CN108359621A - One plant height imitates rhizobium and its application of adsorption of Low Concentration copper ion - Google Patents
One plant height imitates rhizobium and its application of adsorption of Low Concentration copper ion Download PDFInfo
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- CN108359621A CN108359621A CN201810211184.4A CN201810211184A CN108359621A CN 108359621 A CN108359621 A CN 108359621A CN 201810211184 A CN201810211184 A CN 201810211184A CN 108359621 A CN108359621 A CN 108359621A
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- rhizobium
- low concentration
- copper ion
- adsorption
- copper
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- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 title claims abstract description 50
- 229910001431 copper ion Inorganic materials 0.000 title claims abstract description 49
- 241000589180 Rhizobium Species 0.000 title claims abstract description 35
- 238000001179 sorption measurement Methods 0.000 title claims abstract description 31
- 241000894006 Bacteria Species 0.000 claims abstract description 33
- 230000001580 bacterial effect Effects 0.000 claims abstract description 33
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229910052802 copper Inorganic materials 0.000 claims abstract description 29
- 239000010949 copper Substances 0.000 claims abstract description 29
- 239000002351 wastewater Substances 0.000 claims abstract description 29
- 238000010521 absorption reaction Methods 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 13
- 238000009472 formulation Methods 0.000 claims description 7
- 239000000725 suspension Substances 0.000 claims description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 abstract description 12
- 239000010802 sludge Substances 0.000 abstract description 9
- 238000002955 isolation Methods 0.000 abstract description 8
- 239000010865 sewage Substances 0.000 abstract description 6
- 230000006872 improvement Effects 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 26
- 239000002609 medium Substances 0.000 description 22
- 229910001868 water Inorganic materials 0.000 description 19
- 239000000243 solution Substances 0.000 description 17
- 230000000694 effects Effects 0.000 description 15
- 229910001385 heavy metal Inorganic materials 0.000 description 14
- 238000011081 inoculation Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 229910052500 inorganic mineral Inorganic materials 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000011707 mineral Substances 0.000 description 6
- 235000010755 mineral Nutrition 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 229940088594 vitamin Drugs 0.000 description 5
- 239000011782 vitamin Substances 0.000 description 5
- 235000013343 vitamin Nutrition 0.000 description 5
- 229930003231 vitamin Natural products 0.000 description 5
- 150000003722 vitamin derivatives Chemical class 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 238000005374 membrane filtration Methods 0.000 description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- AAWZDTNXLSGCEK-WYWMIBKRSA-N (-)-quinic acid Chemical compound O[C@@H]1C[C@](O)(C(O)=O)C[C@@H](O)[C@H]1O AAWZDTNXLSGCEK-WYWMIBKRSA-N 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 229910004619 Na2MoO4 Inorganic materials 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 238000011953 bioanalysis Methods 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000000205 computational method Methods 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 2
- 229910052603 melanterite Inorganic materials 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000011684 sodium molybdate Substances 0.000 description 2
- 229940074404 sodium succinate Drugs 0.000 description 2
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 description 2
- GNBVPFITFYNRCN-UHFFFAOYSA-M sodium thioglycolate Chemical compound [Na+].[O-]C(=O)CS GNBVPFITFYNRCN-UHFFFAOYSA-M 0.000 description 2
- 229940046307 sodium thioglycolate Drugs 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- ILJSQTXMGCGYMG-UHFFFAOYSA-N triacetic acid Chemical compound CC(=O)CC(=O)CC(O)=O ILJSQTXMGCGYMG-UHFFFAOYSA-N 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- 239000011686 zinc sulphate Substances 0.000 description 2
- 108010023063 Bacto-peptone Proteins 0.000 description 1
- 208000018152 Cerebral disease Diseases 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 101100117236 Drosophila melanogaster speck gene Proteins 0.000 description 1
- 208000012661 Dyskinesia Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 241000589187 Rhizobium sp. Species 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229960004050 aminobenzoic acid Drugs 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- FAPWYRCQGJNNSJ-UBKPKTQASA-L calcium D-pantothenic acid Chemical compound [Ca+2].OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O.OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O FAPWYRCQGJNNSJ-UBKPKTQASA-L 0.000 description 1
- 229960002079 calcium pantothenate Drugs 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- -1 hydrochloric acid sulphur Amine Chemical class 0.000 description 1
- 239000010842 industrial wastewater Substances 0.000 description 1
- AGBQKNBQESQNJD-UHFFFAOYSA-M lipoate Chemical compound [O-]C(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-M 0.000 description 1
