CN109851059A - The method for preparing biological magnetic iron ore removal heavy metal chromium using dissimilatory iron reduction bacterium - Google Patents
The method for preparing biological magnetic iron ore removal heavy metal chromium using dissimilatory iron reduction bacterium Download PDFInfo
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Abstract
The invention belongs to field of environment microorganism, and in particular to a method of biological magnetic iron ore removal heavy metal Cr (VI) is prepared using dissimilatory iron reduction bacterium.The present invention prepares magnetic iron ore using dissimilatory iron reduction bacterium, can both reduce influence of the variation to removal efficiency of heavy metals of condition of culture, and the generation that also can prevent heavy metal from poisoning to bacterial strain itself improves heavy metal Cr (VI) removal efficiency；Acid condition go down removing heavy metals Cr (VI) when, the removal rate of heavy metal Cr (VI), up to 100% can be effectively improved；Biological magnetic iron ore overcomes heavy metal and inhibits to the murder by poisoning that strain cell is grown, and has the advantages that processing Cr (VI) is highly concentrated.
Description
Technical field:
The invention belongs to field of environment microorganism, and in particular to a kind of to prepare biological magnetic iron ore using dissimilatory iron reduction bacterium
The method for removing heavy metal Cr (VI).
Background technique:
Pollution of chromium is to be only second to second of heavy metal pollution type of lead contamination.Usually there are two types of valence state Cr for heavy metal chromium
(VI) and Cr (III), wherein Cr (VI) is soluble easily in water, and oxidisability is strong, toxicity be Cr (III) toxicity 100 times (Gao little Peng,
It opens and owes, Xu Ping waits progress [J] the microbiology of micro-reduction Cr (VI) to be notified to, 2008,35 (5): 820-
824.).Cr (VI) is reduced to Cr (III), is the key technology means for solving heavy metal Cr (VI) pollution.
Chemical method and the physical method of comparing directly or indirectly restore Cr (VI) using microorganism except chromium, have in-situ treatment,
The advantages that at low cost and without secondary pollution.
Dissimilatory iron reduction bacterium be it is a kind of with Fe (III) for electron acceptor, and Fe (III) is reduced into micro- life of Fe (II)
Object general name.Dissimilatory reduction Fe (III) process of microorganism can influence the valence state form of some heavy metal elements, conversion pollution
Object achievees the purpose that administer heavy metal pollution.Raw he etc. is using Cr (VI) in dissimilatory iron reduction removal of bacteria waste water, and principle is just
It is that Cr (VI) is reduced to Cr (III) by the Fe (II) that generates during dissimilatory iron reduction of electron acceptor of the bacterium, while Fe
(II) be reoxidized into Fe (III) (raw to congratulate, Yu Jinqiu, Liu Dengfeng, etc..Prosparol strengthens Cr (VI) in underground water
Biogeochemistry reduction research [J] China Environmental Science, 2015,35 (6): 1693-1699.).But the culture of microorganism
Condition is stringenter, limits it in the scale application process in heavy metals removal field.
Therefore, a kind of method for removing heavy metal Cr (VI) using biology-abiotic reduction process is developed, is passed for breaking through
The high energy consumption of the culture limitation and physico-chemical process of system microorganism, improves heavy metal Cr (VI) removal efficiency, is of great significance.
Summary of the invention:
Biological magnetic iron ore removal heavy metal Cr is prepared using dissimilatory iron reduction bacterium the object of the present invention is to provide a kind of
(VI) method provides technical support to administer heavy metal pollution.
