CN108265013A - One strain of sulfate reduction bacteria and its technique for recycling valuable metal in zinc smelting wastewater - Google Patents
One strain of sulfate reduction bacteria and its technique for recycling valuable metal in zinc smelting wastewater Download PDFInfo
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- CN108265013A CN108265013A CN201611252246.3A CN201611252246A CN108265013A CN 108265013 A CN108265013 A CN 108265013A CN 201611252246 A CN201611252246 A CN 201611252246A CN 108265013 A CN108265013 A CN 108265013A
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Abstract
The present invention provides a strain of sulfate reduction bacteria, the taxology of the bacterial strain is named as Clostridium sartagoforme GRINMds1, and depositary institution is：China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, preservation date：On December 20th, 2016, deposit number：CGMCC No.13369.The bacterial strain has chlorine-resistant, resistance to zinc ion ability, it can be achieved that the efficient process and valuable metal recovery of zinc smelting wastewater, so as to fulfill Heavy Metal Pollution Control.Available for the sulfate reduction reparation of the high zinc Tailings Dam of high chlorine, great market development prospect.
Description
Technical field
The invention belongs to microbe subject fields, are related to the bacterial strain of a strain of sulfate reduction ability, which has
Chlorine-resistant, resistance to zinc ion ability and its technique for recycling valuable metal in zinc smelting wastewater.
Background technology
China is smelted heavy metal wastewater thereby and is often handled at present using lime neutralisation, generates a large amount of neutralization waste residue,
Processing cost is high and is also easy to produce secondary pollution.There is larger application prospect to such wastewater treatment using sulfate reducing bacteria, it can
The valuable metal in waste water is recycled with offset operation cost.However, existing sulfate reducing bacteria in anaerobic fermentation to restore sulfuric acid
The bioprocess of root is more sensitive to chlorion and heavy metal ion such as zinc ion concentration etc., and sulfate reduction rate is low, limits
This application of biological water treatment technology in zinc smelting wastewater processing.
Invention content
First purpose of the present invention is to provide a strain of sulfate reduction bacteria, the bacterium can high Cl- concentration and high zinc from
In the presence of sub- concentration, efficiently sulphate reducing.
Second object of the present invention is to provide a kind of technique of valuable metal in reclaiming smelting waste water, using the technique,
50% processing cost can be reduced, the heavy metal ion such as removal zinc, lead, arsenic, cadmium, copper are stablized and ensure pollutant concentration in water outlet
Less than the requirement of national standard.
To achieve these goals, the present invention provides following technical solution：
The present invention provides a kind of sulfate reducing bacteria, and the Classification And Nomenclature of the bacterium is Clostridium sartagoforme
GRINMds1, depositary institution are：China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are：Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, preservation date：On December 20th, 2016, preservation are compiled
Number：CGMCC No.13369.
The present invention sulfate reducing bacteria (Clostridium sartagoforme) GRINMds1 colony characteristics be：Solid
35 DEG C of growths are very fast on body culture medium, and colony diameter is 3mm after 3 days, and bacterium colony is in regular circle shapes, and quality is fine and close.
The sample of extraction bacterium used of the invention comes from the bed mud in river on Guangxi mineral products periphery, after screening detaches,
Obtain one plant of chlorine-resistant, resistance to zinc ion sulfate reduction bacterial strain, number 40#Bacterial strain.By Morphological Identification, the 16S of bacterial strain
RDNA Sequence Identifications are identified and are named as Clostridium sartagoforme GRINMds1 bacterial strains, after testing the bacterial strain
With preferable sulfate reduction ability.
