CN104998899A - Microbial remediation method capable of mineralizing heavy metal ions based on glyphosate degradation - Google Patents
Microbial remediation method capable of mineralizing heavy metal ions based on glyphosate degradation Download PDFInfo
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- CN104998899A CN104998899A CN201510393343.3A CN201510393343A CN104998899A CN 104998899 A CN104998899 A CN 104998899A CN 201510393343 A CN201510393343 A CN 201510393343A CN 104998899 A CN104998899 A CN 104998899A
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- bacterium liquid
- glyphosate
- heavy metal
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- degradation
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Abstract
The invention discloses a microbial remediation method capable of mineralizing heavy metal ions based on glyphosate degradation. The microbial remediation method comprises the following steps of: separately inoculating bacillus subtilis, saccharomycetes and bacillus mucilaginosus into corresponding culture mediums for cultivation so as to prepare concentrated bacterial liquids with the cell concentration of 10<6>-10<7>/mL; uniformly mixing concentrated bacillus subtilis bacterial liquid, concentrated saccharomycete bacterial liquid and concentrated bacillus mucilaginosus bacterial liquid in proportion so as to prepare compound concentrated bacterial liquid; and adding the compound concentrated bacterial liquid to a solution containing glyphosate and heavy metal ions, uniformly stirring the mixture, allowing the mixture to stand for 4-5 hours to generate precipitates, and washing, filtering and drying the precipitates to obtain minerals which are phosphate and carbonate. The result shows that the compound concentrated bacterial liquid is capable of effectively degrading the glyphosate and mineralizing the heavy metals to form the stable phosphate and carbonate minerals.
Description
Technical field
The invention belongs to the interdisciplinary science technology of microbiological art and environmental science, relate to the microbial process that a kind of heavy metal ion mineralising based on glyphosate degradation is repaired.
Background technology
In recent years, heavy metal pollution is day by day serious, and contaminated area constantly expands, pollution level is constantly deepened, the extent of injury continues to increase, and the economic loss caused also increases year by year.Meanwhile, a large amount of uses of agricultural chemicals glyphosate, residue of glyphosate is accumulated year by year, causes the secondary pollution by heavy-metal contaminated soil, lake, there is huge potential safety hazard.The arable land that current China has 2,667 ten thousand hectares is subject to the residual pollution of heavy metal in various degree, glyphosate, and contaminated arable land accounts for 20% of national total cultivated area.Traditional restorative procedure mainly adopts physical method, chemical method, but energy consumption is high, investment is large, complicated operation, easily produce secondary pollution, is not suitable for large-area pollution amelioration.
Repairing method of microorganism utilizes naturally occurring multiple-microorganism in soil, is phosphate radical, carbon dioxide, water etc. by glyphosate degradation, wherein carbon dioxide by antimicrobial absorbent, be converted into carbonate; The phosphate radical that glyphosate degradation produces, carbonate are used for mineralized heavy metal ion, form stable mineral phosphate, carbonate; End product is mostly harmless, stable material, does not destroy the environment needed for plant growth, and pollutant can be made to remove from environment completely, and the processing time is short, and small investment, can not secondary pollution be produced, simple to operate.
Summary of the invention
Technical problem: the invention provides the microbial process that a kind of heavy metal ion mineralising based on glyphosate degradation is repaired, repairing method of microorganism cost is low, Be very effective, environmental friendliness, can not produce secondary pollution, and eco-compatibility is good.
Technical scheme: the microbial process that the heavy metal ion mineralising that the present invention is based on glyphosate degradation is repaired, comprises the following steps:
1.) respectively bacillus subtilis, saccharomycete, colloid bacillus cereus are seeded in corresponding culture medium and are cultivated, preparing cell concentration is 10
6~ 10
7the concentrated bacterium liquid of individual/mL;
2.) bacillus subtilis is concentrated bacterium liquid, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrate bacterium liquid mixing preparation compound concentrate bacterium liquid;
3.) compound is concentrated in the solution that bacterium liquid adds containing glyphosate, heavy metal ion, stir, glyphosate is degraded and is produced phosphate radical, small organic molecule under bacillus subtilis effect, the small organic molecule produced by glyphosate degradation fast degradation under saccharomycetic effect is carbon dioxide and water, and the carbon dioxide produced by glyphosate degradation absorbs, is converted into carbonate under colloid bacillus cereus effect;
4.) leave standstill 4 ~ 5 hours, after producing precipitation, sediment is fully washed, filter, dry, obtain mineral phosphate, carbonate.
Described compound concentrates that bacterium liquid concentrates bacterium liquid by bacillus subtilis, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrates bacterium liquid and prepares according to volume ratio 1 ﹕ 0.1 ~ 0.2 ﹕ 0.1 ~ 0.2.
