CN102965302A - Composite microbial agent for leaching oil shale and leaching method - Google Patents
Composite microbial agent for leaching oil shale and leaching method Download PDFInfo
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- CN102965302A CN102965302A CN2012103760017A CN201210376001A CN102965302A CN 102965302 A CN102965302 A CN 102965302A CN 2012103760017 A CN2012103760017 A CN 2012103760017A CN 201210376001 A CN201210376001 A CN 201210376001A CN 102965302 A CN102965302 A CN 102965302A
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Abstract
The invention relates to a composite microbial agent for leaching oil shale and a leaching method. The active components of the composite mineral-leaching microbial agent for leaching oil shale are Thiobacillus ferrooxidans and Bacillus mucilaginosus. The leaching method falls into two stages that: in the first stage, a Thiobacillus ferrooxidans working solution made by fermentation and dilution is employed to leach acid soluble ingredients in the oil shale; and in the second stage, a Bacillus mucilaginosus working solution made by fermentation and dilution is employed to leach silicate ingredients in the oil shale. An SEM is adopted to observe the oil shale subjected to mineral leaching, and the results show that inorganic minerals are effectively removed from the oil shale, the mineral structure changes obviously, and the phenomenon of detachment from the oil shale main body is significant. XRD determines that crystal forms of inorganic matters in the oil shale also change obviously. An infrared spectrum is utilized to detect that after mineral leaching, organic matters are effectively accumulated, especially the accumulation of chain aliphatic hydrocarbons. The weight is decreased by about 18% that that before mineral leaching. By an aluminum retort dry distillation method, it is measured that the shale oil yield is increased by about 20% after mineral leaching.
Description
Technical field:
The present invention relates to a kind of bioleaching technology, the especially biological method of soaking the composite fungus agent of molten resinous shale and soaking molten resinous shale
Background technology:
Resinous shale (oil shale) claim again oil shale, is a kind of important energy source mineral products that contain organic matter (common about＞3.5%), and its stock number is huge, is important supplement and the alternate resources of oil and natural gas.Resinous shale is a kind of important energy minerals, and its stock number is huge, is important supplement and the alternate resources of oil and natural gas.China's oil shale resources are than horn of plenty, whole nation oil shale resources are 7199.37 hundred million t, and the shale oil resource is 476.44 hundred million t, and the recyclable resource of shale oil is 119.79 hundred million t, the oil length of China's resinous shale belongs to medium level in general, and inorganic mineral content is generally higher.Reasonable development comprehensive utilization oil shale resources; must obtain good economic and social benefit; more to resinous shale research both at home and abroad in the last few years; main investigative technique and patent concentrate on the physics and chemistry aspect; have the advantages such as equipment is simple, easy to operate, cost is low, pollution is few and utilize microbial technique that shale oil is soaked the ore deposit processing; be beneficial to comprehensive utilization and environment protection, to realizing resource and economic Sustainable development important role and meaning.
The microbial leaching technology all has practical application in the extraction of some metals such as copper, gold, nickel, uranium etc., and especially to the extraction of the metallic element in the black cat, black cat itself is also destroyed when extracting metallic element.We utilize microbial leaching technical finesse resinous shale at imagination based on this, remove wherein inorganic mineral under the precondition that organism is not destroyed in keeping resinous shale.
The microbial leaching technology is applied to the processing of resinous shale, mainly is to utilize the microbial cells direct effect, or by its meta-bolites indirect action the inorganic mineral in the resinous shale is decomposed, gets rid of, and do not destroy organism in the resinous shale.Be applied at present the bacterial classification that resinous shale soaks the ore deposit and mainly be divided into two large classes.
One class is that the chemosynthetic autotroph bacterium mainly is: thiobacillus thiooxidans (T.thiooxidans) and iron protoxide Ferrobacillus (Ferrobacillus ferrooxidans), they have ferrous oxidase or oxidation of reduced sulfur enzyme, ferrous iron and the rapid oxidation of reduced sulphur in these enzymes energy catalysis mineral, the result causes the valuable metal in the mineral to be converted into ion in the solution, and sulphur is converted into SO4 at a low price
2-And discharge H
+, the carbonate in the resinous shale, pyrite and other inorganic mineral that is dissolved in acid can be removed the organism of surplus next porous and the silicon cheese that inorganics is combined closely.
