CN109777839B - Method for removing cadmium in water and generating nano material by using citrobacter freundii - Google Patents
Method for removing cadmium in water and generating nano material by using citrobacter freundii Download PDFInfo
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- CN109777839B CN109777839B CN201711104973.XA CN201711104973A CN109777839B CN 109777839 B CN109777839 B CN 109777839B CN 201711104973 A CN201711104973 A CN 201711104973A CN 109777839 B CN109777839 B CN 109777839B
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Abstract
The invention relates to the technical field of microbial culture, water purification and nano material synthesis, in particular to a method for removing heavy metal cadmium in water in situ and generating nano material precipitate by culturing citrobacter freundii, and particularly relates to a method for decomposing nutrient substances around a culture solution by citrobacter freundii so as to generate a large amount of acid radical ions, wherein part of the acid radical ions such as carbonate and phosphate react with the heavy metal cadmium to form cadmium nano-scale precipitate, so that the heavy metal cadmium in water is removed and simultaneously corresponding nano-scale metal material is synthesized.
Description
Technical Field
The invention relates to the technical field of microbial culture, water purification and nano material synthesis, in particular to a method for removing heavy metal cadmium in water in situ and generating nano material precipitate by culturing citrobacter freundii, and particularly relates to a method for decomposing nutrient substances around a culture solution by citrobacter freundii so as to generate a large amount of acid radical ions, wherein part of the acid radical ions such as carbonate and phosphate react with the heavy metal cadmium to form cadmium nano-scale precipitate, so that the heavy metal cadmium in water is removed and simultaneously corresponding cadmium nano metal material is synthesized.
Background
At present, the heavy metal pollution in water bodies in China is particularly serious, the heavy metal pollution in the water bodies is cadmium pollution, and most of the heavy metal pollution is caused by the exploitation of metal resources, the discharge of production wastewater of chemical plants, the excessive use of farmland fertilizers and tail gas of automobiles. Since heavy metals are mostly present in ionic form in the liquid phase, such form can be absorbed into the body by plants, animals and humans, and continuously accumulate in the body, eventually endangering human health. The prior method for removing heavy metals by activated sludge mostly removes the heavy metals through the strong adsorbability of zooglea and the complexation of protein generated by microorganisms and the heavy metals. The method has the problem of incomplete removal of heavy metals, mainly because zoogloea disappears due to the reduction of nutrient substances around bacteria, and heavy metals adsorbed by the zoogloea are released into water again after the zoogloea disappears; the chelate of the heavy metal and protein complexing process can be decomposed by microorganisms, no complex exists after decomposition, and the heavy metal is released into water again. The invention aims to provide a new degradation way for heavy metals, which converts the heavy metals into stable sediment or mineral substances, so that the removed heavy metals do not enter water again along with the death of microorganisms, but exist in a stable sediment form, the sediment can be collected to be reused as nano materials, and the removal way can be popularized to soil to solve the problem that the heavy metals in the soil are difficult to remove.
Disclosure of Invention
The invention aims to provide a new idea and a new method for degrading metals, and the new idea and the new method can be applied to the treatment of water containing heavy metals or the remediation of heavy metal contaminated soil.
In order to achieve the purpose, the invention adopts the following scheme:
(1) the starting strain was derived from a standard strain of Citrobacter freundii (ATCC43864) purchased at national bacterial banks.
(2) Weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride and 40mg of cadmium chloride, dissolving the peptone, the beef extract and the cadmium chloride in 1000ml of deionized water, adding sodium hydroxide to adjust the pH value of the solution to be neutral, preparing a peptone and beef extract culture medium (hereinafter referred to as a metal phosphate-free culture medium) containing 40mg/L of cadmium chloride, placing the peptone and the beef extract culture medium into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract culture medium for 30min at 121 ℃, taking out the peptone and the beef extract culture medium, placing the beef extract culture medium in a sterile operating platform, cooling the beef extract culture medium to room temperature, and inoculating the beef.
