Preparation method of plant carbonic anhydrase induced carbon absorption mineralizer

Abstract

The invention discloses a preparation method of a plant carbonic anhydrase induced carbon absorption mineralizer, and belongs to the technical field of soil body improvement and reinforcement. The plant carbonic anhydrase induced carbon absorption mineralizer is prepared from the following raw materials in percentage by volume: 20 to 30 percent of plant carbonic anhydrase complex liquid, 20 to 40 percent of carbon sequestration agent A and 30 to 60 percent of carbon sequestration agent B, the plant carbonic anhydrase compound liquid comprises a plant carbonic anhydrase crude extracting solution; the carbon sequestration agent A comprises a soluble salt solution capable of forming carbonate minerals with carbonate; the carbon sequestration agent B comprises a carbon dioxide solution. The plant carbonic anhydrase-induced carbon absorption mineralizer can induce generation of carbonate crystals in a soil body through a biochemical reaction, cement soil body particles and improve the strength of the soil body.

Description

Technical Field

[0001] The invention relates to the technical field of soil improvement and reinforcement, in particular to a method for preparing a plant carbonic anhydrase-induced carbon absorption mineralizer. Background Art

[0002] Natural soil and artificial fill often cannot meet the requirements of strength, bearing capacity, permeability, etc. in engineering. Therefore, it is necessary to artificially process and improve the soil that does not meet the requirements to improve its physical and mechanical properties. Common soil improvement and reinforcement methods include mechanical densification methods such as strong tamping and vibration impaction, and cementitious material injection methods such as high-pressure jet grouting, deep mixing, and grouting. Mechanical densification methods require large machinery, and the energy generated by construction has a great impact on surrounding buildings. The cementitious materials used in the cementitious material injection method are mainly cement, and there are some chemical cementitious materials such as water glass and polymer materials. These cementitious materials are highly viscous and difficult to inject into the soil. They also require the help of large machinery to complete. In addition, some chemical cementitious materials have a relatively large impact on the environment and their own stability is also poor. At the same time, the production of these cementitious materials requires the emission of a large amount of carbon dioxide into the atmosphere, which is not conducive to environmental protection.

[0003] In recent years, biological induction methods have been gradually applied to soil improvement and reinforcement. With the help of biochemical processes existing in nature, inorganic or organic crystals are produced. Unlike chemical precipitation, the types and properties of the reaction products of these processes can be controlled by adjusting the reaction rate of the biochemical process. The produced crystals have a gelling effect, which can fill the pores in the soil particles, cement the soil particles or increase the roughness of the soil, thereby improving the mechanical properties of the soil. The urea hydrolysis process is a commonly used biological induction method, but a large amount of ammonium chloride will remain in the soil after the reaction, which will aggravate the salinization of the soil and the corrosion to the building. Some ammonium roots will be converted into ammonia, affecting the air quality of the construction environment. Therefore, it is urgent to find a green and safe method for soil improvement and reinforcement. Summary of the invention

[0004] The purpose of the present invention is to provide a method for preparing a plant carbonic anhydrase-induced carbon absorption mineralizer to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is a plant carbonic anhydrase-induced carbon absorption mineralizer, which comprises the following raw materials in volume percentage:

[0007] Plant carbonic anhydrase complex solution 20-30%, carbon fixer A 20-40% and carbon fixer B 30-60%;

[0008] The components of the plant carbonic anhydrase complex solution include a plant carbonic anhydrase crude extract;

[0009] The carbon fixer A includes a soluble salt solution that can form carbonate minerals with carbonate ions;

[0010] The carbon fixer B includes a carbon dioxide solution.

[0011] Furthermore, the method for preparing the crude plant carbonic anhydrase extract comprises the following steps:

[0012] The plant tissue containing the plant carbonic anhydrase is crushed and then filtered to obtain a coarse filtrate;

[0013] Add water and pH buffer solution to the crude filtrate, adjust the pH to 8-10, and centrifuge to obtain the crude extract of plant carbonic anhydrase (the crude extract of plant carbonic anhydrase can be stored in a refrigerator at 4° C. for long-term use later);

[0014] The ratio of the plant tissue to the crude plant carbonic anhydrase extract is 5-80 g / L.

