CN102965302B - 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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- CN102965302B CN102965302B CN201210376001.7A CN201210376001A CN102965302B CN 102965302 B CN102965302 B CN 102965302B CN 201210376001 A CN201210376001 A CN 201210376001A CN 102965302 B CN102965302 B CN 102965302B
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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 containing organic (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 compared with horn of plenty, whole nation oil shale resources are 7199.37 hundred million t, and 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 physics and chemistry aspect; and utilize microbial technique to soak ore deposit processing to shale oil, there is the advantages such as equipment is simple, easy to operate, cost is low, pollution is few; be beneficial to comprehensive utilization and environment protection, to realizing resource and economic Sustainable development important role and meaning.
Microbial leaching technology all has practical application, the especially extraction to the metallic element in black cat in the extraction of some metals as copper, gold, nickel, uranium etc., and when extracting metallic element, black cat itself is also destroyed.Based on this, we utilize microbial leaching technical finesse resinous shale at imagination, under the precondition that organism is not destroyed in keeping resinous shale, remove wherein inorganic mineral.
Microbial leaching technology is applied to the processing of resinous shale, is mainly to utilize microbial cells direct effect, or by its meta-bolites indirect action, the inorganic mineral in resinous shale is decomposed, got rid of, and do not destroy the organism in resinous shale.Be applied at present the bacterial classification that resinous shale soaks ore deposit and be mainly divided into two large classes.
One class be chemosynthetic autotroph bacterium mainly: thiobacillus thiooxidans (T.thiooxidans) and iron protoxide Ferrobacillus (Ferrobacillus ferrooxidans), they have ferrous oxidase or oxidation of reduced sulfur enzyme, ferrous iron and reduced sulphur in these enzyme energy catalysis mineral are oxidized rapidly, result causes the valuable metal in mineral to be converted into the ion in solution, and sulphur is converted into SO4 at a low price
2-and discharge H
+, the carbonate in resinous shale, pyrite and other can be dissolved in to sour inorganic mineral and remove, the organism of surplus next porous and the silicon cheese that inorganics is combined closely.
Another kind of be chmosynthetic heterotrophs bacterium mainly: bacillusmusilaginosiengineering (Bacillus mucilaginosu), bacillusmusilaginosiengineering etc.The multiple meta-bolites that silicate bacteria produces in process of growth has flotation effect to mineral, make aluminium silicate mineral particulate in ore particularly quartz be distributed in leach liquor, can discharge the elements such as phosphorus in silicate moiety, potassium, silicon simultaneously, reach the object of analysing silicon hydrochlorate.
China's resinous shale generally has the silicates mineral substance of high level, silicate stable in properties is difficult for removing and combining closely with kerogen, and directly can destroy kerogen with chemical process removal, cause energy loss, reduce the energy output value of resinous shale, utilize microbial leaching technical finesse resinous shale can successfully avoid this problem, therefore studying and how utilizing microorganism removal silicate is the core content of this technological method.
Summary of the invention:
Object 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 resinous shale that can keep again when can effectively remove in resinous shale inorganic mineral is provided;
Another object of the present invention is to provide a kind of bacillusmusilaginosiengineering and thiobacillus ferrooxidant screening and acclimation method;
A further object of the present invention is to provide preservation and the preparation method of a kind of bacillusmusilaginosiengineering and two kinds of ore immersion bacterial cultures of thiobacillus ferrooxidant;
Also object of the present invention is to provide two kinds of a kind of bacillusmusilaginosiengineering and thiobacillus ferrooxidant and soaks the method that ore deposit microbial inoculum soaks molten resinous shale
The object 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, comprises the following steps:
A, screening, in a certain outdoor colliery with resinous shale associated minerals, gather the seriously resinous shale of weathering and be placed in the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivate again 5 days, 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, with the selective silicon hydrochlorate substratum of sterilizing, wash 2 times, resinous shale mineral-leaching bacteria colony A is inoculated in the fresh selective silicon hydrochlorate training liquid nutrient medium that contains oil shale powder, at 30 ℃, cultivates 7 days in 120rpm shaking table, again go down to posterity, tame altogether 5 times;
After c, domestication finish, gained is tamed to the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying that the efficient single bacterium of this strain silicate is: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
Described in step b: selective silicon hydrochlorate nutrient solution formula is: sucrose 10.0g, Na
2hPO
42.0g, MgSO
47H
2o0.2g, NaCl0.1g, distilled water 1000ml;
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 sand pipe to preserve, by soaking ore deposit microbial inoculum, in containing the substratum of oil shale powder, activate; the bacterial classification hidden of going bail for is placed in the 250ml shaking flask of the silicate substratum that 100ml contains oil shale powder, room temperature 120rpm shaking table activation 1-3 days.
The preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, comprises the following steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in claim 2 is inoculated in silicate substratum and is activated, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after activation is soaked to ore deposit microbial inoculum and is re-seeded into fresh silicate substratum, shaking culture is to logarithmic phase;
G, shaking culture in step f to the bacterial classification of logarithmic phase is contained to silicate bacteria fermention medium according to the inoculum size access of 5%-8% fermentor tank in, leavening temperature is 30 ℃, setting D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
H, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
Fermentative medium formula in 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 molten resinous shale is soaked screening and the domestication of ore deposit microbial inoculum, comprises the following steps:
1., screening, get a certain copper mine tailings acid waste water and be placed in the 250ml triangular flask that 90ml distilled water is housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours; Be inoculated in 9K substratum, then cultivate 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 sterilizing 2 times, in resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed to 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 ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for is placed in 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 molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, comprises the following steps:
5., thiobacillus ferrooxidant is inoculated in 9K substratum and is activated, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after activation soaked to ore deposit bacterium be re-seeded in 9K substratum, shaking culture is to logarithmic phase;
7., by step 6. in shaking culture to the bacterial classification of logarithmic phase according to the inoculum size access fermentor tank of 5%-8%, in fermentor tank, contain the fermention medium that acid-producing bacteria is corresponding, leavening temperature is 30 ℃, setting 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 of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
The 9K culture medium prescription of step described in is 5.:
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 aperture, and a liquid after sterilizing and b liquid are mixed in the ratio of 4:1;
The formula of the fermention medium in 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 adjusted pH2～3, underground water preparation.
The method of soaking molten resinous shale with bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans), comprises the following steps: carry out in two stages:
First stage: 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, by solid-to-liquid ratio, be 1:1, aeration rate is 4-5L/min, with diluting the molten resinous shale of thiobacillus ferrooxidant work immersion of preparing after fermentation and through liquid, to remove solubility in acid component in resinous shale, 4 days, continuous dip ore deposit, soaking ore deposit finishes with underground water purge oil shale, to remove and to soak acidic culture residual in molten process and to soak ore deposit bacterium afterwards, sampling analysis soaks ore deposit effect, weightless 8% left and right of resinous shale, aluminium rice steamer dry distillation is measured shale oil output and has been improved 7% left and right.
Subordinate phase: at ambient temperature, by solid-to-liquid ratio, be 1:1, aeration rate is 4-5L/min, with after fermentation and through the bacillusmusilaginosiengineering working fluid continuous dip ore deposit of dilution preparation 6 days, soak the silicate component in molten resinous shale, silicate bacteria soaks ore deposit to be finished sampling analysis and comprehensively soaks ore deposit effect, weightless 18% left and right of resinous shale, and aluminium rice steamer dry distillation is measured shale oil output and has been improved 20% left and right.
Beneficial effect: utilize two kinds of bacterium preparations that screen to soak ore deposit microbial inoculum and divide two sections to soak ore deposit, soak the relative prior art of ore deposit effect very obvious, soaking ore deposit finishes the rear SEM of utilization observation resinous shale surface and occurs obviously can effectively remove the inorganic mineral in resinous shale, 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 XRD determining resinous shale inorganics crystalline form that considerable change also occurs under microbial process simultaneously; And the result of utilizing infrared chromatography to detect to obtain is, soaks the organism of resinous shale behind ore deposit and obtained effective accumulation, the especially accumulation of chain aliphatic hydrocarbon.Soak ore deposit and finish rear resinous shale and become loose, have the appearance of demixing phenomenon, weight with soak ore deposit before compare and reduced by 18% left and right.After utilizing aluminium rice steamer dry distillation mensuration to soak ore deposit, yield of the product has improved 20% left and right.
