CN105199990B - A kind of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 and its application - Google Patents
A kind of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 and its application Download PDFInfo
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- CN105199990B CN105199990B CN201510674270.5A CN201510674270A CN105199990B CN 105199990 B CN105199990 B CN 105199990B CN 201510674270 A CN201510674270 A CN 201510674270A CN 105199990 B CN105199990 B CN 105199990B
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Abstract
The invention discloses a kind of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM Z4 and its applications.Acidithiobacillus ferrooxidans strain GF, Acidithiobacillus thiooxidans, the mixed culture FIM Z4 of acidophilic heterotrophic bacteria, deposit number are:CGMCC No.11190.It can significantly improve Leach reaction dynamics by synergistic effect, accelerate reaction speed, shorten leaching cycle, improve the leaching rate and leaching rate of metal ion (copper and mickel) in target ore deposit.The autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM Z4 are a compatible entirety, and Population Ecological keeps relative stability during long-term domestication.It is applied in biological metallurgy, and preferable leaching effect is respectively provided with to the nickel ion in the copper ion and low grade nickel ore in low-grade copper sulfide ores.
Description
Technical field:
The invention belongs to field of metallurgy, and in particular to a kind of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 and its should
With.
Background technology:
Biohydrometallurgy be with microorganism to ore it is direct, indirectly or both collective effect leach ore in useful gold
A kind of technique belonged to.Compared with conventional metallurgical technology, biological metallurgy have it is at low cost, technological process is simple, environment amenable
Advantage, and can handle traditional metallurgical method can not or reluctant poor value, raw ore and tailing etc..In order to adapt to
Different ore type and the efficiency for improving bacterial leaching find, gather and screens that efficient, adaptable, selectivity is good
Ore leaching bacteria be the key that carry out biological metallurgy basic research and engineering application.
Microorganism currently used for soaking ore deposit is mainly acidophilus, aerobic autotrophic type Thiobacillus, wherein Thiobacillus ferrooxidans
(Acidi thiobacillus ferrooxidans, A.f) is most study and be most heavy in current Bioleaching correlative study
A kind of leaching microbacteria wanted, A.f obtain the energy and fixed CO with ferrous oxide and (or) reduced form sulfide2It is sought as carbon source
Chemoautotrophic growth.Acidithiobacillus thiooxidans (Acidithiobacillus thiooxidans, A.t) are micro- for obligate autotrophy
Biology, can four sulfate of oxidation of elemental sulfur, thiosulfate and chain.In the bioleaching process of sulfide ore, A.t is in
Subordinate status, the A.f that generally accompanys play a role jointly.
Autotrophic type Thiobacillus is generally slow-growing, this is also a low main cause of mineral leaching efficiency.Research shows to soak ore deposit
Some metabolites of microorganism have inhibitory action to the growth of its own.
The content of the invention:
The object of the present invention is to provide one kind can significantly improve Leach reaction dynamics, accelerate reaction speed, shorten leaching
Go out the cycle, improve the autotrophic type of leaching rate of metal ion and heterotroph composite mineral-leaching bacteria colony FIM-Z4-acidophilus oxygen in target ore deposit
Change ferrous Thiobacillus (Acidithiobacillus ferrooxidans), Acidithiobacillus thiooxidans
The mixed culture of (Acidithiobacillus thiooxidans), acidophilic heterotrophic bacteria (Acidiphiliu acidophilum)
Object FIM-Z4 was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 09 08th, 2015
(CGMCC), address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, deposit number are:
CGMCC No.11190。
The present invention also provides Acidithiobacillus ferrooxidans strain GF (Acidithiobacillus ferrooxidans), acidophilus
Thiobacillus thioxidans (Acidithiobacillus thiooxidans), acidophilic heterotrophic bacteria (Acidiphiliu
Acidophilum applications of the mixed culture FIM-Z4) in ore deposit is soaked.Leached preferably when soaking ore deposit in copper or nickel should
With.
The present invention from the ore that Fujian copper-sulphide ores mine gathers by enrichment culture obtain it is a kind of by autotrophic type and
The autotrophic type of heterotroph ore leaching bacteria composition and heterotroph composite mineral-leaching bacteria colony FIM-Z4, the autotrophic type and heterotroph composite mineral-leaching bacterium
The dominant bacteria of group FIM-Z4 is mainly Acidithiobacillus ferrooxidans strain GF (Acidithiobacillus ferrooxidans), acidophilus
Thiobacillus thioxidans (Acidithiobacillus thiooxidans), pseudomonas aeruginosa (pseudomonas sp.),
Acidophilic heterotrophic bacteria (Acidiphilium acidophilum) and Sphingol single-cell (Sphingomonas sp.).With autotrophic type
Leaching ore deposit Mixed Microbes are compared, and the autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 can significantly improve Leach reaction dynamics,
Accelerate reaction speed, shorten leaching cycle, improve target ore deposit (copper and mickel) Bioleaching rate and leaching rate.The autotrophy of the present invention
Type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 are a compatible entirety, and Population Ecological keeps phase during long-term domestication
To stabilization.It is applied in biological metallurgy, is had to the nickel ion in the copper ion and low grade nickel ore in low-grade copper sulfide ores
There is preferable leaching effect.