Another kind of is that the chmosynthetic heterotrophs bacterium mainly is: bacillusmusilaginosiengineering (Bacillus mucilaginosu), bacillusmusilaginosiengineering etc.The multiple meta-bolites that silicate bacteria produces in process of growth has the flotation effect to mineral, make in the ore the aluminium silicate mineral particulate particularly quartz be distributed in the leach liquor, can discharge simultaneously the elements such as phosphorus in the silicate moiety, potassium, silicon, reach the purpose of analysing silicon hydrochlorate.
China's resinous shale generally has the silicates mineral substance of high level, the silicate stable in properties is difficult for removing and combining closely with kerogen, cause energy loss and directly can destroy kerogen with the chemical process removal, reduce the energy output value of resinous shale, utilize microbial leaching technical finesse resinous shale then can successfully avoid this problem, therefore studying and how utilizing removal of microorganisms silicate is the core content of present technique method.
Summary of the invention:
Purpose of the present invention is exactly for above-mentioned the deficiencies in the prior art, and a kind of stable composite mineral-leaching microbial inoculum of organism in the resinous shale that can keep again when can effectively remove in the resinous shale inorganic mineral is provided;
Another object of the present invention provides a kind of bacillusmusilaginosiengineering and thiobacillus ferrooxidant screening and acclimation method;
A further object of the present invention provides preservation and the preparation method of a kind of bacillusmusilaginosiengineering and two kinds of ore immersion bacterial cultures of thiobacillus ferrooxidant;
Also purpose of the present invention provides two kinds of a kind of bacillusmusilaginosiengineering and thiobacillus ferrooxidant and soaks the method that the ore deposit microbial inoculum soaks molten resinous shale
The objective of the invention is to be achieved through the following technical solutions:
Soak the composite mineral-leaching microbial inoculum of molten resinous shale, its activeconstituents is thiobacillus ferrooxidant (Thiobacillus ferrooxidans) and bacillusmusilaginosiengineering (Bacillus mucilaginosus).
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks screening and the acclimation method of ore deposit microbial inoculum, may further comprise the steps:
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, get the mixed bacterium bacterium liquid 5ml of resinous shale mineral-leaching bacteria colony A of step a, the centrifugal 5min of 8000rpm, selective silicon hydrochlorate substratum with sterilization washs 2 times, resinous shale mineral-leaching bacteria colony A is inoculated into the fresh selective silicon hydrochlorate that contains oil shale powder and trains in the liquid nutrient medium, at 30 ℃, cultivates 7 days in the 120rpm shaking table, again go down to posterity, tame altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
Described in the step b: selective silicon hydrochlorate nutrient solution prescription is: sucrose 10.0g, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, distilled water 1000ml;
The silicate culture medium prescription is: sucrose 10.0g, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, distilled water 1000ml;
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 1-3 days.
Soak the preparation method of the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, may further comprise the steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of the logarithmic phase inoculum size access according to 5%-8% is contained in the fermentor tank of silicate bacteria fermention medium, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains bacillusmusilaginosiengineering in the fermented liquid reaches 10
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Fermentative medium formula in the step g is: starch 2.88g/L; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L prepares with underground water.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked screening and the domestication of ore deposit microbial inoculum, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 1-3 days.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, may further comprise the steps:
5., thiobacillus ferrooxidant be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of the logarithmic phase inoculum size access fermentor tank according to 5%-8%, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
The 9K culture medium prescription of step described in 5. is:
A liquid: (NH
4)
2SO
43.0g, K
2HPO
40.5g, KCl0.1g, MgSO
4.7H
2O0.5g, Ca (NO
3)
20.01g; Deionized water 800mL, H
2SO
4Regulate pH to 2.0,121 ℃ of autoclaving 20min;
B liquid: Fe SO
4.7H
2O44.78g/L, deionized water 200mL, H
2SO
4Regulating pH to 2.0, is the membrane filtration degerming of 0.22 μ m with the aperture, with a liquid after sterilizing and the b liquid ratio mixing in 4:1;
The prescription of the fermention medium in the steps d is:
Oil shale powder 20g/L, pyrite 20g/L, (NH
4)
2SO
43.0g/L, K
2HPO
40.5g/L, KCl0.1g/L, MgSO
4.7H
2O0.5g/L, Ca (NO
3)
20.01g/L sulfuric acid is transferred pH2～3, the underground water preparation.
