CN108795991A - A kind of method that pyrite promotes dissimilatory iron reduction bacterium reduction chromium as electron shuttle carrier - Google Patents
A kind of method that pyrite promotes dissimilatory iron reduction bacterium reduction chromium as electron shuttle carrier Download PDFInfo
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- CN108795991A CN108795991A CN201810536768.9A CN201810536768A CN108795991A CN 108795991 A CN108795991 A CN 108795991A CN 201810536768 A CN201810536768 A CN 201810536768A CN 108795991 A CN108795991 A CN 108795991A
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- pyrite
- chromium
- iron reduction
- dissimilatory iron
- reduction bacterium
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide
Abstract
The invention discloses a kind of methods that pyrite restores chromium as electron shuttle carrier promotion dissimilatory iron reduction bacterium, belong to pollutant biologic treating technique field.The pyrite of 0.1-1.5g/L is added into the basic ion culture medium containing Cr (VI), is transferred in serum bottle, dissimilatory iron reduction bacterium is added thereto, is subsequently placed in 160-220rpm, 40-60h is protected from light in 25-35 DEG C of constant-temperature table.Pyrite is passivated in neutral conditions, to Cr (VI) substantially without removing rates, the treatment effect that dissimilatory iron reduction bacterium individually handles Cr (VI) is also bad, but when being combined dissimilatory iron reduction bacterium with pyrite reduction Cr (VI), the reduction rate of Cr (VI) is obviously improved, 2-5 times higher than dissimilatory iron reduction bacterium reduction Cr (VI) is used alone.Pyrite is derived from a wealth of sources with dissimilatory iron reduction bacterium and is easy to get, and is low, the efficient and easy to spread method of the cost of a processing pollution of chromium.
Description
Technical field
The invention belongs to pollutant biologic treating technique fields, and in particular to a kind of pyrite promotees as electron shuttle carrier
The method for restoring chromium into dissimilatory iron reduction bacterium.
Background technology
Many industrial productions such as stainless steel, leather, plating etc. are widely used chromium and generate a large amount of chromate waste waters, if processing
It is improper, huge pollution will be caused to environment, and threat is brought to human health.The common valence state of chromium is Cr (VI) and Cr
(Ⅲ).Cr (VI) is soluble good in water, strong toxicity, and important pollutant is classified as by Environmental Protection Agency USA.And Cr (III) can shape
At precipitation, macroscopic property is good, and toxicity is low, thus the mode of common processing pollution of chromium is to convert Cr (VI) to Cr (III).
Alienation metal reducing miroorganisms are under anaerobic that electron acceptor is breathed using high price variable valent metal, are had good
Good high price polluted heavy metals administer foreground.Pyrite, which derives from a wealth of sources, to be easy to get, its Fe (II) and S- ion pairs Cr
(VI) there is certain reducing power, but it is passivated in neutral conditions, cannot play its effect.It is combined the reduction of alienation metal
Bacterium and pyrite, electronics caused by alienation metal reducing miroorganisms can activate pyrite, ferrikinetics and sulfur cycle be formed, to Cr
(VI) it is restored, efficiency compares exclusive use alienation metal reducing miroorganisms reduction Cr (VI) and improves 2-5 times.
Pyrite used in the present invention is derived from a wealth of sources with dissimilatory iron reduction bacterium to be easy to get, and the two is used processing
Pollution of chromium is low, the efficient and easy to spread method of the cost of a processing pollution of chromium.
Invention content
The purpose of the present invention is the supplement to existing the relevant technologies, a kind of pyrite is provided and is promoted as electron shuttle carrier
The method that dissimilatory iron reduction bacterium restores chromium.