Contain the heterotroph using organic substrates as energy substance in leaching ore body system, it is possible to releasing pair to a certain extent
The inhibitory action of autotrophic bacterium is conducive to the growth of main ore leaching bacteria, gives full play to it and soaks ore deposit effect, improves leaching rate.Mixing leaching
Heterotroph can make full use of nutrients mostly using the metabolite of autotrophic bacterium, dead thalline as the growth energy in ore deposit flora
Matter releases inhibition of the metabolite to autotrophic bacterium, increases the proliferative speed of autotrophic bacterium, and then improves Bioleaching rate.Autotrophic type
It is played an important role with heterotroph ore leaching bacteria by synergistic effect to improving metal leaching rate and leaching rate.
Acidithiobacillus ferrooxidans strain GF (Acidithiobacillus ferrooxidans), the acidophilus sulfur oxide of the present invention
Thiobacillus (Acidithiobacillus thiooxidans), acidophilic heterotrophic bacteria (Acidiphiliu acidophilum) it is mixed
Culture FIM-Z4 is closed to be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on 09 08th, 2015
The heart (CGMCC), address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, deposit number
For:CGMCC No.11190.
Specific embodiment:
Following embodiment is the further explanation to the present invention rather than limitation of the present invention.
Embodiment 1:The enrichment and domestication of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4
The low-grade copper sulfide ores sample ore that Fujian copper-sulphide ores mine is taken to gather, is added in 9K-S culture mediums, 32 DEG C,
150rpm shake cultures treat that culture solution becomes brownish red, after microscopy observation is with the presence of a large amount of movement bacteriums, obtain 1st generation enrichment training
Support object.1st generation enrichment culture object is turned to be commissioned to train foster, method is same as above, but bacterial load will be reduced by generation, turns to be commissioned to train foster 8~10
Dai Hou obtains final enrichment culture object.
The 9K-S culture medium preparation methods are as follows:A liquid:By (NH4)2SO43.0g、K2HPO40.5g、KCl 0.1g、
MgSO4.7H2O 0.5g、Ca(NO3) 0.01g, it is added in 800mL deionized waters, H2SO4PH to 2.0 is adjusted, 121 DEG C of high pressures are gone out
Bacterium 20min obtains A liquid;B liquid:By FeSO4.7H2O 44.78g are added in 200mL deionized waters, H2SO4PH to 2.0 is adjusted,
Aperture is 0.22 μm of membrane filtration degerming;C:Simple substance sulphur powder 5g, is sterilized using batch process.A liquid after 800ml is sterilized with
200ml B liquid is added in C, and 9K-S culture mediums are obtained after mixing.
Carrying out ore domestication culture to above-mentioned enrichment culture object, the culture medium of Initial culture is 9K-S culture mediums, 32 DEG C,
In 150rpm shake cultures, domestication 6 generations of culture, reduce the content of ferrous sulfate and sulphur powder in above-mentioned 9K-S culture mediums, simultaneously by generation
Increase the content of low-grade copper sulfide ores sample ore as energy substance, so as to improve adaptability of the enriched substance to target ore deposit.Often
The secondary Shaking culture time is 7~10d, and microscopy is it is observed that largely movement bacterium exists.No longer add during last time domestication
Add ferrous sulfate and sulphur powder, ore becomes unique energy substance.Finally with the low-grade vulcanization of addition in 9K culture mediums (no Fe)
Copper mine sample ore flora is carried out repeatedly to turn to be commissioned to train as unique energy source substance it is foster, be derived from it is stable by ore domestication from
The type of supporting and heterotroph composite mineral-leaching bacteria colony are autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4.