Method with bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans) are soaked molten resinous shale may further comprise the steps: carry out in two stages:
Fs: first resinous shale is crushed to 1-5cm, 5-10cm, 10-15cm, 15-20cm, the sample quality ratio of every kind of particle size range is 25%, at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, with soaking molten resinous shale after the fermentation and through the thiobacillus ferrooxidant working fluid of liquid dilution preparation, to remove solubility in acid component in the resinous shale, continuous dip ore deposit 4 days is soaked and is soaked acidic culture residual in the molten process and soak the ore deposit bacterium with the removal of underground water purge oil shale after finish in the ore deposit, and sampling analysis soaks the ore deposit effect, resinous shale weightlessness about 8%, aluminium rice steamer dry distillation have been measured the shale oil output increased about 7%.
Subordinate phase: at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, with after the fermentation and through the bacillusmusilaginosiengineering working fluid continuous dip ore deposit of dilution preparation 6 days, soak the silicate component in the molten resinous shale, silicate bacteria soaks the ore deposit to be finished sampling analysis and comprehensively soaks the ore deposit effect, and resinous shale weightlessness about 18%, aluminium rice steamer dry distillation have been measured the shale oil output increased about 20%.
Beneficial effect: utilize two kinds of bacterium preparations that screen to soak the ore deposit microbial inoculum and divide two sections to soak the ore deposit, it is very obvious to soak the relative prior art of ore deposit effect, soak and utilize the surperficial inorganic mineral that occurs obviously can effectively remove in the resinous shale of SEM observation resinous shale after finish in the ore deposit, keep organism stability, it is loose that resinous shale itself also becomes.Mineral substance structure considerable change under microbial process, disengaging resinous shale main body phenomenon is very remarkable; Utilize simultaneously XRD determining resinous shale inorganics crystalline form that considerable change also occurs under microbial process; And the result who utilizes infrared chromatography to detect to obtain is, soaks that the organism of resinous shale has obtained effective accumulation, the especially accumulation of chain aliphatic hydrocarbon behind the ore deposit.Soak the ore deposit finish after resinous shale become loose, the appearance of demixing phenomenon is arranged, weight with soak the ore deposit before compare and reduced about 18%.Yield of the product has improved about 20% after utilizing aluminium rice steamer dry distillation mensuration to soak the ore deposit.
Description of drawings
Accompanying drawing 1 is the Sample Scan electromicroscopic photograph that the ferrous thiobacillus of not oxidised is processed
Accompanying drawing 2 is stereoscan photographs after the sample that thiobacillus ferrooxidant is processed soaks ore deposit 4d
Accompanying drawing 3a is the Sample Scan electromicroscopic photograph of processing without bacillusmusilaginosiengineering
Accompanying drawing 3b is the sample energy spectrogram of processing without bacillusmusilaginosiengineering
Accompanying drawing 4a is the Sample Scan electromicroscopic photograph of processing through bacillusmusilaginosiengineering
Accompanying drawing 4b is the sample energy spectrogram of processing through bacillusmusilaginosiengineering
Accompanying drawing 5 is that bacillusmusilaginosiengineering soaks ore deposit process silicate dissolving figure
Accompanying drawing 6 is that XRD analysis is soaked each component relative content variation in the process of ore deposit
Accompanying drawing 7 is that IR analyzes and to soak in the process of ore deposit organic enrichment and stability in the resinous shale
Classification And Nomenclature: bacillusmusilaginosiengineering
Latin literary fame: Bacillus mucilaginosus
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center
Address: Datun Road, Chaoyang District, Beijing City
Preservation date: on 09 07th, 2012
Deposit number: CGMCC No.6542
Classification And Nomenclature: thiobacillus ferrooxidant
Latin literary fame: Thiobacillus ferrooxidans
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center
Address: Datun Road, Chaoyang District, Beijing City
Preservation date: on 09 07th, 2012
Deposit number: CGMCC No.6543
Embodiment:
The present invention is described in further detail below in conjunction with example:
Soak the composite mineral-leaching microbial inoculum of molten resinous shale, its activeconstituents is bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans).