- 235000019136 lipoic acid Nutrition 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 238000002161 passivation Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 1
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 1
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 1
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 229960002663 thioctic acid Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/41—Rhizobium
-
- 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
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Water Supply & Treatment (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Environmental & Geological Engineering (AREA)
- Hydrology & Water Resources (AREA)
- Biodiversity & Conservation Biology (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses the rhizobium that a plant height imitates adsorption of Low Concentration copper ion（Rhizobiumsp.）S2, which is characterized in that the bacterial strain was preserved in Guangdong Province's Culture Collection on the 24th in September in 2017, and culture presevation number is GDMCC NO：60247.The bacterial strain is to be screened from the activated sludge that Guangzhou drips cellar for storing things sewage plant using bacterium isolation medium, is isolated, can efficiently, the copper ion of rapidly adsorption of Low Concentration, 90% can reach in 15min to the absorption of the copper ion of a concentration of 2mg/L, removal rate, thalline is 7.6mg/g wet cells to the maximal absorptive capacity of copper ion, it can be used for the improvement of low concentration copper-containing wastewater to reach wastewater discharge standard, there is larger application prospect.
Description
Technical field
The invention belongs to technical field of environmental microorganism.More particularly, to a strain capable of high-efficiency adsorption of Low Concentration copper ion
Rhizobium and its application.
Background technology
With city and industrial expansion, metallic wastewater is on the rise to the pollution of environment.Difficult to degrade in industrial wastewater,
The heavy metal contaminants of strong toxicity accumulate in water body, and the water plant of water body, animal system are generated serious harm, and can be led to
Cross the health that food chain influences the mankind.Although the heavy metal wastewater thereby of current industrial discharge can substantially reduce it through processing and contain
Amount, but the concentration of heavy metal ion in the heavy metal wastewater thereby of many industrial discharges is difficult that discharge standard is fully achieved.Wherein copper
Just heavy metal contaminants are important.
Copper is a kind of indispensable element of animals and plants, and human body, which lacks copper, can cause anaemia, and paratrichosis, bone and artery are abnormal,
So that cerebral disorders.Although copper is important essential trace element, misapplication also easily causes toxic reaction.Excess intake
Copper can lead to the damage of liver cell and red blood cell, further cause hepatic sclerosis, diarrhea, vomiting, dyskinesia and sensory nerve barrier
Hinder.
Currently, the administering method of heavy metal pollution includes mainly three categories：Physical, chemical method and bioanalysis.Physical
It takes effect with chemical method relatively fast, but high expensive, is also easy to produce secondary pollution.Bioanalysis mainly utilizes plant and microorganism pair
The absorption of heavy metal and passivation.Wherein microbial treatment heavy metal pollution has at low cost, high efficiency, to environmental disruption
Small, non-secondary pollution is suitable for the advantages such as large area repair and receives favor, has a good application prospect.
But the presently found microorganism about low concentration copper ion in absorption and removal heavy metal wastewater thereby but rarely has report
Road.
Invention content
The technical problem to be solved by the present invention is to overcome to lack copper in absorption and removal heavy metal wastewater thereby in the prior art
The defect and deficiency of the microorganism of ion, provide the rhizobium of plant height effect adsorption of Low Concentration copper ion, and the bacterial strain is to utilize
Bacterium isolation medium screens from the activated sludge that Guangzhou drips cellar for storing things sewage plant, is isolated, can efficiently, rapidly adsorb low
The copper ion of concentration can be used for the improvement of low concentration copper-containing wastewater to reach wastewater discharge standard, have larger application prospect.
The object of the present invention is to provide the rhizobium that a plant height imitates adsorption of Low Concentration copper ion.
It is a further object of the present invention to provide above-mentioned rhizobium to abolish the application in administering in heavy metal.
The above-mentioned purpose of the present invention is to give realization by the following technical programs：
One plant height imitates the rhizobium of adsorption of Low Concentration copper ion（Rhizobiumsp.）S2 bacterial strains, the bacterial strain belong to rhizobium
Belong to, which was preserved in Guangdong Province's Culture Collection on the 24th in September in 2017, and culture presevation number is GDMCC NO：
60247.Classification And Nomenclature number isRhizobiumsp.；Preservation address is No. 59 building 5 of compound of GuangZhou, China city martyr Road 100
Building.