Technical solution of the present invention includes the following steps:
(1) dissimilatory iron reduction bacterium the culture of dissimilatory iron reduction bacterium: is inoculated in basal medium by 1-5% inoculum concentration
In, under anaerobic, 25-35 DEG C, 100-150r/min cultivates 2-5d；
Basic media components (g/L): glucose 5-15, yeast extract 1-3, tryptone 0.5-2, NaCl 20-
40, K2HPO41-2, NaHCO31-2, Fe (OH)3300-500mg/L, remaining is water, pH 7.0 ± 0.2；
Preferably, the NaHCO3First configuration concentration is that the solution of 50g/L is added using filtration method degerming by 20-40mL/L
Add to basal medium；
Preferably, the dissimilatory iron reduction bacterium is Klebsiella sp.KB52；
Preferably, incubation time 4-5d；
(2) preparation of biological magnetic iron ore: after culture 2-5d, being aseptically collected by centrifugation insoluble matter for bacterium solution, will not
Molten object dries to obtain biological magnetic iron ore；
Preferably, after cultivating 2-5d, bacterium solution is transferred to sterile centrifugation tube and is separated by solid-liquid separation, centrifugation (9000r/min,
1min).Insoluble matter is gathered in centrifugation bottom of the tube in thallus and culture solution, in thick, is transferred to culture dish, drying (100 DEG C,
5h), biological magnetic iron ore sample is in powdered.
(3) Cr (VI) is removed: Cr (VI) 10-60:1 in mass ratio in biological magnetic iron ore and liquid to be processed is added to
In liquid to be processed containing concentration not higher than 80mg/L Cr (VI), adjusting pH is 2.0-8.0, after reacting 6-12h, Cr's (VI)
Removal rate can reach 100%；
Preferably, Cr (VI) mass ratio in biological magnetic iron ore and liquid to be processed is 25-50:1；
It is highly preferred that Cr (VI) mass ratio in biological magnetic iron ore and liquid to be processed is 50:1；
Preferably, reaction pH is 2.0-6.0；
It is highly preferred that reaction pH is 2.0-5.0；
Preferably, reaction time 8-10h；
It is highly preferred that the reaction time is 10h；
Preferably, the concentration of Cr (VI) is not higher than 60mg/L in liquid to be processed；
More preferably, the concentration of Cr (VI) is not higher than 40mg/L in liquid to be processed.
The utility model has the advantages that
1, compared with commonly throwing bacterium method, magnetic iron ore is prepared using dissimilatory iron reduction bacterium, can both reduce condition of culture
Change the influence to removal efficiency of heavy metals, the generation that also can prevent heavy metal from poisoning to bacterial strain itself improves heavy metal Cr
(VI) removal efficiency.In addition, by containing biological magnetic iron ore is directly added in Cr (VI) waste water, it can be to avoid microculture mistake
Superfluous organic culture solution remains in waste water in journey, increases subsequent wastewater treatment cost.
2, the present invention using dissimilatory iron reduction bacterium Klebsiella sp.KB52 preparation magnetic iron ore in pH 7.0 hereinafter,
I.e. acid condition go down removing heavy metals Cr (VI) when, compared in neutral conditions, heavy metal Cr (VI) can be effectively improved
Removal rate, up to 100%.
3, method provided by the invention, compared with using traditional microbiological processing heavy metal Cr (VI), biological magnetic iron ore gram
It has taken heavy metal to inhibit the murder by poisoning that strain cell is grown, has had the advantages that processing Cr (VI) is highly concentrated.Based on biological magnetic iron ore
Partial size is small, the big feature of response area, it can be achieved that Cr (VI) high removal rate, can be used for (VI) containing Cr within the shorter release time
Waste water emergency processing.
Detailed description of the invention:
The scanning electron microscope (SEM) of Fig. 1 biology magnetic iron ore observes figure
Wherein, (a) (c) (e) is the Fe (OH) without biological treatment3SEM figure under different multiples；(b)(d)(f)
Fe (OH) is utilized for bacterial strain KB523SEM figure of the biological magnetic iron ore of synthesis under different multiples；
Influence of Fig. 2 incubation time to Cr (VI) removal rate；
Influence of Fig. 3 initial ph value to Cr (VI) removal rate；
Fig. 4 biology magnetic iron ore injected volume influences Cr (VI) removal rate；
Fig. 5 biology magnetic iron ore release time influences Cr (VI) removal rate；
Fig. 6 various concentration Cr (VI) influences removal rate.