Technical scheme of the present invention is summarized as follows：
A kind of method that pyrite promotes dissimilatory iron reduction bacterium reduction chromium as electron shuttle carrier, includes the following steps：
(1) culture and enrichment of dissimilatory iron reduction bacterium：Dissimilatory iron reduction bacterium is accessed in LB liquid medium and is cultivated to logarithm
Growth period, culture solution is centrifuged, twice washing and settling flux, obtain bacteria suspension;
(2) bacteria suspension obtained in step (1) is accessed into the bases 100mL ion culture medium in anaerobic operation case, and to its
In add electron donor, pyrite and Cr (VI) solution;
(3) culture solution is taken to be filtered separation, detection in different time intervals;
It is bicarbonate buffer to be washed twice in above method step (1) with buffer solution used in settling flux, PIPES delays
Fliud flushing or HEPES buffer solution;
The pH value of LB culture mediums used in above method step, basic ion culture medium and buffer solution is 6.5-7.5;
Final concentration of 1 × 108-6 × 108/mL after basal medium is added in bacteria suspension in above method step (2);
Electron donor includes lactic acid, lactose, formic acid and sucrose, final concentration of 16-30mM in above method step (2);
The granularity of iron content sulphide ore is 325 mesh of > (44 μm) in above method step (2), and training is added after ultraviolet irradiation sterilizing
Support base;
Cr's (VI) adds a concentration of < 55mg/L in above method step (2);
In above method step (3) depending on reaction speed, 0.5-8h of interval is extracted reaction solution, and filter method is 0.22 μm
Membrane filtration.
Pyrite of the present invention promotes the method that dissimilatory iron reduction bacterium restores chromium as electron shuttle carrier, specifically includes following
Step：
(1) domestication and culture of dissimilatory iron reduction bacterium：Dissimilatory iron reduction bacterium is accessed into LB liquid medium (10g/L tryptoses
Peptone, 5g/L yeast extracts, 10g/L NaCl) in carry out enrichment culture, cultivate initial pH 7,30 DEG C of temperature, shaking table turns
Fast 170-200rpm.Culture collects culture solution and carries out centrifugation receipts under the conditions of 6000rpm, 8min, 20 DEG C to exponential phase
Collection.The thalline of collection is washed twice with bicarbonate buffer (2.5 g/L NaHCO3,2.5 g/L NaCl, pH 7)
Wash, after thalline be suspended in buffer solution obtain bacteria suspension;
(2) bacteria suspension in step (1) is added in basic ion culture medium under anaerobic.Basic ion culture medium
Composition is as follows：1.2 g/L(NH4)2SO4, 1 g/L K2HPO4, 0.4 g/L KH2PO4, 0.17 g/LNaHCO3, 0.25
G/L MgSO4 7H2O, 0.072 g/L CaCl2 2H2O, and trace element：1 mg/L FeSO4•7H2O, 0.2 mg/
LMnCO3, 1 mg/L CoCl2•6H2O, 0.28 mg/L ZnCl2, 3.5 mg/L H3BO3, 1 mg/L NiCl2•
6H2O, 0.05 mg/L CuSO4•5H2O, 22 mg/L Na2-EDTA.Final concentration of 4 × 108/mL of bacterial strain in system.
And sodium lactate, iron content sulphide ore and Cr (VI) solution is added into system under anaerobic, pyrite contains admittedly in system
Rate adds a concentration of < 55mg/L for 0.01 ﹪ -0.1 ﹪'s, Cr (VI);
(3) it is extracted reaction solution in different time intervals, separation is filtered using 0.22 μm of filter membrane, to detaching obtained reaction
Liquid carries out the detection of Cr (VI) content.
The advantage of the present invention is as follows：
A kind of pyrite disclosed by the invention as electron shuttle carrier promote dissimilatory iron reduction bacterium restore chromium method have with
Lower advantage：Alienation metal reducing miroorganisms used in the present invention are derived from a wealth of sources with pyrite to be easy to get, and the two is used place
Pollution of chromium is managed, is low, the efficient and easy to spread method of the cost of a processing pollution of chromium.
Description of the drawings
Fig. 1 is that Shewanella is under pyrite assistance in the present invention, to the reduction effect that initial concentration is 15.6mg/LCr (VI)
Figure;
Fig. 2 is that Shewanella is under pyrite assistance in the present invention, to the reduction effect figure that initial concentration is 26mg/LCr (VI);
Fig. 3 is that Shewanella is under pyrite assistance in the present invention, to the reduction effect figure that initial concentration is 36mg/LCr (VI);
Fig. 4 is that Shewanella is under pyrite assistance in the present invention, to the reduction effect figure that initial concentration is 50mg/LCr (VI);
Fig. 5 is that Shewanella restores 26mg/L Cr (VI) and imitate in the case where the pyrite of 1g/L and 0.1g/L is assisted in the present invention
Fruit is schemed;
Fig. 6 is that Shewanella is in the case where the pyrite of 0. 1g/L is assisted in the present invention, to the chromium reduction effect figure of chromium slag soil.