Specifically, the 16S rDNA of the rhizobium such as SEQ ID NO：Shown in 1.
The rhizobium of the present invention（Rhizobiumsp.）S2 bacterial strains are to drip cellar for storing things dirt from Guangzhou using bacterium isolation medium
It is screened in the activated sludge of water factory, is isolated；Specifically, the separation of the bacterial strain and cultural method include the following steps：
（1）It takes activated sludge and has been subjected to sterilizing and be down to the enriched medium of room temperature by 1：3 volume ratio is uniformly mixed, and is put into wide
In mouth bottle, bottleneck is sealed with sterilized gauze, replaces half culture medium every 2d, 30 DEG C of standings are protected from light culture 30d.
（2）After enrichment culture 30d, the water sample at water sludge interface is taken, utilizes isolation medium culture 3d.After 3d, richness is taken
Collection culture solution is suitably diluted, and is taken 1mL to be inoculated in solid medium, is placed in 30 DEG C of constant incubators, and culture 3d is protected from light.It waits for
After growing bacterium colony, chooses the different bacterium colony such as shape, size, color and distinguish streak inoculation in corresponding LB culture medium flat plates, directly
It is fallen to no miscellaneous bacteria.Then the inoculation of acquisition is put into 4 DEG C of refrigerators and is saved backup in LB medium slants.
The formula of the enriched medium is：Sodium succinate 1.00g, NaNO30.10g, NaAc0.20g, sodium thioglycolate
0.05g, 0.01M chinic acid ferrous solution 2.0ml, Wolfe ' s vitamin mixtures 10.0mL, Wolfe ' s minerals mixed liquors
5.0mL, deionized water 983mL.
The formula of isolation medium is：KH2PO4 0.68g, NaNO30.12g, Wolfe ' s vitamin mixture 10.0mL,
Wolfe ' s minerals mixing 5.0ml, 0.01M chinic acid ferrous solution 2.0mL, resazurin 2mg, NaAc 0.20g, deionized water
983mL adjusts pH to 7.0 with 10M NaOH, and solid medium then separately adds 1.5% agar.
Wherein, the minerals mixture formula is：Ammonia (base) triacetic acid 1.5g, MgSO4·7H2O 3.0g, MnSO4·
H2O 0.5g, NaCl 1.0g, FeSO4·7H2O 0.1g, CoCl2·6H2O 0.1g, CaCl20.1g, ZnSO4·7H2O 0.1g,
CuSO4·5H2O0.01g, AlK (SO4)2·12H2O 0.01g, H3BO30.01g, Na2MoO42H2O 0.01g, deionized water
1.0L, with KOH tune pH to 7.0.
Rhizobium of the present invention（Rhizobiumsp.）The copper ion of S2 bacterium strain capable of high-efficiency, rapidly adsorption of Low Concentration,
It can be used for the improvement of low concentration copper-containing wastewater to reach wastewater discharge standard, there is larger application prospect.
Therefore, rhizobium of the present invention administer in low concentration copper-containing wastewater or are preparing the micro- of adsorption of Low Concentration copper ion
Application in biological agent all falls in the scope of protection of the present invention.
Preferably, in the low concentration copper-containing wastewater copper ion a concentration of 2~30mg/L.
Preferably, the pH of the low concentration copper-containing wastewater is 4~7（It is preferred that 5~6）.
Specifically, the application is that first to adjust the pH of low concentration copper-containing wastewater be 4~7, then by rhizobium of the present invention
Or its zymotic fluid is added in waste water and is handled.
Meanwhile the present invention also provides a kind of bacterial strain suspension of efficient absorption low concentration copper ion, the bacteria suspension includes
Above-mentioned rhizobium.
A kind of microorganism formulation of efficient absorption low concentration copper ion, the microorganism formulation include above-mentioned rhizobium or its
The zymotic fluid of bacterial strain.
In addition, the application of above-mentioned bacterial strains suspension or microorganism formulation in administering low concentration copper-containing wastewater is also in the present invention
In protection domain.
Specifically, the application is that above-mentioned bacterial strains suspension or microorganism formulation are added into copper-containing wastewater containing low concentration.