[0015] Furthermore, the plant tissue includes one or more of food crop leaves, weed leaves and vegetable leaves.

[0016] From the perspective of availability and economy, it is best to use plant tissues that have no commercial value.

[0017] Furthermore, the food crop leaves include one or more of corn, sorghum and rice leaves; the vegetable leaves include one or more of spinach, water spinach, rapeseed and Chinese cabbage leaves.

[0018] Furthermore, the crushing method includes mechanical crushing or ultrasonic crushing.

[0019] Furthermore, the enzyme activity of the crude plant carbonic anhydrase extract is 20 to 60 U / mL.

[0020] Furthermore, the carbon-fixing agent A is a calcium salt and / or magnesium salt solution.

[0021] Furthermore, the calcium salt in the calcium salt solution includes one or more of calcium chloride, calcium nitrate and calcium acetate; the magnesium salt in the magnesium salt solution includes magnesium chloride and / or magnesium sulfate.

[0022] Furthermore, the concentration of ions that can form carbonate minerals with carbonate ions in the soluble salt solution is 0.06 to 0.6 mol / L.

[0023] Furthermore, the components of the plant carbonic anhydrase complex solution also include a surfactant.

[0024] Furthermore, the dosage of the surfactant is 0.025-0.25% of the volume of the plant carbonic anhydrase-induced carbon absorption mineralization agent.

[0025] Furthermore, the method for preparing the carbon dioxide solution includes: injecting gas containing carbon dioxide into water to obtain the solution.

[0026] Furthermore, the carbon dioxide solution is a carbon dioxide saturated solution.

[0027] Furthermore, the gas containing carbon dioxide includes pure carbon dioxide gas and / or air.

[0028] Furthermore, the pH buffer solution includes Hepes or Tris.

[0029] The use of pH buffer solution can provide a relatively stable pH environment for the plant carbonic anhydrase-induced carbon absorption mineralizer, ensuring the stable progress of the biochemical process.

[0030] The second technical solution of the present invention is a method for preparing the above-mentioned plant carbonic anhydrase-induced carbon absorption mineralizer, comprising the following steps:

[0031] The plant carbonic anhydrase complex liquid, the carbon fixer A and the carbon fixer B are uniformly mixed and the pH is adjusted to 8-10 to obtain the plant carbonic anhydrase inducing carbon absorption mineralization agent.

[0032] The third technical solution of the present invention: an application of the above-mentioned plant carbonic anhydrase-induced carbon absorption mineralizer in soil improvement and reinforcement.

[0033] The plant carbonic anhydrase-induced carbon absorption mineralizer of the present invention can induce carbonate crystals in the soil through its biochemical reaction, cement soil particles, and improve soil strength. When used, different effects (different strengths and permeabilities) can be achieved by applying the plant carbonic anhydrase-induced carbon absorption mineralizer multiple times, and different purposes can be achieved. The degree of strength improvement and the degree of permeability reduction can be controlled by adjusting the application amount, the number of applications, etc.

[0034] At the same time, when improving and reinforcing the soil surface, carbon dioxide in the atmosphere can be used as a reaction substrate to achieve the formation of carbonate crystals, thereby achieving the triple effects of carbon fixation, soil improvement and cost reduction.

[0035] The present invention discloses the following technical effects:

[0036] (1) The plant carbonic anhydrase-induced carbon absorption mineralizer of the present invention can induce carbonate crystals in the soil through biochemical reactions, cement soil particles, and improve soil strength.

[0037] (2) The improved principle of the plant carbonic anhydrase-induced carbon absorption mineralizer of the present invention is that biochemical reactions exist naturally in nature, especially in the ecological environment of soil. Compared with artificial injection of chemical cementitious materials, it is more stable and will not cause pollution to the environment.

[0038] (3) The plant carbonic anhydrase-induced carbon absorption mineralizer of the present invention requires less reaction substrate, the reaction product is in a crystalline form, and the amount of by-product salt after the reaction is terminated is small. Compared with other biological induction methods, it has less impact on the chemical environment of the soil itself and reduces the cost of use.