The present invention also provides a kind of culture medium for amplifying the above-mentioned sulfate reducing bacteria of culture, which is：It should
The preparation method of culture medium is：Basal medium：1.0g/L ammonium chlorides, 2.0g/L sodium sulphate, 1.0g/L zinc sulfate, 2.0g/L seven
Water magnesium sulfate, 0.5g/L potassium dihydrogen phosphates, 0.1g/L calcium chloride, 3.0g/L peptones, 2.8mL/L sodium lactates, pH7.0~7.5,
121 DEG C of sterilizing 20min coolings are spare；Iron ammonium sulfate mother liquor：10g/L iron ammonium sulfates, 0.1g/L ascorbic acids, 4g/L L-
Cysteine hydrochloride, filtration sterilization；By basal medium and iron ammonium sulfate mother liquor according to volume ratio 20:1 ratio mixing
Uniformly.
The present invention also provides a kind of techniques for recycling valuable metal in zinc smelting wastewater, include the following steps：
1) zinc smelting wastewater tune pH is neutralized to pH5.0-6.0；
2) right to use profit is inoculated in bioreactor and requires sulfate described in the claim 1 of 2 medium cultures
Reducing bacteria, bacteria concentration 108A/mL, inoculation volume make a living the 1% of object reactor volume, carbon source then are passed through bioreactor；
3) zinc smelting wastewater for making step 1) neutralized enters contact reactor progress selective precipitation according to sequence and obtains
Heavy metal precipitation object and supernatant, heavy metal precipitation object enter zinc as raw material and select smelting system, and supernatant enters bioreactor,
And pipeline is set to flow back between contact reactor and bioreactor；
4) bioreactor outflow waste water is passed through aerobic reactor to react, reaction solution discharge or reuse, reaction production
Bioreactor is returned after raw sludge condensation as carbon source.
Preferably, step 1) step is：Zinc smelting wastewater is flowed into neutralization reactor, milk of lime is added in and adjusts pH, fully
It is stood after stirring, obtains sediment and supernatant, sediment obtains filter residue and filtrate after press filtration, and supernatant and filtrate are merged
As neutralized zinc smelting wastewater.
Preferably, carbon source described in step 2) is final concentration of：Glycerine 9%, sodium lactate 7%.
Preferably, the pH of contact reactor described in step 3) is 5.0-6.0.
Preferably, the HRT of contact reactor described in step 3) is 2-3h.
Preferably, the bioreactor is up flow anaerobic sludge blanket reactor, and internal filling glass microballon, glass is micro-
A diameter of 1~the 2mm of pearl, bulk density are 270 ± 30kg/m3, filling rate 20%.
Preferably, the HRT of the bioreactor is 14-18h, and temperature is 28~35 DEG C in bioreactor.
Preferably, the HRT of the aerobic reactor is 2-6h.
HRT is hydraulic detention time, refers to mean residence time of the treatment sewage in reactor.
The advantage of the invention is that：
1. the sulfate reducing bacteria used is under faintly acid pH (pH5.0-6.0), there are high-concentration chlorine ions and zinc ion
In the case of, there is sulfate-reducing activity, improve the adaptability of wastewater treatment.
2. technique provided by the invention uses the design of split-phase, suppression of the metal ion to microorganism in bioreactor is reduced
System；Simultaneously because microorganism growing environment maintains pH6-7 in bioreactor so that sulfate reduction rate rise to 80% with
On.
3. bioreactor uses upflow anaerobic sludge blanket process (UASB) reactor, the quick separating of muddy water can be realized,
Reflux only withdrawing fluid, and sludge microbe will not be refluxed；Pass through the S generated in the bioreactor that flows back2-Ion energy
Enough selective precipitation ponds for returning to front in real time, reduce S2-The inhibition of microorganism in ion pair bioreactor.
4. biological sulfate-reducing process production alkaline consumption acid, the basicity of generation can be used for being promoted the pH value in selective precipitation pond,
It is popped one's head in online by pH and water pump can realize that the control of pH in selective precipitation pond simultaneously realizes the selection of valuable heavy metal
Property precipitation.
5. the aerobic reaction tank of postposition can make full use of carbon source remaining in water outlet to generate sludge, while reach water outlet
Mark；The bioreactor that can flow back after the sludge condensation of generation does carbon source, reduces the consumption of carbon source.