Compared with prior art, the invention has the advantages that：
Present invention finds the rhizobium that a plant height imitates adsorption of Low Concentration copper ion（Rhizobiumsp.）S2 bacterial strains, the bacterial strain
It is preserved in Guangdong Province's Culture Collection within 24th in September in 2017（GDMCC）, culture presevation number is GDMCC NO：
60247；The bacterial strain is to be screened, detached from the activated sludge that Guangzhou drips cellar for storing things sewage plant using bacterium isolation medium
Arrive, can efficiently, the copper ion of rapidly adsorption of Low Concentration, to the absorption of the copper ion of a concentration of 2mg/L, removal rate in 15min
90% is can reach, thalline is 7.6mg/g wet cells to the maximal absorptive capacity of copper ion, can be used for low concentration copper-containing wastewater
It administers to reach wastewater discharge standard, there is larger application prospect.
Description of the drawings
Fig. 1 is rhizobium of the present invention（Rhizobiumsp.）S2 strain development dendrograms；
Fig. 2 is rhizobium of the present invention（Rhizobiumsp.）The growth curve chart of S2 bacterial strains；
Fig. 3 is rhizobium of the present invention（Rhizobiumsp.）S2 bacterial strains change over time figure to the absorption of copper；
Fig. 4 is rhizobium of the present invention（Rhizobiumsp.）Influence of the S2 bacterial strains throwing bacterium amount to the adsorption effect of copper；
Fig. 5 is rhizobium of the present invention（Rhizobiumsp.）S2 bacterial strains removal effect figure in the MBR that actual waste water is run；
Fig. 6 is rhizobium of the present invention（Rhizobiumsp.）Microscopic comparison of the S2 bacterial strains before and after absorbing copper.
Specific implementation mode
It is further illustrated the present invention below in conjunction with Figure of description and specific embodiment, but embodiment is not to the present invention
It limits in any form.Unless stated otherwise, the present invention uses reagent, method and apparatus routinely try for the art
Agent, method and apparatus.
Unless stated otherwise, following embodiment agents useful for same and material are purchased in market.
The separation and identification of 1 bacterial strain of embodiment
1, bacterium source
Guangzhou drip cellar for storing things sewage plant activated sludge.
2, culture medium
（1）Enriched medium：Sodium succinate 1.00g, NaNO30.10g, NaAc0.20g, sodium thioglycolate 0.05g, 0.01M Kui
Buddhist nun acid ferrous solution 2.0ml, Wolfe ' s vitamin mixtures 10.0mL, Wolfe ' s minerals mixed liquor 5.0mL, deionized water
983mL。
（2）Isolation medium：KH2PO4 0.68g, NaNO30.12g, Wolfe ' s vitamin mixture 10.0mL,
Wolfe ' s minerals mixing 5.0ml, 0.01M chinic acid ferrous solution 2.0mL, resazurin 2mg, NaAc 0.20g, deionized water
983mL adjusts pH to 7.0 with 10M NaOH, and solid medium then separately adds 1.5% agar.
（3）Wolfe ' s vitamin mixtures：Biotin 2.0mg, folic acid 2.0mg, pyridoxine hydrochloride 10.0mg, hydrochloric acid sulphur
Amine element 5.0mg, riboflavin 5.0mg, niacin 5.0mg, calcium pantothenate 5.0mg, vitamin B120.1mg, p-aminobenzoic acid 5.0mg,
Lipoic acid 5.0mg, deionized water 1.0L.
（4）Minerals mixed liquor：Ammonia (base) triacetic acid 1.5g, MgSO4·7H2O 3.0g, MnSO4·H2O 0.5g, NaCl
1.0g, FeSO4·7H2O 0.1g, CoCl2·6H2O 0.1g, CaCl20.1g, ZnSO4·7H2O 0.1g, CuSO4·
5H2O0.01g, AlK (SO4)2·12H2O 0.01g, H3BO30.01g, Na2MoO42H2O 0.01g, deionized water 1.0L are used
KOH tune pH to 7.0.
（5）Fermentation medium：Bacto peptone 10g, yeast extract powder 5g, NaCl 5g, glucose 1g and pure water
1000mL, pH are adjusted to 7.0 ± 0.2.
