CN103173356A - Intermediate-temperature mineral leaching composite microbial system for leaching copper pyrites - Google Patents

Intermediate-temperature mineral leaching composite microbial system for leaching copper pyrites Download PDF

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CN103173356A
CN103173356A CN2011104396324A CN201110439632A CN103173356A CN 103173356 A CN103173356 A CN 103173356A CN 2011104396324 A CN2011104396324 A CN 2011104396324A CN 201110439632 A CN201110439632 A CN 201110439632A CN 103173356 A CN103173356 A CN 103173356A
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leaching
composite microbial
microbial system
chalcopyrite
temperature
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CN103173356B (en
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刘文彦
陈勃伟
刘兴宇
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GRINM Resources and Environment Technology Co Ltd
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Beijing General Research Institute for Non Ferrous Metals
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

The invention provides an intermediate-temperature mineral leaching composite microbial system for leaching copper pyrites. The composite microbial system is named Sulfobacillus thermosulfidooxidans 6Y-1, is preserved in the China center for type culture collection (CCTCC) in Wuhan University in November 10th 2010, and has a preservation number of CCTCC M2010297. The composite microbial system consists of 77% of Sulfobacillus thermosulfidooxidans, 5% of Acidithiomicrobium sp, 12% of Acidithiobacillus caldus and 6% of Ferroplasma sp. The microbial system has high copper pyrite leaching efficiency.