Embodiment 2:The composition analysis of autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 bacterial populations
Using the autotrophic type after the analysis domestication of Illumina MiSeq high throughput sequencing technologies and heterotroph composite mineral-leaching bacterium
The biological community structure of group FIM-Z4.It mainly comprises the following steps:Autotrophic type and heterotroph composite mineral-leaching are extracted using CTAB/NaCl methods
The total DNA of flora FIM-Z4 using the genomic DNA integrality of 1% agarose gel electrophoresis detection extracting, expands bacterial 16 S
The V3 areas of rRNA carry out high-flux sequence by Illumina Miseq microarray datasets, extract high quality sequence, sequence is carried out
Bioinformatic analysis obtains the situation of structure of community and abundance.The result shows that the autotrophic type and heterotroph composite mineral-leaching bacteria colony
The dominant bacteria of FIM-Z4 is mainly that (abundance accounts for Acidithiobacillus ferrooxidans strain GF (Acidithiobacillus ferrooxidans)
Than 80.03%), Acidithiobacillus thiooxidans (Acidithiobacillus thiooxidans) (abundance accounting 9.02%), copper
Green pseudomonad (pseudomonas sp.) (abundance accounting 7.07%), acidophilic heterotrophic bacteria (Acidiphiliu
Acidophilum) (abundance accounting 1.80%) and Sphingol single-cell (Sphingomonas sp.) (abundance accounting 1.11%).
This autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 are named as:Acidithiobacillus ferrooxidans strain GF
(Acidithiobacillus ferrooxidans), Acidithiobacillus thiooxidans (Acidithiobacillus
Thiooxidans), the mixed culture FIM-Z4 of acidophilic heterotrophic bacteria (Acidiphiliu acidophilum), in 2015 09
The moon is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address on the 08th:Beijing's southern exposure
The institute 3 of area North Star West Road 1, Institute of Microorganism, Academia Sinica, deposit number are:CGMCC No.11190.
Embodiment 3:The leaching test of low-grade copper sulfide ores
Using certain low-grade copper sulfide ores as sample ore, the granularity ﹤ 0.075mm of sample ore, essential mineral composition is chalcopyrite, spot copper
Ore deposit, pyrite and galena, copper grade 0.47%.
By autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 (i.e. above-mentioned Acidithiobacillus ferrooxidans strain GFs
(Acidithiobacillus ferrooxidans), Acidithiobacillus thiooxidans (Acidithiobacillus
Thiooxidans), the mixed culture FIM-Z4 of acidophilic heterotrophic bacteria (Acidiphiliu acidophilum)) and control bacterium
(Mixed Microbes of Acidithiobacillus ferrooxidans strain GF and Acidithiobacillus thiooxidans, the two content in Mixed Microbes and autotrophic type and different
The Acidithiobacillus ferrooxidans strain GF supported in type composite mineral-leaching bacteria colony FIM-Z4 is consistent with the content of Acidithiobacillus thiooxidans) respectively
It accesses in 9K-S culture mediums (with embodiment 1), 32 DEG C, the observation of 150rpm microscopies and cell count are found, autotrophic type and heterotroph
Composite mineral-leaching bacteria colony FIM-Z4 passes through of short duration lag phase, just quickly enters exponential phase.By bacterium solution under 8000r/min from
Heart 20min abandons supernatant and is precipitated, and washs precipitation with the sulfuric acid solutions sterilized in advance of pH 2.0, is then centrifuged on reject
Clear liquid so operates three times, repeatedly to reach Fe in bacteria-removing liquid3+Purpose, iron-free cell suspension is obtained, in biomicroscope
Under counted, adjustment cell concentration be 1 × 109cell·mL-1.9K culture mediums are taken (without Fe2+, i.e., in conventional 9K culture mediums
In be not added with FeSO4.7H2O, other compositions are constant) 180mL is placed in 500mL conical flasks, weighs 20g sample ores and be put into conical flask,
Sulfuric acid adjusts slurry pH, and control initial pH value is 1.8~2.0.Above-mentioned cell concentration of adjusting is added in as 1 × 109cell·
mL-1Autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 (5% inoculum concentration of volume fraction), 32 DEG C, 150rpm shake cultures.
Simultaneously not add ore leaching bacteria as blank control, to add above-mentioned Acidithiobacillus ferrooxidans strain GF and Acidithiobacillus thiooxidans
Mixed Microbes according to same treatment conduct compare.Acid consumption in leaching process is supplemented by the sulfuric acid of 10mol/L, evaporation loss
Moisture by add distilled water supplement.The Cu in leachate is measured after leaching 20d2+Concentration.Blank control group Cu2+Leaching rate
For 17%, using Acidithiobacillus ferrooxidans strain GF and the Mixed Microbes of oxidation sulfurous Thiobacillus, Cu2+Leaching rate is 52%, and is used
The autotrophic type of the present invention and heterotroph composite mineral-leaching bacteria colony FIM-Z4, Cu2+Leaching rate is up to 73%.Show to rely solely on acidleach
Effect, Cu2+Leaching effect is weaker, and passes through the effect of ore leaching bacteria, Cu2+Leaching effect is remarkably reinforced.With autotrophic type is used alone
The leaching microbacteria Mixed Microbes of Acidithiobacillus thiooxidans (Acidithiobacillus ferrooxidans strain GF with) is compared, autotrophic type and heterotroph leaching
Ore deposit compound bacteria (autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4) passes through synergistic effect, Cu2+Leaching effect significantly increases.