The beneficial effects of the present invention are：
The present invention provides a kind of sulfate reducing bacteria and the technique using valuable metal in the recycling zinc smelting wastewater of the bacterium,
Contact reactor selective precipitation is controlled to recycle valuable metal using reflux, technique provided by the invention can compare common process
Cost reduction 30-50%.
Description of the drawings
Fig. 1 is the scanning electron microscope (SEM) photograph of sulfate reducing bacteria strain provided by the present invention.
Fig. 2 is the process flow diagram of the technique of valuable metal in recycling zinc smelting wastewater provided by the invention.
Specific embodiment
The present invention includes the concentration and separation of the organic matter degradation bacterial strain of enduring high-concentration heavy metal ion, screening purifying, drop
Solution ability detects and identification, is described as follows：
Sulfate reducing bacteria used in the present invention, the taxology of the bacterium are named as Clostridium sartagoforme
GRINMds1, depositary institution are：China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are：Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, preservation date：On December 20th, 2016, preservation are compiled
Number：CGMCC No.13369.
Embodiment 1
The concentration and separation of sulfate reducing bacteria and purifying
(1) culture medium
Basic solid medium：1.0g/L ammonium chlorides, 2.0g/L sodium sulphate, 1.0g/L zinc sulfate, seven water sulfuric acid of 2.0g/L
Magnesium, 0.5g/L potassium dihydrogen phosphates, 0.1g/L calcium chloride, 3.0g/L peptones, 2.8mL/L sodium lactates, agar powder 20g, pH7.4~
7.6。
Basal liquid medium：1.0g/L ammonium chlorides, 2.0g/L sodium sulphate, 1.0g/L zinc sulfate, seven water sulfuric acid of 2.0g/L
Magnesium, 0.5g/L potassium dihydrogen phosphates, 0.1g/L calcium chloride, 3.0g/L peptones, 2.8mL/L sodium lactates, pH7.0~7.5.
121 DEG C of sterilizing 20min coolings are spare in high-pressure sterilizing pot respectively for above-mentioned basal medium.It is another to prepare ferrous sulfate
Ammonium mother liquor：10g/L iron ammonium sulfates, 0.1g/L ascorbic acids, 4g/L L-cysteine hydrochlorides, per 100mL after filtration sterilization
Basal medium adds in the 5mL iron ammonium sulfate mother liquors, and solid medium and fluid nutrient medium is made.
(2) sample collection and preparation
The sample of separation of bacterial used in this experiment comes from the tailing sample in Guangxi somewhere, 10 under surface of tailings reservoir~
1kg tailings sample is taken to be packed into sterile sealed bag kind at 20cm, sample is taken back by laboratory by cold chain transportation and places interior for 24 hours
It is saved backup in 4 DEG C of refrigerators.
(3) enrichment culture of sulfate reduction bacterial strain
The tailing sample of precise 20g is put into the 250mL conical flasks equipped with 100mL fluid nutrient mediums, uses magnetic agitation
Device stirs 40min, stratification with the rotating speed of 300r/min.Supernatant is taken to observe, bacterium amount is less.
20mL supernatants are taken in the 250mL conical flasks equipped with 200mL aforesaid liquid culture mediums, seal bottleneck in 35 DEG C of trainings
It supports case and stands enrichment culture, the Lead acetate paper of a moistening is placed in advance in bottle mouth position.When the complete blackening of culture medium, lead acetate
Test paper blackening, the gas of apparent rotten egg smell can be smelt by opening bottleneck, illustrate sulfate reducing bacteria amount reproduction, bacterium solution
Concentration reaches 108A/mL.Conical flask is taken out, supernatant is taken to carry out 10 respectively-1、10-2、10-3、10-4、10-5、10-6, six ladders
The dilution of degree.Sterilized solid medium is poured into plate, thickness probably accounts for 1/3rd of plate.After agar cooling,
The 20 μ L of bacteria suspension of each dilution gradient are added in into corresponding culture dish respectively, it is uniform with coating rod coating.Treat that bacterium solution infiltration is several
After minute, import the solid medium of a certain amount of cooling again in plate, make 2/3rds of the entire tablet of culture fiduciary point, bacterium solution
It is clipped between two layers of culture medium, causes the environment of anaerobism.The tablet made in triplicate, is placed on 35 DEG C of incubators by each concentration
In, quiescent culture 3~4 days observes the variation of flat-plate bacterial colony daily.
