CN114292882A - Method for promoting microorganism to induce carbonate precipitation by using attapulgite clay - Google Patents
Method for promoting microorganism to induce carbonate precipitation by using attapulgite clay Download PDFInfo
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Abstract
The invention discloses a method for promoting microbial induced carbonate precipitation by using attapulgite clay, belonging to the technical field of microbial induced carbonate precipitation. The method provides a material basis for Microbial Induced Carbonate Precipitation (MICP) by utilizing calcium ions in the attapulgite clay, and promotes the microbial induced carbonate precipitation. The method comprises the steps of inoculating the microbial strain liquid into a liquid culture medium containing the attapulgite clay leaching liquor, simultaneously adding urea, carrying out constant-temperature shaking culture, promoting the microbial induction to generate white precipitates, carrying out characterization identification on the generated precipitates, and determining the feasibility of the precipitates. The method has the advantages of low economic cost, simple extraction, no secondary pollution and good application prospect in the aspects of water heavy metal removal and soil heavy metal passivation and stabilization.
Description
Technical Field
The invention relates to the technical field of microbial induced carbonate precipitation and attapulgite clay application, in particular to a method for promoting microbial induced carbonate precipitation by using attapulgite clay.
Background
The phenomenon of soil microorganism-induced calcium carbonate precipitation (MICP) was first discovered by Boquet et al as early as 1973. Microorganism in natural environment, because the surface of bacteria is charged with negative charge, metal ions such as calcium ions, magnesium ions and the like in the environment can be enriched on the surface of cells, and then the metal ions are combined with carbonate generated by decomposing urea by urease, so that the metal ions are precipitated into the environment in the form of carbonate or attached to the surface of the microorganism. Part of heavy metal ions and Ca2+Has chemical similarity, and can replace Ca in calcium carbonate crystal lattice during the process of microorganism-induced carbonate precipitation2+The metal ions are precipitated into calcite crystals in a coprecipitation mode, or heavy metals are adsorbed on the surfaces of the calcite, and even some heavy metals are mutually complexed with extracellular polymers generated by microorganisms and are wrapped by the calcite, so that the metal ions are immobilized. MICP can be widely used in the fields of biological cement production, soil solidification, heavy metal passivation, crack repair and the like, but the MICP process has not been applied in various fields on a large scale due to the high cost of nutrient components and calcium sources required by microorganisms and the like. Thus, reducing the economic cost of MICP is one of the important issues to be addressed.
CN108718586A discloses a method for solidifying sandy soil by using oyster shells as a regenerative calcium source through carbonate precipitation induced by microorganisms. Wherein 8% -15% of nitric acid is used for dissolving calcium ions in oyster shells, but the extraction cost of a calcium source is high and the risk of acid pollution is existed.
The microbial induction of carbonate precipitation requires a large amount of free cations, particularly calcium ions, and a large number of studies have shown that among the existing calcium salts, calcium chloride is the most precipitated as a calcium source, and the formed carbonate is mainly calcite. Calcium chloride is the most preferred calcium source without accounting for economic costs. However, the economic cost of calcium chloride is high, so that the popularization and application of the technology are limited. Therefore, finding a low-cost calcium source has important significance for popularization and application of the MICP technology.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The present invention is directed to overcoming the above-mentioned drawbacks of the prior art and providing a method for promoting microbial induced carbonate precipitation by using attapulgite clay.
The invention is realized by the following steps:
in a first aspect, the invention provides a method for promoting microbial induced carbonate precipitation by using attapulgite clay, which provides a material basis for microbial induced carbonate precipitation by using calcium ions in the attapulgite clay to promote microbial induced carbonate precipitation.
In a second aspect, the invention also provides an application of the method for promoting microorganism to induce carbonate precipitation by using the attapulgite clay in the aspects of water heavy metal removal and soil heavy metal passivation stabilization.