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks screening and the acclimation method of ore deposit microbial inoculum, may further comprise the steps:
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, get the mixed bacterium bacterium liquid 5ml of resinous shale mineral-leaching bacteria colony A of step a, the centrifugal 5min of 8000rpm, selective silicon hydrochlorate substratum with sterilization washs 2 times, resinous shale mineral-leaching bacteria colony A is inoculated in the fresh selective silicon hydrochlorate nutrient solution that contains oil shale powder, at 30 ℃, cultivates 7 days in the 120rpm shaking table, again go down to posterity, tame altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
Described in the step b: selective silicon hydrochlorate nutrient solution prescription is: sucrose 10.0g, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, distilled water 1000ml;
Selective silicon hydrochlorate culture medium prescription is: sucrose 10.0g, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, distilled water 1000ml;
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 1-3 days.
Soak the preparation method of the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, may further comprise the steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of the logarithmic phase inoculum size access according to 5%-8% is contained in the fermentor tank of silicate bacteria fermention medium, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains bacillusmusilaginosiengineering in the fermented liquid reaches 10
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Fermentative medium formula in the step g is: starch 2.88g/L; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L prepares with underground water.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked screening and the domestication of ore deposit microbial inoculum, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 1-3 days.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, may further comprise the steps:
5., thiobacillus ferrooxidant be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of the logarithmic phase inoculum size access fermentor tank according to 5%-8%, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
The 9K culture medium prescription of step described in 5. is:
A liquid: (NH
4)
2SO
43.0g, K
2HPO
40.5g, KCl0.1g, MgSO
4.7H
2O0.5g, Ca (NO
3)
20.01g; Deionized water 800mL, H
2SO
4Regulate pH to 2.0,121 ℃ of autoclaving 20min;
B liquid: Fe SO
4.7H
2O44.78g/L, deionized water 200mL, H
2SO
4Regulating pH to 2.0, is the membrane filtration degerming of 0.22 μ m with the aperture, with a liquid after sterilizing and the b liquid ratio mixing in 4:1;
The prescription of the fermention medium in the steps d is:
Oil shale powder 20g/L, pyrite 20g/L, (NH
4)
2SO
43.0g/L, K
2HPO
40.5g/L, KCl0.1g/L, MgSO
4.7H
2O0.5g/L, Ca (NO
3)
20.01g/L sulfuric acid is transferred pH2～3, the underground water preparation.
Method with bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans) are soaked molten resinous shale may further comprise the steps: carry out in two stages:
Fs: first resinous shale is crushed to 1-5cm, 5-10cm, 10-15cm, 15-20cm, the sample quality ratio of every kind of particle size range is 25%, at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, and thiobacillus ferrooxidant working fluid rear and that prepare through the liquid dilution soaks the molten solubility in acid component in the resinous shale of having removed, continuous dip ore deposit 4 days with fermentation, soak and soak acidic culture residual in the molten process and soak the ore deposit bacterium with the removal of underground water purge oil shale after finish in the ore deposit, sampling analysis soaks the ore deposit effect, and resinous shale weightlessness 8%, aluminium rice steamer dry distillation have been measured the shale oil output increased 7%.