Description

The temperature that is used for Leaching of chalcopyrite is soaked the ore deposit composite microbial system
Technical field
The present invention relates to a temperature that is used for Leaching of chalcopyrite and soak the ore deposit composite microbial system.
Background technology
Copper is important meals raw material indispensable in development of modern industry.Along with the progress of Economic development and society, improve day by day the status of Copper Processing Industry in the national economic development.At present, the normal temperature biological dump leaching technology of secondary copper sulfide mineral has been widely used in commercial production.Yet, studies have shown that, the normal temperature Bioleaching is for the main body of copper resource---but is difficult to prove effective chalcopyrite.Since nineteen ninety-nine, the experiment situation of two relevant chalcopyrite thermophile bacteria of the continuous issue of mining industry major company of BHP Billiton and Mintek/Bactech agitation leach.Result shows, extreme thermoacidophile agitation leach chalcopyrite concentrate is practical, can obtain very high copper recovery in the short period of time.But thermoacidophile is extremely responsive to high pulp density, gas-liquid limited transmission under high temperature, and in immersion liquid, oxygen and carbon dioxide content are low, and leaching efficiency is had a negative impact.
Summary of the invention
The object of the invention is to, provide in one temperature to soak ore deposit bacterium compound system, this fungus strain has higher leaching efficiency to chalcopyrite.
For achieving the above object, the present invention is by the following technical solutions:
A kind of temperature for Leaching of chalcopyrite is soaked the ore deposit composite microbial system, its name is called Sulfobacillus thermosulfidooxidans 6Y-1, this composite microbial system has been deposited in Chinese Typical Representative culture collection center C CTCC, the address is positioned at Wuhan University, preservation date is on November 10th, 2010, and deposit number is CCTCC NO:M2010297.
Composite microbial system as above, its working temperature are 55 ℃,
The bacterial classification of this composite microbial system consists of Sulfobacillus thermosulfidooxidans 77%, Acidithiomicrobium sp 5%, Acidithiobacillus caldus 12% and Ferroplasma sp 6%.
Composite microbial system as above, its preparation method be,
The sample of described composite microbial system is from certain chalcopyrite dump leaching ore deposit heap;
Adopt traditional method enrichment culture and tame chalcopyrite and obtain described composite microbial system from the sample that gathers;
Adopt 16S rDNA clone library technology that the Microbial community structure in above-mentioned composite microbial system is analyzed, technological line is: total DNA extraction → 16S rDNA fragment amplification → be connected → transform intestinal bacteria → screening positive clone → enzyme cutting type → mensuration sequence → Blast comparative analysis with carrier;
Adopt traditional method to investigate this composite microbial system to the leaching effect of chalcopyrite, method is: add 5-10% chalcopyrite breeze in the 200mL ordinary tap water, acid balance is to pH 1.6-1.75; After adding the yeast powder of 0.1-0.2%, with 10% the described composite microbial system nutrient solution of inoculum size access, 55 ℃ of lower 140rpm cultivate, and leach 7-15 after day, detect copper and iron leaching efficient.
Beneficial effect of the present invention is:
Mesophilic bacteria provided by the invention system, its higher temperatures bacterium has better environmental compatibility, is easier to cultivate, and working temperature is lower, and more convenient operation especially has extremely strong commercial application prospect to low-grade copper sulfide ores such as chalcopyrite in the application of heap leaching method.
The present invention relates to a composite microbial system, its name is called Sulfobacillus thermosulfidooxidans 6Y-1, this composite microbial system has been deposited in Chinese Typical Representative culture collection center C CTCC, the address is positioned at Wuhan University, preservation date is on November 10th, 2010, and deposit number is CCTCC NO:M2010297.
Embodiment
The invention will be further described below in conjunction with embodiment, and following illustrated embodiment is for ease of understanding better the present invention, but be not used for limiting the present invention.Experimental technique in following embodiment if no special instructions, is ordinary method.Experiment material used in following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.
Embodiment 1: middle temperature is soaked the acquisition of ore deposit composite microbial system 6Y-1
1. enrichment:
Get the 200mL ordinary tap water, regulating its pH is 1.7, adds the 0.20g yeast powder, adds 300g to take from the ore sample of the discarded dump leaching of Dexing copper mine ore deposit heap, 55 ℃ of insulations 24 hours.
2. cultivate:
By following formulated 100mL culture medium A: K 2HPO 41g, (NH 4) 2CO 32g, NaHCO 30.5g, CaCl 20.2g, MgSO 40.8g, distilled water 1L; Adjusting pH is 1.6-1.7.Configure substratum B by following compound method: in the 100mL ordinary tap water, add 10g chalcopyrite breeze, preferably, the particle diameter of chalcopyrite breeze is 200 orders, regulates pH with the vitriol oil and carries out acid balance, until pH is stabilized in 1.6-1.7.Before inoculation, culture medium A and B are mixed.During inoculation, first add the yeast powder of 0.1-0.15% in mixed culture medium, then add the enrichment culture thing, inoculum size is 20～30% of cumulative volume for the first time; 55 ℃ of 140rpm shaking culture.
3. domestication:
Utilize identical mixed culture medium to add the brass breeze of 5-10%, inoculate 10% culture under pH 1.6-1.7 condition, 55 ℃ are carried out bacterial leaching adaptability domestication cultivation, and obtaining bacteria concentration is 10 7～10 9The stable ore deposit composite microbial system that soaks of individual/mL.
Embodiment 2: middle temperature is soaked the analysis of community structure of bacteria in the composite microbial system L-A of ore deposit
1, the extraction of total DNA