The invention has the following beneficial effects:
the invention provides a method for promoting microbial induced carbonate precipitation by using attapulgite clay, which is a method for providing a material basis for Microbial Induced Carbonate Precipitation (MICP) by using calcium ions in the attapulgite clay to promote the microbial induced carbonate precipitation. The method comprises the steps of inoculating the microbial strain liquid into a liquid culture medium containing the attapulgite clay leaching liquor, simultaneously adding urea, carrying out constant-temperature shaking culture, promoting the microbial induction to generate white precipitates, carrying out characterization identification on the generated precipitates, and determining the feasibility of the precipitates. The method has the advantages of low economic cost, simple extraction, no secondary pollution and good application prospect in the aspects of water heavy metal removal and soil heavy metal passivation and stabilization.
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 embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a schematic view of a method for promoting microbial induced carbonate precipitation using attapulgite clay;
FIG. 2 shows the presence of Bacillus fusiformis (LF) in CaCl2SEM pictures of the resulting precipitate under induction;
FIG. 3 is an SEM image of the precipitation of Bacillus fusiformis (LF) under the induction of a water-soluble leaching solution of silver attapulgite clay;
FIG. 4 shows enterococcus (LZU-1) in CaCl2SEM pictures of the resulting precipitate under induction;
FIG. 5 is an SEM image of the precipitation of enterococcus (LZU-1) induced by water soluble leaching solution of silver attapulgite clay;
FIG. 6 is an XRD pattern of precipitates generated by two strains under induction of different calcium sources (B-ATP: silver attapulgite clay water-soluble leaching liquor);
FIG. 7 shows the removal efficiency of two strains induced by different calcium sources for removing As solutions with the same concentration;
FIG. 8 shows the As remediation efficiency for soil remediation by attapulgite clay;
FIG. 9 is a graph of the remediation efficiency of As in soil for different urea concentrations;
FIG. 10 shows the contents of calcium and magnesium ions in the attapulgite clay extracted at different solid-to-liquid ratios.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The invention aims to provide a method for promoting microorganism to induce carbonate precipitation by utilizing attapulgite clay. The method is an environmental application method of the attapulgite clay, provides a material basis for the microbial induction of carbonate precipitation by utilizing calcium ions in the attapulgite clay, and can optimize the environmental benefit and the economic benefit of the microbial induction of carbonate precipitation.
In order to achieve the above object of the present invention, the following solutions are adopted:
the embodiment of the invention provides a method for promoting microbial induced carbonate precipitation by using attapulgite clay, which provides a material basis for microbial induced carbonate precipitation by using calcium ions in the attapulgite clay and promotes the microbial induced carbonate precipitation.
In an alternative embodiment, the method for promoting microbial-induced carbonate precipitation using attapulgite clay comprises the steps of: inoculating the enriched bacterial liquid into a liquid culture medium containing the attapulgite clay leaching liquor, simultaneously adding urea, and carrying out constant-temperature shaking culture to promote the microorganism to induce and generate white precipitated calcium carbonate.
In an alternative embodiment, the preparation of the broth comprising attapulgite clay leaching solution comprises the steps of: adding 9-11g of peptone, 4-6g of sodium chloride, 2-4g of beef extract and 1050ml of attapulgite clay leaching liquor into a culture medium, adjusting the pH to 6.8 +/-0.2, and sterilizing at the high temperature of 121 ℃ and under high pressure for 20-30 min;
preferably, adding 10g of peptone, 5g of sodium chloride, 3g of beef extract and 1000ml of attapulgite clay leaching liquor into the culture medium, adjusting the pH to 6.8 +/-0.2, and sterilizing at 121 ℃ for 20min under high temperature and high pressure to obtain a liquid culture medium containing the attapulgite clay leaching liquor.
In an alternative embodiment, the preparation of the attapulgite clay leachate comprises the following steps: mixing attapulgite clay and water, and oscillating at constant temperature to extract calcium ions in the attapulgite clay;
preferably, the attapulgite clay and distilled water are mixed in a solid-to-liquid ratio of 1: 2-10, shaking at room temperature and 180rpm for 2-3h, centrifuging, filtering, and storing the filtrate to obtain attapulgite clay leaching solution;
preferably, the attapulgite clay and distilled water are mixed in a solid-to-liquid ratio of 1:10, mixing at room temperature, shaking at 200rpm for 2h, centrifuging at 4000rpm for 20min, filtering, and storing the filtrate to obtain attapulgite clay leaching liquor;
more preferably, the attapulgite clay is air-dried, ground, and sieved with 0.15mm sieve before use.
