CN110589987A - Preparation method of denitrifying bacteria nano-iron composite material for underground water denitrification - Google Patents
Preparation method of denitrifying bacteria nano-iron composite material for underground water denitrification Download PDFInfo
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Abstract
The invention aims to prepare a denitrifying bacteria nano-iron composite material by utilizing the biological characteristic reduction of denitrifying bacteria and a method for removing nitrate nitrogen by utilizing the material, ferric iron is introduced by using anhydrous ferric chloride, continuous culture is carried out for three months, the reaction temperature is controlled at room temperature, the pH value is controlled at 7.45, and synthesized nano-iron particles are polydispersed on the surface of bacteria and are better dispersed. The method has the advantages of simple and convenient operation, rapid reaction, environmental protection and low energy consumption, and can realize continuous reaction.
Description
Technical Field
The invention belongs to a method for preparing a denitrifying bacteria nano-iron composite material by utilizing the biological characteristic reduction of denitrifying bacteria and removing nitrate nitrogen in underground water by utilizing the material.
Background
The traditional preparation of nano-iron particles mainly comprises physical methods, chemical methods and electrochemical methods, such as physical vapor deposition methods, high-energy ball milling methods, solid-phase reduction methods and the like. But the physical method preparation has low purity and uneven particle distribution, other impurities are easily introduced in the chemical method preparation, and the nano material prepared by the microbial reduction method has the advantages of low cost, mild reaction conditions, no need of adding a chemical reducing agent and a protective agent, environmental protection, good stability of nano particles and the like; the microorganism is used as a biological template, and the microorganism can be used for regulating and preparing the shape and the size of the nano material by means of the regular epidermal layer of the surface of the microorganism or the rich functional groups in the epidermal layer. The synthesized nano-iron can be directly loaded on the denitrifying bacteria to form the denitrifying bacteria nano-iron composite material.
Nitrate contamination has been a significant environmental problem in groundwater quite common. Early in the 60 s of this century, both the united states and europe have reported nitrate pollution of groundwater due to the application of chemical nitrogen fertilizers. The investigation shows that the water quality of the American drinking water is nearly 1/4 in the case of exceeding the standard of nitrate nitrogen; in the welsh area of the uk, the nitrate content in the groundwater for 180 million people to drink at 125 is greatly exceeded. In the next decades, nitrate pollution of underground water has been reported in successive countries and regions such as the United states, Europe, Japan, etc. Such as: denmark has increased groundwater nitrate content by a factor of 3 over the last 30 years, and also has a tendency to continue to increase. In the early 80 s, the investigation result of the pollution condition of underground water in China shows that the content of nitrate nitrogen in underground water in cities of Xian, Changchun, Chengdu and the like is greatly overproof. Therefore, research on the nitrate pollution of underground water and the control thereof become hot spots of research at home and abroad at present.
The existing treatment technology of nitrate nitrogen in underground water can be mainly divided into three types of physical chemical method, chemical method and biological method. The physical and chemical repairing technology mainly comprises electrodialysis, reverse osmosis, distillation, ion exchange and the like[4 6]In which electrodialysis, reverse osmosis, distillation, etc. are due to the presence of NO3Non-selectivity, and simultaneously removes nitrateHe has inorganic salt beneficial to human body, and has lower removal efficiency and higher cost; although the ion exchange method can selectively remove nitrate, it requires regeneration of high-concentration salts or acids, and thus generates wastewater containing high-concentration nitrate, sulfate, and the like, which makes post-treatment difficult. The chemical method can be divided into an active metal reduction method and a catalytic denitrification method[7]Two major types, the reducing agents which are researched more in the current active metal reduction method comprise Fe, Cd, Al, Zn and some alloys[8 10]And the like. Since many of the metal reductants themselves or the reduction by-products are toxic to the human body, their use in groundwater remediation is severely limited. The use of the denitrifying bacteria nano-iron composite material can achieve higher removal efficiency and low cost, and can reduce the removal of N2Besides, the byproducts provide theoretical technology and method for developing underground water nitrate nitrogen composite materials with high reaction activity and high stability.
Disclosure of Invention
The invention aims to research a method for preparing a nano-iron composite material by biological reduction of a synthetic denitrifying bacterium and denitrification performance of the nano-iron composite material. We utilize the biological reducibility of denitrifying bacteria to reduce Fe3+And loading the composite material on denitrifying bacteria to form the denitrifying bacteria nano-iron composite material for removing nitrate nitrogen and the like in water.
