN 114381377B
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- 241000228212 Aspergillus Species 0.000 title claims abstract description 84
- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 48
- 150000002500 ions Chemical class 0.000 title claims abstract description 43
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims description 13
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 33
- 239000002351 wastewater Substances 0.000 claims description 25
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 18
- 229910052799 carbon Inorganic materials 0.000 claims description 18
- 239000001963 growth medium Substances 0.000 claims description 17
- 229910052757 nitrogen Inorganic materials 0.000 claims description 17
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- 238000012258 culturing Methods 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 12
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- 239000008103 glucose Substances 0.000 claims description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 229930006000 Sucrose Natural products 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
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- 230000001580 bacterial effect Effects 0.000 description 11
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- 230000002378 acidificating effect Effects 0.000 description 10
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
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- 238000010521 absorption reaction Methods 0.000 description 6
- 229910052782 aluminium Inorganic materials 0.000 description 6
- 238000005070 sampling Methods 0.000 description 6
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 4
- 206010064930 age-related macular degeneration Diseases 0.000 description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 241000228257 Aspergillus sp. Species 0.000 description 3
- 238000001502 gel electrophoresis Methods 0.000 description 3
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- 241000894006 Bacteria Species 0.000 description 2
- 241001477352 Bengalia Species 0.000 description 2
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- -1 aluminum ion Chemical class 0.000 description 2
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- 229910052793 cadmium Inorganic materials 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
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- 229910001431 copper ion Inorganic materials 0.000 description 2
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- 239000010842 industrial wastewater Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
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- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 241000605222 Acidithiobacillus ferrooxidans Species 0.000 description 1
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- 238000007400 DNA extraction Methods 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000003914 acid mine drainage Methods 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
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- 229940041514 candida albicans extract Drugs 0.000 description 1
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- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
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- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000000464 low-speed centrifugation Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000012803 optimization experiment Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
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- 239000001632 sodium acetate Substances 0.000 description 1
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- 229910052569 sulfide mineral Inorganic materials 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- 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
- C12N1/14—Fungi; Culture media therefor
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/06—Contaminated groundwater or leachate
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the field of environmental protection, in particular to aspergillus MF1 for removing heavy metal ions, a microbial inoculum, a heavy metal degradation agent and application thereof. The invention provides Aspergillus (Aspergillus sp) MF1 for removing heavy metal ions, wherein the preservation number of the Aspergillus MF1 is GDMCC NO.61350. The strain provided by the invention can realize the effect of removing various heavy metal ions under the condition of low pH.
Description
Technical Field
The invention relates to the field of environmental protection, in particular to aspergillus MF1 for removing heavy metal ions, a microbial inoculum, a preparation method and application thereof.
Background
In the mining process, the sulfide minerals undergo a series of physical and chemical reactions such as leaching, oxidation, hydrolysis and the like under the action of air, water and microorganisms to form sulfuric acid-ferric sulfate solution with lower pH value, generally 2.5-5.5, and various metal ions such as Cu in the ore can be dissolved under the condition 2+ 、Zn 2+ 、Pb 2+ 、Mn 2+ Cd and Cd 2+ And the like, the heavy metal ions have high toxicity, not only can seriously pollute water resources and influence the yield and quality of crops, but also are extremely easy to enrich and expand in biological chains, and finally cause chronic poisoning in the accumulation of certain organs of human bodies, so that the health of human beings is endangered. Thus, according to the pollution characteristics of acid mine wastewater (AMD)Solving the problems of wide attention of governments and social communities, namely, solving the problems of economical and practical treatment methods, eliminating the harm and ensuring the development sustainability of mineral resources.
Among the methods for treating AMD, the microorganism method is a relatively wide treatment technology applied to AMD treatment due to the characteristics of strong applicability, low investment, environmental friendliness, small pollution and the like, and currently applied microorganisms comprise algae, fungi and bacteria, but heavy metal ions are mainly removed under neutral conditions or acidic conditions, and reports of fungus strains screened to obtain heavy metal ion resistance in extremely acidic environments are relatively limited. In addition, acidic industrial wastewater, particularly acidic mine wastewater, generally contains a plurality of metal ions at the same time, and therefore, it is necessary to screen out strains which are acid-resistant and capable of removing a plurality of heavy metals at the same time.
Disclosure of Invention
In order to solve the problems, the invention provides aspergillus MF1 for removing heavy metal ions, a microbial inoculum, a preparation method and application thereof. The strain provided by the invention can realize the effect of removing various heavy metal ions under the condition of low pH.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides Aspergillus (Aspergillus sp) MF1 for removing heavy metal ions, wherein the preservation number of the Aspergillus MF1 is GDMCC NO.61350.