Specific embodiment:
In order to which the objects, technical solutions and advantages of this patent are more clearly understood, below in conjunction with specific embodiment to this
Patent is further elaborated.It should be appreciated that specific embodiment described herein is only used to explain this patent, not
For limiting the present invention.
Dissimilatory iron reduction bacterium Klebsiella sp.KB52 used in the present invention is one plant and is isolated from Sediments of Bohai Sea
Dissimilatory iron reduction bacterium, bacterial strain GenBank KM233642, in scientific and technical literature, (Liu Hongyan, Wang Hongyu, Xie Lixia wait
One plant of iron-reducing bacteria separation and Fe (III) reducing property [J] Marine Sciences, 2016,40 (3): 65- in marine sediment
70.) it is announced in, the public can obtain from University Of Science and Technology Of Tianjin ocean and Environmental Studies Institute.
The method of Cr (VI) removal rate is measured in following embodiment are as follows:
1.0mL solution to be measured is taken, is centrifuged (4500rpm, 5min), supernatant 0.5mL is collected, with 0.2mL H2SO4
(0.68mol L-1), the mixing of 0.2mL diphenylcarbazide, then plus distilled water is settled to 5mL, and reaction solution is measured at 540nm
Trap calculates Cr in solution (VI) concentration using standard curve.
The calculating of Cr (VI) removal rate is according to formula: removal rate=(Ci-Cf)/Ci × 100%, wherein Ci represents initial Cr
(VI) concentration (mg/L), Cf terminate Cr (VI) concentration (mg/L).
Specific embodiment the present invention will be further explained explanation will be passed through below.
Embodiment 1: the physical features of biological magnetic iron ore
(1) dissimilatory iron reduction bacterium Klebsiella sp.KB52 is inoculated in 150mL anaerobism bottle by 1% inoculum concentration and (contains
100mL basal medium), inflated with nitrogen 5min, guarantee anaerobic culture environment, 30 DEG C, 120r/min, shaking table culture；
Basic media components (g/L): glucose 10, yeast extract 2.0, tryptone 1.0, NaCl 30.0,
K2HPO41.5, NaHCO31.5, Fe (OH)3400mg/L, pH 7.0 ± 0.2；NaHCO3Configuration concentration is the molten of 50g/L first
Liquid is added to basal medium by 30mL/L using filtration method degerming；
Above-mentioned with artificial synthesized Fe (OH)3For electron acceptor, glucose is the redox cultivating system of electron donor
In, bacterial strain KB52 can carry out dissimilatory iron reduction using insolubility Fe (III) form, and accumulation generates Fe (II) after cultivating 96h
Concentration is 37.23 ± 4.12mg/L.All do not have in culture solution in identical incubation time for the blank group of not inoculating strain
Determine Fe (II) concentration.This shows to accumulate Fe (II) concentration in culture solution, different for electron acceptor with Fe (III) by bacterial strain KB52
Change to accumulate in iron reduction process and generate.
(2) preparation of biological magnetic iron ore: after culture 96h, bacterium solution is transferred to sterile centrifugation tube and is separated by solid-liquid separation, centrifugation
(9000r/min, 1min)；Insoluble matter is gathered in centrifugation bottom of the tube in thallus and culture solution, in thick, be transferred to culture dish, dry
Dry (100 DEG C, 5h), sample are obtained containing Fe (III) and Fe (II) biology magnetic iron ore in powdered.
It is observed using pattern of the scanning electron microscope (SEM) to the biological magnetic iron ore of synthesis, (a) (c) in Fig. 1
(e) for without the Fe (OH) of biological treatment3SEM figure under different multiples, (b) (d) (f) is that bacterial strain KB52 utilizes Fe (OH)3
SEM figure of the biological magnetic iron ore of synthesis under different multiples.From Fig. 1 (a) (b) as can be seen that when amplification factor is 800,
Random without the sample shape of biological treatment, surface is more smooth, and the sample Jing Guo biological treatment equally present it is random
Shape, surface is rougher.When they are amplified to 1.50k (c, d) times, the institute after biological treatment can be significantly found out
The hole that there are many biological magnetic iron ore surfaces of formation, increases its surface area, and then enhance the suction-operated to heavy metal.
