CN100368531C - Acidophilic leptospirillumferrooxidans and its uses - Google Patents
Acidophilic leptospirillumferrooxidans and its uses Download PDFInfo
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- CN100368531C CN100368531C CNB2005101277417A CN200510127741A CN100368531C CN 100368531 C CN100368531 C CN 100368531C CN B2005101277417 A CNB2005101277417 A CN B2005101277417A CN 200510127741 A CN200510127741 A CN 200510127741A CN 100368531 C CN100368531 C CN 100368531C
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The present invention discloses leptospiriLLum ferriphiLum and the application thereof. The leptospiriLLum ferriphiLum UBK03 of the present invention is preserved in the common microorganism center of China Committee for Culture Collections of Microorganisms (CCCCM) in january 12, 2005, and is called as CGMCC for short, and the preserving number is CGMCC No. 1290. The bacterial strain can growth in environmental conditions of low pH value, high temperature and high salinity, and has strong F-resistant capability. The maximal fluorine ion tolerance concentration can reach 1.2 to 1.4 g/L, and the bacterial strain of the present invention can be widely used for the microorganism metallurgical industry.
Description
Technical field
The present invention relates to an acidophilic leptospirillumferrooandans and application thereof.
Background technology
The microbial metallurgy technology is usually used in the extraction and the recovery of metal in the middle low-grade metal ore of Containing Sulfur mineral.Ordinary person changed into solvable state with metal from the indissoluble state to make it enter solution, thereby is reclaimed for adding oxygenant and sulfuric acid during chemical method leached under higher acidity condition.But its oxygenant and sulfuric acid large usage quantity improve the technology cost greatly.And that the biological metallurgy technology can utilize the acidophilia action of microorganisms to overcome is above-mentioned unfavorable.The acidophilia chemoautotrophic bacteria mainly contains ThiobaciLLus ferrooxidans (thiobacillus ferrooxidant), ThiobaciLLus thiooxidans (thiobacillus thiooxidans), FerrobaciLLus ferrooxidans (ferric oxide Ferrobacillus), LeptospiriLLum ferrooxidans (iron protoxide hook end spirobacteria) etc., their optimal pH scope is between 2.0-3.0.In the biological metallurgy process, the pH value just has the ferric iron precipitation when above 2.0 and produces, and partly precipitated is wrapped in ore particles surface (US 5089412 for RaLph P.HackL et aL., Bacteria for oxidizing muLtimetaLLic suLphide ores); Just there is yellow potassium ferrovanadium class material to produce when total concentration of iron is 5～8g/L in the bacterium leaching system of pH value＞1.6, therefore causes oxygenate content to reduce, influence the leaching yield of metal.The bacterium leaching process is all attempted to carry out under low pH environment mostly at present.Screening and separating has high oxygenation efficiencies under extremely low pH envrionment conditions metallurgical bacterium has great application prospect.
Summary of the invention
The purpose of this invention is to provide the sour iron oxidation hook end spirobacteria of having a liking for that a strain can grow in low pH, high-temperature and high salinity environment.
Provided by the present inventionly have a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 and be deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on January 12nd, 2005, it abbreviates CGMCC as, and deposit number is CGMCC No.1290.
This is had a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 and is Gram-negative bacteria, and thalline is shape in the shape of a spiral, and juvenile stage is an arc, and is short and small; The aged phase is a 3-5 circle spirrillum, and elongated, form is similar to LeptospiriLLum ferrooxidans.
Another object of the present invention provides the purposes that the present invention has a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLumferriphiLum) UBK03 CGMCC № 1290.
Of the present inventionly have a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 and have good iron oxidation activity, can be at Fe
2+Be applied in the oxidation.
In addition, the present invention has a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 and has the strong ore deposit ability of soaking, and this is one of its key property, and the biology that can be used for sulfur compound ore and fluorine-containing ore leaches.
The present invention filters out a strain by steps such as enrichment, uv irradiating, purifying and has the sour iron oxidation of having a liking for of strong iron oxidation capacity hook end spirobacteria (LeotospiriLLum ferriphiLum) UBK03 CGMCC № 1290, this bacterial strain can and be grown under the high salinity envrionment conditions under low pH value, comparatively high temps, and, have strong anti-F
-Ability, maximum fluorion tolerance concentration can reach 1.2-1.4g/L, can be used widely in microbial metallurgy industry.
Description of drawings
Fig. 1 is UBK03 and 19 hours Fe of BK02 strain culturing under the condition of different pH
2+Oxidation ratio is (Fe relatively
2+Oxidation ratio refers to Fe in the certain hour
2+Be oxidized to Fe
3+Percentage, surplus with);
Fig. 2 is UBK03 and 18 hours Fe of BK02 strain culturing under the condition of different temperatures
2+Oxidation ratio relatively;
Fig. 3 is UBK03 and BK02 bacterial strain Fe under the optimum condition
2+Oxidation ratio changes comparison in time;
Fig. 4 be UBK03 with the BK02 bacterial strain at different F
-Complete oxidation Fe during concentration
2+Required time relatively;
Fig. 5 is accumulative total uranium leaching yield change curve.