Subordinate phase: at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, with after the fermentation and through the bacillusmusilaginosiengineering working fluid continuous dip ore deposit of dilution preparation 6 days, soak the silicate component in the molten resinous shale, silicate bacteria soaks the ore deposit to be finished sampling analysis and comprehensively soaks the ore deposit effect, and resinous shale weightlessness about 18%, aluminium rice steamer dry distillation have been measured the shale oil output increased about 20%;
Embodiment 1
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in the molten resinous shale is soaked the ore deposit microbial inoculum, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 2 days.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, may further comprise the steps:
5., thiobacillus ferrooxidant be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of logarithmic phase according to 6% inoculum size access fermentor tank, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time 50h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, the resinous shale mineral-leaching bacteria colony A that gets step a mixes bacterium bacterium liquid 5ml, and the centrifugal 5min of 8000rpm is with the sucrose 10.0g of sterilization, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, the selective silicon hydrochlorate substratum washing of distilled water 1000ml preparation 2 times, resinous shale mineral-leaching bacteria colony A is inoculated into the fresh sucrose 10.0g that contains oil shale powder, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g in the selective silicon hydrochlorate nutrient solution of distilled water 1000ml preparation, at 30 ℃, cultivated 7 days in the 120rpm shaking table, again went down to posterity, and tamed altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 2 days.
Soak the preparation method of the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, may further comprise the steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of logarithmic phase is contained starch 2.88g/L according to 5% inoculum size access; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L, in the fermentor tank with the silicate bacteria fermention medium of underground water preparation, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time 48h is cultured to logarithmic phase, and the concentration of bacillusmusilaginosiengineering reaches 10 in the acquisition fermented liquid
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Step 3, choose Huadian underground mining oil shale ore, choose particle diameter at 1-5cm, 5-10cm, 10-15cm, the sample of 15-20cm, the sample quality ratio of every kind of particle size range are 25% proportioning and mix.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, is 1:1 by solid-to-liquid ratio, adds the working fluid 40L of Fermentation of Thiobacillus ferrooxidans liquid preparation; At ambient temperature, aeration rate is 4L/min, and thiobacillus ferrooxidant working fluid rear and that prepare through the liquid dilution soaks the molten solubility in acid component in the resinous shale of having removed with fermentation, the control temperature is at 30 ℃, and pH=3 is under the saturated aeration status, molten 4 days of continuous dip finishes the fs and soaks the ore deposit.
Step 5, remove to put into after soaking acidic culture residual in the molten process and soaking the ore deposit bacterium with underground water purge oil shale and soak the ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering liquid 40L, at ambient temperature, aeration rate is 5L/min, the control temperature is at 30 ℃, pH=7, under the saturated aeration status, molten 6 days of continuous dip, remove silicate component, finish subordinate phase and soak the ore deposit.
All samples soak after the ore deposit finishes, and soak after tested the ore deposit result, after the destructive distillation shale oil output increased 22%.
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in the molten resinous shale is soaked the ore deposit microbial inoculum, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 3 days.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, may further comprise the steps:
5., thiobacillus ferrooxidant be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of logarithmic phase according to 8% inoculum size access fermentor tank, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time 49h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, the resinous shale mineral-leaching bacteria colony A that gets step a mixes bacterium bacterium liquid 5ml, and the centrifugal 5min of 8000rpm is with the sucrose 10.0g of sterilization, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, the selective silicon hydrochlorate substratum washing of distilled water 1000ml preparation 2 times, resinous shale mineral-leaching bacteria colony A is inoculated into the fresh sucrose 10.0g that contains oil shale powder, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g in the selective silicon hydrochlorate nutrient solution of distilled water 1000ml preparation, at 30 ℃, cultivated 7 days in the 120rpm shaking table, again went down to posterity, and tamed altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 3 days.
Soak the preparation method of the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, may further comprise the steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of logarithmic phase is contained starch 2.88g/L according to 8% inoculum size access; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L, in the fermentor tank with the silicate bacteria fermention medium of underground water preparation, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time 50h is cultured to logarithmic phase, and the concentration of bacillusmusilaginosiengineering reaches 10 in the acquisition fermented liquid
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Step 3, choose farming peace oil shale ore, choose particle diameter at 1-5cm, 5-10cm, 10-15cm, the sample of 15-20cm, the sample quality ratio of every kind of particle size range are 25% proportioning and mix.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, is 1:1 by solid-to-liquid ratio, adds the working fluid 40L of Fermentation of Thiobacillus ferrooxidans liquid preparation; At ambient temperature, aeration rate is 4L/min, and thiobacillus ferrooxidant working fluid rear and that prepare through the liquid dilution soaks the molten solubility in acid component in the resinous shale of having removed with fermentation, the control temperature is at 28 ℃, and pH=3 is under the saturated aeration status, molten 4 days of continuous dip finishes the fs and soaks the ore deposit.