3, the separation, screening and identification of bacterial strain
（1）It takes activated sludge and has been subjected to sterilizing and be down to the enriched medium of room temperature by 1：3 volume ratio is uniformly mixed, and is put into wide
In mouth bottle, bottleneck is sealed with sterilized gauze, replaces half culture medium every 2d, 30 °C of standings are protected from light culture 30d.Work as enrichment
After cultivating 30d, the water sample at water sludge interface is taken, isolation medium culture 3d is utilized.After 3d, enrichment culture liquid is taken to carry out appropriate dilute
It releases, takes 1mL to be inoculated in solid medium, be placed in 30 DEG C of constant incubators, be protected from light culture 3d.After growing bacterium colony, choose shape
The different bacterium colony such as shape, size, color distinguishes streak inoculation in corresponding LB culture medium flat plates, up to no miscellaneous bacteria falls.Then will
The inoculation of acquisition is put into 4 °C of refrigerators and is saved backup in LB medium slants.
（2）The bacterial strain determined after screening and culturing is cultivated into 1~2d in 30 °C, then carries out 16S rDNA sequencing identifications.It is logical
Cross the 16S rDNA sequences that PCR obtains the bacterial strain（Such as SEQ ID NO：Shown in 1）, through sequencing and BLAST homology alignments
It is analyzed with phylogenetic evolution（Shown in Fig. 1）, the results showed that it is rhizobium in isolated strain classification meaning
（Rhizobiumsp.）, it is named asRhizobiumSp.S2 bacterial strains, and growth curve is measured with colony counting method,
The results are shown in Figure 2：4-12h is exponential phase, and 12h-36h is slow growth period, is decline phase after 36h.By the bacterium
Strain was preserved in Guangdong Province's Culture Collection on the 24th in September in 2017, and culture presevation number is GDMCC NO：60247；Point
Class name number isRhizobiumsp.；Preservation address is 5 building, the building of compound the 59th of GuangZhou, China city martyr Road 100.
Study on adsorption properties of 2 bacterial strain of embodiment to copper
1, influence of the adsorption time to adsorption effect
（1）By the inoculation of embodiment 1 in liquid fermentation medium, 30 DEG C of 2 d of 150rpm shaking table cultures.It is collected by centrifugation
Wet bacterium is washed with deionized 3 times.The wet bacterium for collecting clean is weighed into 0.06g, is put into equipped with 100mL cuprics（2mg/L）It is molten
Liquid（pH=6.0±0.2）In, it puts and shakes up absorption into shaking table（30 DEG C, 150rpm）.It is identical dense to be not added with containing for thalline simultaneously
The aqueous solution of degree copper ion concentration compares, and is respectively 0,15,30,60,90,120,180min to sample successively in adsorption time
Afterwards, 10min is centrifuged with 5000rpm rotating speeds, with 0.22 micron of membrane filtration, filtrate is taken to measure copper ion in the front and back solution of absorption
The variation of concentration.
Removal rate（R）Computational methods：
R（%）=（C0-Ct）/C0*100%
Wherein, C0For the initial concentration of copper ion in solution, the dense of the copper ion in solution after thalline adsorbs t minutes is added in Ct
Degree.
（2）The results are shown in Figure 3, and the rhizobium strains of embodiment 1 is fast to being adsorbed between 0~15min for copper ion
Fast absorption phase and adsorption efficiency depend primarily on the stage, enter absorption phase at a slow speed after 15min, reach in 90min
Balance.
2, influence of the copper ion initial concentration to adsorption effect
（1）By the inoculation of embodiment 1 in liquid fermentation medium, 30 DEG C of 2 d of 150rpm shaking table cultures.It is collected by centrifugation
Wet bacterium is washed with deionized 3 times.The wet bacterium for collecting clean is weighed into 0.06g, is respectively put into equipped with 100mL cuprics（2,5,
10,15,20,30mg/L）Solution（pH=6.0±0.2）In, it puts and shakes up absorption 2h into shaking table（30 DEG C, 150rpm）.Simultaneously
The aqueous solution containing same concentrations copper ion concentration to be not added with thalline compares, and is sampled successively after adsorbing 2h, with 5000rpm
Rotating speed centrifuges 10min, with 0.22 micron of membrane filtration, filtrate is taken to measure the variation of copper ion concentration in the front and back solution of absorption.
The computational methods of adsorbance：
Q=（C0-Ct）/Cb*100%
Wherein, C0For the initial concentration of copper ion in solution, Ct is the dense of the copper ion in solution after addition thalline adsorbs t minutes
Degree, CbFor the concentration of bacterium in solution.
