CN110294534B - Microbial functional material and method for removing pentavalent vanadium in underground water - Google Patents

Microbial functional material and method for removing pentavalent vanadium in underground water Download PDF

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Publication number
CN110294534B
CN110294534B CN201910613802.2A CN201910613802A CN110294534B CN 110294534 B CN110294534 B CN 110294534B CN 201910613802 A CN201910613802 A CN 201910613802A CN 110294534 B CN110294534 B CN 110294534B
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CN
China
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pentavalent vanadium
underground water
vanadium
lactococcus
raffinose
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CN201910613802.2A
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Chinese (zh)
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CN110294534A (en
Inventor
张宝刚
李依娜
成玉彤
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中国地质大学(北京)
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2305/00Use of specific compounds during water treatment
    • C02F2305/06Nutrients for stimulating the growth of microorganisms

Abstract

The embodiment of the specification provides a microbial functional material and a method for removing pentavalent vanadium in underground water by using the microbial functional material. Wherein the material comprises: lactococcus raffinose and a carbon source, wherein lactococcus raffinose is used for reducing pentavalent vanadium in groundwater, and the carbon source is used as a nutrient of lactococcus raffinose and an electron donor when lactococcus raffinose is used for reducing pentavalent vanadium in groundwater. The method comprises the following steps: and putting the microbial functional material into underground water containing the pentavalent vanadium in an ionic state, so that the microorganisms reduce the pentavalent vanadium into precipitable tetravalent vanadium by taking a microbial supported carbon source as an electron donor. Therefore, the quinquevalent vanadium in the underground water is removed by using the microbial functional material, and meanwhile, an excellent removing effect is obtained.

Description

Microbial functional material and method for removing pentavalent vanadium in underground water
Technical Field
One or more embodiments of the present disclosure relate to the technical field of groundwater treatment, and more particularly, to a microbial functional material and a method for removing pentavalent vanadium from groundwater using the microbial functional material.
Background
At present, methods for removing pentavalent vanadium from groundwater mainly comprise physical methods and chemical methods, wherein the physical methods comprise soil improvement methods, electrolysis methods and the like. Specifically, the soil improvement method is to replace, mix and dilute or deeply bury the vanadium-contaminated soil by means of engineering machinery to remove or reduce the toxicity of vanadium and reduce the risk of soil contamination, and the method is high in cost and easy to cause secondary pollution. The electrolysis method is that a direct current power supply is introduced into an electrolytic bath filled with waste water, metal ions are separated out at the anode, and H is separated out at the cathode2The pentavalent vanadium can be reduced, and the precipitate of the tetravalent vanadium is generated along with the increase of the pH value of the solution, so that the method has high energy consumption and low efficiency. In addition, the chemical method includes chemical leaching, chemical adsorption, and the like. In particular, chemical leaching efficiently and rapidly removes heavy metal contaminants from soil by reacting a leaching agent with heavy metals in the soil to form more stable metal complexes or soluble metal ions, which may alter the physicochemical properties of the soil and introduce new contaminants. In the adsorption method, because molecules on the solid surface of the adsorption material are in an unbalanced or unsaturated state, vanadium ions in contact with the molecules can be adsorbed to the surface of the material to balance the molecular force of the vanadium ions so as to achieve the aim of removing the vanadium ions.
The microbial reduction and immobilization of pentavalent vanadium gradually becomes a research hotspot because of the economy, high efficiency, low cost and easy operation compared with the methods. Specifically, the microorganism functional bacteria play a key role in the process of reducing and fixing pentavalent vanadium. Therefore, there is an urgent need to propose an improved scheme based on functional bacteria of microorganisms for optimizing the treatment effect on groundwater.
Disclosure of Invention
One or more embodiments of the present specification describe a microbial functional material and a method for removing pentavalent vanadium from groundwater using the microbial functional material, for removing pentavalent vanadium and optimizing the removal effect.
According to a first aspect, there is provided a microbial functional material comprising:
lactococcus raffinose for the reduction of pentavalent vanadium in groundwater.
A carbon source for use as a nutrient for said lactococcus raffinose and for use as an electron donor when said lactococcus raffinose reduces pentavalent vanadium in groundwater.
In one possible implementation, the carbon source comprises one or more of:
glucose, acetate, soluble starch, sodium citrate and lactate.