The invention also provides a microbial inoculum, and the active ingredients of the microbial inoculum comprise the aspergillus MF1.
Preferably, the number of spores of aspergillus MF1 per Kg or per L of said inoculum is 8×10 10 And each.
The invention also provides a preparation method of the microbial inoculum, which comprises the following steps: mixing and culturing the strain and a culture medium to obtain the microbial inoculum; the culture medium comprises a nitrogen source and a carbon source; the carbon source comprises glucose and/or sucrose; the nitrogen source comprises yeast and/or peptone.
Preferably, the weight ratio of the carbon source to the nitrogen source is (7.5-15): 1.
The invention also provides application of the aspergillus MF1 or the microbial inoculum prepared by the preparation method in removing heavy metal ions.
The invention also provides application of the aspergillus MF1 or the microbial inoculum prepared by the preparation method in removing heavy metal ions in water.
Preferably, the water comprises wastewater or groundwater.
Preferably, the wastewater comprises an acidic wastewater.
Preferably, the pH value of the acid wastewater is more than or equal to 3.5.
The beneficial effects are that: the invention provides Aspergillus (Aspergillus sp) MF1 for removing heavy metal ions, wherein the preservation number of the Aspergillus MF1 is GDMCC NO.61350. The strain selected by the invention is screened from acid mine wastewater in the eastern part of Anhui, can not only endure low pH, but also can effectively remove various heavy metal ions; and the strain is used for Cu 2+ The maximum tolerance concentration is 770.18mg/g, when Cu 2+ At concentrations of 50mg/L and 100mg/L, aspergillus MF1 against Cu 2+ The removal rate of (2) can reach 78.40% and 64.47%, respectively.
Description of biological preservation
Aspergillus sp MF1 is deposited in the Guangdong province microbiological bacterial collection center (GDMCC for short), and has a deposit address of 59 th floor 5 of the university of Hirscht 100 in Guangzhou city, a deposit date of 2020, 12 months and 7 days, and a deposit number of GDMCC NO.61350.
Drawings
FIG. 1 is a morphological identification map of Aspergillus MF1, wherein the morphology map is a colony map of Aspergillus MF1 and the morphology map is a colony map of Aspergillus MF1, respectively;
FIG. 2 is a diagram of PCR results of 18S rDNA of Aspergillus MF1, wherein the left picture is a characteristic map, and the right picture is a gel electrophoresis diagram of a PCR product of Aspergillus strain;
FIG. 3 is a phylogenetic tree of Aspergillus MF 1;
FIG. 4 shows the effect of different pH on the growth of Aspergillus MF 1;
FIG. 5 shows the biomass of Aspergillus MF1 under different carbon source conditions;
FIG. 6 shows the biomass of Aspergillus MF1 under different nitrogen source conditions;
FIG. 7 shows the growth process Cu of Aspergillus MF1 2+ Concentration variation and Aspergillus MF1 vs. Cu 2+ The removal rate;
FIG. 8 shows Mn during the growth of Aspergillus MF1 2+ Concentration variation;
FIG. 9 shows the ratio of MF1 to Mn of Aspergillus 2+ The removal rate;
FIG. 10 is a graph showing the effect of Aspergillus MF1 on the concentration of various metal ions at pH4 when the metal ions coexist;
FIG. 11 shows the removal of various metal ions by Aspergillus MF1 in the presence of multiple metal ions at pH 4;
FIG. 12 is a graph showing the effect of the coexistence of various metal ions on the concentration of various metal ions in Aspergillus MF1 at pH 5.5;
FIG. 13 shows the removal of various metal ions by Aspergillus MF1 in the presence of various metal ions at pH 5.5.
Detailed Description
The invention provides Aspergillus (Aspergillus sp) MF1 for removing heavy metal ions, wherein the preservation number of the Aspergillus MF1 is GDMCC NO.61350.