(4) purifying of sulfate reduction bacterial strain
The variation of bacterium colony on solid plate is observed, with connecing the bacterium colony of the single color blackening of collarium picking to being equipped with 10ml liquid
In the test tube of culture medium, quiescent culture obtains single pure bacterium for three days under the conditions of 35 DEG C.
Embodiment 2
The identification of strain
The colonial morphology on solid medium tablet is observed, and the form of bacterial strain is observed with light microscope, by gained
The morphological feature of bacterial strain with《Fungal identification handbook compares (Wei Jingchao works)》It compares, obtains the Preliminary Identification knot of wanted bacterial strain
Fruit.Then with the 16S rDNA sequences of PCR amplification bacterial strain and sequencing analysis analysis is carried out again, the following SEQ ID of sequencing data
Shown in No.1.
By the 16S sequences that sequencing obtains to NCBI public databases (https://blast.ncbi.nlm.nih.gov/
Blast.cgiPROGRAM=blastn＆PAGE_TYPE=Blast Search＆LINK_LOC=blasthome) compared
Right, comparison result is Clostridium sartagoforme, is named as GRINMds1, scanning electron microscope (SEM) photograph is as shown in Figure 1.
Embodiment 3
The technological process of valuable metal in certain zinc smelting wastewater is recycled with reference to shown in figure 2.
Step 1：Certain zinc smelting wastewater flows into neutralization reactor (neutralization pond), adds in milk of lime and adjusts pH, control pH is
5.7, it is stood after being sufficiently stirred, obtains sediment and supernatant, sediment obtains filter residue (i.e. gypsum tailings) and filtrate after press filtration,
Supernatant and filtrate are combined into neutralized zinc smelting wastewater；
Step 2：Sulfate reducing bacteria GRINMds1 is added in fluid nutrient medium 35 DEG C of quiescent cultures to bacteria concentration
108A/mL is inoculated into bioreactor for the 1% of reaction system according to inoculation volume, then adds in glycerine, sodium lactate is extremely
Final concentration (mass percent) is glycerine 9%, and sodium lactate 7% is used as carbon source；
Bioreactor used is upflow anaerobic sludge blanket process (UASB) reactor, and internal filling glass microballon, glass is micro-
A diameter of 1~the 2mm of pearl, bulk density are 270 ± 30kg/m3, filling rate 20%.
Step 3：The neutralized zinc smelting wastewater that step 1 is obtained enters haptoreaction with bioreactor water outlet
Device is stood after being sufficiently stirred, and obtains heavy metal precipitation object and the supernatant based on zinc sulphide, the heavy metal based on zinc sulphide
Sediment can enter zinc as raw material and select smelting system, wherein, the pH of contact reactor is 5.7, the HRT (waterpower of contact reactor
Residence time) for 2.5h, the outlet port of pipeline that contact reactor connection bioreactor is discharged should be arranged on haptoreaction
1.5m under liquid level in device；
Under this pH, the Zn in solution2+Ion can be with the S in bioreactor phegma2-Ionic reaction generates
ZnS realizes the precipitation of the heavy metals such as zinc.