(3) Weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride, 40mg of cadmium chloride and 116mg of disodium phosphoglycerate pentahydrate (the molar ratio of the disodium phosphoglycerate to the cadmium is 1: 2), dissolving the peptone, the beef extract and a culture medium (hereinafter referred to as a culture medium with metal phosphate) containing 40mg/L of cadmium chloride and 116mg/L of disodium phosphoglycerate after the peptone and the beef extract are dissolved in 1000ml of deionized water and added with sodium hydroxide to adjust the pH of the solution to be neutral, placing the peptone and the beef extract into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract at 121 ℃ for 30min, taking out the peptone and the beef extract, placing the beef extract in a sterile operation table, cooling the peptone and the beef extract to room temperature.
(4) A culture medium containing metal phosphate and metal phosphate-free was poured into 150ml Erlenmeyer flasks of 250ml in a sterile operating table, several of which were selected from among the standard Citrobacter freundii strains, and inoculated into the poured culture medium containing metal phosphate and metal phosphate-free.
(5) Placing the inoculated culture medium with metal phosphate and metal phosphate-free ester in a shaking table at the temperature of 35 ℃, culturing for 6 days, then extracting the precipitate, and taking out the bacterial liquid.
(6) And taking out the domesticated Citrobacter freundii and the cadmium phosphate synthesized by the Citrobacter freundii under the condition of no disodium glycerophosphate pentahydrate and the cadmium phosphate synthesized by the Citrobacter freundii under the condition of the disodium glycerophosphate pentahydrate, wherein the domestication can be repeated for a plurality of times, and the adaptability of the Citrobacter freundii to cadmium is stronger as the times are more.
Drawings
FIG. 1A screenshot of Citrobacter freundii under an electron microscope (magnification 1000)
FIG. 235 ℃ is a graph showing the effect of metal phosphate and metal phosphate-free medium on cadmium removal
FIG. 3 XRD search pattern of medium precipitation with metal phosphate-free
FIG. 4 XRD search pattern of medium precipitates with metal phosphate
FIG. 5 a diagram of a cadmium carbonate electron lens
FIG. 6 a diagram of a cadmium phosphate electron lens
Detailed Description
1. Weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride and 40mg of cadmium chloride, dissolving the peptone, the beef extract and the cadmium chloride in 1000ml of deionized water, adding sodium hydroxide to adjust the pH value of the solution to be neutral, preparing a peptone and beef extract culture medium (hereinafter referred to as a metal phosphate-free culture medium) containing 40mg/L of cadmium chloride, placing the peptone and the beef extract culture medium into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract culture medium for 30min at 121 ℃, taking out the peptone and the beef extract culture medium, placing the beef extract culture medium in a sterile operating platform, cooling the beef extract culture medium to room temperature, and inoculating the beef.
2. Weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride, 40mg of cadmium chloride and 116mg of disodium phosphoglycerate pentahydrate (the molar ratio of the disodium phosphoglycerate to the cadmium is 1: 2), dissolving the peptone, the beef extract and a culture medium (hereinafter referred to as a culture medium with metal phosphate) containing 40mg/L of cadmium chloride and 116mg/L of disodium phosphoglycerate after the peptone and the beef extract are dissolved in 1000ml of deionized water and added with sodium hydroxide to adjust the pH of the solution to be neutral, placing the peptone and the beef extract into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract at 121 ℃ for 30min, taking out the peptone and the beef extract, placing the beef extract in a sterile operation table, cooling the peptone and the beef extract to room temperature.
3. A culture medium containing metal phosphate and metal phosphate-free was poured into 150ml Erlenmeyer flasks of 250ml in a sterile operating table, several of which were selected from among the standard Citrobacter freundii strains, and inoculated into the poured culture medium containing metal phosphate and metal phosphate-free.
4. Placing the inoculated culture medium with metal phosphate and metal phosphate-free ester in a shaking table at the temperature of 35 ℃, culturing for 6 days, then extracting the precipitate, and taking out the bacterial liquid.