The composite microbial system that obtains in embodiment 1 is seeded in above-mentioned mixed culture medium, and inoculum size is volume percent 5%, cultivates 20 hours for 40-50 ℃.
The 1mL nutrient solution is put into 1.5mL Eppendorf pipe, 12000rpm, 4 ℃ of centrifugal 5 minutes collection thalline wash thalline 2-3 time with 1 * TES.The thalline Eddy diffusion is broken up thalline in 435 μ L TES, add the N,O-Diacetylmuramidase 125 μ L of 20mg/mL, the Proteinase K 2.5 μ L of 10mg/mL, 37 ℃ of water-baths are gently shaken and are incubated 4 hours, add 50 μ L 20%SDS, gently shaking in 37 ℃ of water-baths is incubated 30 minutes, adds the NaClO of 5mol/L 4125 μ L occur without protein film to the interface for 3-4 time with isopyknic phenol-chloroform-primary isoamyl alcohol (three's volume ratio is 25: 24: 1) extracting.Draw the upper strata water and be placed in the NaAc-EDTA-Na that ice bath adds the 3mol/L of 1/10 volume 2, add 2 times of cold dehydrated alcohols of volume, upset mixing, ice bath 5 minutes.Centrifugal 20 minutes precipitation DNA of 12000rpm.The ethanol desalination that adds volume percent 70%, 4 ℃ of placements of spending the night, centrifugal 10 minutes of 7000rpm, sucking-off ethanol.Air-dry, add the second distillation water dissolution.Transferring DNA concentration after electrophoresis detection is 25 μ g/mL.
2, pcr amplification 16S rDNA partial sequence
Universal primer: bacterium primer 2 7F and 1492R
27F(SEQ?ID：No.1)：5’-AGAGTTTGATCCTGGCTCAG-3’
1492R(SEQ?ID：No.2)：5’-TACGGTTACCTTGTTACGACTT-3’
Ancient bacterium primer 2 5F and 1492R
25F(SEQ?ID：No.3)：5’-TCYGGTTGATCCYGCCRG-3’
1492R(SEQ?ID：No.2)：5’-TACGGTTACCTTGTTACGACTT-3’
Figure BDA0000124202900000031
The PCR reaction conditions:
Denaturation: 95 ℃ of 5min;
Circulation: 95 ℃ of 1min, 55 ℃ of 2min, 72 ℃ of 1min, 30 circulations;
Extend: 72 ℃ of 5min.
Through electrophoresis detection, prove the PCR product that obtains about 1000bp.
3, PCR product purification
Adopt the Wizard SV Gel and PCR Clean-up System product (catalog number: A9281) carry out purifying of Promega company.Concrete operations are: after gel electrophoresis, required band is downcut and put into the 1.5mL centrifuge tube; The weight of weighing adhesive tape, and add the film binding soln with the amount of 1 μ L/mg, 50-65 ℃ of insulation dissolving; Pouring the mixture after dissolving into the pillar room temperature placed after 1 minute with 16000 rotating speeds centrifugal 1 minute; Adding 700 μ L to wash coating solution adds 500 μ L to wash coating solution centrifugal 5 minutes again after centrifugal 1 minute with same rotating speed; Pillar is transferred to gently added 50 μ L nuclease free water in clean centrifuge tube, rearmounted 4 ℃ of preservations in centrifugal 1 minute.
4, the PCR product is connected with carrier
Connection carrier select Promega company pGEM T-easy carrier (catalog number: A1360), linked system such as following table:
4 ℃ of placements of spending the night after mixing.
Wherein, provide in the pGEM T-easy carrier of Control DNA for Promega company.
5, transform
The connection product of step 4 is transformed competent escherichia coli cell DH5 α (Beijing thing technology limited liability company of vast safe Hang Seng product).Get respectively 50 μ L, 100 μ L, 200 μ L are coated with flat board, cultivate 12-16 hour in 37 ℃ of incubators.According to the principle picking white colony dibbling of blue hickie screening on flat board, 50 bacterium colonies of each dull and stereotyped dibbling, the position mark number 1-50 of each bacterium colony cultivated 12 hours for 37 ℃.
6, the screening of positive colony
Adopt the bacterium colony round pcr.With the toothpick picking thalline of totally 60 different bacterium colonies add respectively in the PCR pipe that 50 μ L sterilized waters are housed in advance, carry out mark on the PCR pipe, this mark is corresponding one by one with the lable number of getting bacterium colony on flat board.With the PCR pipe in 95 ℃ the heating 5 minutes, after being chilled to room temperature, centrifugal 5 minutes of 15000rpm.Getting 10 μ L supernatants is that template is carried out bacterium colony PCR, primer is SP6 and T7, wherein the SP6 sequence is (SEQ ID:No.4) 5 '-CATACGATTTAGGTGACACTATAG-3 ', and the T7 sequence is (SEQ ID:No.5) 5 '-TAATACGACTCACTATAGGG-3 '.Bacterium colony PCR composing system and the reaction conditions all amplification condition with above-mentioned 16S rDNA are identical.Get 10 μ L PCR products and determine positive colony through gel detection.
7, positive colony order-checking and Phylogenetic Analysis
On LB substratum solid plate, send order-checking company to check order after 37 ℃ of incubated overnight the positive colony colony inoculation.Institute calling sequence input NCBI website is compared with existing sequence in Blast program and Genebank.
8, middle temperature is soaked ore deposit composite microbial system microbial species group structure composition:
Through above-mentioned analysis, result shows: this fungus strain is comprised of Sulfobacillus thermosulfidooxidans (77%), Acidithiomicrobium sp (5%), Acidithiobacillus caldus (12%) and Ferroplasma sp (6%).
Should middle temperature soak ore deposit composite microbial system called after Sulfobacillus thermosulfidooxidans 6Y-1, on November 10th, 2010, it is stored in Chinese Typical Representative culture collection center C CTCC (address: in Wuhan University), deposit number is CCTCC NO:M2010297.
Embodiment 3: in this, temperature is soaked the ore deposit composite microbial system to the leaching effect analysis of chalcopyrite
At first the composite microbial system Sulfobacillus thermosulfidooxidans 6Y-1 with cryopreservation activates in the triangular flask that the 200mL substratum is housed; Described substratum is for to form according to following formulated: K 2HPO 41g, (NH 4) 2CO 32g, NaHCO 30.5g, CaCl 20.2g, MgSO 40.8g, yeast powder 1g, distilled water 1L.After bacterial classification after activation is centrifugal, bacterium mud is transferred 200mL in acid balance, ordinary tap water that contain 5% chalcopyrite breeze, 55 ℃ cultivate weeks after, get the 50mL leach liquor, detect wherein copper and iron concentration after filtering, calculate copper and iron leaching efficient.Result shows, the leaching efficiency of copper is 34.78%, and iron leaching efficient is 30.3%.This leaching efficiency is higher than the efficient of existing mesophilic bacteria kind.
Figure IDA0000124202980000011
Figure IDA0000124202980000021