Embodiment
Embodiment 1, have a liking for the screening of sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290
Of the present inventionly have a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 and leach from the diafiltration of China's nuclear industry " 721 uranium ore " uranium ore microorganism that separation and purification obtains in the recrement.With the 9K substratum of slag samples with pH3.0-3.5, the enrichment culture of under 30-35 ℃ of condition, vibrating, repeatedly switching is treated the stable back of its oxidation activity purifying after ultraviolet mutagenesis is cultivated in the high salinity ore substratum of pH1.2-1.5, promptly obtains having a liking for sour iron oxidation hook end spirobacteria UBK03.
This UBK03 bacterial strain is typical acidophilia chemosynthetic autotroph bacterium, can efficient oxidation Fe when pH1.2-1.5
2+, the sulfide in again can the efficient oxidation ore; Can pass through Oxidation of Fe
2+, pyrite (FeS
2) and obtain the energy growth, its product is Fe
3+, sulfuric acid and vitriol; Utilize CO
2Be carbon source; Its optimum growth temperature is between 35-40 ℃.The UBK03 bacterial strain can be in the uranium leach liquor of tool high salinity Oxidation of Fe fast
2+, uranium and fluorion are had strong tolerance.16SrRNA gene order to this UBK03 bacterial strain is carried out the systematics analysis revealed, and it and LeptospiriLLumferriphiLum bacterium homology reach more than 90%, so but called after is had a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum).
In each of the embodiments described below, be contrast all with thiobacillus ferrooxidant (AcidobaciLLus ferrooxidans) BK02 bacterial strain.The BK02 bacterial strain obtains from Commission of Science, Technology and Industry for National Defence's " 721 uranium ore " shoal mining area uranium ore separation, and at 30-35 ℃, with improvement 9K substratum enrichment culture, repeatedly also purifying gained is cultivated in switching under the condition of pH2.0-2.5.The BK02 bacterial strain is the obligate autotrophy bacterium, and optimum growth temperature is 30-35 ℃, and best pH is 2.0-2.5.
In each of the embodiments described below, used ore substratum:, use 5g/L-8g/LH by liquid-solid ratio 10: 1,5: 1,2: 1,1: 1 (volume (L)/weight (kg)) respectively with each 1000g of particle diameter 1-10mm uranium ore
2SO
4Acidified aqueous solution 5-8d, it is constant to make liquid pH keep 24h at 1.3-1.4, and this acidizing fluid is the ore substratum, and (time spent is added FeSO
47H
2O is to [Fe
2+]=5g/L), wherein solid-to-liquid ratio is that 1: 1 ore substratum composition is as shown in table 1; Test is as shown in table 2 with the ore composition.
Table 1 ore substratum (liquid-solid ratio is 1: 1) composition and the g/L of ionic concn unit
| Composition | F - | CL - | SO 4 2- | NO 3 - | PO 4 3- | Li | Na | K | Mg | Ca | NH 4 + | U 6+ | Fe * | Salinity |
| Concentration | 1.58 | 0.56 | 36.62 | 0.84 | 1.07 | 2.70 | 0.99 | 0.21 | 0.29 | 0.85 | 0.39 | 1.25 | 5.05 | 52.4 |
* the total Tie Tong of solubility excessively manually adds FeSO in this substratum
47H
2O maintains about 5g/L.
Table 2 test ore composition
| Ore sample number | Analysis project and analytical results ω (B)/% | ||||||||
| SiO 2 | AL 2O 3 | Fe 2O 3 | FeO | CaO | MgO | K 2O | Na 2O | MnO | |
| BSZ | 69.46 | 13.44 | 0.96 | 1.38 | 4.42 | 0.63 | 2.48 | 3.16 | 0.08 |
| Ore sample number | Analysis project and analytical results ω (B)/% | ||||||||
| TiO 2 | P 2O 5 | nnn | S 2- | S 6+ | F | U(IV) | U(VI) | ∑U | |
| BSZ | 0.30 | 0.53 | 2.76 | 0.70 | 0.07 | 1.05 | 0.366 | 0.058 | 0.424 |
*BSZ is three mining area ore compound samples.(nnn is a loss on ignition, the amount that is reduced when promptly ore heats under 950 ℃)
Used cultural method is: all using solid-to-liquid ratio in the example 2,3,4 is 5: 1 ore substratum; It is the different multiples diluent of 1: 1 ore substratum that example 5 uses liquid-solid ratio respectively, fluorine concentration be zero to use the 9K substratum in the same old way; Inoculum size is 20%; The UBK03 bacterial strain places under 35-40 ℃, pH1.4 condition and cultivates, and contrast BK02 bacterial strain places under 30-35 ℃, pH2.0 condition and cultivates; But the strain growth testing of characteristic uses temp .-regulating type gas bath vibrator (Chinese Jiangsu, BS-IE type) to shake a bottle shaking culture, and rotating speed 140rpm-160rpm uranium ore leaching test uses the aerator pump inflation, and air flow is 5.47L/min.