Amplification factor be 6.5k (e, f) when, more can significantly find out the sample after biological treatment compared with untreated, surface
It is coarse there are many attachment and having apparent hole, see Fig. 1.
Influence of 2 incubation time of embodiment to Cr (VI) removal rate
(1) dissimilatory iron reduction bacterium Klebsiella sp.KB52 is inoculated in 150mL anaerobism bottle by 1% inoculum concentration and (contains
100mL basal medium), inflated with nitrogen 5min, guarantee anaerobic culture environment, 30 DEG C, 120r/min, shaking table culture；
Basic media components (g/L): glucose 10, yeast extract 2.0, tryptone 1.0, NaCl 30.0,
K2HPO41.5, NaHCO31.5, Fe (OH)3400mg/L, pH 7.0 ± 0.2；NaHCO3Configuration concentration is the molten of 50g/L first
Liquid is added to basal medium by 30mL/L using filtration method degerming；
During incubation is shone, during using the identical mode of embodiment 1 to collect bacterial strain KB52 dissimilatory iron reduction for 24 hours
The biological magnetic iron ore sediment of formation, is dried into powder, and biological magnetic iron ore is added in 2g/L ratio containing 40mg/L Cr
(VI) in liquid to be processed, pH is adjusted after 5.0,10h, to measure the removal efficiency of Cr (VI) (not add magnetic iron ore as control
Group), as a result such as Fig. 2.
As shown in Fig. 2, bacterial strain KB52 growth was slower at first 2 days of culture, accumulation generation Fe (II) concentration is low,
This culture period collects thallus, prepares biological magnetic iron ore, also there was only 38% and 44% to Cr (VI) removal efficiency.When culture 3 days
When, strain cell density obviously increases, and shows that cell grows into logarithmic growth phase.The biological magnetic iron ore pair of this culture period preparation
Cr (VI) removal efficiency can reach 80%.When culture enters the 4th and 5 day, strain cell density reaches highest, and Cr (VI) is gone
Except rate nearly reaches 100%, show this coated magnetic iron ore of film institute by synthesizing in microbial cell, to heavy metal Cr
(VI)) with the feature of high reaction activity.The biological magnetic iron ore of synthesis has significant Cr (VI) removal efficiency.
Influence of 3 pH value of embodiment to Cr (VI) removal rate
When bacterial strain KB52 being cultivated 4 days in redox cultivating system by 1 method of embodiment, centrifugation, precipitating drying, system
Standby biology magnetic iron ore.Biological magnetic iron ore is added to the different initial pH values containing 40mg/L Cr (VI) in 2g/L ratio to wait locating
It manages in liquid, handles 10h, as a result such as Fig. 3.
From the figure 3, it may be seen that the biological magnetic iron ore prepared by bacterial strain KB52 is in different pH value to Cr (VI) removal efficiency
It has differences.When pH value < 6.0, biological magnetic iron ore is relatively high to the removal efficiency of Cr (VI), and removal effect is obvious.Cr(VI)
When solution ph is 2.0,3.0,4.0 and 5.0, biological magnetic iron ore reaches 100% to the removal rate of Cr (VI)；Cr (VI) pH value of solution
When value is 6.0, biological magnetic iron ore is 78% to Cr (VI) removal rate, and when pH value is 7.0 and 8.0, biological magnetic iron ore is to Cr
(VI) removal rate is 43% and 37% respectively.As it can be seen that the biological magnetic iron ore prepared by bacterial strain KB52 is acid containing Cr in processing
(VI) clear superiority is shown in terms of waste water.