Then supernatant fraction is made to flow into bioreactor, influx 100%, bioreactor water outlet overflow (is returned
Stream) to contact reactor, wherein, bioreactor HRT is 15h, and water temperature is maintained at 33 DEG C in bioreactor；
Bioreactor can Growth of Sulfate Reducing Bacteria, sulfate reducing bacteria using carbon source reduction water body in sulfate radical be
S2-Ion, by containing generation S2-The reaction solution of ion is back to contact reactor, can make S2-Ion and contact reactor into
Zn in water2+Reaction, generation ZnS precipitations, for recycling.
Step 4：Remaining supernatant, which enters in aerobic reactor, after step 3 reflux reacts, and aerobic reactor HRT is
4h, aerobic reactor water outlet can direct emission, react and bioreactor returned after the sludge condensation of generation as carbon source.
Contact reactor is to the gravity overflow that bioreactor to aerobic reactor is by high process control, in biological respinse
The process setting water pump of device return contact reactor flows back.
Heavy metal in being discharged after zinc smelting wastewater and processing is detected, testing result is shown in Table 1.Heavy metal is obtained to sink
The content of Zn-ef ficiency is 15.3% in starch.
Table 1 (unit mg/L)
| Zn | As | Cu | Cd | Pb | Cl | |
| Raw water | 123.4 | 50.4 | 0.3 | 11.6 | 70.4 | 200 |
| Water outlet | 0.97 | 0.11 | 0.09 | 0.01 | 0.12 | 140 |
| Emission limit a) | 1.5 | 0.5 | 0.5 | 0.05 | 0.3 | - |
a)GB25466-2010《Lead Zn Cr coating pollutant emission standard》
From above-described embodiment as can be seen that sulfate reducing bacteria provided by the invention not only have very strong chlorine-resistant, resistance to zinc from
Sub- ability, and heavy metal can be effectively removed, realize qualified discharge.
SEQUENCE LISTING
<110>Beijing Non-Ferrous Metal Research General Academy
<120>One strain of sulfate reduction bacteria and its technique for recycling valuable metal in zinc smelting wastewater
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1311
<212> DNA
<213> Clostridium sartagoforme
<400> 1
ctagcggcgg acgggtgagt aacacgtggg caacctgcct tatagagggg aatagccttc 60
cgaaaggaag attaataccg cataacatta cattttcgca tgaagaagta attaaaggag 120
taatccgcta taagatgggc ccgcggcgca ttagctagtt ggtgaggtaa cggctcacca 180
aggcgacgat gcgtagccga cctgagaggg tgatcggcca cattgggact gagacacggc 240
ccagactcct acgggaggca gcagtgggga atattgcaca atgggggaaa ccctgatgca 300
gcaacgccgc gtgagtgatg aaggtcttcg gatcgtaaag ctctgtcttc agggacgata 360
atgacggtac ctgaggagga agccacggct aactacgtgc cagcagccgc ggtaatacgt 420
aggtggcgag cgttgtccgg atttactggg cgtaaaggga gcgtaggcgg atttttaagt 480
gagatgtgaa atacccgggc tcaacttggg tgctgcattt caaactggaa gtctagagtg 540
caggagagga gagtggaatt cctagtgtag cggtgaaatg cgtagagatt aggaagaaca 600
ccagtggcga aggcgactct ctggactgta actgacgctg aggctcgaaa gcgtggggag 660
caaacaggat tagataccct ggtagtccac gccgtaaacg atgaatacta ggtgtagggg 720
ttgtcatgac ctctgtgccg ccgcaaacgc attaagtatt ccgcctgggg agtacggtcg 780
caagattaaa actcaaagga attgacgggg gcccgcacaa gcagcggagc atgtggttta 840
attcgaagca acgcgaagaa ccttacctag acttgacatc tcctgaatta ctcttaatcg 900
aggaagtccc ttcggggaca ggaagacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt 960
gagatgttgg gttaagtccc gcaacgagcg caacccttat tgttagttgc taccattaag 1020
ttgagcactc tagcaagact gcccgggtta accgggagga aggtggggat gacgtcaaat 1080
catcatgccc cttatgtcta gggctacaca cgtgctacaa tggcaagtac aaaaagatgc 1140
aataccgcaa ggtggagcca aacttaaaaa cttgtctcag ttcggattgt aggctgaaac 1200
tcgcctacat gaagctggag ttgctagtaa tcgcgaatca gaatgtcgcg gtgaatacgt 1260
tcccgggcct tgtacacacc gcccgtcaca ccatgagagt tggcaatacc t 1311
Claims (10)
- A 1. strain of sulfate reduction bacteria, which is characterized in that the taxology of the bacterium is named as Clostridium sartagoforme GRINMds1, depositary institution are：China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are：Beijing The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, preservation date：On December 20th, 2016, preservation are compiled Number：CGMCC No.13369.