In an alternative embodiment, the attapulgite clay is selected from at least one of the attapulgite clays of silver in Gansu province and Linzee in Gansu province.
In an alternative embodiment, the conditions of the constant temperature shaking culture are as follows: adding urea accounting for 4-10% of the total mass of the liquid culture medium, and performing shaking culture in a constant-temperature shaker at 25-30 ℃ and at 120-150rpm for 3-5 d;
preferably, the conditions of the constant temperature shaking culture are as follows: adding urea which accounts for 6 percent of the total mass of the liquid culture medium, and carrying out shake culture in a constant temperature shaker at 30 ℃ and 150rpm for 3d-5 d.
In an alternative embodiment, the microorganism is a urease-producing strain.
In alternative embodiments, the urease producing strain is selected from lysinibacillus fusiformis LF and/or enterococcus LZU-1;
preferably, the lysine bacillus fusiformis LF is lysine bacillus fusiformis with the number of preservation library BNCC210664 of Beijing Beinan biological strains in China;
preferably, the enterococcus LZU-1 is enterococcus with a preservation number of CGMCC 22622 in China general microbiological culture Collection center.
The embodiment of the invention provides a method for promoting microorganism to induce carbonate precipitation by utilizing attapulgite clay, wherein the used enterococcus LZU-1 has a preservation unit as follows: the China general microbiological culture Collection center has the following preservation addresses: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, with the deposit number: CGMCC 22622, the preservation date is: 31/5/2021, latin names by taxonomic nomenclature: enterococcus gallinarum.
In an alternative embodiment, the microorganism is expanded before being inoculated into a liquid culture medium containing attapulgite clay leaching liquor;
preferably, the expanding culture of the microorganism comprises the following steps: inoculating the preserved strain in a nutrient broth culture medium in a super clean bench with an inoculation amount of 1% -2%Culturing at 25-30 deg.C under 150rpm in a constant temperature oscillator, and periodically detecting its light absorption value OD600Up to the absorbance OD 6001 to 1.5;
preferably, the preserved strain is inoculated in 50ml of nutrient broth in a super clean bench, the inoculum size is 1% -2%, the strain is cultured in a constant temperature oscillator at 30 ℃ and 150rpm, and the light absorption value OD is detected periodically600Up to the absorbance OD 6001, completing the expanding culture of the microorganisms;
preferably, the nutrient broth medium consists of: 9-11g/L of peptone, 2-4g/L of beef extract, 4-6g/L of sodium chloride and pH of 7 +/-0.2.
In a second aspect, an embodiment of the present invention further provides an application of the method for promoting microbial induced carbonate precipitation by using attapulgite clay in removal of heavy metals in water and passivation and stabilization of heavy metals in soil.
The embodiment of the invention provides a method for promoting microorganism to induce carbonate precipitation by using attapulgite clay. Compared with the prior method, the method has the advantages that:
(1) the attapulgite clay reserves are abundant in China and the economic cost is low, and the attapulgite clay leachate generated in the production process of the existing large quantity of attapulgite clay industries or the wastewater generated by modifying the attapulgite clay needs to be treated, so that the economic cost of microorganism-induced carbonate precipitation can be reduced by taking the attapulgite clay leachate as a calcium source, and the waste is recycled;
(2) the method for extracting calcium ions in the attapulgite clay is simple and environment-friendly, and does not cause secondary pollution; the attapulgite clay and the MICP can reduce the emission of greenhouse gases during the process of passivating heavy metal pollutants.
(3) Magnesium ions in the attapulgite clay have a promoting effect on the generation of carbonate precipitates.
The features and properties of the present invention are described in further detail below with reference to examples.