In one possible implementation, the initial concentration of the carbon source is between 1 and 10 mM.
According to a second aspect, there is provided a method for removing pentavalent vanadium from groundwater using a microbial functional material, the microbial functional material being in any one of the above-mentioned implementations of the first aspect, the method comprising:
and putting the microorganism functional material into underground water containing the pentavalent vanadium in an ionic state, so that the microorganism reduces the pentavalent vanadium into precipitable tetravalent vanadium by relying on the carbon source as an electron donor.
In one possible embodiment, the method further comprises:
and under the condition that the pH value of the underground water is less than 7, adding an alkaline substance into the underground water so that the difference between the pH value of the underground water and 7 is less than a preset threshold value.
In one possible embodiment, the method further comprises:
and under the condition that the pH value of the underground water is more than 7, adding an acidic substance into the underground water so that the difference between the pH value of the underground water and 7 is less than a preset threshold value.
In one possible embodiment, the ionic pentavalent vanadium includes (VO)3)-And/or (VO)4)3-
The microbial functional material and the method for removing pentavalent vanadium by using the same provided by the embodiment of the specification can efficiently reduce and fix the pentavalent vanadium in the groundwater.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 shows a reactor physical picture according to one embodiment. Wherein BS represents a blank system for killing microorganisms at high temperature, CFS represents a carbon-free system, and AS is a microbial activity system having a carbon source.
Figure 2 shows the concentration of pentavalent vanadium in the AS, CFS, BS system with sodium citrate AS the carbon source.
FIG. 3 shows the concentration profile of pentavalent vanadium in five sets of reaction solutions with different carbon sources.
FIG. 4 shows the curve of the variation of the concentration of pentavalent vanadium in reaction solutions having different initial concentrations of pentavalent vanadium.
FIG. 5 shows the curve of the variation of the concentration of pentavalent vanadium in reaction solutions with different initial concentrations of sodium citrate.
FIG. 6 shows the concentration of pentavalent vanadium in reaction solutions having different pH values.
FIG. 7 shows the curve of the concentration of pentavalent vanadium in reaction solutions having different amounts of bacteria.
FIG. 8 shows a bar graph of the VFAs content of the reaction solution as a function of the number of days of reaction.
FIG. 9 shows a bar graph of the changes of NADH and cytochrome C with the number of days of reaction.
FIG. 10 shows a bar graph of EPS of lactococcus raffinose as a function of days of reaction.
FIG. 11 shows a bar graph of glycogen in lactococcus raffinose as a function of the number of days of reaction.
FIG. 12 is a bar graph showing the change of ATP content in the reaction solution with the number of days of reaction.
FIG. 13 shows the gene copy number of a functional gene involved in cytochrome.
FIG. 14 shows the gene copy number associated with denitrification determined in lactococcus raffinose.
Detailed Description
It is to be understood that the scope of the invention is not to be limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
As mentioned above, the microorganism functional bacteria play a key role in the process of reducing and fixing pentavalent vanadium. At present, common microorganisms reported to have vanadium reduction include Clavibacterium thioredoxins (Geobacter meteriurus), Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans) and Thiobacillus thiooxidans (Acidithiobacillus thiooxidans), Shewanella oneidensis MR-1, Pseudomonas aeruginosa (Pseudomonas aeruginosa). Based on this, the inventors have creatively proposed the reduction of pentavalent vanadium by Lactococcus raffinosus (Lactococcus raffinosus). Experiments show that the reduction process is very efficient, and meanwhile, the research on the obtained reduction mechanism has guiding significance.
The technical solutions of the embodiments of the present invention will be described in more detail with reference to specific examples.
Example 1 feasibility study experiment of lactococcus raffinose for reducing pentavalent vanadium
The experimental operation steps are as follows:
1. and (5) culturing lactococcus raffinose.
In one example, lactococcus raffinose was inoculated on sterilized LB medium for growth.
2. Extracting lactococcus raffinose.
In one example, an overnight aliquot of the culture was centrifuged from the culture vessel, the supernatant was decanted, and the lower layer was rinsed twice with sterile water and centrifuged. Further, in a specific operation example, 40ml of the certain amount of the bacterial suspension was taken out. In a specific example of operation, wherein each centrifugation is involved, the setting of the rotation speed 4000rpm for 20min may be included.