Claims (2)

1. a temperature that is used for Leaching of chalcopyrite is soaked the ore deposit composite microbial system, it is characterized in that,
Its name is called Sulfobacillus thermosulfidooxidans 6Y-1, this composite microbial system has been deposited in Chinese Typical Representative culture collection center C CTCC, the address is positioned at Wuhan University, and preservation date is on November 10th, 2010, and deposit number is CCTCC NO:M2010297.
2. fungus strain according to claim 1, is characterized in that,
The bacterial classification of this composite microbial system consists of Sulfobacillus thermosulfidooxidans 77%, Acidithiomicrobium sp 5%, Acidithiobacillus caldus 12% and Ferroplasma sp 6%.
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CN106635854A (en) * 2015-10-29 2017-05-10 北京有色金属研究总院 Oligographic bacterium composition and application of oligographic bacterium composition in inhibition on generation of jarosite in biological leaching
CN106906360A (en) * 2015-12-22 2017-06-30 北京有色金属研究总院 A kind of technique of synthetical recovery cupric cobalt sulfate slag
CN106282551B (en) * 2016-09-18 2018-03-13 深圳市如茵生态环境建设有限公司 A kind of granulation heap leaching method of shale and low-grade difficult fine ore

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105802869A (en) * 2014-12-30 2016-07-27 北京有色金属研究总院 Medium-temperature ore-leaching composite microbial system used for leaching of chalcopyrite and ore leaching process
CN105802869B (en) * 2014-12-30 2019-10-01 有研工程技术研究院有限公司 A kind of medium temperature leaching mine composite microbial system and deposit impregnating technology for Leaching of chalcopyrite
CN106635854A (en) * 2015-10-29 2017-05-10 北京有色金属研究总院 Oligographic bacterium composition and application of oligographic bacterium composition in inhibition on generation of jarosite in biological leaching
CN106906360A (en) * 2015-12-22 2017-06-30 北京有色金属研究总院 A kind of technique of synthetical recovery cupric cobalt sulfate slag
CN106906360B (en) * 2015-12-22 2018-06-29 北京有色金属研究总院 A kind of technique of synthetical recovery cupric cobalt sulfate slag
CN106282551B (en) * 2016-09-18 2018-03-13 深圳市如茵生态环境建设有限公司 A kind of granulation heap leaching method of shale and low-grade difficult fine ore

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