The aspergillus MF1 according to the invention preferably has the following properties:
(1) The bacterial colony is gray yellow, the surface is dry and uneven, the middle part is raised, and the edge is flat;
(2) Hypha grows vigorously, one end of the hypha is in a long column shape, the other end of the hypha is in a crown shape, a circle of spores grow outside, and the spores are green and oval;
(3) The strain has strong acid resistance and can survive in an environment with the pH value more than or equal to 3.5;
(4) The nucleotide sequence of the 18SrDNA of the aspergillus MF1 is shown as SEQ ID NO:1 is shown as follows: TGCGGAAGGATCATTACCGAGTGAGGGCCCTCTGGGTCCAACCTCCCCACCCGTGTCTATCGTACCTTGTTGCTTCGGCGGGCCCGCCGTTTCGACGGCCGC CGGGGAGGCCTTGCGCCCCCGGGCCCGCGCCCGCCGAAGACCCCAACATG AACGCTGTTCTGAAAGTATGCAGTCTGAGTTGATTATCGTAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGAAGAACGCAGCGAAA TGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAGTCTTTGAACGCACATTGCGCCCCCTGGTATTCCGGGGGGCATGCCTGTCCGAGCGTCATT GCTGCCCTCAAGCACGGCTTGTGTGTTGGGCCCCCGTCCCCCTCTCCCGGG GGACGGGCCCGAAAGGCAGCGGCGGCACCGCGTCCGGTCCTCGAGCGTATGGGGCTTTGTCACCTGCTCTGTAGGCCCGGCCGGCGCCAGCCGACACCCA ACTTTATTTTTCTAAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACT TAAGCATA. The strain selected by the invention is screened from acid mine wastewater in the eastern part of Anhui, is screened under the extremely acidic condition, and can grow normally under the condition that the pH is more than or equal to 3.5, so that the strain can not only endure low pH, but also remove heavy metals simultaneously, and has higher removal rate under the relatively neutral condition.
The invention provides a microbial inoculum, and the active ingredients of the microbial inoculum comprise the aspergillus MF1. In the present invention, the spore count of Aspergillus MF1 per Kg or per L of the microbial inoculum is preferably 8X 10 10 And each. The microbial inoculum disclosed by the invention can be used for effectively removing various heavy metal ions, and has high removal rate.
The invention also provides a preparation method of the microbial inoculum, which comprises the following steps: mixing and culturing the strain and a culture medium to obtain the microbial inoculum; the culture medium comprises a nitrogen source and a carbon source; the carbon source comprises glucose and/or sucrose; the nitrogen source comprises yeast and/or peptone.
In the present invention, the carbon source preferably includes glucose; the nitrogen source preferably comprises yeast; the weight ratio of the carbon source to the nitrogen source is preferably (7.5 to 15): 1, more preferably 10:1. The source of the carbon source or nitrogen source is not particularly limited in the present invention, and commercially available products known to those skilled in the art may be used. The invention optimizes the culture medium, so that the strain cultured in a certain time has high biomass and low culture cost.
The strain disclosed by the invention has the advantages of low pH tolerance, effective removal of various heavy metal ions, high removal rate and low culture cost, so that the strain, the microbial inoculum containing the strain and the heavy metal degradation agent containing the strain can be applied to removal of heavy metal ions.
The invention providesThe application of the aspergillus MF1 or the microbial inoculum prepared by the preparation method in removing heavy metal ions, and further in removing heavy metal ions in water, wherein the heavy metal ions comprise Fe 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ And Zn 2+ One or more of the following; the water preferably includes wastewater and groundwater; the wastewater comprises preferably an acidic wastewater, further preferably an acidic industrial wastewater, and more preferably an acidic mine wastewater. The acid mine wastewater is preferably acid mine wastewater (Acid mine drainage, AMD for short) with extreme environment, which is formed by exposing sulfide ores hidden underground through the combined action of water, oxygen, thiobacillus ferrooxidans and other factors in mining, road construction and other large-scale excavation activities, and has the characteristics of low pH, high concentration sulfate radical and high concentration heavy metal ion.
The pH value of the acid mine wastewater is preferably 2.5-6.5; the concentration of sulfate radical in the acid mine wastewater is preferably 500-5000 mg/L; the concentration of different heavy metal ions in the acid mine wastewater is preferably 5-500 mg/L, wherein Fe 3+ The concentration of (C) is preferably 5-500 mg/L, al 3+ The concentration of (C) is preferably 5-1000 mg/L, mn 2+ The concentration of Cu is preferably 5-1000 mg/L 2+ The concentration of Zn is preferably 5-100 mg/L 2+ The concentration of (C) is preferably 5 to 100mg/L.