Microorganism is more sensitive to the variation of pH value in culture solution, and when pH value is less than 6.0, microbe survival can meet with huge
Big challenge is difficult to grow.Therefore, Cr (VI) waste water is contained for acidity, is using traditional microbiological processing heavy metal Cr (VI)
Difficult to realize, the present invention has well solved this problem.
Influence of the 4 magnetic iron ore injected volume of embodiment to Cr (VI) removal efficiency
It is 40mg/L, pH value 5.0, the same embodiment of other conditions that setting Cr (VI) solution ph, which is 5.0, Cr (VI) concentration,
1, add the biological magnetic iron ore of different injected volumes respectively, influence of the analysis of magnetic iron ore injected volume to Cr (VI) removal efficiency, as a result
See Fig. 4.
From fig. 4, it can be seen that increasing with biological magnetic iron ore injected volume, the removal efficiency of Cr (VI) is obvious in Cr (VI) solution
It improves, i.e., when injected volume is 0.5g/L, 1.0g/L and 2g/L, Cr (VI) removal rate is increased to 100% from 38%, 86%.When after
Continuous to improve magnetic iron ore injected volume (4g/L, 6g/L and 8g/L), the variation of Cr (VI) removal rate less, has reached 100%.
Influence of the biological magnetic iron ore release time of embodiment 5 to Cr (VI) removal rate
Other conditions are added to containing 40mg/L Cr (VI) in 2g/L ratio wait locate with embodiment 1, by biological magnetic iron ore
It manages in liquid, pH value 5.0, the release time that biological magnetic iron ore is thrown to Cr (VI) solution is set as 12h, and interval 2h sampling is surveyed
Cr in solution (VI) removal rate is determined, as a result such as Fig. 5.
From fig. 5, it can be seen that the removal effect of Cr (VI) has been shown in Cr (VI) solution when biological magnetic iron ore launches 4h
Existing, Cr (VI) removal rate is 34%；When biological magnetic iron ore launches 8h, 80% can reach to Cr (VI) removal rate；Launch 10h
When, biological magnetic iron ore has reached 100% to Cr (VI) removal rate.
Biological magnetic iron ore mainly utilizes it to accumulate during the preparation process, and generation Fe (II) restores Cr (VI) and bacterial strain is thin
The rich magnetic absorption Cr (VI) of mucilage secretion and magnetic iron ore itself reaches dual removal Cr (VI) purpose.By biological magnet
Mine is directly launched in Cr (VI) solution, mainly happens is that electronation and physisorption.Therefore, bacterium method is thrown with common
It compares, biological magnetic iron ore can be such that the removal time of Cr (VI) is obviously shortened.
The biological magnetic iron ore of embodiment 6 is to various concentration Cr (VI) removal rate
It is 5mg/L, 10mg/L, 20mg/L, 40mg/L, 60mg/L and 80mg/L, Cr respectively that Cr (VI) solution concentration, which is arranged,
(VI) solution ph is 5.0, and biological magnetic iron ore injected volume is 2g/L, and release time 10h, other conditions are with embodiment 1, as a result such as
Fig. 6.
It will be appreciated from fig. 6 that biological magnetic iron ore all shows significant removal effect to various concentration Cr (VI).When Cr (VI) is molten
Liquid concentration is relatively low, i.e. when 5mg/L and 10mg/L, biological magnetic iron ore is 100% to Cr (VI) removal efficiency；When Cr (VI) solution
When concentration is improved to 20mg/L and 40mg/L, biological magnetic iron ore remains to reach 100% to Cr (VI) removal efficiency.
And Cr (VI) is used as a kind of heavy metal contaminants, and it is serious to inhibit cell growth, microorganism generation is poisoned and inhibits to make
With big, many microorganisms are less than 20mg/L to Cr (VI) concentration tolerance range.When Cr (VI) solution concentration is 60mg/L, biology
Magnetic iron ore is still able to maintain 80% to Cr (VI) removal efficiency.As it can be seen that being removed in water body by biological magnetic iron ore prepared by bacterial strain KB52
Cr (VI) contains Cr (VI) waste water especially for wide scope and high concentration, has removal efficiency high, is not poisoned and inhibited by Cr (VI)
The advantages of.