Used analysis test method is: soluble iron (Fe in the culturing bacterium process
2+And Fe
3+) employing EDTA volumetry (indicator is a methyl violet, and sulphosalicylic acid developer, ammonium persulphate are oxygenant); PH value and redox potential adopt pH meter (Mettler Toledo, 320-S type precision acidity meter, Switzerland) to measure in the bacteria growth process; [F
-] measure with fluoride ion electrode and pHS-3C potentiometer; Ore culture medium solution intermediate ion concentration utilizes DX-600 type ion chromatograph (U.S.) to measure; Uranium concentration NH
4V0
3Titration measuring.
The research of embodiment 2, UBK03 strain growth pH scope
Use the ore substratum, with Fe
2+Be the energy, temperature is 40 ℃ (are 35 ℃ to BK02 temperature in the same old way), at pH0.6-2.5 scope shaking culture simultaneously respectively, Fe
2+Oxidation ratio is represented the growing state of bacterium, the Fe during to microbial culture 19h with the pH value
2+The oxidation ratio mapping, the result as shown in Figure 1.
As seen from Figure 1, the UBK03 bacterial strain all can be grown in pH0.6-2.5, and optimal pH is 1.2-1.8, and for avoiding the sedimentary generation of siderotil, optimal pH is 1.2-1.5 when advising its application.Prolong incubation time to 44 hour under the pH0.6 condition, the UBK03 bacterial strain can be with 5g/L Fe
2+Complete oxidation shows that this bacterium still can adapt to lower pH condition.Compare with UBK03, control strain BK02 then has narrower pH growth scope (pH1.4-2.5), and its ability of growing under extremely low pH condition can not show a candle to bacterial strain UBK03, and obvious sediment is all arranged under the pH2.5 condition.
The research of embodiment 3, UBK03 strain growth temperature range
Use the ore substratum, with Fe
2+Be the energy, pH is 1.4 (is 2.0 to BK02pH in the same old way), shaking culture simultaneously respectively in temperature is 10-50 ℃ of scope, Fe
2+Oxidation ratio is represented the growing state of bacterium, the Fe during to microbial culture 18h with temperature
2+The oxidation ratio mapping, the result as shown in Figure 2.
As seen from Figure 2, the optimum growth temperature of UBK03 bacterial strain is 35-40 ℃; Prolong incubation time, can be higher than 50 ℃ and then not grow being lower than slowly growth under 10 ℃ of conditions; Control strain BK02 does not then grow when being higher than 45 ℃.
Embodiment 4, UBK03 bacterial strain Fe
2+Oxidation capacity research
Use the ore substratum, with Fe
2+Be the energy, 40 ℃ of temperature, pH1.4 (to BK02 in the same old way 35 ℃ of temperature, pH2.0) shaking culture under the condition, with incubation time to Fe
2+The oxidation ratio mapping, the result as shown in Figure 3.
As seen from Figure 3, when the UBK03 bacterial strain was cultivated under optimum growing condition, it can be with 5g/LFe in 19 hours
2+Complete oxidation, the control strain BK02 stronger with oxidation activity compares, its Fe
2+The time of complete oxidation has shifted to an earlier date more than 7 hour.Show that this bacterial strain has strong iron oxidation capacity.
Embodiment 5, UBK03 bacterial strain are to F
-Tolerance research
Use different F
-The ore substratum of concentration is with Fe
2+Be the energy, (to BK02 is 35 ℃ of temperature in the same old way, pH1.8) shaking culture under the condition is with different F at 40 ℃ of temperature, pH1.4
-Concentration is to Fe
2+The mapping of complete oxidation required time, the result as shown in Figure 4.
As seen from Figure 4, F
-To all influential (F of the growth of two bacterial strains
-When concentration is very low, two bacterial strain complete oxidation Fe
2+Required time is all about 24 hours), along with F
-Concentration increases, UBK03 bacterial strain complete oxidation Fe
2+Required time constantly increases, but amplification is little, at F
-Required time was 3.5 days when concentration was 1480mg/L, and the BK02 bacterial strain was not then seen growth within 15 days.Suzuki etc. studies show that, when pH2.3, the biological activity of acidophilia thiobacillus thiooxidans (Acidithiobacillus thiooxidans) was suppressed (Suzuki fully when fluoride concn was 1.9mg/L in the nutrient solution, I., Lee.D., MacKay B.etc., Appl.Environ.Microbiol., 65 (1999), 5163-5168.).Show that this bacterial strain is to F
-Have stronger tolerance, the biology that can be applicable to fluorine-containing ore leaches.