In order to further illustrate the present invention, the aspergillus MF1 strain for removing heavy metal ions, the microbial inoculum, the preparation method and the application thereof provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Enrichment culture of aspergillus MF 1: 50mL of acid mine wastewater at the eastern part of Anhui is taken, and 3000 r.min of a desk centrifuge is used -1 Removing impurities by low-speed centrifugation; transferring 1mL of the supernatant, centrifuging, inoculating into 50mL of modified Martin liquid culture medium (formula shown in Table 1), and rotating at 28deg.C and 120r.min -1 Enrichment culture is carried out for 72h under the condition, and the inoculation amount is 2% according to the volume ratio every 3 daysTransferring and culturing once, transferring and culturing 2 times for later use;
separation and purification of aspergillus MF 1: sterilizing the Bengalia red culture medium, the culture dish, the coating rod, the dilution pipe, the pure water and the like in a high-temperature sterilization pot at 121 ℃ for 20min before the experiment, pouring 25-30 mL of flat plate in a sterile environment, and cooling for later use; taking 1mL of bacterial liquid after 3 times of transfer culture enrichment, and placing the bacterial liquid into a dilution tube according to l0 -1 、10 -2 、10 -3 、 10 -4 、10 -5 Sequentially diluting the culture solution in a gradient way, respectively absorbing 0.2mL of diluted bacterial solutions with different gradients in a sterile environment, coating the diluted bacterial solutions on a plate of a Bengalia red culture medium (the formula is shown in table 1), culturing the culture solution in a constant temperature incubator at 28 ℃ in an inverted way, and culturing the culture solution for 2 to 3 days until macroscopic colonies grow on the plate; and selecting a typical single colony growing on the flat plate, repeatedly scribing until the microscopic bacteria form is uniform, and stopping scribing and separating. Inoculating the obtained pure bacterial colony into a bacteria-retaining tube, preserving the bacterial colony as a bacterial strain in a refrigerator at 4 ℃, and transferring the bacterial colony to a refrigerator at-81 ℃ for preservation after one day;
table 1 culture medium formulation
Example 2
Morphological identification: the strains obtained in example 1 were observed for colony, mycelium, spore and other structures by high-power fluorescence microscopy, and the fungus color, size, shape were recorded, and identified by reference to "fungus identification handbook (Wei Jingchao, 1979) and" Chinese fungus Saint "(Zhang Zhongyi, 2014), and the identification results are shown in FIG. 1: the bacterial colony is gray yellow, has dry and uneven surface, raised middle part and smooth edge; hypha grows vigorously, one end of the hypha is in a long column shape, the other end of the hypha is in a crown shape, a circle of spores grow outside, and the spores are green and oval; the primary identification result is Aspergillus.
Example 3
Molecular biology identification: extracting DNA of the strain screened in example 1 according to Ezup column type fungus genome DNA extraction kit (SK 8259) provided by Shanghai bioengineering Co., ltd., and performing PCR amplification on the extracted DNA, wherein the primers are 18SrDNA universal primer NS1 (shown as SEQ ID NO: 2: 5'-GTAGTCATATGCTTGTCTC-3') and NS6 (shown as SEQ ID NO: 3: 5'-GCATCACAGACCTGTTATTGCCTC-3'); the result of PCR amplification was observed by gel electrophoresis (1% agarose electrophoresis, 150V, 100mA 20min electrophoresis, see FIG. 2), and the PCR product was sequenced by Shanghai Biotechnology Co., ltd, and the sequencing result was shown as SEQ ID NO:1 is shown in the specification; the sequencing results of 18SrDNA were blast aligned at NCBI and phylogenetic tree was constructed using software MEGA-X, see FIG. 3.
As can be seen from FIGS. 2 and 3, the results of the gel electrophoresis pattern and phylogenetic tree of the PCR products show that the similarity between the strain sequences and Aspergillus after blast comparison on NCBI reaches 99.77%, which proves that a pure strain of Aspergillus is obtained after screening.
Example 4
The biological quantity of Aspergillus (Aspergillus sp) MF1 is used as an evaluation index, pH gradients of 1.5, 2.5, 3.5, 4.5, 5.5 and 7.0 are set, modified Martin culture medium (5.00 g of peptone, 2.00g of yeast extract powder, 20.00g of glucose, 1.00g of monopotassium phosphate, 0.50g of magnesium sulfate, 0.10g of chloramphenicol and 1000mL of distilled water) is selected, and 100 mu L of Aspergillus (Aspergillus sp) MF1 spore suspension is added, wherein the spore concentration in the spore suspension is 8 multiplied by 10 10 CFU/L, and culturing at 28deg.C for 120r.min -1 Culturing for 144h in a constant temperature shaking incubator, sampling every 16h, placing in an overspeed low temperature centrifuge, and regulating the rotating speed to 8000 r.min -1 Centrifuging for 5min, discarding supernatant, and measuring mycelium dry weight, wherein the result is shown in figure 4, and the strain can normally grow under the condition that the pH is 3.5-7.0; at a pH of 2.5, the strain can grow, but the growth amount is low; at a pH of 1.5, the strain can grow abnormally.