Accompanying drawing explanation
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 of thiobacillus ferrooxidant processing 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 resinous shale
Classification And Nomenclature: bacillusmusilaginosiengineering
Latin literary fame: Bacillus mucilaginosus
Depositary institution: China Committee for Culture Collection of Microorganisms's 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's common micro-organisms center
Address: Datun Road, Chaoyang District, Beijing City
Preservation date: on 09 07th, 2012
Deposit number: CGMCC No.6543
Embodiment:
Below in conjunction with example, the present invention is described in further detail:
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, comprises the following steps:
A, screening, in a certain outdoor colliery with resinous shale associated minerals, gather the seriously resinous shale of weathering and be placed in the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivate again 5 days, 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, with the selective silicon hydrochlorate substratum of sterilizing, wash 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 120rpm shaking table, again go down to posterity, tame altogether 5 times;
After c, domestication finish, gained is tamed to the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying that the efficient single bacterium of this strain silicate is: bacillusmusilaginosiengineering (Bacillus mucilaginosus).
Described in step b: selective silicon hydrochlorate nutrient solution formula 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 sand pipe to preserve, by soaking ore deposit microbial inoculum, in containing the substratum of oil shale powder, activate; the bacterial classification hidden of going bail for is placed in the 250ml shaking flask of the silicate substratum that 100ml contains oil shale powder, room temperature 120rpm shaking table activation 1-3 days.
The preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, comprises the following steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in claim 2 is inoculated in silicate substratum and is activated, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after activation is soaked to ore deposit microbial inoculum and is re-seeded into fresh silicate substratum, shaking culture is to logarithmic phase;
G, shaking culture in step f to the bacterial classification of logarithmic phase is contained to silicate bacteria fermention medium according to the inoculum size access of 5%-8% fermentor tank in, leavening temperature is 30 ℃, setting D0 value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
H, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
Fermentative medium formula in 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 molten resinous shale is soaked screening and the domestication of ore deposit microbial inoculum, comprises the following steps:
1., screening, get a certain copper mine tailings acid waste water and be placed in the 250ml triangular flask that 90ml distilled water is housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours; Be inoculated in 9K substratum, then cultivate 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 sterilizing 2 times, in resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed to 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 ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for is placed in 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 molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, comprises the following steps:
5., thiobacillus ferrooxidant is inoculated in 9K substratum and is activated, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after activation soaked to ore deposit bacterium be re-seeded in 9K substratum, shaking culture is to logarithmic phase;
7., by step 6. in shaking culture to the bacterial classification of logarithmic phase according to the inoculum size access fermentor tank of 5%-8%, in fermentor tank, contain the fermention medium that acid-producing bacteria is corresponding, leavening temperature is 30 ℃, setting 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 of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
The 9K culture medium prescription of step described in is 5.:
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 aperture, and a liquid after sterilizing and b liquid are mixed in the ratio of 4:1;
The formula of the fermention medium in 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 adjusted pH2～3, underground water preparation.
The method of soaking molten resinous shale with bacillusmusilaginosiengineering (Bacillus mucilaginosus) and thiobacillus ferrooxidant (Thiobacillus ferrooxidans), comprises the following steps: carry out in two stages:
First stage: 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, by solid-to-liquid ratio, be 1:1, aeration rate is 4-5L/min, with having removed solubility in acid component in resinous shale after fermentation and through the thiobacillus ferrooxidant work immersion of liquid dilution preparation is molten, 4 days, continuous dip ore deposit, soaking ore deposit finishes with underground water purge oil shale, to remove and to soak acidic culture residual in molten process and to soak ore deposit bacterium afterwards, sampling analysis soaks ore deposit effect, resinous shale weightlessness 8%, aluminium rice steamer dry distillation is measured shale oil output and has been improved 7%.