Specific implementation mode
Following embodiment or embodiment are intended to further illustrate the present invention, rather than limitation of the invention.Following reality
Example 1,2,3 is applied to be handled with the 1000mg/L potassium bichromate solutions manually prepared.
Embodiment 1：
This example the method mainly includes the following steps that：
(1) enrichment culture will be carried out in Shewanella oneidensis MR-1 access LB liquid mediums, culture is initial
PH 7,30 DEG C of temperature, shaking speed 170rpm.It cultivates after 12h to exponential phase, collects culture solution in 6000rpm, 8min,
It is collected by centrifugation under the conditions of 20 DEG C.The thalline of collection is washed twice with bicarbonate buffer, after thalline is suspended
Bacteria suspension is obtained in buffer solution;
(2) 100mL basal mediums are added in bacteria suspension in step (1) in anaerobic operation case, the bacterial strain in system is dense eventually
Degree is 4 × 108/mL.And 20mM sodium lactates, 0.01g pyrite is added into system in anaerobic operation case, and add respectively
Cr (VI) solution (15.6,25,36,50 mg/L) of various concentration.Blank control group is set simultaneously：Basal medium+Cr
(VI) solution；Biological control group：Basal medium+Shewanella oneidensis MR-1+ Cr (VI) solution；Chemistry is right
According to group：Basal medium+pyrite+Cr (VI) solution.System, which is placed in serum bottle, carries out Anaerobic Treatment, and anaerobism bottle is placed in 30
DEG C, cultivate in 200 rpm shaking tables;
(3) 0.5mL culture solutions are taken out at interval in different times, and separation is filtered with 0.22 μm of filter membrane, are obtained to separation
Culture solution carry out the detection of Cr (VI) content, testing result is as shown in Figs 1-4;
(4) result：In neutral conditions, pyrite is passivated, the effect of no reduction Cr (VI).But Shewanella
The electron activable pyrite of oneidensis MR-1 releases forms Fe (II)/Fe (III) cycles and sulphur member in mineral surfaces
Element recycles to promote the reduction of Cr (VI), in the system, goes back iron ore and serves as electron shuttle carrier, promotes the reduction of Cr (VI), also
Former rate is far above exclusive use Shewanella oneidensis MR-1 removal efficiency of chrome, in four chromium concn gradient tests
The conclusion is obtained for confirmation.
Embodiment 2：
This example the method mainly includes the following steps that：
(1) enrichment culture will be carried out in Shewanella oneidensis MR-1 access LB liquid mediums, culture is initial
PH 7,30 DEG C of temperature, shaking speed 170rpm.It cultivates after 12h to exponential phase, collects culture solution in 6000rpm, 8min,
It is collected by centrifugation under the conditions of 20 DEG C.The thalline of collection is washed twice with bicarbonate buffer, after thalline is suspended
Bacteria suspension is obtained in buffer solution;
(2) 100mL basal mediums are added in bacteria suspension in step (1) in anaerobic operation case, the bacterial strain in system is dense eventually
Degree is 4 × 108/mL.And Cr (VI) solution of 20mM sodium lactates, 25 mg/L is added into system in anaerobic operation case, with
And the pyrite (0.1g/L, 1g/L) of various concentration.Blank control group is set simultaneously：Basal medium+Cr (VI) solution；It is raw
Object control group：Basal medium+Shewanella oneidensis MR-1+ Cr (VI) solution；Chemical reference group：Basis training
Support base+pyrite (0.1g/L, 1g/L)+Cr (VI) solution.System, which is placed in serum bottle, carries out Anaerobic Treatment, and anaerobism bottle is placed in
30 DEG C, cultivate in 200 rpm shaking tables;
(3) 0.5mL culture solutions are taken out at interval in different times, and separation is filtered with 0.22 μm of filter membrane, are obtained to separation
Culture solution carry out Cr (VI) content detection.Testing result is as shown in Figure 5;
(4) result：In neutral conditions, pyrite is passivated, the effect of no reduction of hexavalent chromium.But Shewanella
The electron activable pyrite of oneidensis MR-1 releases forms Fe (II)/Fe (III) cycles and sulphur member in mineral surfaces
Element cycle is come the pyrite concentration difference for promoting the reduction of Cr (VI), and adding, and chromium rate of reduction is different, 1g/L pyrite
Efficiency is promoted to be far above the promotion efficiency of 0.1g/L pyrite.As seen from the figure, the efficiency that pyrite reduction chromium is added in system is remote
Higher than the efficiency that exclusive use Shewanella removes chromium.