Example 5
Culture medium optimization experiment
Under the acidic condition with the pH of 3.5, the culture conditions such as carbon-nitrogen source type carbon-nitrogen ratio and the like are optimized by taking the aspergillus biomass and the carbon-nitrogen source utilization rate as evaluation indexes.
The carbon source is glucose, sucrose, starch, ethylene glycol, and sodium acetate, and the nitrogen source is yeast, peptone, peptone+yeast, urea, and nitrateSodium sulfate and ammonium sulfate are used as nitrogen sources, the carbon-nitrogen ratios are 5.0, 7.5, 10.0, 12.5 and 15.0 respectively, a modified Martin culture medium (the culture medium is the same as that of example 4) is selected, 100 mu L of Aspergillus (Aspergillus sp) MF1 spore suspension is added, wherein the spore concentration in the spore suspension is 8 multiplied by 10 10 CFU/L, and culturing at 28deg.C for 120r.min -1 Culturing for 144h in a constant temperature shaking incubator, sampling every 16h, placing in an overspeed low temperature centrifuge, and regulating the rotating speed to 8000 r.min -1 Centrifuging for 5min, discarding supernatant, and measuring mycelium dry weight, wherein the measurement results are shown in tables 2-4, FIG. 5 and FIG. 6.
TABLE 2 biomass and specific growth rate of Aspergillus sp.MF1 under different carbon source conditions
| Optimizing conditions | Optimized formulation | biomass/(g.L) -1 ) | Specific growth rate of 10 -2 /h -1 |
| Glucose | 8.96±0.26 | 5.64±0.01 | |
| Sucrose | 8.03±0.06 | 5.52±0.01 | |
| Carbon source | Starch | 6.38±0.09 | 5.28±0.02 |
| Acetic acid sodium salt | 4.16±0.05 | 4.84±0.01 | |
| Ethylene glycol | 2.25±0.08 | 4.20±0.04 |
TABLE 3 biomass and specific growth rate of Aspergillus sp.MF1 under different carbon source conditions
TABLE 4 biomass and specific growth rate of Aspergillus sp.MF1 at different carbon-to-nitrogen ratios
As is clear from tables 2 to 4, FIGS. 5 and 6, when the carbon source is glucose or sucrose, the nitrogen source is yeast and/or peptone, and the carbon-nitrogen ratio is (7.5 to 15): 1, the biomass of the strain cultured in a certain period of time is high, and the culture cost is low; and when the carbon source is glucose, the nitrogen source is yeast, the carbon-nitrogen ratio is 10:1, and the biomass is highest.
Example 6
CuSO was added to the modified Martin medium (medium same as example 4) separately 4 Solution to Cu 2+ The concentration is 50mg/L respectively,100mg/L, 300mg/L, 500mg/L, 1000mg/L of Aspergillus sp MF1 spore suspension was added, wherein the spore concentration in the spore suspension was 8X 10 10 CFU/L, adjusting pH=3.5, culturing at 28deg.C under 120r/min for 144 hr, sampling at intervals of 48 hr for measuring fungal biomass, and measuring Cu in sample supernatant by flame atomic absorption (atomic absorption spectrometer AA240FS, VAR-IAN, USA) 2+ Concentration was calculated and Cu after 144h of incubation 2+ The removal rate and the measurement results are shown in Table 5, table 6 and FIG. 7.
TABLE 5 different Cu at pH3.5 2+ Concentration treatment of Cu at different times 2+ Concentration (unit: g/L)
Note that: cu at 0h in Table 5 2+ Concentration as determined by flame atomic absorption spectrophotometer, and formulated Cu 2+ There is some error in concentration, but the error is within the allowable range of measurement.
TABLE 6 different Cu at pH3.5 2+ After 144h of concentration treatment of Cu 2+ Removal rate of
| Cu 2+ Treatment concentration (mg/L) | 50 | 100 | 300 | 500 | 1000 |
| Removal rate (%) | 78.40 | 64.47 | 20.44 | 14.33 | 13.36 |
Note that: removal rate = (measurement value of 0 h-measurement value of 144 h)/measurement value of 144h×100% in table 6.
As can be seen from tables 5, 6 and 7, at pH3.5, aspergillus MF1 is specific for Cu 2+ After 48h, the Aspergillus MF1 is in logarithmic growth phase to propagate great amount of thallus and to eliminate great amount of Cu 2+ The method comprises the steps of carrying out a first treatment on the surface of the And Aspergillus MF1 for Cu 2+ The removal rate is along with Cu 2+ The concentration is increased and decreased, and Cu is as follows 2+ The removal rate is as high as 78.40% when the concentration is 50mg/L, and the removal rate is as high as Cu 2+ The concentration is increased to 1000mg/L, and the removal rate is reduced to 13.36%.