Described heavy metal ion is zinc ion.
Beneficial effect: the present invention compared with prior art, has the following advantages:
1, compared with traditional physical method, chemical method, the initiative product of glyphosate degradation that utilizes carries out the mineralising of heavy metal ion, realizes the disposable reparation of two kinds of pollution sources simultaneously.
2, under the effect of bacillus subtilis, glyphosate degradation product is phosphate radical and straight chain small organic molecule; Straight chain small organic molecule is under saccharomycetic effect, and end product is carbon dioxide, water etc.; Carbon dioxide is absorbed by colloid bacillus cereus, is converted into carbonate; Carbonate, phosphate radical mineralized heavy metal ion, form mineral phosphate, carbonate.
3, the microbial process that adopts of the present invention, have the features such as efficient, environmental protection, the mineralogical property of formation is stable, durability is strong, and the carbon dioxide produced in process effectively can catch utilization, slows down greenhouse effects.
Accompanying drawing explanation
Fig. 1 is the mineralization product XRD figure adopting the method to be formed,
Fig. 2 is the mineralization product energy spectrogram adopting the method to be formed,
Fig. 3 a is that the mineralization product adopting the method to be formed amplifies 10000 times of SEM figure,
Fig. 3 b is that the mineralization product adopting the method to be formed amplifies 20000 times of SEM figure,
Fig. 3 c is that the mineralization product adopting the method to be formed amplifies 30000 times of SEM figure,
Fig. 3 d is that the mineralization product adopting the method to be formed amplifies 100000 times of SEM figure,
Fig. 4 is the mineralization product IR figure adopting the method to be formed.
Detailed description of the invention
Bacillus subtilis of the present invention, saccharomycete, colloid bacillus cereus all derive from Chinese industrial Culture Collection.
The microbial process that the heavy metal ion mineralising that the present invention is based on glyphosate degradation is repaired, method step is as follows:
(1) obtain bacillus subtilis and concentrate bacterium liquid: bacillus subtilis is inoculated in the culture medium solution after sterilizing, often liter of culture medium contains peptone 5 ~ 8g, beef leaching thing 3 ~ 5g, NaCl 5 ~ 6g, agar 15 ~ 20g, MgSO
40.4 ~ 0.6g, and control pH is 7 ~ 8, shaken cultivation 24h at 35 ~ 38 DEG C, obtain the bacterium liquid containing bacillus subtilis, at 4 DEG C after 6000 ~ 8000rpm high speed centrifugation, 10 ~ 15min, add deionized water after the culture medium nutriment of removing upper strata, in concentrated bacterium liquid, contained cell concentration is 10
6~ 10
7individual/mL.
(2) obtain saccharomycete and concentrate bacterium liquid: saccharomycete is inoculated in the culture medium solution after sterilizing, culture medium solution is 5 ° of brewer's wort 1000ml, containing agar 15 ~ 20g, shaken cultivation 24h at 28 ~ 30 DEG C, obtain containing saccharomycetic bacterium liquid, at 4 DEG C after 6000 ~ 8000rpm high speed centrifugation, 10 ~ 15min, add deionized water after the culture medium nutriment of removing upper strata, in concentrated bacterium liquid, contained cell concentration is 10
6~ 10
7individual/mL.
(3) obtain colloid bacillus cereus and concentrate bacterium liquid: colloid bacillus cereus is inoculated in the culture medium solution after sterilizing, often liter of culture medium contains sucrose 8 ~ 12g, Na
2hPO
412H
2o 2 ~ 3g, MgSO
40.4 ~ 0.6g, CaCO
30.5 ~ 1.5g, KCl 0.1 ~ 0.2g, (NH
4)
2sO
40.4 ~ 0.6g, and control pH is 7 ~ 8, shaken cultivation 24h at 30 ~ 35 DEG C, obtain the bacterium liquid containing colloid bacillus cereus, at 4 DEG C after 6000 ~ 8000rpm high speed centrifugation, 10 ~ 15min, add deionized water after the culture medium nutriment of removing upper strata, in concentrated bacterium liquid, contained cell concentration is 10
6~ 10
7individual/mL.
(4) compound concentrates bacterium liquid: bacillus subtilis is concentrated bacterium liquid, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrate bacterium liquid according to volume ratio 1 ﹕ 0.1 ~ 0.2 ﹕ 0.1 ~ 0.2 prepare compound concentrate bacterium liquid.
(5) compound is concentrated in the solution that bacterium liquid adds containing glyphosate, heavy metal ion, stir, make compound concentrate bacterium liquid in the solution dispersed, leave standstill 4 ~ 5 hours.After producing precipitation, sediment fully washed, avoids the impurity such as organic matter to be wrapped in sediment, impact analysis test result, filter, dry rear sample analysis.