FIG. 1 shows the morphology of Citrobacter freundii under 1000 times magnification, which is obviously rod-shaped and many; FIG. 2 shows that the degradation is continued for 6 days at 35 ℃, the concentration of cadmium chloride with metal phosphate is 0, the degradation efficiency reaches 100%, the concentration of cadmium chloride with metal phosphate is 5mg/L, and the degradation effect reaches 87.5%; FIG. 3 is a result of a search performed by XRD after drying the precipitate containing metal phosphate, and the search results show that the peak of cadmium carbonate, a standard substance, has a high matching degree with the unknown substance, so that the precipitate containing metal phosphate is determined to be cadmium carbonate; FIG. 4 is a result of a search performed by XRD after drying the precipitate with metal phosphate, and the search result is obtained, from which it is apparent that the peak of cadmium phosphate as a standard substance has a high matching degree with the unknown substance, and the main peaks correspond one-to-one, so that the precipitate with metal phosphate is determined to be cadmium carbonate; FIG. 5 is a TEM test chart of the precipitate with metal phosphate-free ester, and it can be seen that the formed cadmium carbonate is in a vertical stripe shape, similar to the texture of wood, and has a regular shape; FIG. 6 is a TEM test of a precipitate with metal phosphate, showing that the cadmium phosphate in the run is spherical and closely arranged, and uniform in size, with the diameter of the spheres being 40 nm.
In conclusion, the method removes heavy metal cadmium in the water body, and also prepares nano cadmium materials of cadmium carbonate and cadmium phosphate with different shapes, so that the cadmium is recycled, and toxic substances are converted into non-toxic cadmium metal nano materials.
Claims (4)
1. A method for removing heavy metal cadmium in water and generating a cadmium nano metal material by using Citrobacter freundii is characterized in that the method comprises the steps of respectively culturing the Citrobacter freundii in a liquid culture medium containing cadmium and no disodium glycerophosphate, and containing cadmium and disodium glycerophosphate, placing bacteria in a shaking table after inoculation, and culturing for 6 days at 35 ℃ so as to achieve the purpose of removing the heavy metal cadmium in situ and generate the cadmium-containing metal nano material;
the cadmium-containing metal nano material is cadmium carbonate in a culture medium with metal or without disodium glycerophosphate, and cadmium phosphate in a culture medium with metal or with disodium glycerophosphate.
2. The method according to claim 1, wherein the Citrobacter freundii is a standard strain purchased from national bacteria banks, Citrobacter freundii (ATCC 43864).
3. The method of claim 1, wherein the metal-free disodium phosphoglyceride medium is formulated and inoculated by: weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride and 40mg of cadmium chloride, dissolving the peptone, the beef extract and the cadmium chloride in 1000ml of deionized water, adding sodium hydroxide to adjust the pH of the solution to be neutral, preparing a peptone and beef extract culture medium (hereinafter referred to as a metal phosphate-free culture medium) containing 40mg/L of cadmium chloride, placing the peptone and the beef extract culture medium into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract culture medium for 30min at 121 ℃, taking out the peptone and the beef extract culture medium, placing the beef extract culture medium into a sterile operating platform, cooling the beef extract culture medium to room temperature, and inoculating the beef extract; the formula of the culture medium containing metal and disodium glycerophosphate comprises the following steps: weighing 10g of peptone, 5g of beef extract, 5g of sodium chloride, 40mg of cadmium chloride and 116mg of disodium phosphoglycerate pentahydrate (the molar ratio of the disodium phosphoglycerate to the cadmium is 1: 2), dissolving the peptone, the beef extract and a culture medium (hereinafter referred to as a culture medium with metal phosphate) containing 40mg/L of cadmium chloride and 116mg/L of disodium phosphoglycerate after the peptone and the beef extract are dissolved in 1000ml of deionized water and added with sodium hydroxide to adjust the pH of the solution to be neutral, placing the peptone and the beef extract into a high-pressure steam sterilization pot, sterilizing the peptone and the beef extract at 121 ℃ for 30min, taking out the peptone and the beef extract, placing the beef extract in a sterile operation table, cooling the peptone and the beef extract to room temperature.
4. The method of claim 3, wherein the medium formulations with disodium metal-or-phosphate and disodium metal-or-phosphate are increased or decreased by the same factor at the same time, facilitating use in small or large scale cultivation.
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