Example 7
MnSO was added to the modified Martin medium (medium same as example 4) respectively 4 ·H 2 O solution to Mn 2+ The concentration of the suspension is 100mg/L, 300mg/L, 500mg/L, 1000mg/L and 2000mg/L respectively, 100. Mu.L of Aspergillus sp MF1 spore suspension is added, wherein the spore concentration in the spore suspension is 8×10 10 CFU/L, adjusting pH=3.5, culturing at 28deg.C under 120r/min for 144 hr, sampling at intervals of 48 hr for measuring fungus biomass, and measuring Mn of supernatant by flame atomic absorption method (atomic absorption spectrometer AA240FS, VAR-IAN, USA) 2+ Concentration and Mn after 144h of culture was calculated 2+ The removal rate and the measurement results are shown in Table 7, table 8, FIG. 8 and FIG. 9.
TABLE 7 different Mn at pH3.5 2+ Concentration treatment of Mn at different times 2+ Concentration (unit: g/L)
Note that: mn of 0h in Table 7 2+ Concentration as determined by flame atomic absorption spectrophotometer, and formulated Mn 2+ There is some error in concentration, but the error is within the allowable range of measurement.
TABLE 8 different Mn at pH3.5 2+ Mn after 144h of concentration treatment 2+ Removal rate of
| Mn 2+ Treatment concentration (mg/L) | 100 | 300 | 500 | 1000 | 2000 |
| Removal rate (%) | 92.69 | 43.05 | 39.24 | 20.89 | 12.35 |
Note that: removal rate = (measurement value of 0 h-measurement value of 144 h)/measurement value of 144h×100% in table 8.
As can be seen from tables 7 and 8, at pH3.5, aspergillus vs. Mn 2+ After 48h, the Aspergillus is in logarithmic growth phase, so that the thallus is largely propagated and a large amount of Mn is removed 2+ The method comprises the steps of carrying out a first treatment on the surface of the Aspergillus pairIn Mn 2+ Removal rate with Mn 2+ The concentration is increased and reduced, and Mn is as follows 2+ The removal rate is as high as 92.69% when the concentration is 100mg/L, and the removal rate is along with Mn 2+ The concentration is increased to 2000mg/L, and the removal rate is reduced to 12.35%
Example 8
Fe was added to a modified Martin medium (Medium same as example 4) having a pH of 4 3+ 、Al 3+ 、 Mn 2+ 、Cu 2+ 、Zn 2+ Obtaining a stock solution regulating culture medium, wherein the stock solution regulating culture medium contains Fe 3+ The concentration of (C) is 100 mg.L -1 ，Al 3+ The concentration of (C) is 500 mg.L -1 ，Mn 2+ The concentration of (C) is 500 mg.L -1 ，Cu 2+ The concentration of (C) is 100 mg.L -1 ，Zn 2+ The concentration of (C) is 100 mg.L -1 100. Mu.L of Aspergillus sp MF1 spore suspension was then added, wherein the spore concentration in the spore suspension was 8X 10 10 CFU/L, noted as experimental group; 100. Mu.L of a spore suspension of Aspergillus sp MF1 was added to a modified Martin medium at pH4, wherein the spore concentration in the spore suspension was 8X 10 10 CFU/L, recorded as control group;
placing the experimental group and the control group under the culture condition of 28 ℃ and 120 r.min respectively -1 Continuously culturing for 112h in a constant temperature shaking incubator, sampling, placing in an overspeed low-temperature centrifuge, and regulating the rotation speed to 8000 r.min -1 Centrifuging for 5min, and reserving supernatant for determining Fe therein 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ 、Zn 2+ Each set of experiments was repeated 3 times, the concentration change results are shown in table 10 and fig. 10, the heavy metal ion removal effect is shown in fig. 11, hyphae were used for measurement of dry weight, and the dry weight results are shown in table 9.
TABLE 9 influence of the coexistence of various metal ions at pH4 on Aspergillus sp MF1 growth
| Biomass (g/L) | Error of | |
| Control group | 9.4382 | 2.01％ |
| Experimental group | 9.3423 | 0.76％ |
As is clear from Table 9, the growth of the Aspergillus MF1 experimental group was less affected by the coexistence of various heavy metal ions than the control group, and the biomass was 98.98% of that in the absence of heavy metal ions.