Step 5, remove to put into after soaking acidic culture residual in the molten process and soaking the ore deposit bacterium with underground water purge oil shale and soak the ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering working fluid 40L, at ambient temperature, aeration rate is 5L/min, the control temperature is at 28 ℃, pH=7, under the saturated aeration status, molten 6 days of continuous dip, remove silicate component, finish subordinate phase and soak the ore deposit.
All samples soak after the ore deposit finishes, and soak after tested the ore deposit result, after the destructive distillation shale oil output increased 21%.
Embodiment 3
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in the molten resinous shale is soaked the ore deposit microbial inoculum, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 2 days.
The thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, may further comprise the steps:
5., thiobacillus ferrooxidant be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of logarithmic phase according to 7% inoculum size access fermentor tank, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time 48h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, the resinous shale mineral-leaching bacteria colony A that gets step a mixes bacterium bacterium liquid 5ml, and the centrifugal 5min of 8000rpm is with the sucrose 10.0g of sterilization, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, the selective silicon hydrochlorate substratum washing of distilled water 1000ml preparation 2 times, resinous shale mineral-leaching bacteria colony A is inoculated into the fresh sucrose 10.0g that contains oil shale powder, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g in the selective silicon hydrochlorate nutrient solution of distilled water 1000ml preparation, at 30 ℃, cultivated 7 days in the 120rpm shaking table, again went down to posterity, and tamed altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
The bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 2 days.
Soak the preparation method of the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, may further comprise the steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of logarithmic phase is contained starch 2.88g/L according to 7% inoculum size access; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L, in the fermentor tank with the silicate bacteria fermention medium of underground water preparation, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time is 49h, is cultured to logarithmic phase, and the concentration of bacillusmusilaginosiengineering reaches 10 in the acquisition fermented liquid
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Step 3, choose the Fushun oil shale ore, choose particle diameter at 1-5cm, 5-10cm, 10-15cm, the sample of 15-20cm, the sample quality ratio of every kind of particle size range are 25% proportioning and mix.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, is 1:1 by solid-to-liquid ratio, adds the working fluid 40L of Fermentation of Thiobacillus ferrooxidans liquid preparation; At ambient temperature, aeration rate is 4L/min, and thiobacillus ferrooxidant working fluid rear and that prepare through the liquid dilution soaks the molten solubility in acid component in the resinous shale of having removed with fermentation, the control temperature is at 25 ℃, and pH=3 is under the saturated aeration status, molten 4 days of continuous dip finishes the fs and soaks the ore deposit.
Step 5, remove to put into after soaking acidic culture residual in the molten process and soaking the ore deposit bacterium with underground water purge oil shale and soak the ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering working fluid 40L, at ambient temperature, aeration rate is 4L/min, the control temperature is at 25 ℃, pH=7, under the saturated aeration status, molten 6 days of continuous dip, remove the silicate group, finish subordinate phase and soak the ore deposit.
All samples soak after the ore deposit finishes, and soak after tested the ore deposit result, after the destructive distillation shale oil output increased 19%.
Claims (10)
1. a composite mineral-leaching microbial inoculum that soaks molten resinous shale is characterized in that, its activeconstituents is bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxifans).