3. One experimental group and two control groups were prepared.
In one example, a sterile serum bottle was used as a reaction vessel, and the reaction vessel was covered with a foil paper to protect it from light, and an equal amount of lactococcus raffinose was taken out from the lactococcus raffinose after washing and centrifugation, and inoculated in an equal amount of artificial simulated groundwater, respectively. Further, a carbon source was added to one of the reaction vessels AS an experimental group with a carbon source system (AS). For the other two reaction vessels, one of them was directly used as a control group of a carbon source-free microorganism system (CFS), and the other was used as a control group of a system (BS) after killing microorganisms at high temperature after adding carbon source and killing microorganisms at high temperature. In a specific example of operation, the artificial simulated groundwater initially charged in each reaction vessel was 100 ml.
The experimental results are as follows:
fig. 1 shows experimental pictures of one experimental group and two control groups, in which foil paper originally wrapped in a sterile serum bottle was torn off after 10 days.
FIG. 2 shows the concentration variation curve of pentavalent vanadium in AS, CFS, BS system using sodium citrate AS carbon source.
Through test analysis, AS can be seen from fig. 2, in the AS experimental group, the reduction efficiency to pentavalent vanadium within 10 days is 86.46%, and the reduction efficiencies of the CFS control group and the BS control group are 24.23% and 2%, respectively.
Therefore, the lactococcus raffinose has the capability of reducing and fixing pentavalent vanadium, and the removal of the pentavalent vanadium in the groundwater by utilizing the lactococcus raffinose is feasible and can be realized efficiently.
Example 2 optimization of carbon sources
The experimental steps are as follows:
1. five different carbon sources were selected, including glucose, acetate, soluble starch, citrate and lactate. In one working example, the acetate, citrate and lactate salts are specifically sodium acetate, sodium citrate and sodium lactate, respectively.
2. Five groups of reaction solutions are correspondingly configured, wherein the reaction solutions respectively comprise the same amount of lactococcus raffinose, artificial simulated underground water and the five different carbon sources with the same concentration. In one working example, the concentration of the carbon source is 1 to 10 mM. In a specific example of operation, the carbon source concentration is 5 mM.
The experimental results are as follows:
FIG. 3 shows the concentration profile of pentavalent vanadium in five sets of reaction solutions with different carbon sources.
As can be seen from FIG. 3, the reduction efficiencies of the different carbon sources, i.e., glucose, sodium acetate, soluble starch, sodium citrate and sodium lactate, were 80.58%, 72.40%, 69.52%, 86.46% and 80.59%, respectively, after 10 days. Therefore, the carbon sources all have good reduction effect, and particularly, the sodium citrate has the highest reduction efficiency and the best removal effect. Therefore, in subsequent experiments, sodium citrate was chosen as the carbon source.
Example 3 experiment of influencing factors on reduction efficiency
The following characteristics are selected as influencing factors for experimental study: the initial concentration of pentavalent vanadium, the initial concentration of sodium citrate, the pH value of the reaction solution and the volume of the liquid of lactococcus raffinose in the underground water.
1. The initial concentration of pentavalent vanadium is taken as an influence factor
Four sets of reaction solutions were prepared, wherein the initial concentrations of pentavalent vanadium in each set of reaction solutions were 25mg/L, 50mg/L, 75mg/L and 100mg/L, respectively, and the reduction rates of pentavalent vanadium in each set of reaction solutions after 10 days were 81.30%, 86.46%, 84.51% and 79.37%, respectively, and in particular, see fig. 4, which shows the concentration change curves of pentavalent vanadium in reaction solutions having different initial concentrations of pentavalent vanadium.
Analysis shows that the microbial functional material provided by the invention has a good treatment effect on groundwater with different concentrations of pentavalent vanadium, can efficiently reduce the pentavalent vanadium therein, and particularly has a removal efficiency of 86.46% for the pentavalent vanadium when the pentavalent vanadium concentration is 50mg/L, and then the reduction rate is slightly reduced with the increase of the initial concentration of the pentavalent vanadium.
2. The initial concentration of sodium citrate is taken as the influence factor
Four sets of reaction solutions were prepared, in which the initial concentrations of sodium citrate in the respective sets of reaction solutions were 1mM, 2.5mM, 5mM and 10mM, and the reduction rates of pentavalent vanadium in the respective sets of reaction solutions after 10 days were 60.05%, 75.41%, 86.46% and 80.01%, respectively, and in particular, see fig. 5, which is a graph showing the change in the concentration of pentavalent vanadium in reaction solutions having different initial concentrations of sodium citrate.