The 9K fluid nutrient medium preparation methods are as follows:A liquid:(NH4)2SO43.0g,K2HPO40.5g, KCl 0.1g,
MgSO4.7H2O 0.5g,Ca(NO3), 0.01g, 800mL deionized water dissolvings, H2SO4Adjust pH to 2.0,121 DEG C of high pressure sterilization
20min;B liquid:FeSO4.7H2O 44.78g, deionized water 200mL dissolve, H2SO4PH to 2.0 is adjusted, aperture is 0.22 μm
Membrane filtration degerming.It is for use by being dispensed after the A liquid after sterilizing and B liquid mixings.
Embodiment 4:The leaching test of low-grade nickel sulfide ore
Using certain low-grade nickel sulfide ore as sample ore, the granularity ﹤ 0.075mm of sample ore, essential mineral composition for nickel pyrrhotite,
Pentlandite and chalcopyrite.Ore nickel grade is 0.73%.
By autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 (i.e. above-mentioned Acidithiobacillus ferrooxidans strain GFs
(Acidithiobacillus ferrooxidans), Acidithiobacillus thiooxidans (Acidithiobacillus
Thiooxidans), the mixed culture FIM-Z4 of acidophilic heterotrophic bacteria (Acidiphiliu acidophilum)) and control bacterium
(Mixed Microbes of Acidithiobacillus ferrooxidans strain GF and Acidithiobacillus thiooxidans, the two content in Mixed Microbes and autotrophic type and different
It is consistent with Acidithiobacillus thiooxidans to support the Acidithiobacillus ferrooxidans strain GF in type composite mineral-leaching bacteria colony FIM-Z4) it is respectively connected to
In 9K-S culture mediums (with embodiment 1), 32 DEG C, 150rpm cultures, microscopy observation and cell count are found, autotrophic type and heterotroph
Composite mineral-leaching bacteria colony FIM-Z4 passes through of short duration lag phase, just quickly enters exponential phase.By bacterium solution under 8000r/min from
Heart 20min abandons supernatant and is precipitated, and washs precipitation with the sulfuric acid solutions sterilized in advance of pH 2.0, is then centrifuged on reject
Clear liquid so operates three times, repeatedly to reach Fe in bacteria-removing liquid3+Purpose, iron-free cell suspension is obtained, in biomicroscope
Under counted, adjustment cell concentration be 1 × 109cell·mL-1.9K culture mediums are taken (without Fe2+) 180mL be placed in 500mL cone
It in shape bottle, weighs 20g sample ores and is put into conical flask, sulfuric acid adjusts slurry pH, and control initial pH value is 1.8~2.0.In addition
It states and adjusts cell concentration as 1 × 109cell·mL-1Autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 (volume fractions
5% inoculum concentration), 32 DEG C, 150rpm shake cultures.Simultaneously not add ore leaching bacteria as blank control, to add above-mentioned acidophilus
The Mixed Microbes of Thiobacillus ferrooxidans and Acidithiobacillus thiooxidans are compareed according to the conduct equally handled.Acid in leaching process
Consumption is supplemented by adding the sulfuric acid of 10mol/L, and the moisture of evaporation loss is supplemented by adding 9K culture mediums.After leaching 20d, blank
Control group Ni2+Leaching rate is 12%, using Acidithiobacillus ferrooxidans strain GF and the Mixed Microbes of oxidation sulfurous Thiobacillus, Ni2+It leaches
Rate is 51%, and autotrophic type using the present invention and heterotroph composite mineral-leaching bacteria colony FIM-Z4, Ni2+Leaching rate is up to 70%.With
Above the result shows that relying solely on the effect of acidleach, Ni2+Leaching effect is faint, and passes through the effect of ore leaching bacteria, Ni2+Leaching effect
It is remarkably reinforced.Be used alone autotrophic type leaching microbacteria (Acidithiobacillus ferrooxidans strain GF and Acidithiobacillus thiooxidans it is mixed
Close bacterium) it compares, autotrophic type and heterotroph leaching ore deposit compound bacteria (autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4) pass through collaboration
Effect, Ni2+Immersion Zn enhancing with obvious effects.
Claims (3)
1. a kind of separated autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4, deposit number are:CGMCC No.11190.
2. the application of autotrophic type described in claim 1 and heterotroph composite mineral-leaching bacteria colony FIM-Z4 in ore deposit is soaked.
3. application according to claim 2, it is characterised in that autotrophic type and heterotroph composite mineral-leaching bacteria colony FIM-Z4 are soaking
The application in copper or nickel is leached during ore deposit.
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CN111808773B (en) * | 2020-07-22 | 2021-02-05 | 东华理工大学 | Acidithiobacillus ferrooxidans and application thereof and ore leaching method |
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