Subordinate phase: at ambient temperature, by solid-to-liquid ratio, be 1:1, aeration rate is 4-5L/min, with after fermentation and through the bacillusmusilaginosiengineering working fluid continuous dip ore deposit of dilution preparation 6 days, soak the silicate component in molten resinous shale, silicate bacteria soaks ore deposit to be finished sampling analysis and comprehensively soaks ore deposit effect, weightless 18% left and right of resinous shale, and aluminium rice steamer dry distillation is measured shale oil output and has been improved 20% left and right;
Embodiment 1
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in molten resinous shale is soaked ore deposit microbial inoculum, comprise the following steps:
1., screening, get a certain copper mine tailings acid waste water and be placed in the 250ml triangular flask that 90ml distilled water is housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours; Be inoculated in 9K substratum, then cultivate 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 sterilizing 2 times, in resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed to 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 ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for is placed in 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 molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, comprises the following steps:
5., thiobacillus ferrooxidant is inoculated in 9K substratum and is activated, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after activation soaked to ore deposit bacterium be re-seeded in 9K substratum, shaking culture is to logarithmic phase;
7., by step 6. in shaking culture to the bacterial classification of logarithmic phase according to 6% inoculum size access fermentor tank, in fermentor tank, contain the fermention medium that acid-producing bacteria is corresponding, leavening temperature is 30 ℃, setting 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 of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, in a certain outdoor colliery with resinous shale associated minerals, gather the seriously resinous shale of weathering and be placed in the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivate again 5 days, 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, with the sucrose 10.0g of sterilizing, K
2hPO
42.0g, MgSO
47H
2o0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2sO
42g, selective silicon hydrochlorate substratum washing prepared by distilled water 1000ml 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 selective silicon hydrochlorate nutrient solution prepared by distilled water 1000ml, at 30 ℃, cultivates in 120rpm shaking table 7 days, again goes down to posterity, and tames altogether 5 times;
After c, domestication finish, gained is tamed to the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying that the efficient single bacterium of this strain silicate is: 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 sand pipe to preserve, by soaking ore deposit microbial inoculum, in containing the substratum of oil shale powder, activate; the bacterial classification hidden of going bail for is placed in the 250ml shaking flask of the silicate substratum that 100ml contains oil shale powder, room temperature 120rpm shaking table activation 2 days.
The preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, comprises the following steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in claim 2 is inoculated in silicate substratum and is activated, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after activation is soaked to ore deposit microbial inoculum and is re-seeded into fresh silicate substratum, shaking culture is to logarithmic phase;
G, shaking culture in step f to the bacterial classification of logarithmic phase is contained to 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 of the silicate bacteria fermention medium of preparing with underground water, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time 48h, is cultured to logarithmic phase, and the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
H, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of 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 is 25% proportioning and mixes.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, by solid-to-liquid ratio, is 1:1, adds working fluid 40L prepared by Fermentation of Thiobacillus ferrooxidans liquid; At ambient temperature, aeration rate is 4L/min, with having removed solubility in acid component in resinous shale after fermentation and through the thiobacillus ferrooxidant work immersion of liquid dilution preparation is molten, control temperature at 30 ℃, pH=3, under saturated aeration status, molten 4 days of continuous dip, finishes the first stage and soaks ore deposit.
Step 5, with underground water purge oil shale, remove after soaking acidic culture residual in molten process and soaking ore deposit bacterium and put into and soak ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering liquid 40L, at ambient temperature, aeration rate is 5L/min, control temperature at 30 ℃, pH=7, under saturated aeration status, molten 6 days of continuous dip, remove silicate component, complete subordinate phase and soak ore deposit.
All samples soak after ore deposit finishes, and soak after tested ore deposit result, and after destructive distillation, shale oil output has improved 22%.
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in molten resinous shale is soaked ore deposit microbial inoculum, comprise the following steps:
1., screening, get a certain copper mine tailings acid waste water and be placed in the 250ml triangular flask that 90ml distilled water is housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours; Be inoculated in 9K substratum, then cultivate 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 sterilizing 2 times, in resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed to 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 ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for is placed in 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 molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, comprises the following steps:
5., thiobacillus ferrooxidant is inoculated in 9K substratum and is activated, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after activation soaked to ore deposit bacterium be re-seeded in 9K substratum, shaking culture is to logarithmic phase;
7., by step 6. in shaking culture to the bacterial classification of logarithmic phase according to 8% inoculum size access fermentor tank, in fermentor tank, contain the fermention medium that acid-producing bacteria is corresponding, leavening temperature is 30 ℃, setting 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 of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, in a certain outdoor colliery with resinous shale associated minerals, gather the seriously resinous shale of weathering and be placed in the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivate again 5 days, 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, with the sucrose 10.0g of sterilizing, K
2hPO
42.0g, MgSO
47H
2o0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2sO
42g, selective silicon hydrochlorate substratum washing prepared by distilled water 1000ml 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 selective silicon hydrochlorate nutrient solution prepared by distilled water 1000ml, at 30 ℃, cultivates in 120rpm shaking table 7 days, again goes down to posterity, and tames altogether 5 times;
After c, domestication finish, gained is tamed to the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying that the efficient single bacterium of this strain silicate is: 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 sand pipe to preserve, by soaking ore deposit microbial inoculum, in containing the substratum of oil shale powder, activate; the bacterial classification hidden of going bail for is placed in the 250ml shaking flask of the silicate substratum that 100ml contains oil shale powder, room temperature 120rpm shaking table activation 3 days.
The preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, comprises the following steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in claim 2 is inoculated in silicate substratum and is activated, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after activation is soaked to ore deposit microbial inoculum and is re-seeded into fresh silicate substratum, shaking culture is to logarithmic phase;
G, shaking culture in step f to the bacterial classification of logarithmic phase is contained to 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 of the silicate bacteria fermention medium of preparing with underground water, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time 50h, is cultured to logarithmic phase, and the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
H, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
Step 3, choose agriculture 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 is 25% proportioning and mixes.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, by solid-to-liquid ratio, is 1:1, adds working fluid 40L prepared by Fermentation of Thiobacillus ferrooxidans liquid; At ambient temperature, aeration rate is 4L/min, with having removed solubility in acid component in resinous shale after fermentation and through the thiobacillus ferrooxidant work immersion of liquid dilution preparation is molten, control temperature at 28 ℃, pH=3, under saturated aeration status, molten 4 days of continuous dip, finishes the first stage and soaks ore deposit.
Step 5, with underground water purge oil shale, remove after soaking acidic culture residual in molten process and soaking ore deposit bacterium and put into and soak ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering working fluid 40L, at ambient temperature, aeration rate is 5L/min, control temperature at 28 ℃, pH=7, under saturated aeration status, molten 6 days of continuous dip, remove silicate component, complete subordinate phase and soak ore deposit.
All samples soak after ore deposit finishes, and soak after tested ore deposit result, and after destructive distillation, shale oil output has improved 21%.
Embodiment 3
Step 1, soak screening and domestication that the thiobacillus ferrooxidant of solubility in acid composition in molten resinous shale is soaked ore deposit microbial inoculum, comprise the following steps:
1., screening, get a certain copper mine tailings acid waste water and be placed in the 250ml triangular flask that 90ml distilled water is housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours; Be inoculated in 9K substratum, then cultivate 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 sterilizing 2 times, in resinous shale mineral-leaching bacteria colony B inoculation 9K substratum, at 30 ℃, cultivates 7d in 120rpm shaking table, again goes down to posterity, and tames altogether 5 times;
3., after domestication finishes, gained is tamed to 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 ore deposit acid-producing bacteria and be: thiobacillus ferrooxidant Thiobacillus ferrooxidans.
The thiobacillus ferrooxidant of soaking solubility in acid composition in molten resinous shale is soaked the store method of ore deposit microbial inoculum, and bacterial classification adopts sand pipe to preserve, and the bacterial classification of the Tibetan of going bail for is placed in 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 molten resinous shale is soaked the preparation method of ore deposit microbial inoculum, comprises the following steps:
5., thiobacillus ferrooxidant is inoculated in 9K substratum and is activated, and go down to posterity 2 times;
6., the ferrous sulphur bar of the change after activation soaked to ore deposit bacterium be re-seeded in 9K substratum, shaking culture is to logarithmic phase;
7., by step 6. in shaking culture to the bacterial classification of logarithmic phase according to 7% inoculum size access fermentor tank, in fermentor tank, contain the fermention medium that acid-producing bacteria is corresponding, leavening temperature is 30 ℃, setting 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 of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching; Stand-by.