A method for promoting microorganism to induce carbonate precipitation by using attapulgite clay comprises the following steps:
(1) pretreating the attapulgite clay: the attapulgite clay adopted by the invention is silver attapulgite clay (B-ATP), and the cost is low. Naturally air drying the collected attapulgite clay, and sieving with a 0.15mm sieve for later use.
(2) Extracting water-soluble calcium ions in the attapulgite clay: the attapulgite clay sample and distilled water are mixed according to a solid-to-liquid ratio of 1: mixing at a ratio of 10(m/V), shaking at constant temperature oscillator at 25 deg.C and 200rpm for 2 hr, centrifuging at 4000rpm in high speed centrifuge for 20min, and filtering to obtain attapulgite clay leaching solution.
(3) Preparation of liquid medium: the invention uses Nutrient Broth (NB): 10g of peptone, 5g of sodium chloride, 3g of beef extract and 1000ml of attapulgite clay leaching liquor, adjusting the pH to 6.8 +/-0.2, and sterilizing at 121 ℃ for 20min under high temperature and high pressure. The physicochemical properties and water-soluble calcium and magnesium ion contents of the above silver attapulgite clay (B-ATP) are shown in Table 1.
(4) Expanding culture of the strain: inoculating the preserved strain in 50ml Nutrient Broth (NB) culture medium in a super clean bench with an inoculum size of 1% -2%, culturing at 30 deg.C and 150rpm in a constant temperature oscillator, and periodically detecting its light absorption value OD600(Optical sensitivity 600) until its absorbance OD600Around 1.
(5) And (3) generation and identification of calcite precipitation induced by microorganisms: and (3) inoculating the enriched bacterial liquid in the step (4) into the culture medium in the step (3) in a super clean bench, adding 6% of urea, culturing for 3d-5d at 30 ℃ and 150rpm in a constant-temperature oscillator to generate white precipitates, centrifuging, and carrying out cold drying. The main components of the precipitated product were then determined by Scanning Electron Microscopy (SEM) and X-ray diffraction (XRD) analysis.
In order to verify the applicability of the method, four different attapulgite clays are respectively selected to stabilize heavy metal-like arsenic (As) in aqueous solution and soil. The four attapulgite clays are respectively: the attapulgite clay comprises silver attapulgite clay (B-ATP), Linze attapulgite clay 1(L-ATP1), Linze attapulgite clay 2(L-ATP2) and Linze attapulgite clay 3(L-ATP3), and has physical and chemical properties and water solubility of Ca2+、Mg2+The contents are shown in table 1, and the total rock mineral composition and the clay mineral composition are shown in tables 2 and 3.
TABLE 1 physicochemical Properties of four Attapulgite ClaysAnd water-soluble Ca2+、Mg2+Content (wt.)
Table 2 whole rock mineral composition (%) -of four attapulgite clays
TABLE 3 Clay mineral composition of the four Attapulgite Clays
Note: s is montmorillonite, I/S is a mixed layer of illite and montmorillonite, I is illite, K is kaolinite, C is chlorite, C/S is a mixed layer of chlorite and montmorillonite, and Pa is attapulgite, i.e., palygorskite. The mix layer ratio (% S) is the percentage content of montmorillonite.
Example 1
The four attapulgite clay water-soluble extracting solutions provide material basis for MICP to remove heavy metal-like arsenic (As) in the water solution;
firstly, preparing a liquid culture medium by using a water-soluble extracting solution of attapulgite clay (ATP): weighing 35g of silver attapulgite clay, and mixing the materials according to a solid-liquid ratio of 1: adding 350ml distilled water into 10, shaking for 2h (200rpm, 25 deg.C), centrifuging (4000rpm, 10min), and filtering to obtain 300ml extractive solution.
Second, blank (CK) and control were prepared with distilled water to obtain Na having an As concentration of 20m g/L3AsO4Mixing the solution with nutrient broth, and preparing Na with As concentration of 20m g/L from attapulgite extract3AsO4Solution and nutrient broth mix. The three parallel experiments were performed in 50ml Erlenmeyer flasks. Sterilizing at 121 deg.C under high temperature and high pressure for 20min, cooling to room temperature, inoculating 1ml of LF strain enriched bacterial liquid in sterile table, and adding urea solution into conical flask (0.22 μ for urea solution)m water system filter head filtration, final urea concentration 6%).