[0039] (4) The plant carbonic anhydrase-induced carbon absorption mineralizer of the present invention can absorb carbon dioxide in the atmosphere. The absorbed carbon dioxide can be used as a reaction substrate, so that the reaction can occur continuously without the input of additional reaction substrates, and carbon dioxide can be fixed, thereby reducing the carbon dioxide content in the atmosphere. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0041] Figure 1 The schematic diagram of the preparation process and use of the plant carbonic anhydrase-induced carbon absorption mineralizer;

[0042] Figure 2 This is a SEM image of calcium carbonate particles produced by the plant carbonic anhydrase group;

[0043] Figure 3 SEM image of calcium carbonate particles produced by a commercial carbonic anhydrase set. DETAILED DESCRIPTION

[0044] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0045] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0046] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0047] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0048] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0049] The first aspect of the present invention provides a plant carbonic anhydrase-induced carbon absorption mineralizer, which comprises the following raw materials, measured by volume percentage: 20-30% of a plant carbonic anhydrase complex solution, 20-40% of a carbon fixer A (carbon absorption mineralizer A) and 30-60% of a carbon fixer B (carbon absorption mineralizer B).

[0050] In a second aspect, the present invention provides a method for preparing the above-mentioned plant carbonic anhydrase-induced carbon absorption mineralizer, comprising the following steps:

[0051] (1) Extraction of plant carbonic anhydrase: water is added to spinach leaves, rice leaves, corn leaves, water spinach leaves, and purslane leaves, and the ratio of leaves to plant carbonic anhydrase crude extract is controlled to be 5-80 g / 1L, and then mechanical or ultrasonic crushing is performed and filtered, and pH buffer solution is added to the filtrate to control the extraction pH to 8-10, and centrifugation is performed to obtain a plant carbonic anhydrase solution (i.e., a plant carbonic anhydrase crude extract, and the activity of the plant carbonic anhydrase in the plant carbonic anhydrase crude extract is 20-60 U / mL).

[0052] Wherein, the pH buffer solution includes Hepes or Tris.

[0053] (2) The crude extract of plant carbonic anhydrase and a surfactant (the surfactant is Triton X-100, and the amount of the surfactant is 0.025-0.25% of the volume of the plant carbonic anhydrase inducing carbon absorption mineralization agent) are mixed uniformly to obtain a plant carbonic anhydrase complex solution.

[0054] (3) Preparation of ionic salt solution: Weigh a certain amount of soluble salt that can form a precipitate (carbonate mineral) with carbonate ions and add it to water to form a solution. The concentration of ions that can form carbonate minerals with carbonate ions in the solution is controlled to be 0.06-0.6 mol / L to obtain carbon fixation agent A (carbon absorption mineralizer A).

[0055] Among them, the soluble salt that can form a precipitate with carbonate is a calcium salt and / or a magnesium salt; the calcium salt includes one or more of calcium chloride, calcium nitrate and calcium acetate; the magnesium salt includes magnesium chloride and / or magnesium sulfate.

[0056] (4) Carbon dioxide is added to water until saturated to obtain a carbon fixation agent B (carbon absorption mineralization agent B).

[0057] (5) After the plant carbonic anhydrase complex solution, carbon fixer A (carbon absorption mineralizer A) and carbon fixer B (carbon absorption mineralizer B) are uniformly mixed, the pH is adjusted to 8-10 using a pH buffer solution to obtain a plant carbonic anhydrase-induced carbon absorption mineralizer (i.e., a carbonic anhydrase carbon absorption mineralizer).

[0058] The third aspect of the present invention is the use of the above-mentioned plant carbonic anhydrase-induced carbon absorption mineralizer in soil improvement and reinforcement.

[0059] In the present invention, the plant carbonic anhydrase-induced carbon absorption mineralizer can absorb carbon dioxide in the atmosphere and can also absorb carbon dioxide waste gas to improve and reinforce the soil.

[0060] The specific preparation process and schematic diagram of the plant carbonic anhydrase-induced carbon absorption mineralizer are shown in Figure 1 .