Embodiment 6, UBK03 bacterial strain soak the ore deposit capability study
Get liquid-solid ratio two parts of each 1000g of caproic acid uranium ore, insert respectively in the plastic tank of 2L tool inflation mechanism, and add [Fe respectively by 1: 1 (v/m)
3+UBK03 of]＞75% (pH 1.2-1.4) and BK02 (pH 1.6-2.0) nutrient solution 800ml, inflation is soaked under 35-40 ℃ and 30-35 ℃ of condition respectively, and every 24h changes liquid once.To leach fate to the mapping of accumulative total uranium leaching yield, the result as shown in Figure 5.
As seen from Figure 5, through 7 days, bacterial strain UBK03 accumulative total uranium leaching yield was up to 87.0%, and control strain BK02 accumulative total uranium leaching yield only is 64.7%.This shows that the UBK03 bacterial strain has the strong ore deposit ability of soaking, can be applicable in the leaching of sulfur compound ore biology.
Claims (6)
1. have a liking for sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290.
2. have a liking for the application of sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 in the oxidation ferro-compound.
3. application according to claim 2 is characterized in that: be that ferro-compound is oxidized to the solubility ferric iron.
4. have a liking for the application of sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 in the sulfur compound ore biology leaches.
5. application according to claim 4 is characterized in that: described biological the leaching is the uranium that extracts in the sulfur compound ore.
6. have a liking for the application of sour iron oxidation hook end spirobacteria (LeptospiriLLum ferriphiLum) UBK03 CGMCC № 1290 in fluorine-containing ore biology leaches.
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538540B (en) * | 2008-03-21 | 2011-05-25 | 中国黄金集团公司技术中心 | Composite mineral-leaching bacteria colony and use of same in biological metallurgy |
| CL2009001474A1 (en) * | 2008-06-29 | 2009-11-20 | Consolidated Nominees Pty Limited | Procedure to treat sulfurized mineral that includes bioleaching in solution of cl- ions with a consortium of mixed cultures of leptosprillium ferriphilum and sulfur, halophilic or halotolerant oxidizing microorganism, where the solution contains 1500-30000 ppm of cl- and also contains al, mg or na, at> 10 degrees and ph 1-3. |
| CN102002588B (en) * | 2010-12-30 | 2013-05-22 | 南华大学 | Bioleaching method-uranium leaching from fungus |
| CN103642732A (en) * | 2013-12-03 | 2014-03-19 | 东华理工大学 | Acidithiobacillus ferrooxidans strain with high resistance to fluorine |
| CN104130962B (en) * | 2014-07-31 | 2017-02-01 | 黑龙江八一农垦大学 | Leptospirillum ferriphilum strain and application in nickel sulfide ore leaching |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1056646C (en) * | 1997-07-04 | 2000-09-20 | 云南大姚铜矿 | Microbe and its application |
| WO2002070671A1 (en) * | 2001-03-01 | 2002-09-12 | Wisconsin Alumni Research Foundation | Acidophile archaeal organism ferroplasma acidarmanus, fer1 |
| CN1475586A (en) * | 2002-08-15 | 2004-02-18 | 北京有色金属研究总院 | Acid resistant mutagenic bacteria and its dump leaching process used for ore |
-
2005
- 2005-12-05 CN CNB2005101277417A patent/CN100368531C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1056646C (en) * | 1997-07-04 | 2000-09-20 | 云南大姚铜矿 | Microbe and its application |
| WO2002070671A1 (en) * | 2001-03-01 | 2002-09-12 | Wisconsin Alumni Research Foundation | Acidophile archaeal organism ferroplasma acidarmanus, fer1 |
| CN1475586A (en) * | 2002-08-15 | 2004-02-18 | 北京有色金属研究总院 | Acid resistant mutagenic bacteria and its dump leaching process used for ore |
Non-Patent Citations (2)
| Title |
|---|
| Molecular relationship between two groups of the genusleptospirillum and the finding that leptospirillum ferriphilum sp.nov. dominates south african commercial biooxidation tanksthat operate at 40℃. Nicolette J. Coram, Douglas E. Rawlings.Applied and Environmental Microbiology,Vol.68 No.2. 2002 * |
| 耐高矿化度浸矿菌的驯化及浸铀效果. 刘艳，刘亚洁，陈功新，郑志宏，李林.有色金属（冶炼部分），第5期. 2005 * |
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