And thirdly, culturing for 20 days at 25 ℃ and 150rpm, finally measuring the change of As concentration before and after removal, and calculating the removal efficiency. Wherein, the blank group (CK) is added with 20m g/L arsenic, 6% urea and 1ml enriched bacterial liquid; a control group containing calcium chloride as calcium source was supplemented with 20m g/L arsenic, 6% urea and 60mM CaCl2Inoculating 1ml of LF bacterial liquid; the experimental group using the water-soluble attapulgite clay extract as the calcium source was supplemented with 20m g/L arsenic, 6% urea and 1ml of LF bacterial liquid.
All treatments of LZU-1 strain were identical to LF strain.
Example 2
The attapulgite clay provides a material base for MICP in soil and solidifies heavy metal arsenic (As);
step one, enriching bacterial liquid: performing amplification culture on the LF strain by using NB liquid medium according to the amount required by the experiment (the inoculation amount is 1-5%); and (3) pretreating the soil sample, naturally drying the collected soil sample, grinding, and sieving by using a 2mm sieve for later use.
Step two, culturing the bacteria liquid to be enriched for 18-20h, and comparing the bacteria liquid with a reference group: soil, urea, LF (Bacillus Calmette-Guerin) bacteria and CaCl2And the bacteria liquid is mixed according to the solid-liquid ratio of 7: 3, adding 12 percent of urea and CaCl2The amount of (2) was 60mM, and LF was added as a bacterial suspension. Experimental groups: the addition amounts of the soil, the urea, the LF bacteria and the attapulgite clay are the same as those of a control group, the addition amount of the attapulgite clay is 10%, and the attapulgite clay added in the example is silver attapulgite clay (B-ATP), Linze attapulgite clay 1(L-ATP1), Linze attapulgite clay 2(L-ATP2) and Linze attapulgite clay 3(L-ATP 3).
It can be concluded from examples 1 and 2 that attapulgite clay, either as such or as leachate, can provide a material basis for the MICP process. When the attapulgite clay and the urease-producing microorganisms are used for jointly remedying the heavy metal pollution of the water body or the soil, the leaching liquor or the modified wastewater of the attapulgite clay can be utilized, and the attapulgite clay can also be utilized as it is.
As shown in fig. 2 to 5, the morphology of calcium carbonate precipitates induced by the two are slightly different from each other as shown by SEM analysis results: the calcium chloride-induced precipitate is of a regular rhomboid structure, and the silver attapulgite-induced precipitate is of an irregular rhomboid structure.
As shown in fig. 6, XRD analysis revealed that the precipitate generated using calcium chloride as a calcium source was mainly calcite, and the precipitate generated using silver attapulgite clay (B-ATP) as a calcium source was also mainly calcite.
As can be seen from the analysis of FIG. 7, for the aqueous solution with the same As concentration, the attapulgite clay extract was used As the calcium source and CaCl2The former has higher removal efficiency as a calcium source. In the As experiment in passivated soil, though CaCl2The removal efficiency as a calcium source is higher, but attapulgite clay is preferable in view of economic cost and environmental significance (prevention of soil salinization). CaCl2Needs to be processed and purified, and the attapulgite clay only needs to be simply dissolved in water or crude ore, and researches show that Mg is contained2+The MICP mineralizing crystallinity can be improved, and Table 4 shows that the removal rate of As by magnesium ions and LZU-1 is in positive correlation.
TABLE 4 removal rate of As from the extract of attapulgite clay in aqueous solution and the ratio of Ca to attapulgite clay2+、 Mg2+Correlation analysis of content
| Species of calcium source | As passivation rate LF | As passivation rate LZU-1 | Calcium ion | Magnesium ion | |
| Species of calcium source | 1.00 | -0.20 | 0.00 | -0.67 | -0.33 |
| As passivation rate LF | -0.20 | 1.00 | 0.40 | 0.67 | 0.33 |
| As passivation rate LZU-1 | 0.00 | 0.40 | 1.00 | 0.00 | 1.000* |
| Calcium ion | -0.67 | 0.67 | 0.00 | 1.00 | 0.00 |
| Magnesium ion | -0.33 | 0.33 | 1.000** | 0.00 | 1.00 |
Note: *. at the 0.05 level (two-tailed), the correlation was significant. At the 0.01 scale (double tail), the correlation was significant.