The key steps of the method are obtained on the basis of a large number of experimental tests, documents and analyses, and comprise the following points:
1. adding about 5ml of screened denitrifying bacteria (with other miscellaneous bacteria eliminated) into 5 conical flasks respectively, preparing a liquid culture medium suitable for the growth of the denitrifying bacteria, dissolving the liquid culture medium in a 1L beaker, preparing 100ml of trace elements, adding 1ml of trace elements into the liquid culture medium, adjusting the pH value to be within the range of 7-7.5, adding the mixture into 5 conical flasks respectively to obtain a total volume of about 150ml, and adding a small amount of Fecl3The solution in the conical flask changes from transparent to light yellow. The medium was not changed every 3 to 4 days and the procedure was repeated.
2. Repeating the operation 1 for several times, and measuring the growth curve of the denitrifying bacteria in a complete growth cycle by using an ultraviolet spectrophotometerMeasuring the absorbance at a fixed wavelength of 600nm every 3 hours, and measuring the addition of Fecl3The growth curve of the denitrifying bacteria after the solution is nearly in accordance with the growth curve of the microorganism, and then the Fecl is increased3The amount of the solution.
3. Repeating the operations 1 and 2 for a plurality of times, and finding out that the Fecl is accurately weighed3When the water content is 0.3g and 100ml, the growth curve of the denitrifying bacteria basically conforms to the trend of the growth curve of microorganisms, and by utilizing a TEM means, the nano iron particles loaded on the denitrifying bacteria can be seen to be distributed on the surfaces of cell membranes, cell walls and flagella, and the dispersibility is good.
4. Dividing cultured denitrifying bacteria loaded with nano-iron into a plurality of gradients to study the optimal synthesis conditions, and controlling the addition of Fecl3The volume of the solution was such that, after repeated 1.2.3. operations, it was found that Fecl was added when it was added3When the solution is 4.5ml, the load of denitrifying bacteria is the most, and the dispersive growth condition is the best.
5. Weighing 0.287g of sodium nitrate powder, dissolving the sodium nitrate powder in 100ml of water, dissolving 2ml of the sodium nitrate powder in a 1L volumetric flask for constant volume, uniformly mixing, standing for 5min, taking 10ml of the denitrifying bacteria solution cultured for 2-3 days, placing the solution in a clean conical flask which is dried for 5 hours and is at 100 ℃, weighing 240ml of the sodium nitrate solution by using a 100ml volumetric flask, adding the sodium nitrate solution into the conical flask, at the moment, weighing 250ml of the solution in the conical flask, and measuring the initial pH value by using a pH meter.
6. Taking 25ml of water sample and a colorimetric tube every 1 hour, adding 1ml of hydrochloric acid by a pipette, and fixing the volume to 5 ml. And measuring the absorbance at the wavelength of 220nm and 275nm by using an ultraviolet spectrophotometer, substituting the absorbance into a previously prepared nitrate nitrogen standard curve, and calculating to obtain accurate nitrate nitrogen data. The results show that Fecl is not added3The bacterial liquid of (A) and the added Fecl3Longer than the degradation time and slower rate. When the pH rises to about 9.5, no degradation takes place.
Drawings
FIG. 1 and FIG. 2 show the composite material of denitrifying bacteria nano-iron synthesized by the present invention.
Detailed Description
The invention is further illustrated by the following examples.
Example 1:
we tested a: denitrifying bacteria 10ml +240ml nitrate solution b: denitrifying bacteria +1.5ml Fecl3The solution totaled 10ml +240ml nitrate solution c: denitrifying bacterium +2.5ml Fecl3The solution totaled 10ml +240ml nitrate solution, d: denitrifying bacteria +3.5ml Fecl3Solution 10ml +240ml nitrate solution, e: denitrifying bacteria +4.5ml Fecl3The total solution of 10ml +240ml nitrate solution was compared to remove nitrate, and the results show that: the removal rate of nitrate nitrogen of the denitrifying bacteria solution and the nano-iron composite material of the nano-denitrifying bacteria can reach 99% within 8 hours, and the degrading rate of the nano-iron composite material of the denitrifying bacteria is higher, and the degrading time is about 7 hours.
Example 2:
we tested a: denitrifying bacteria 10ml +240ml nitrate solution b: denitrifying bacteria +1.5ml Fecl3The solution totaled 10ml +240ml nitrate solution c: denitrifying bacterium +2.5ml Fecl3The solution totaled 10ml +240ml nitrate solution, d: denitrifying bacteria +3.5ml Fecl3The solution totaled 10ml +240ml nitrate solution, e: denitrifying bacteria +4.5ml Fecl3The total solution of 10ml +240ml nitrate solution was compared to remove nitrate, and the results show that: the removal rate of nitrate nitrogen in 4 hours of the denitrifying bacteria solution and the nano-scale iron composite material of the nano-denitrifying bacteria can reach 99 percent, the degradation rate of the nano-scale iron composite material of the denitrifying bacteria is higher, and 4.5ml of Fecl is added3The solution degradation rate is fastest.