Embodiment 3：
This example the method mainly includes the following steps that：
(1) enrichment culture will be carried out in Shewanella oneidensis MR-1 access LB liquid mediums, cultivates initial pH
7,30 DEG C of temperature, shaking speed 170rpm.To exponential phase after culture 12h, culture solution is collected in 6000rpm, 8min, 20
It is collected by centrifugation under the conditions of DEG C.The thalline of collection is washed twice with bicarbonate buffer, after thalline is suspended in
Bacteria suspension is obtained in buffer solution.The chromium slag soil that chromium content is 530 mg/kg is air-dried, it is levigate, cross 0.25 mm soil sieves;
(2) 100mL basal mediums are added in bacteria suspension in step (1) in anaerobic operation case, the bacterial strain in system is dense eventually
Degree is 4 × 108/mL.And 20mM sodium lactates, 6 g chromium slags soil and 0.01g Huang iron is added into system in anaerobic operation case
Mine.Blank control group is set simultaneously：Basal medium+chromium slag soil；Biological control group：Basal medium+Shewanella
Oneidensis MR-1+ chromium slag soil.Chemical reference group：Basal medium+pyrite (0.1g/L)+chromium slag soil.System
It is placed in serum bottle and carries out Anaerobic Treatment, anaerobism bottle is placed in 30 DEG C, cultivates in 200 rpm shaking tables;
(3) 0.5mL culture solutions are taken out at interval in different times, and separation is filtered with 0.22 μm of filter membrane, are obtained to separation
Culture solution carry out Cr (VI) content detection.Testing result is as shown in Figure 6;
(4) result：As seen from the figure, in the system, treatment effect of the pyrite for Cr VI in chromium slag soil is used alone
Bad, the effect that Shewanella oneidensis MR-1 are used alone is also general, but Shewanella oneidensis
MR-1 has preferable reduction rate with the system of combining of pyrite to the Cr VI in chromium slag soil, than Shewanella is used alone
The also commercial weight of oneidensis MR-1 reduction Cr (VI) is about 2 times high.The system has good answer for the processing of chromium slag soil
Use foreground.
Claims (8)
1. a kind of pyrite promotes the method that dissimilatory iron reduction bacterium restores chromium as electron shuttle carrier, which is characterized in that including
Following steps：
(1) culture and enrichment of dissimilatory iron reduction bacterium：Dissimilatory iron reduction bacterium is accessed culture to logarithm in LB liquid medium to give birth to
For a long time, thalline is collected, bacteria suspension is obtained;
(2) bacteria suspension obtained in step (1) in anaerobic operation case is connected to basic ion culture medium, and added thereto
Electron donor, pyrite and Cr (VI) solution;
(3) culture solution is taken to be filtered separation, detection in different time intervals.
2. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that collect in step (1) buffer solution used in thalline be bicarbonate buffer, PIPES buffer solutions or
HEPES buffer solution.
3. a kind of pyrite according to claim 1 or 2 promotes the reduction of dissimilatory iron reduction bacterium as electron shuttle carrier
The method of chromium, which is characterized in that the pH value of LB culture mediums used, basic ion culture medium and buffer solution is 6.5-7.5.
4. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that the bacterial concentration in step (2) after the neutral basic ion culture medium of bacteria suspension addition is 1 × 108-6
×108A/mL.
5. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that electron donor includes lactic acid, lactose, formic acid and sucrose, final concentration of 16-30mM in step (2).
6. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that in step (2) granularity of pyrite be 325 mesh of > (44 μm), ultraviolet irradiation sterilizing after add into
Culture medium.
7. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that Cr's (VI) adds a concentration of < 55mg/L in step (2).
8. promoting dissimilatory iron reduction bacterium to restore chromium as electron shuttle carrier according to a kind of pyrite described in claim 1
Method, which is characterized in that in step (3) depending on reaction speed, 0.5-8h of interval is extracted reaction solution, with 0.22 μm of filter membrane
It is filtered separation.
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Cited By (9)
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| CN109574407A (en) * | 2018-12-29 | 2019-04-05 | 中南民族大学 | A kind of method that bioanalysis efficiently removes butyl xanthate and heavy metal Cr (VI) in flotation waste water |
| CN109851059A (en) * | 2019-01-24 | 2019-06-07 | 天津科技大学 | The method for preparing biological magnetic iron ore removal heavy metal chromium using dissimilatory iron reduction bacterium |
| CN110143673A (en) * | 2019-05-24 | 2019-08-20 | 中国地质大学（北京） | A kind of method of the fixed Cr VI of the microorganism that iron ore is supported |
| CN111659729A (en) * | 2020-05-27 | 2020-09-15 | 北京化工大学 | Method for restoring chromium-polluted soil by Shewanella |
| CN112157119A (en) * | 2020-08-12 | 2021-01-01 | 中南大学 | Restoration method for controlling microorganisms to reduce heavy metals by iron-containing minerals |
| CN112608922A (en) * | 2020-12-30 | 2021-04-06 | 中国科学技术大学 | Method for regulating and controlling electron current of dissimilatory metal reducing bacteria |
| CN114921388A (en) * | 2022-04-11 | 2022-08-19 | 河北工业大学 | Iron reducing flora and application thereof |
| CN115490338A (en) * | 2022-10-21 | 2022-12-20 | 太原理工大学 | Method for treating waste water containing perrhenate |
| CN115504572A (en) * | 2022-10-21 | 2022-12-23 | 太原理工大学 | Method for treating wastewater containing pertechnetate |
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| CN109574407A (en) * | 2018-12-29 | 2019-04-05 | 中南民族大学 | A kind of method that bioanalysis efficiently removes butyl xanthate and heavy metal Cr (VI) in flotation waste water |
| CN109574407B (en) * | 2018-12-29 | 2021-09-03 | 中南民族大学 | Method for efficiently removing butyl xanthate and heavy metals Cr (VI) in flotation wastewater by biological method |
| CN109851059A (en) * | 2019-01-24 | 2019-06-07 | 天津科技大学 | The method for preparing biological magnetic iron ore removal heavy metal chromium using dissimilatory iron reduction bacterium |
| CN109851059B (en) * | 2019-01-24 | 2022-04-08 | 天津科技大学 | Method for removing heavy metal chromium from biological magnetite prepared by using dissimilatory iron reducing bacteria |
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| CN112157119A (en) * | 2020-08-12 | 2021-01-01 | 中南大学 | Restoration method for controlling microorganisms to reduce heavy metals by iron-containing minerals |
| CN112608922A (en) * | 2020-12-30 | 2021-04-06 | 中国科学技术大学 | Method for regulating and controlling electron current of dissimilatory metal reducing bacteria |
| CN114921388A (en) * | 2022-04-11 | 2022-08-19 | 河北工业大学 | Iron reducing flora and application thereof |
| CN114921388B (en) * | 2022-04-11 | 2023-09-15 | 河北工业大学 | Iron-reducing flora and application thereof |
| CN115490338A (en) * | 2022-10-21 | 2022-12-20 | 太原理工大学 | Method for treating waste water containing perrhenate |
| CN115504572A (en) * | 2022-10-21 | 2022-12-23 | 太原理工大学 | Method for treating wastewater containing pertechnetate |
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Application publication date: 20181113 |