2. a bacillusmusilaginosiengineering that soaks molten resinous shale mesosilicic acid salt soaks screening and the acclimation method of ore deposit microbial inoculum, it is characterized in that, may further comprise the steps:
A, screening, having gathered seriously in a certain colliery that has the resinous shale associated minerals in the open, the resinous shale of weathering places the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivated again 5 days, and be resinous shale mineral-leaching bacteria colony A;
B, domestication, get the mixed bacterium bacterium liquid 5ml of resinous shale mineral-leaching bacteria colony A of step a, the centrifugal 5min of 8000rpm, selective silicon hydrochlorate substratum with sterilization washs 2 times, resinous shale mineral-leaching bacteria colony A is inoculated in the fresh selective silicon hydrochlorate nutrient solution that contains oil shale powder, at 30 ℃, cultivates 7 days in the 120rpm shaking table, again go down to posterity, tame altogether 5 times;
After c, domestication finish, gained is tamed the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying the efficient single bacterium of this strain silicate be: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
Described in the step b: selective silicon hydrochlorate nutrient solution prescription is: sucrose 10.0g, Na
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, distilled water 1000ml;
The silicate culture medium prescription is: sucrose 10.0g, K
2HPO
42.0g, MgSO
47H
2O0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2SO
42g, distilled water 1000ml.
3. a colloid bacillus cereus that soaks molten resinous shale mesosilicic acid salt soaks the store method of ore deposit microbial inoculum, it is characterized in that, bacterial classification adopts the sand pipe to preserve, to soak the ore deposit microbial inoculum activates in containing the substratum of oil shale powder; the bacterial classification of hiding of going bail for places 100ml to contain the 250ml shaking flask of the silicate substratum of oil shale powder, and room temperature 120rpm shaking table activates 1-3 days.
4. a preparation method who soaks the colloid bacillus cereus immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt is characterized in that, may further comprise the steps:
E, the colloid bacillus cereus immersion ore bacterium liquid in the claim 2 is inoculated in the silicate substratum activates, and go down to posterity 2 times;
F, the colloid bacillus cereus after will activating soak the ore deposit microbial inoculum and are re-seeded into fresh silicate substratum, and shaking culture is to logarithmic phase;
G, shaking culture among the step f to the bacterial classification of the logarithmic phase inoculum size access according to 5%-8% is contained in the fermentor tank of silicate bacteria fermention medium, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains colloid bacillus cereus in the fermented liquid reaches 10
8/ ml;
H, the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
Fermentative medium formula in the step g is: starch 2.88g/L; Maltose 2.08g/L; Ammonium nitrate 0.24g/L; Sodium Nitrite 0.47g/L; Urea 0.18g/L prepares with underground water.
5. screening and the domestication that the thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the ore deposit microbial inoculum is characterized in that, may further comprise the steps:
1., the screening, get a certain copper mine tailings acid waste water and place the 250ml triangular flask that 90ml distilled water is housed, shaking culture is 24 hours in 30 ℃ of constant temperature oscillators; Be inoculated in the 9K substratum, cultivated again 5 days, be resinous shale mineral-leaching bacteria colony B;
2., domestication, get the mixed bacterium bacterium liquid 5ml of B of step a, the centrifugal 5min of 8000rpm with the 9K substratum washing of sterilization 2 times, in the resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in the 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed the separation and purification of flora plate streaking, respectively obtain 1 plant height effect and soak the single bacterium of mineral products acid, 4 ℃ of Refrigerator stores;
4., through identifying that this plant height effect soaks the ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
6. a thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the store method of ore deposit microbial inoculum, it is characterized in that, bacterial classification adopts the sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for places the 250ml shaking flask of 100ml9K substratum, room temperature 120rpm shaking table activation 1-3 days.
7. a thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, it is characterized in that, may further comprise the steps:
5., the thiobacillus ferrooxidant in the claim 5 be inoculated in the 9K substratum activate, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after will activating soaks the ore deposit bacterium and is re-seeded in the 9K substratum, shaking culture is to logarithmic phase;
7., with step 6. in shaking culture to the bacterial classification of the logarithmic phase inoculum size access fermentor tank according to 5%-8%, contain fermention medium corresponding to acid-producing bacteria in the fermentor tank, leavening temperature is 30 ℃, setting the D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, obtain fermented liquid, the fermented liquid bacteria concentration of thiobacillus ferrooxidant reaches 10
7/ ml;
8., the fermented liquid that obtains is soaked mining working fluid with namely making after ten times of the underground water dilutions, be directly used in bioleaching.