A, screening, in a certain outdoor colliery with resinous shale associated minerals, gather the seriously resinous shale of weathering and be placed in the 250ml triangular flask that granulated glass sphere and 90ml distilled water are housed, in 30 ℃ of constant temperature oscillators, shaking culture is 24 hours, be inoculated in the selectivity nutrient solution that contains oil shale powder potassium deficiency, nitrogen, cultivate again 5 days, 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, with the sucrose 10.0g of sterilizing, K
2hPO
42.0g, MgSO
47H
2o0.2g, NaCl0.1g, yeast extract paste 0.5g, (NH
4)
2sO
42g, selective silicon hydrochlorate substratum washing prepared by distilled water 1000ml 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 selective silicon hydrochlorate nutrient solution prepared by distilled water 1000ml, at 30 ℃, cultivates in 120rpm shaking table 7 days, again goes down to posterity, and tames altogether 5 times;
After c, domestication finish, gained is tamed to the separation and purification of flora plate streaking, obtain the efficient single bacterium of 1 strain silicate, 4 ℃ of Refrigerator stores.
D, through identifying that the efficient single bacterium of this strain silicate is: 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 sand pipe to preserve, by soaking ore deposit microbial inoculum, in containing the substratum of oil shale powder, activate; the bacterial classification hidden of going bail for is placed in the 250ml shaking flask of the silicate substratum that 100ml contains oil shale powder, room temperature 120rpm shaking table activation 2 days.
The preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, comprises the following steps:
E, the bacillusmusilaginosiengineering immersion ore bacterium liquid in claim 2 is inoculated in silicate substratum and is activated, and go down to posterity 2 times;
F, the bacillusmusilaginosiengineering after activation is soaked to ore deposit microbial inoculum and is re-seeded into fresh silicate substratum, shaking culture is to logarithmic phase;
G, shaking culture in step f to the bacterial classification of logarithmic phase is contained to 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 of the silicate bacteria fermention medium of preparing with underground water, leavening temperature is 30 ℃, and setting D0 value is 6mg/L, and fermentation time is 49h, is cultured to logarithmic phase, and the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
H, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
Step 3, choose 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 is 25% proportioning and mixes.
What step 4, the resinous shale 40kg that proportioning is mixed joined soaks in the groove of ore deposit, by solid-to-liquid ratio, is 1:1, adds working fluid 40L prepared by Fermentation of Thiobacillus ferrooxidans liquid; At ambient temperature, aeration rate is 4L/min, with having removed solubility in acid component in resinous shale after fermentation and through the thiobacillus ferrooxidant work immersion of liquid dilution preparation is molten, control temperature at 25 ℃, pH=3, under saturated aeration status, molten 4 days of continuous dip, finishes the first stage and soaks ore deposit.
Step 5, with underground water purge oil shale, remove after soaking acidic culture residual in molten process and soaking ore deposit bacterium and put into and soak ore deposit groove, solid-to-liquid ratio is 1:1, add bacillusmusilaginosiengineering working fluid 40L, at ambient temperature, aeration rate is 4L/min, control temperature at 25 ℃, pH=7, under saturated aeration status, molten 6 days of continuous dip, remove silicate group, complete subordinate phase and soak ore deposit.
All samples soak after ore deposit finishes, and soak after tested ore deposit result, and after destructive distillation, shale oil output has improved 19%.
Claims (3)
1. soak molten resinous shale and soak an ore deposit microbial inoculum, it is characterized in that, its activeconstituents is bacillusmusilaginosiengineering (Bacillus mucilaginosus), and described bacillusmusilaginosiengineering deposit number is: CGMCC No.6542.
2. a preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt, is characterized in that, comprises the following steps:
A by deposit number is: the bacillusmusilaginosiengineering of CGMCC No.6542 is inoculated in silicate substratum and activates, and goes down to posterity 2 times;
Described 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;
B is re-seeded into fresh silicate substratum by the bacillusmusilaginosiengineering after activation, and shaking culture is to logarithmic phase;
In the fermentor tank that c contains silicate bacteria fermention medium by shaking culture in step b to the bacterial classification of logarithmic phase according to the inoculum size access of 5%-8%, leavening temperature is 30 ℃, setting DO value is 6mg/L, fermentation time is 48-50h, be cultured to logarithmic phase, the concentration that obtains bacillusmusilaginosiengineering in fermented liquid reaches 10
8/ ml;
D, the fermented liquid of acquisition is soaked to mining working fluid with making after ten times of underground water dilutions, be directly used in bioleaching.
3. according to the preparation method who soaks the bacillusmusilaginosiengineering immersion ore bacterium liquid of molten resinous shale mesosilicic acid salt claimed in claim 2, it is characterized in that, the fermentative medium formula described in step c 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.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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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