(6) performance test: analysis result shows, compound concentrates bacterium liquid can effectively degradation of glyphosate, mineralising heavy metal, forms stable mineral phosphate, carbonate.
Fig. 1 shows the mineralising reparation adopting the present invention can realize the heavy metal ion based on glyphosate degradation, forms mineral phosphate, the carbonate of stable in properties.
Fig. 2 shows the EDAX results of mineralization product
Fig. 3 a, b, c, d show the microscopic pattern of different amplification mineralization product
Fig. 4 shows the functional group of mineralization product
Example:
(1) peptone 6g, beef leaching thing 4g is taken, NaCl 5g, agar 18g, MgSO
40.5g is dissolved in 1000ml deionized water, and control pH is that under 7,125 DEG C of high temperature, sterilizing is taken out to be cooled after 30 minutes; Be seeded to by bacillus subtilis in cooling culture medium solution, shaken cultivation at 37 DEG C, frequency of oscillation is 170r/min, incubation time 24h; By cultured bacterium liquid high speed centrifugation 10min, centrifuge speed is 8000rpm, and temperature is 4 DEG C, removes upper strata culture medium nutriment, adds deionized water 100mL and make concentrated bacterium liquid.
(2) taking agar 18g is dissolved in 1000ml5 ° of brewer's wort, and under 125 DEG C of high temperature, sterilizing is taken out to be cooled after 30 minutes; Be seeded to by saccharomycete in cooling culture medium solution, shaken cultivation at 30 DEG C, frequency of oscillation is 170r/min, incubation time 24h; By cultured bacterium liquid high speed centrifugation 10min, centrifuge speed is 8000rpm, and temperature is 4 DEG C, removes upper strata culture medium nutriment, adds deionized water 100mL and make concentrated bacterium liquid.
(3) sucrose 10g, Na is taken
2hPO
412H
2o 3g, MgSO
40.5g, CaCO
31g, KCl 0.1g, (NH
4)
2sO
40.4g is dissolved in 1000ml deionized water, and control pH is that under 7,125 DEG C of high temperature, sterilizing is taken out to be cooled after 30 minutes; Colloid bacillus cereus is inoculated in cooled culture medium solution, shaken cultivation at 35 DEG C, frequency of oscillation is 170r/min, incubation time 24h; By cultured bacterium liquid high speed centrifugation 10min, centrifuge speed is 8000rpm, and temperature is 4 DEG C, removes upper strata culture medium nutriment, adds deionized water 100mL and make concentrated bacterium liquid.
(4) bacillus subtilis is concentrated bacterium liquid, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrate bacterium liquid according to volume ratio 10ml, 1ml, 1ml mixing preparation compound concentrate bacterium liquid.
(5) 12ml compound is concentrated in the 100ml solution that bacterium liquid adds containing glyphosate, heavy metal zinc ion (glyphosate concentration 500mg/L, heavy metal zinc ion concentration 150mg/L), stir, make compound concentrate bacterium liquid in the solution dispersed, leave standstill 4 hours.After producing precipitation, by deionized water, three times washed to sediment, filter, dry rear sample analysis.Analysis result shows, compound concentrates bacterium liquid can effectively degradation of glyphosate, mineralising heavy metal, forms stable mineral Zn
3(PO
4)
24H
2o, Zn (CO
3) (OH)
6h
2o.
Claims (3)
1., based on the microbial process that the heavy metal ion mineralising of glyphosate degradation is repaired, it is characterized in that, the method comprises the following steps:
1) respectively bacillus subtilis, saccharomycete, colloid bacillus cereus are seeded in corresponding culture medium and are cultivated, preparing cell concentration is 10
6~ 10
7the concentrated bacterium liquid of individual/mL;
2) bacillus subtilis is concentrated bacterium liquid, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrate bacterium liquid mixing preparation compound concentrate bacterium liquid;
3) compound is concentrated in the solution that bacterium liquid adds containing glyphosate, heavy metal ion, stir, glyphosate is degraded and is produced phosphate radical, small organic molecule under bacillus subtilis effect, the small organic molecule produced by glyphosate degradation fast degradation under saccharomycetic effect is carbon dioxide and water, and the carbon dioxide produced by glyphosate degradation absorbs, is converted into carbonate under colloid bacillus cereus effect;
4) leave standstill 4 ~ 5 hours, after producing precipitation, sediment is fully washed, filter, dry, obtain mineral phosphate, carbonate.