As shown in FIG. 8, in the soil, calcium chloride, B-ATP, L-ATP1, L-ATP2 and L-ATP3 are respectively used As calcium sources to stabilize As in the soil, and the repairing effect is As follows: the repair efficiency of the attapulgite clay serving As a calcium source for As in the soil is slightly lower than that of the calcium chloride serving As a calcium source, but the cost performance of the attapulgite clay serving As the calcium source is higher by comprehensively considering the difference between the price and the manufacturing process of the calcium chloride and the attapulgite clay.
Comparative example 1
Under the condition of quantifying the concentration of calcium chloride by 60mM, the concentration of urea is respectively 0%, 2%, 4%, 6%, 8% and 10%, LZU-1 and LF strains are added into soil, meanwhile, required nutrient broth culture media are added, and the bacteria liquid is mixed according to the solid-liquid ratio of 7: and 3, adding the urea, and evaluating the optimal urea concentration through the passivation rate of As in the soil after remediation.
According to FIG. 9, when the amount of urea added was 6%, the inactivation rate of As in soil was the highest by both strains, and thus the optimum amount of urea added in the medium was 6%. Among them, the effect of urea concentration on inactivation of As in soil by LZU-1 strain was more significant than that of LF strain.
Comparative example 2
Extracting the four attapulgite clays respectively at different solid-to-liquid ratios, wherein the solid-to-liquid ratios are respectively 1: 2. 1: 5. 1: weighing a certain amount of attapulgite clay, adding distilled water according to different solid-to-liquid ratios, shaking for 2-3h (200rpm, 25 ℃), centrifuging (4000rpm, 10min), filtering to obtain extract solutions under different solid-to-liquid ratios, and measuring the contents of calcium and magnesium by flame atomic absorption spectrophotometry.
According to the graph shown in fig. 10, four kinds of attapulgite clay are respectively mixed at a solid-to-liquid ratio of 1: 2. 1: 5. 1: when the content of calcium and magnesium in the extracting solution is 10 hours, the content of calcium extracted from the L-ATP3 is reduced greatly along with the reduction of the solid-to-liquid ratio, and the concentration of calcium ions in the rest attapulgite clay extracting solution is reduced slightly. The volume of the extract and the calcium ion concentration are obtained at equal mass while taking into account different solid-to-liquid ratios, resulting in a preferred solid-to-liquid ratio of 1: 10.
In summary, the embodiment of the present invention provides a method for promoting microbial induced carbonate precipitation by using attapulgite clay, which provides a material basis for Microbial Induced Carbonate Precipitation (MICP) by using calcium ions in attapulgite clay, so as to promote microbial induced carbonate precipitation. The key point is that the MICP process is carried out by utilizing calcium ions in the attapulgite clay. By pretreating the attapulgite clay, the soluble calcium ions in the attapulgite clay are utilized to provide a material basis for the MICP process. Extracting calcium ions in the attapulgite clay by distilled water, culturing strains by using the extract, adding required urea, and performing characterization and identification on the generated precipitate to determine the feasibility of the precipitate. The method has the advantages of low economic cost, simple extraction, no secondary pollution and good application prospect in the aspects of water heavy metal removal and soil heavy metal passivation and stabilization.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The method for promoting microbial induction of carbonate precipitation by using the attapulgite clay is characterized by providing a material basis for microbial induction of carbonate precipitation by using calcium ions in the attapulgite clay to promote microbial induction of carbonate precipitation.
2. The method of claim 1, comprising the steps of: inoculating the enriched bacterial liquid into a liquid culture medium containing the attapulgite clay leaching liquor, simultaneously adding urea, and carrying out constant-temperature shaking culture to promote the microorganism to induce and generate white precipitated calcium carbonate.