（2）The results show that with the raising of copper ion concentration, adsorption efficiency declines.Because of single bacterial adsorption copper ion
Ability be limited, throwing bacterium amount is fixed, and with the increase of copper ion concentration, removal rate reduces.The unit adsorbance of bacterium increases,
Maximum value is 7.6mg/g wet cells, reaches an equilibrium state.
3, influence of the throwing bacterium amount to adsorption effect
（1）By the inoculation of embodiment 1 in liquid fermentation medium, 30 DEG C of 150rpm shaking table cultures 2d.Bacterium is collected by centrifugation
Body is washed with deionized 3 times.The thalline for collecting clean is weighed into 0.02,0.04,0.06,0.08,1.0,1.2g wet bacterium respectively
Body is respectively put into equipped with 100mL cuprics（2mg/L）Solution（pH=6.0±0.2）In, it puts and shakes up absorption 2h into shaking table（30
DEG C, 150rpm）.Simultaneously compareed with the aqueous solution containing same concentrations copper ion concentration for being not added with thalline, after adsorbing 2h according to
Sub-sampling centrifuges 10min with 5000rpm rotating speeds, with 0.22 micron of membrane filtration, filtrate is taken to measure copper in the front and back solution of absorption
The variation of ion concentration.
（2）The results are shown in Figure 4, and with the increase of throwing bacterium amount, the removal rate of copper ion gradually rises, and is in throwing bacterium amount
After 0.6g/L, removal rate increasing degree tapers into.
4, influences of the pH to adsorption effect
（1）By the inoculation of embodiment 1 in liquid fermentation medium, 30 DEG C of 2 d of 150rpm shaking table cultures.It is collected by centrifugation
Thalline is washed with deionized 3 times.The thalline for collecting clean is weighed into 0.06g wet thallus, it is 1,2,3,4,5 to be respectively put into pH,
6,7 100mL copper ion solutions（2mg/L）In, it puts and shakes up absorption 2h into shaking table（30 DEG C, 150rpm）.Simultaneously to be not added with bacterium
The aqueous solution containing same concentrations copper ion concentration of body compares, and is sampled successively after adsorbing 2h, is centrifuged with 5000rpm rotating speeds
10min takes filtrate to measure the variation of copper ion concentration in the front and back solution of absorption with 0.22 micron of membrane filtration.
（2）The results show that bacterial strain is maximum in pH=6 to the unit adsorbance of copper ion, and when pH is 1~3, does not almost have
There is any adsorption effect, when pH is 3~6, adsorption effect enhances as its pH value increases, and adsorption effect is again with pH later
Value is increased and is reduced.
In conclusion bacterium to the removal effect optimal conditions of copper be its PH=6, throwing bacterium amount be 1.2g/L when to initial
In 2mg/L copper ions, the removal rate after adsorbing 15min reaches 90% or more to concentration.
Application of 3 bacterial strain of embodiment on actual industrial tail water
1, the application on actual industrial tail water
Zhongshan City, Guangdong Province electroplating processes sewage plant is to found the factory for 2011, a little heavy metal containing sewages is mainly handled, because in recent years
The raising of environmental requirement, the original treatment facility of water factory have been unable to meet wastewater treatment requirement up to standard, and the said firm is quasi- to carry out skill
Change.The factory and office manage tail water detection, copper ion concentration in 2~5mg/L or so, PH between 4~5, COD 20~30mg/L it
Between, take the tail water to carry out rhizobium adsorption experiment in laboratory.
MBR system, system volume 9L, two groups of systems, respectively blank group and experimental group are built in laboratory.System connects
Reforwarding row（8 hours of hydraulic detention time）, using sewage plant tail water as water inlet, rhizobium are added, after observing its good biofilm,
Detection water outlet copper ion concentration.
Its treatment effect as shown in figure 5, influent concentration is 2.2mg/L or so, 2 as a child outlet effect drop is remarkably reinforced
Low, 6 as a child tended towards stability, and MBR system is 76% or so to the removal rate of copper in actual waste water.