Example 3:
we tested a: denitrifying bacteria 10ml +240ml nitrate solution b: denitrifying bacteria +4.5ml Fecl3The solutions were 10ml +240ml nitrate solutions, the same control was set at each temperature, and it was found that the degradation rate was the best at room temperature, the removal rate was 99% in each case, and it was found that 4.5ml Fecl was added3The solution has faster degradation rate, and when the temperature is too high, the enzyme activity is higher than 50 DEG CThe sexual activity is reduced and no degradation is carried out.
Example 4:
we tested a: denitrifying bacteria 20ml +130ml nitrate solution b: denitrifying bacteria +4.5ml Fecl3The total solution of 20ml +130ml nitrate solution is introduced, and the same control group is set under the condition of nitrogen with different durations, so that the result shows that the degradation rate is the best under the conditions of 0min and 2min, probably because the denitrifying bacteria are aerobic heterotrophic denitrifying bacteria, and in addition, nitrogen with less time is introduced to play a role in protection, so that the nano zero-valent iron is prevented from being oxidized.
Example 5:
we tested a: denitrifying bacteria 10ml +240ml nitrate solution b: denitrifying bacteria +1.5ml Fecl3The solution totaled 10ml +240ml nitrate solution c: denitrifying bacteria +2.5ml Fecl3The solution totaled 10ml +240ml nitrate solution, d: denitrifying bacteria +3.5ml Fecl3The solution totaled 10ml +240ml nitrate solution, e: denitrifying bacteria +4.5ml Fecl3The solution is 10ml +240ml nitrate solution, the sample in the group a has almost no degradation, which indicates that the denitrifying bacteria have been basically inactivated, and the gradient change relationship appears in the groups b, c, d and e, probably because of the difference of the nano iron amount.
Claims (6)
1. A method for synthesizing nano-iron by utilizing the biological reduction characteristic of denitrifying bacteria and loading the nano-iron on the denitrifying bacteria and utilizing the composite material to degrade nitrate nitrogen is characterized in that 0.1g of anhydrous ferric chloride is dissolved in 50ml of distilled water under the culture conditions of room temperature, pH 7.45 and 100 revolutions of a shaking table, the anhydrous ferric chloride is added into a denitrifying bacteria culture medium after being fully dissolved, the culture medium is replaced after three days of culture, the amount of the anhydrous ferric chloride is increased to 0.3g and is also dissolved in 50ml of distilled water after being cultured for one month according to the method, the anhydrous ferric chloride is added into the culture medium after being fully dissolved, and the nano-iron composite material for the denitrifying bacteria is synthesized when the amount of the ferric chloride is continuously increased to 0.5g after being cultured for two months at room temperature. The method for degrading nitrate nitrogen by using the material is characterized in that 10ml of bacterial liquid is added into 240ml of nitrate solution with the concentration of 0.287g, the reaction pH is 8.3, and the degradation rate reaches over 99 percent after the reaction is carried out for 7 hours at the room temperature.
2. The method for synthesizing nano-iron and loading the nano-iron on denitrifying bacteria by using the biological reduction characteristic of the denitrifying bacteria and degrading nitrate nitrogen by using the composite material as claimed in claim 1, wherein the synthesizing process is performed at room temperature and under the condition that the pH value is 7.45, and the whole culturing process is performed at the rotation speed of the shaking table of 100.
3. The method for synthesizing nano-iron and loading the nano-iron on denitrifying bacteria by utilizing the biological reduction characteristic of the denitrifying bacteria and utilizing the composite material to degrade nitrate nitrogen as claimed in claim 1, wherein the ferric ions are introduced in the form of ferric chloride, and the ferric ions introduced in other forms have different characteristics.
4. The method for synthesizing nano-iron and loading the nano-iron on denitrifying bacteria by utilizing the biological reduction characteristic of the denitrifying bacteria and degrading nitrate nitrogen by utilizing the composite material as claimed in claim 1, wherein the denitrifying bacteria are used as reducing agents, no additional reducing agent is needed, and the reducing agents exist in a large amount in nature.
5. The method for synthesizing nano-iron and loading the nano-iron on denitrifying bacteria by utilizing the biological reduction characteristic of the denitrifying bacteria and utilizing the composite material to degrade nitrate nitrogen as claimed in claim 1, wherein the nano-iron can be successfully loaded on the denitrifying bacteria only when the anhydrous ferric chloride is added to reach 0.5g, and otherwise, the nano-iron is unloaded.
6. The method for synthesizing nano-iron by utilizing the biological reduction characteristic of denitrifying bacteria and loading the nano-iron on the denitrifying bacteria and utilizing the composite material to degrade nitrate nitrogen as claimed in claim 1, wherein 10ml of bacterial liquid is added into 240ml of nitrate solution with the concentration of 0.287g, the reaction pH is 8.3, and the degradation rate is more than 99% after the reaction is carried out for 7 hours at room temperature.
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