A kind of method for preparing biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium of embodiment 7.
(1) dissimilatory iron reduction bacterium Klebsiella sp.KB52 the culture of dissimilatory iron reduction bacterium: is pressed into 3% inoculum concentration
It is inoculated in basal medium, under anaerobic, 25 DEG C, 150r/min cultivates 4d；
Basic media components (g/L): glucose 15, yeast extract 3, tryptone 2, NaCl 40, K2HPO42,
NaHCO32, Fe (OH)3500mg/L, remaining is water, pH 7.0 ± 0.2；
Wherein, NaHCO3It is that the solution that first configuration concentration is 50g/L is added to base by 40mL/L using filtration method degerming
Basal culture medium；
(2) preparation of biological magnetic iron ore: after culture 4d, bacterium solution is transferred to sterile centrifugation tube and is separated by solid-liquid separation, be centrifuged
(9000r/min, 1min).Insoluble matter is gathered in centrifugation bottom of the tube in thallus and culture solution, in thick, be transferred to culture dish, dry
Dry (100 DEG C, 5h), biological magnetic iron ore sample is in powdered.
(3) Cr (VI) is removed: Cr (VI) 50:1 in mass ratio in biological magnetic iron ore and liquid to be processed is added to containing Cr
(VI) concentration is in the liquid to be processed of 40mg/L, and adjusting pH is 2.0, and after reacting 10h, the removal rate of Cr (VI) is 100%.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
The limitation to the scope of the patents therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art,
Under the premise of not departing from this patent design, the respective embodiments described above can also make several deformations, combination and improve, these all belong to
In the protection scope of this patent.Therefore, the protection scope of this patent should be subject to the claims.
Claims (10)
1. a kind of method for preparing biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium, which is characterized in that
Include the following steps:
(1) culture of dissimilatory iron reduction bacterium: dissimilatory iron reduction bacterium is inoculated in culture medium by 1-5% inoculum concentration, in nothing
Under the conditions of oxygen, 25-35 DEG C, 100-150r/min cultivates 2-5d；
(2) preparation of biological magnetic iron ore: after culture 2-5d, insoluble matter is aseptically collected by centrifugation in bacterium solution, by insoluble matter
Dry to obtain biological magnetic iron ore；
(3) Cr (VI) is removed: by Cr (VI) 10-60:1 in mass ratio in biological magnetic iron ore and liquid to be processed, be added to containing
In liquid to be processed of the concentration not higher than 80mg/L Cr (VI), adjusting pH is 2.0-8.0, reacts 6-12h.
2. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that the dissimilatory iron reduction bacterium is Klebsiella sp.KB52.
3. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that the incubation time of thallus is 4-5d.
4. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that Cr (VI) concentration ratio in biological magnetic iron ore and liquid to be processed is 25-50:1.
5. a kind of as claimed in claim 4 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that Cr (VI) concentration ratio in biological magnetic iron ore and liquid to be processed is 50:1.
6. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that reaction pH is 2.0-6.0.
7. a kind of as claimed in claim 6 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that reaction pH is 2.0-5.0.
8. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that the concentration of Cr (VI) is not higher than 60mg/L in liquid to be processed.
9. a kind of as claimed in claim 8 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that the concentration of Cr (VI) is not higher than 40mg/L in liquid to be processed.
10. a kind of as described in claim 1 prepare biological magnetic iron ore removal heavy metal Cr (VI) using dissimilatory iron reduction bacterium
Method, which is characterized in that culture medium group become ingredient: glucose 5-15g/L, yeast extract 1-3g/L, tryptone
0.5-2g/L, NaCl 20-40g/L, K2HPO4 1-2g/L, NaHCO3 1-2g/L, Fe (OH) 3 300-500mg/L, remaining is
Water, pH 7.0 ± 0.2.
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