2, under microscope before and after microorganism adsorption copper ion form variation
Biomembrane in experimental group and blank group is carried out under scanning electron microscope it has been observed that the results are shown in Figure 6, blank group
The form for seeing bacterium of energy complete display is rod-shaped, and ne ar is full mellow and full；Experimental group cell surface is adhered, by substance packet
It wraps up in, cell shape is imperfect.The cell surface of experimental group can be brighter than blank group, according to scanning electron microscope principle of instrument, heavy metal
Substance is shown as white speck on instrument, a surface part is accumulated in after thalline absorption zinc ion, so having around thalline
Light, it is metallic copper substance to illustrate that cell surface is adhered substance.
From the point of view of application experiment, the isolated rhizobium of the present invention have preferable absorption to the heavy metal wastewater thereby of low concentration
Effect.
Sequence table
<110>Zhongshan University
<120>One plant height imitates rhizobium and its application of adsorption of Low Concentration copper ion
<130> YG18100886AA042
<141> 2018-03-14
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1081
<212> DNA
<213>Rhizobium S2 (Rhizobium sp.S2)
<400> 1
gtgggggcag gcttacacat gcaagtcgaa cgccccgcaa ggggagtggc agacgggtga 60
gtaacgcgtg ggaacatacc ctttcctgcg gaatagctcc gggaaactgg aattaatacc 120
gcatacgccc tacgggggaa agatttatcg gggaaggatt ggcccgcgtt ggattagcta 180
gttggtgggg taaaggccta ccaaggcgac gatccatagc tggtctgaga ggatgatcag 240
ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtgg ggaatattgg 300
acaatgggcg caagcctgat ccagccatgc cgcgtgagtg atgaaggcct tagggttgta 360
aagctctttc accgatgaag ataatgacgg tagtcggaga agaagccccg gctaacttcg 420
tgccagcagc cgcggtaata cgaagggggc tagcgttgtt cggaattact gggcgtaaag 480
cgcacgtagg cggatattta agtcaggggt gaaatcccgc agctcaactg cggaactgcc 540
tttgatactg ggtatcttga gtatggaaga ggtaagtgga attccgagtg tagaggtgaa 600
attcgtagat attcggagga acaccagtgg cgaaggcggc ttactggtcc attactgacg 660
ctgaggtgcg aaagcgtggg gagcaaacag gattagatac cctggtagtc cacgccgtaa 720
acgatgaatg ttagccgtcg ggcagtatac tgttcggtgg cgcagctaac gcattaaaca 780
ttccgcctgg ggagtacggt cgcaagatta aaactcaaag gaattgacgg gggcccgcac 840
aagcggtgga gcatgtggtt taattcgaag caacgcgcag aacccttacc agctcttgac 900
attcggggta tgggcattgg agacgatgtc cttcagttag gctgggcccc agaacaggtg 960
ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt ttaagtcccg cacgagcgca 1020
ccctcgccct tagttgcagc atttagtttg gcacttttaa ggggactgcc gggtgaataa 1080
g 1081
Claims (9)
1. a plant height imitates the rhizobium of adsorption of Low Concentration copper ion（Rhizobiumsp.）S2 bacterial strains, which is characterized in that the bacterium
Strain was preserved in Guangdong Province's Culture Collection on the 24th in September in 2017, and culture presevation number is GDMCC NO：60247.
2. rhizobium according to claim 1, which is characterized in that the 16S rDNA of the rhizobium such as SEQ ID NO：1
It is shown.
3. rhizobium described in claim 1 administer in low concentration copper-containing wastewater or in the microorganisms for preparing adsorption of Low Concentration copper ion
Application in preparation.
4. application according to claim 3, which is characterized in that a concentration of the 2 of copper ion in the low concentration copper-containing wastewater
~30mg/L.
5. application according to claim 3, which is characterized in that first adjust the pH to 4~7 of low concentration copper-containing wastewater, then will
Bacterial strain described in claim 1 or its zymotic fluid are added in waste water.
6. a kind of bacterial strain suspension of efficient absorption low concentration copper ion, which is characterized in that include root nodule described in claim 1
Bacterium.
7. a kind of microorganism formulation of efficient absorption low concentration copper ion, which is characterized in that include rhizobium described in claim 1
Or its zymotic fluid.
8. the application of bacterial strain suspension described in claim 6 or 7 microorganism formulations in administering low concentration copper-containing wastewater.
9. application according to claim 8, which is characterized in that the application is to add power into copper-containing wastewater containing low concentration
Profit requires the 6 bacterial strain suspension or 7 microorganism formulations.
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