[0061] Example 1

[0062] A method for preparing a plant carbonic anhydrase-induced carbon absorption mineralizer:

[0063] (1) 40 g of waste spinach leaves were placed in a precooled crusher, and liquid nitrogen was quickly added to mechanically crush the cells. The solution after cell rupture was coarsely filtered through gauze; then poured into a 2 L beaker, and 1 L of pure water was added to the beaker. The pH of the solution was adjusted to 8 using Hepes buffer, and then placed in a high-speed refrigerated centrifuge for refrigerated centrifugation at a speed of 12000 r / min and a temperature of 0 to 4°C for 15 min to obtain a crude extract of plant carbonic anhydrase.

[0064] The activity of the crude extract of plant carbonic anhydrase was determined by the following method: the pH meter method was used, the test environment was 0°C, and one activity unit U = n*(t 0 / t e -1), where n is the enzyme dilution multiple, t 0 and t eThe time taken by the dead enzyme and the live enzyme to reduce the saturated carbon dioxide solution from 8.2 to 7.2 respectively. After measurement, the activity of the obtained enzyme solution is 60U / mL.

[0065] (2) The crude extract of plant carbonic anhydrase and a surfactant (the surfactant is Triton X-100, and the amount of the surfactant is 0.025-0.25% of the volume of the plant carbonic anhydrase inducing carbon absorption mineralization agent) are mixed uniformly to obtain a plant carbonic anhydrase complex solution.

[0066] (3) Weigh 16.65 g of calcium chloride and dissolve it in 250 mL of pure water to prepare 0.6 mol / L carbon fixer A.

[0067] (4) Place 250 mL of pure water in a 500 mL beaker, connect one end of a thin tube to a carbon dioxide cylinder, and place the other end below the liquid surface of the pure water to aerate the pure water. Adjust the pressure regulating valve of the carbon dioxide cylinder so that the bubbles flow out in uniform small bubbles. During the aeration, the solution is placed in an ice bath at 0 to 4°C, and the aeration time is maintained at more than 30 minutes to obtain a carbon fixer B (i.e., a carbon dioxide saturated solution).

[0068] (5) 200 mL of the above-mentioned carbon fixer A, 600 mL of the carbon fixer B and 200 mL of the plant carbonic anhydrase complex solution were mixed evenly, and the pH of the solution was adjusted to 8 with Hepes buffer to obtain a plant carbonic anhydrase-induced carbon absorption mineralization agent.

[0069] The plant carbonic anhydrase-induced carbon absorption mineralizer was placed in a high-pressure spray pot and the dry density was 1.61 g / cm 3 d 50 =0.35mm, pH 8 desert aeolian sand surface spray treatment, plant carbonic anhydrase induced carbon absorption mineralization agent dosage of 4L / m 2 Desert wind-blown sand.

[0070] The results showed that after 7 days of spraying, the surface particles of the aeolian sand were cemented together by the induced calcium carbonate crystals, which generated strength on the soil surface. At a wind speed of 10m / s, the aeolian sand was almost not lost. In a humid state, the treated surface can further absorb carbon dioxide in the air for mineralization, and the effect can be further enhanced.

[0071] Example 2

[0072] 3g, 10g, 20g and 50g of waste spinach leaves were respectively placed in a precooled crusher, and liquid nitrogen was quickly added to mechanically crush the cells, and the solution after cell rupture was coarsely filtered through gauze; then poured into a 2L beaker, and 1L of pure water was added to the beaker, the pH of the solution was adjusted to 8 using Hepes buffer, and then placed in a high-speed refrigerated centrifuge for refrigerated centrifugation at a centrifugal speed of 12000r / min, a temperature of 0-4°C, and centrifuged for 15min to obtain 3g / L, 10g / L, 20g / L and 50g / L crude extracts of plant carbonic anhydrase.

[0073] Take 20mL of crude extract of plant carbonic anhydrase with concentrations of 3g / L, 10g / L, 20g / L and 50g / L respectively, mix with 20mL of calcium chloride carbon fixer A with concentration of 0.31mol / / L, and then mix with 60mL of carbon fixer B, adjust the pH of the mixed solution to 8 with Hepes buffer, filter and dry the precipitate after complete reaction, and the mass of precipitated calcium carbonate is 0.04g, 0.08g, 0.122g and 0.134g respectively. According to calculation, the carbon fixation rates are 21%, 42%, 64% and 71% respectively.