Analysis can show that the removal rate of pentavalent vanadium is increased and then decreased along with the increase of the initial concentration of sodium citrate, and particularly when the concentration of sodium citrate is 5mM, the removal rate is the highest and reaches 86.46%.
3. The pH value of the reaction solution is taken as the influence factor
Four sets of reaction solutions were prepared, wherein the pH values of the reaction solutions of each set were 5, 6, 7 and 8, respectively, and the reduction rates of pentavalent vanadium in the reaction solutions of each set after 10 days were 66.87%, 77.02%, 86.46% and 79.44%, respectively, and in particular, see fig. 6, which shows the concentration variation curves of pentavalent vanadium in the reaction solutions having different pH values. It is to be noted that, among them, the adjustment of the pH of the reaction solution can be carried out by using an acidic substance or a basic substance.
Analysis shows that when the pH value is 7, the activity of lactococcus raffinose is highest, and the removal effect of pentavalent vanadium is best.
4. The volume of the bacterial liquid of lactococcus raffinose is taken as an influencing factor
The volume of the bacterial suspension is the volume of the bacterial suspension taken out from the culture of lactococcus raffinose.
Specifically, four groups of reaction solutions were prepared, wherein lactococcus raffinose inoculated in each group of reaction solutions was extracted from 10mL, 20mL, 40mL, and 100mL of bacterial solutions, respectively. After 10 days, the removal rate of pentavalent vanadium in each group of reaction solution is 58.48%, 76.02%, 86.46% and 100%, and particularly, see fig. 7.
Analysis can show that the removal rate of pentavalent vanadium increases with the increase of the initial number of lactococcus raffinose, and when the bacterial liquid is increased to 100ml, the reduction rate of V (V) reaches 100% after 5 days.
As described above, it is understood from examples 1 and 2 that lactococcus raffinose has the ability to reduce and immobilize pentavalent vanadium and can achieve high-efficiency removal, and particularly, the reduction efficiency is the best when sodium citrate is used as a carbon source. From example 3, it is understood that the initial concentration of pentavalent vanadium, the initial concentration of sodium citrate, the pH of the reaction solution, and the volume of lactococcus raffinose bacteria in the groundwater all affect the removal rate of pentavalent vanadium. Based on the above, the embodiments of the present disclosure provide a microbial functional material, and a method for removing pentavalent vanadium from groundwater by using the microbial functional material.
Specifically, the microbial functional material comprises: lactococcus raffinose and a carbon source, wherein lactococcus raffinose is used for reducing pentavalent vanadium in underground water; the carbon source is used as a nutrient for the lactococcus raffinose and as an electron donor for the lactococcus raffinose in reducing pentavalent vanadium in groundwater.
In one possible embodiment, the carbon source comprises one or more of the following: glucose, acetate, soluble starch, citrate and lactate.
In one possible embodiment, the initial concentration of the carbon source is 1-10 mM.
The microbial functional material can be used for removing pentavalent vanadium in underground water, and the specific using method comprises the following steps: and (3) putting the microorganism functional material into underground water containing the pentavalent vanadium in an ionic state, so that the microorganism reduces the pentavalent vanadium into precipitable tetravalent vanadium by relying on the carbon source as an electron donor. Wherein the ionic pentavalent vanadium includes (VO)3)-And/or (VO)4)3-
In one possible embodiment, the method further comprises: and in the case that the pH value of the underground water is less than 7, adding alkaline substances into the underground water so that the difference value between the pH value of the underground water and 7 is less than a preset threshold value. In another possible embodiment, the method further comprises: and in the case that the pH value of the underground water is more than 7, adding an acidic substance into the underground water so that the difference between the pH value of the underground water and 7 is less than a preset threshold value. More specifically, the predetermined threshold value may be set according to actual needs, such as 1 or 0.5.
Therefore, by adopting the microbial functional material provided by the embodiment of the specification, the pentavalent vanadium in the underground water can be efficiently removed.
Next, with reference to other examples, the mechanism of reduction of pentavalent vanadium by lactococcus raffinose is studied, and the research result has guiding significance for subsequent experiments and applications.