3. The method of claim 2, wherein the preparation of the liquid medium containing attapulgite clay leaching solution comprises the steps of: adding 9-11g of peptone, 4-6g of sodium chloride, 2-4g of beef extract and 1050ml of attapulgite clay leaching liquor into a culture medium, adjusting the pH to 6.8 +/-0.2, and sterilizing at the high temperature of 121 ℃ and under high pressure for 20-25 min;
preferably, 10g of peptone, 5g of sodium chloride, 3g of beef extract and 1000ml of attapulgite clay leaching liquor are added into the culture medium, the pH is adjusted to 6.8 +/-0.2, and the liquid culture medium containing the attapulgite clay leaching liquor is obtained by high-temperature high-pressure sterilization at 121 ℃ for 20 min.
4. The method according to claim 3, wherein the preparation of the attapulgite clay leaching liquor comprises the following steps: mixing attapulgite clay and water, and oscillating at constant temperature to extract calcium ions in the attapulgite clay;
preferably, the attapulgite clay and distilled water are mixed in a solid-to-liquid ratio of 1: 2-1: 10, mixing, shaking at room temperature and 180rpm for 2-3h, centrifuging, filtering, and storing filtrate to obtain the attapulgite clay leaching liquor;
preferably, the attapulgite clay and distilled water are mixed in a solid-to-liquid ratio of 1:10, at room temperature, shaking at 200rpm for 2h, centrifuging at 4000rpm for 20min, filtering, and storing the filtrate to obtain the attapulgite clay leaching liquor;
more preferably, the attapulgite clay is air-dried, ground and sieved with a 0.15mm sieve before use.
5. The method according to claim 4, wherein the attapulgite clay is selected from at least one of the group consisting of silver attapulgite clay of Gansu province and Lingzhou attapulgite clay of Gansu province.
6. The method according to claim 2, wherein the conditions of the constant temperature shaking culture are as follows: adding urea accounting for 4-10% of the total mass of the liquid culture medium, and performing shaking culture in a constant-temperature shaker at 25-30 ℃ and at 120-150rpm for 3-5 d;
preferably, the conditions of the constant temperature shaking culture are as follows: adding urea which accounts for 6 percent of the total mass of the liquid culture medium, and carrying out shake culture in a constant temperature shaker at 30 ℃ and 150rpm for 3d-5 d.
7. The method of claim 2, wherein the microorganism is a urease-producing strain.
8. The method according to claim 7, wherein the urease producing strain is selected from the group consisting of lysinibacillus fusiformis LF and/or enterococcus LZU-1;
preferably, the lysine bacillus fusiformis LF is lysine bacillus fusiformis with the number of preservation library BNCC210664 of Beijing Beinan biological strains in China;
preferably, the enterococcus LZU-1 is enterococcus with a preservation number of CGMCC 22622 in China general microbiological culture Collection center.
9. The method according to claim 8, wherein the microorganism is expanded before being inoculated into the liquid medium containing attapulgite clay leaching liquor;
preferably, the expanding culture of the microorganism comprises the following steps: inoculating the preserved strain in a nutrient broth culture medium in a super clean bench with an inoculum size of 1% -2%, culturing in a constant temperature oscillator at 25-30 deg.C and 120-600Up to the absorbance OD6001 to 1.5;
preferably, the preserved strain is inoculated in 50ml of nutrient broth in a super clean bench, the inoculum size is 1% -2%, the strain is cultured in a constant temperature oscillator at 30 ℃ and 150rpm, and the light absorption value OD is detected periodically600Up to the absorbance OD6001, completing the expanding culture of the microorganisms;
preferably, the nutrient broth medium consists of: 9-11g/L of peptone, 2-4g/L of beef extract, 4-6g/L of sodium chloride and pH of 7 +/-0.2.
10. The method for promoting microorganism-induced carbonate precipitation by using attapulgite clay according to any one of claims 1-9, wherein the method is applied to the removal of heavy metals in water and the passivation and stabilization of heavy metals in soil.
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