- A kind of 2. culture medium that sulfate reducing bacteria as described in claim 1 is cultivated for amplification, which is characterized in that the culture The preparation method of base is：Basal medium：1.0g/L ammonium chlorides, 2.0g/L sodium sulphate, 1.0g/L zinc sulfate, seven water sulphur of 2.0g/L Sour magnesium, 0.5g/L potassium dihydrogen phosphates, 0.1g/L calcium chloride, 3.0g/L peptones, 2.8mL/L sodium lactates, pH7.0~7.5,121 DEG C sterilizing 20min cooling it is spare；Iron ammonium sulfate mother liquor：10g/L iron ammonium sulfates, 0.1g/L ascorbic acids, half Guangs of 4g/L L- Propylhomoserin hydrochloride, filtration sterilization；By basal medium and iron ammonium sulfate mother liquor according to volume ratio 20:1 ratio is uniformly mixed.
- 3. a kind of technique for recycling valuable metal in zinc smelting wastewater, which is characterized in that include the following steps：1) zinc smelting wastewater tune pH is neutralized to pH5.0-6.0；2) right to use profit is inoculated in bioreactor and requires sulfate reduction described in the claim 1 of 2 medium cultures Bacterium, bacteria concentration 108A/mL, inoculation volume make a living the 1% of object reactor volume, carbon source then are passed through bioreactor；3) zinc smelting wastewater for making step 1) neutralized enters contact reactor progress selective precipitation according to sequence and obtains a huge sum of money Belonging to sediment and supernatant, heavy metal precipitation object enters zinc as raw material and selects smelting system, and supernatant enters bioreactor, and Pipeline is set to flow back between contact reactor and bioreactor；4) bioreactor outflow waste water is passed through aerobic reactor to react, generation is reacted in reaction solution discharge or reuse Bioreactor is returned after sludge condensation as carbon source.
- 4. as claimed in claim 3 in recycling zinc smelting wastewater valuable metal technique, which is characterized in that step 1) step is： Zinc smelting wastewater is flowed into neutralization reactor, milk of lime is added in and adjusts pH, stood after being sufficiently stirred, obtain sediment and supernatant, Sediment obtains filter residue and filtrate after press filtration, and it is neutralized zinc smelting wastewater that supernatant and filtrate, which are merged,.
- 5. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that carbon described in step 2) Source it is final concentration of：Glycerine 9%, sodium lactate 7%.
- 6. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that connect described in step 3) The pH for touching reactor is 5.0-6.0.
- 7. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that connect described in step 3) The HRT for touching reactor is 2-3h.
- 8. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that the bioreactor For up flow anaerobic sludge blanket reactor, internal filling glass microballon, Glass Bead Diameter is 1~2mm, bulk density 270 ±30kg/m3, filling rate 20%.
- 9. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that the bioreactor HRT for 14-18h, temperature is 28~35 DEG C in bioreactor.
- 10. the technique of valuable metal in zinc smelting wastewater is recycled as claimed in claim 3, which is characterized in that the aerobic reaction The HRT of device is 2-6h.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110078209A (en) * | 2019-05-16 | 2019-08-02 | 桂林电子科技大学 | A kind of method of micro-reduction sulfate formation cadmium sulphur mine |
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