Example 4 reduction mechanism study experiment
The experimental contents are as follows:
preparing a reaction solution, wherein the reaction solution comprises the microbial functional material and underground water containing pentavalent vanadium. Samples were then taken daily from the reactor to determine ATP levels, and after filtration through a 0.22 micron filter, VFAs were determined. And, taking equivalent solution of 1, 5 and 10 days of the reaction cycle, centrifuging and washing to determine EPS, cytochrome C, NADH, glycogen and related functional genes of lactococcus raffinose, wherein the control group is a microorganism system without pentavalent vanadium (hereinafter referred to as vanadium-free control group).
After 10 days of reaction, the cell morphology was observed with a JEOL Transmission Electron Microscope (TEM) at 200kV (JEM-2100, Hitachi Limited, Japan), and the intracellular and extracellular distribution of the reduced product was observed with a TEM/EDS combined with a Scanning Transmission Electron Microscope (STEM) in accordance with the ultrathin section (50-60nm) of the obtained sample material.
The experimental results are as follows:
1. STEM showed that many particles distributed on and inside the cells were probably precipitates resulting from the reduction of pentavalent vanadium to tetravalent vanadium, and scans of pentavalent vanadium in lactococcus raffinose showed higher intracellular intensity than extracellular membranes, indicating that most of the pentavalent vanadium is prone to intracellular reduction.
2. FIG. 8 shows the change of VFAs content in the reaction solution with the number of days of reaction, FIG. 9 shows the change of NADH and cytochrome C of lactococcus raffinose with the number of days of reaction, FIG. 10 shows the change of EPS of lactococcus raffinose with the number of days of reaction, FIG. 11 shows the change of glycogen in lactococcus raffinose with the number of days of reaction, and FIG. 12 shows the change of ATP content in the reaction solution with the number of days of reaction.
Analysis shows that the EPS secretion protects cells against invasion of pentavalent vanadium, the NADH content is higher than that of a vanadium-free control group, the ATP content is increased along with the increase of reaction time, NADH generated by microbial metabolism reacts with oxidized phosphate, and a large amount of ATP is generated through electron transfer, so that the microbes keep activity and provide energy for reducing pentavalent vanadium.
3. FIG. 13 shows the gene copy number of a functional gene involved in cytochrome. FIG. 14 shows the gene copy number associated with denitrification determined in lactococcus raffinose.
Analysis has shown that cytochrome C transports electrons from the inner membrane to the periplasm and across the outer membrane to external electron acceptors, and MtrC and OmcB play an important role in this process.
Quantitatively detecting the abundances of nitrate reductase genes napA, narG, nitrite reductase genes nirS, nirK, nitric oxide reductase genes qnorB and nitrous oxide reductase genes nosZ by qPCR, and finding that the gene copy number of the gene nirS for coding the nitrite reductase is most obviously increased along with the increase of reaction time in a microbial system containing pentavalent vanadium compared with a microorganism which is not contacted with the pentavalent vanadium, and the gene copy number reaches 10^12 copy number/ng DNA; the gene copy numbers of the genes napA and narG for coding nitrate reductase reach 10^10 copy number/ng DNA and 10^6 copy number/ng DNA respectively. Therefore, the nitrite reductase gene nirS and the nitrate reductase genes napA and narG are probably involved in the process of reducing pentavalent vanadium by lactococcus raffinose.
The research on the mechanism of reducing pentavalent vanadium by lactococcus raffinose has guiding significance for subsequent experiments and applications.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims (1)

1. An application of a microbial functional material in removing underground water pentavalent vanadium is disclosed, wherein the microbial functional material is put into underground water containing ionic pentavalent vanadium to obtain a reaction solution; the microorganism functional material comprises lactococcus raffinose and sodium citrate; wherein the lactococcus raffinose is used for reducing pentavalent vanadium in underground water; the sodium citrate is used as a nutrient for the lactococcus raffinose and an electron donor for the lactococcus raffinose in reducing pentavalent vanadium in underground water; in the reaction solution, the initial concentration of pentavalent vanadium is 50mg/L, the initial concentration of sodium citrate is 5mM, the pH value of the reaction solution is 7, and the volume of the bacterial liquid of lactococcus raffinose is 100 mL; the reaction time is more than or equal to 5 days.
CN201910613802.2A 2019-07-09 2019-07-09 Microbial functional material and method for removing pentavalent vanadium in underground water CN110294534B (en)

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