TABLE 10 influence of Aspergillus sp MF1 on the concentration of different Metal ions in the coexistence of multiple Metal ions at pH4
| Initial concentration (mg/L) | Post treatment concentration (mg/L) | Error of | Removal rate (%) | |
| Fe 3+ | 100 | 54.58 | 3.62％ | 45.42％ |
| Al 3+ | 500 | 285.44 | 2.79％ | 42.91％ |
| Mn 2+ | 500 | 459.85 | 1.68％ | 8.03％ |
| Cu 2+ | 100 | 61.65 | 9.84％ | 38.35％ |
| Zn 2+ | 100 | 79.15 | 2.86％ | 20.85％ |
As can be seen from table 10, fig. 10 and fig. 11, the strain of the present invention can effectively remove various heavy metal ions under the conditions of coexistence of various heavy metal ions and low pH, and has high ferric iron, aluminum and copper ion removal rates, wherein the ferric iron removal rate reaches 45.42%, the aluminum ion removal rate reaches 42.91%, and the treatment effect on aluminum-containing wastewater at low pH is superior to that of other methods.
Example 9
Fe was added to the modified Martin medium at pH5.5 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ 、Zn 2+ A stock solution regulating medium, wherein the stock solution regulates Fe in the medium 3+ The concentration of (C) is 50 mg.L -1 ，Al 3+ The concentration of (C) is 50 mg.L -1 ，Mn 2+ The concentration of (C) is 50 mg.L -1 ，Cu 2+ The concentration of (C) is 50 mg.L -1 ，Zn 2+ The concentration of (C) is 50 mg.L -1 100. Mu.L of Aspergillus sp MF1 spore suspension was then added, wherein the spore concentration in the steamed stuffed bun suspension was 8X 10 10 CFU/L, noted as experimental group; 100. Mu.L of spore suspension of Aspergillus sp MF1 was added to a modified Martin medium at pH5.5, wherein the spore concentration in the steamed stuffed bun suspension was 8X 10 10 CFU/L, recorded as control group;
placing the experimental group and the control group under the culture condition of 28 ℃ and 120 r.min respectively -1 Continuously culturing for 112h in a constant temperature shaking incubator, sampling, placing in an overspeed low-temperature centrifuge, and regulating the rotation speed to 8000 r.min -1 Centrifuging for 5min, and reserving supernatant for determining Fe in solution 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ 、Zn 2+ Each set of experiments was repeated 3 times, the concentration changes are shown in table 12 and fig. 12, the heavy metal ion removal effect is shown in fig. 13, hyphae were used for measurement of dry weight, and the dry weight results are shown in table 11.
TABLE 11 influence of coexistence of various metal ions at pH5.5 on Aspergillus sp MF1 growth
| Biomass (g/L) | Error of | |
| Control group | 12.4961 | 1.73％ |
| Experimental group | 11.7865 | 0.21％ |
As is clear from Table 11, in the coexistence of various heavy metal ions, the growth of the Aspergillus experiment group was less affected by the growth than the control group, and the biomass was 94.32% of the growth than the growth in the absence of the heavy metal ions.
TABLE 12 influence of Aspergillus sp MF1 on the concentration of different Metal ions in the coexistence of multiple Metal ions at pH5.5
| Initial concentration (mg/L) | Post treatment concentration (mg/L) | Error of | Removal rate (%) | |
| Fe 3+ | 50 | 12.15 | 5.2％ | 75.70％ |
| Al 3+ | 50 | 21.06 | 2.62％ | 57.88％ |
| Mn 2+ | 50 | 46.08 | 0.61％ | 7.84％ |
| Cu 2+ | 50 | 15.69 | 2.34％ | 68.62％ |
| Zn 2+ | 50 | 34.82 | 3.21％ | 30.36％ |
As can be seen from table 12, fig. 12 and fig. 13, the strain of the present invention can effectively remove various heavy metal ions under the conditions of coexistence of various heavy metal ions and low pH, and has high ferric iron, aluminum and copper ion removal rates, wherein the ferric iron removal rate reaches 75.70%, the aluminum ion removal rate reaches 57.88%, and the treatment effect on the aluminum-containing wastewater at low pH is superior to that of other methods.
According to the embodiment described above, the selected strain is screened from acid mine wastewater at the eastern part of Anhui, can not only endure low pH, but also can effectively remove various heavy metal ions; and the strain is used for Cu 2+ The maximum tolerance concentration is 770.18mg/g, when Cu 2+ At concentrations of 50mg/L and 100mg/L, aspergillus MF1 against Cu 2+ The removal rate of (2) can reach 78.40% and 64.47%, respectively.