8. soak the preparation method of ore deposit microbial inoculum according to the thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale claimed in claim 7, it is characterized in that, the 9K culture medium prescription of step described in 5. is:
A liquid: (NH
4)
2SO
43.0g, K
2HPO
40.5g, KCl0.1g, MgSO
4.7H
2O0.5g, Ca (NO
3)
20.01g; Deionized water 800mL, H
2SO
4Regulate pH to 2.0,121 ℃ of autoclaving 20min;
B liquid: Fe SO
4.7H
2O44.78g/L, deionized water 200mL, H
2SO
4Regulating pH to 2.0, is the membrane filtration degerming of 0.22 μ m with the aperture, with a liquid after sterilizing and the b liquid ratio mixing in 4:1.
9. soak the preparation method of ore deposit microbial inoculum according to the thiobacillus ferrooxidant of soaking solubility in acid composition in the molten resinous shale claimed in claim 7, it is characterized in that the prescription of the fermention medium in the steps d is:
Oil shale powder 20g/L, pyrite 20g/L, (NH
4)
2SO
43.0g/L, K
2HPO
40.5g/L, KCl0.1g/L, MgSO
4.7H
2O0.5g/L, Ca (NO
3)
20.01g/L sulfuric acid is transferred pH2～3, the underground water preparation.
10. a bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans) method of soaking molten resinous shale is characterized in that, may further comprise the steps: carry out in two stages:
Fs: first resinous shale is crushed to 1-5cm, 5-10cm, 10-15cm, 15-20cm, the sample quality ratio of every kind of particle size range is 25%, at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, and thiobacillus ferrooxidant working fluid rear and that prepare through the liquid dilution soaks the molten solubility in acid component in the resinous shale of having removed, continuous dip ore deposit 4 days with fermentation, soak and soak acidic culture residual in the molten process and soak the ore deposit bacterium with the removal of underground water purge oil shale after finish in the ore deposit, sampling analysis soaks the ore deposit effect, and resinous shale weightlessness about 8%, aluminium rice steamer dry distillation have been measured the shale oil output increased about 7%.
Subordinate phase: at ambient temperature, be 1:1 by solid-to-liquid ratio, aeration rate is 4-5L/min, with after the fermentation and through the bacillusmusilaginosiengineering working fluid continuous dip ore deposit of dilution preparation 6 days, soak the silicate component in the molten resinous shale, silicate bacteria soaks ore deposit end sampling analysis and comprehensively soaks the ore deposit effect, and resinous shale weightlessness about 18%, aluminium rice steamer dry distillation are measured yield of the product and improved about 20%.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3982995A (en) * | 1975-05-07 | 1976-09-28 | The University Of Southern California | Method of converting oil shale into a fuel |
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2012
- 2012-09-30 CN CN201210376001.7A patent/CN102965302B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3982995A (en) * | 1975-05-07 | 1976-09-28 | The University Of Southern California | Method of converting oil shale into a fuel |
Non-Patent Citations (5)
| Title |
|---|
| 《Organic Geochemistry》 19901231 M.M.Vrvić et al. Demineralization of an oil shale by Bacillus circulans ("siliceous bacteria") 1203-1209 1-10 第16卷, 第4-6期 * |
| J.FINDLEY,M.D. APPLEMAN ET AL.: "Degradation of Oil Shale by Sulfur-Oxidizing Bacteria", 《APPLIED MICROBIOLOGY》, vol. 28, no. 3, 30 September 1974 (1974-09-30), pages 460 - 464 * |
| M.M.VRVIć ET AL.: "Demineralization of an oil shale by Bacillus circulans ("siliceous bacteria")", 《ORGANIC GEOCHEMISTRY》, vol. 16, no. 46, 31 December 1990 (1990-12-31), pages 1203 - 1209 * |
| M.M.VRVIć ET AL.: "Preparation of rich kerogen concentrates：Removal of pyrite with Thiobacillus ferrooxidans", 《ORGANIC GEOCHEMISTRY》, vol. 13, no. 46, 31 December 1988 (1988-12-31), pages 1109 - 1104 * |
| 张兰英等: "微生物浸矿驱提页岩油的技术进展", 《吉林大学学报（地球科学版）》, vol. 41, no. 5, 30 September 2011 (2011-09-30), pages 1562 - 1572 * |
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