2. the microbial process repaired of the heavy metal ion mineralising based on glyphosate degradation according to claim 1, it is characterized in that, described compound concentrates that bacterium liquid concentrates bacterium liquid by bacillus subtilis, saccharomycete concentrates bacterium liquid, colloid bacillus cereus concentrates bacterium liquid and prepares according to volume ratio 1 ﹕ 0.1 ~ 0.2 ﹕ 0.1 ~ 0.2.
3. the microbial process repaired of the heavy metal ion mineralising based on glyphosate degradation according to claim 1, it is characterized in that, described heavy metal ion is zinc ion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510393343.3A CN104998899B (en) | 2015-07-07 | 2015-07-07 | A kind of method that employing microbial degradation glyphosate is used for heavy metal ion mineralising |
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| CN201510393343.3A CN104998899B (en) | 2015-07-07 | 2015-07-07 | A kind of method that employing microbial degradation glyphosate is used for heavy metal ion mineralising |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058189A (en) * | 2017-06-05 | 2017-08-18 | 青岛农业大学 | Bacillus megaterium and its processing heavy metal glyphosate combined pollution in apply |
| CN107090413A (en) * | 2017-06-05 | 2017-08-25 | 青岛农业大学 | Glyphosate degradation bacterium and its processing heavy metal glyphosate combined pollution in apply |
| CN109433816A (en) * | 2018-11-26 | 2019-03-08 | 天津大学前沿技术研究院有限公司 | A kind of restorative procedure of glyphosate-heavy-metal composite pollution agricultural land soil |
| CN109626590A (en) * | 2018-11-30 | 2019-04-16 | 温州大学 | Bacillus subtilis bacterium fine oreization removes nickel plating solution method and knockout drum and test method |
| CN109772871A (en) * | 2019-01-29 | 2019-05-21 | 天津大学 | A method of glyphosate polluted farmland soil is repaired using subcritical low-temperature extraction technology |
| CN112646759A (en) * | 2020-07-17 | 2021-04-13 | 东南大学 | Mineralized microorganism increment preparation method |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108675586A (en) * | 2018-06-26 | 2018-10-19 | 温州大学 | The method of carbonate bacterium and the composite passivated heavy metal of phosphate bacterium and consolidation pollution electroplating sludge |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992019719A1 (en) * | 1991-05-03 | 1992-11-12 | Zeneca Limited | Glyphosate degrading bacteria |
| CN102757896A (en) * | 2012-07-10 | 2012-10-31 | 徐州奇缘生态农业发展有限公司 | Efficient microbial soil remediation agent |
-
2015
- 2015-07-07 CN CN201510393343.3A patent/CN104998899B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992019719A1 (en) * | 1991-05-03 | 1992-11-12 | Zeneca Limited | Glyphosate degrading bacteria |
| CN102757896A (en) * | 2012-07-10 | 2012-10-31 | 徐州奇缘生态农业发展有限公司 | Efficient microbial soil remediation agent |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058189A (en) * | 2017-06-05 | 2017-08-18 | 青岛农业大学 | Bacillus megaterium and its processing heavy metal glyphosate combined pollution in apply |
| CN107090413A (en) * | 2017-06-05 | 2017-08-25 | 青岛农业大学 | Glyphosate degradation bacterium and its processing heavy metal glyphosate combined pollution in apply |
| CN107058189B (en) * | 2017-06-05 | 2019-05-28 | 青岛农业大学 | Bacillus megaterium and its processing heavy metal-glyphosate combined pollution in apply |
| CN107090413B (en) * | 2017-06-05 | 2019-07-09 | 青岛农业大学 | Glyphosate degradation bacterium and its processing heavy metal-glyphosate combined pollution in apply |
| CN109433816A (en) * | 2018-11-26 | 2019-03-08 | 天津大学前沿技术研究院有限公司 | A kind of restorative procedure of glyphosate-heavy-metal composite pollution agricultural land soil |
| CN109626590A (en) * | 2018-11-30 | 2019-04-16 | 温州大学 | Bacillus subtilis bacterium fine oreization removes nickel plating solution method and knockout drum and test method |
| CN109772871A (en) * | 2019-01-29 | 2019-05-21 | 天津大学 | A method of glyphosate polluted farmland soil is repaired using subcritical low-temperature extraction technology |
| CN112646759A (en) * | 2020-07-17 | 2021-04-13 | 东南大学 | Mineralized microorganism increment preparation method |
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| CN104998899B (en) | 2017-03-15 |
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Address after: 210093 Nanjing University Science Park, 22 Hankou Road, Gulou District, Nanjing City, Jiangsu Province Patentee after: Southeast University Address before: 211189 No. 8 East Qimin Road, Xingcheng, Camera, Xigang Office, Qixia District, Nanjing, Jiangsu Province Patentee before: Southeast University |
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