While the invention has been described in terms of preferred embodiments, it is not intended to be limited thereto, but rather to enable any person skilled in the art to make various changes and modifications without departing from the spirit and scope of the present invention, which is therefore to be limited only by the appended claims.
Sequence listing
<110> university of synthetic fertilizer industry
<120> aspergillus MF1 strain for removing heavy metal ions, microbial inoculum, preparation method and application thereof
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ccgggcccgc gcccgccgaa gaccccaaca tgaacgctgt tctgaaagta tgcagtctga 180
gttgattatc gtaatcagtt aaaactttca acaacggatc tcttggttcc ggcatcgatg 240
aagaacgcag cgaaatgcga taagtaatgt gaattgcaga attcagtgaa tcatcgagtc 300
tttgaacgca cattgcgccc cctggtattc cggggggcat gcctgtccga gcgtcattgc 360
tgccctcaag cacggcttgt gtgttgggcc cccgtccccc tctcccgggg gacgggcccg 420
aaaggcagcg gcggcaccgc gtccggtcct cgagcgtatg gggctttgtc acctgctctg 480
taggcccggc cggcgccagc cgacacccaa ctttattttt ctaaggttga cctcggatca 540
ggtagggata cccgctgaac ttaagcata 569
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Claims (7)
1. Aspergillus strain for removing heavy metal ionsAspergillus sp) MF1, characterized in that it is deposited with the collection of microorganisms of the cantonese province under the deposit number GDMCC No.61350.
2. A microbial inoculum, wherein an active ingredient of the microbial inoculum comprises the aspergillus MF1 as claimed in claim 1.
3. The microbial agent according to claim 2, wherein the number of spores of aspergillus MF1 per Kg or per L of said microbial agent is 8 x 10 10 And each.
4. A method for preparing the microbial inoculum according to claim 2 or 3, which comprises: mixing and culturing aspergillus MF1 according to claim 1 with a culture medium to obtain the microbial inoculum; the culture medium comprises a nitrogen source and a carbon source; the carbon source comprises glucose and/or sucrose; the nitrogen source comprises yeast and/or peptone.
5. The method according to claim 4, wherein the weight ratio of the carbon source to the nitrogen source is 7.5 to 15:1.
6. Use of the aspergillus MF1 according to claim 1 or the microbial inoculum according to claim 2 or 3 or the microbial inoculum prepared by the preparation method according to claim 4 or 5 for removing heavy metal ions; the pH value of the reaction system for removing heavy metal ions is 3.5-7.0; the heavy metal ion comprises Fe 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ And Zn 2+ One or more of them.
7. The use of the aspergillus MF1 according to claim 1 or the microbial inoculum according to claim 2 or 3 or the microbial inoculum prepared by the preparation method according to claim 4 or 5 for removing heavy metal ions in water; the water includes wastewater; the wastewater comprises acid wastewater; the pH value of the acid wastewater is more than or equal to 3.5; the heavy metal ion comprises Fe 3+ 、Al 3+ 、Mn 2+ 、Cu 2+ And Zn 2+ One or more of them.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102795712A (en) * | 2012-09-03 | 2012-11-28 | 许继联华国际环境工程有限责任公司 | Method for treating heavy metal-containing pollutant wastewater |
| CN103642703A (en) * | 2013-12-12 | 2014-03-19 | 北京林业大学 | Production method of effective phosphate solubilizing aspergillus japonicus agent with characteristic of tolerance to heavy metals |
| CN107384804A (en) * | 2017-08-08 | 2017-11-24 | 中国科学院烟台海岸带研究所 | Gibberella NT 1 and its application |
| WO2021196693A1 (en) * | 2020-04-03 | 2021-10-07 | 河北科技大学 | Penicillium oxalicum sdf-25 and use thereof |
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| CN102795712A (en) * | 2012-09-03 | 2012-11-28 | 许继联华国际环境工程有限责任公司 | Method for treating heavy metal-containing pollutant wastewater |
| CN103642703A (en) * | 2013-12-12 | 2014-03-19 | 北京林业大学 | Production method of effective phosphate solubilizing aspergillus japonicus agent with characteristic of tolerance to heavy metals |
| CN107384804A (en) * | 2017-08-08 | 2017-11-24 | 中国科学院烟台海岸带研究所 | Gibberella NT 1 and its application |
| WO2021196693A1 (en) * | 2020-04-03 | 2021-10-07 | 河北科技大学 | Penicillium oxalicum sdf-25 and use thereof |
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| Title |
|---|
| "不同重金属负荷下黑曲霉的产酸特性及重金属溶出效果";高云西 等;《江苏农业科学》;第46卷(第11期);第285-287页 * |
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