[0074] Comparative Example 1

[0075] (1) 5 g of waste spinach leaves were placed in a precooled crusher, and liquid nitrogen was quickly added to mechanically crush the cells. The solution after cell rupture was coarsely filtered through gauze; then poured into a 2 L beaker, and 1 L of pure water was added to the beaker. The pH of the solution was adjusted to 8 using Hepes buffer, and then placed in a high-speed refrigerated centrifuge for refrigerated centrifugation at a speed of 12000 r / min and a temperature of 0 to 4°C for 15 min to obtain a crude extract of plant carbonic anhydrase with a concentration of 5 g / L.

[0076] (2) Take 20 mL of 5 g / L plant carbonic anhydrase crude extract and 0.087 g / L commercial carbonic anhydrase (Dulai, extracted from bovine red blood cells, 3000 U / mg) and mix them with 20 mL of 0.31 mol / L calcium chloride carbon fixer A, and then 60 mL of carbon fixer B. Adjust the pH of the mixed solution to 8 with Hepes buffer. Measure the amount of calcium carbonate produced in 5 minutes. The plant carbonic anhydrase group produces 0.013 g, and the commercial carbonic anhydrase group produces 0.005 g. Through SEM test ( Figure 2 and Figure 3 ) The calcium carbonate morphologies produced by the two groups of experiments were also different. The calcium carbonate particles produced by plant carbonic anhydrase were larger, more angular, and irregular in shape than those produced by commercial urease, and contributed more to the strength.

[0077] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A plant carbonic anhydrase-induced carbon absorption mineralizer, characterized in that: In terms of volume percentage, it includes the following raw materials: Plant carbonic anhydrase complex solution 20-30%, carbon fixer A 20-40% and carbon fixer B 30-60%; The components of the plant carbonic anhydrase complex solution include a plant carbonic anhydrase crude extract; The carbon fixer A includes a soluble salt solution that can form carbonate minerals with carbonate ions; The carbon fixer B includes a carbon dioxide solution.

2. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 1, characterized in that: The method for preparing the crude plant carbonic anhydrase extract comprises the following steps: The plant tissue containing the plant carbonic anhydrase is crushed and then filtered to obtain a coarse filtrate; Adding water and pH buffer solution to the crude filtrate, adjusting the pH to 8-10, and centrifuging to obtain the crude extract of the plant carbonic anhydrase; The ratio of the plant tissue to the crude plant carbonic anhydrase extract is 5-80 g / L.

3. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 2, characterized in that: The plant tissue comprises one or more of food crop leaves, weed leaves and vegetable leaves.

4. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 2, characterized in that: The enzyme activity of the crude plant carbonic anhydrase extract is 20-60 U / mL.

5. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 1, characterized in that: The carbon fixer A is a calcium salt and / or magnesium salt solution.

6. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 1, characterized in that: The concentration of ions that can form carbonate minerals with carbonate ions in the soluble salt solution is 0.06-0.6 mol / L.

7. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 1, characterized in that: The components of the plant carbonic anhydrase complex solution also include a surfactant.

8. The plant carbonic anhydrase-induced carbon absorption mineralizer according to claim 7, characterized in that: The dosage of the surfactant is 0.025-0.25% of the volume of the plant carbonic anhydrase-induced carbon absorption mineralization agent.

9. A method for preparing the plant carbonic anhydrase-induced carbon absorption mineralizer according to any one of claims 1 to 8, characterized in that: The following steps are involved: The plant carbonic anhydrase complex liquid, the carbon fixer A and the carbon fixer B are uniformly mixed and the pH is adjusted to 8-10 to obtain the plant carbonic anhydrase inducing carbon absorption mineralization agent.

10. Use of the plant carbonic anhydrase-induced carbon absorption mineralizer according to any one of claims 1 to 8 in soil improvement and reinforcement.

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