Method for controllable microbial deposition of carbonate minerals with different crystal forms and sizes
By constructing a recombinant strain through genetic engineering, the problem of slow rate of microbial catalytic CO2 hydration reaction was solved, and efficient deposition of carbonate minerals with different crystal sizes was achieved. It is suitable for environments such as water and concrete, with low cost and stable performance.
Patent Information
- Application Number
- CN202510905279.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-02
- Publication Date
- 2025-10-17
AI Technical Summary
In the existing technology, the microbial catalysis of CO2 hydration reaction rate is slow, the carbonic anhydrase activity is low, the expression level is small and the stability is poor, making it difficult to achieve large-scale industrial use of microorganisms to deposit carbonate minerals of different crystal sizes.
Through genetic engineering, a recombinant strain was constructed, the CA gene protein sequence and His tag were introduced, the engineered bacteria were constructed, the reaction conditions were regulated, and carbonate minerals with different crystal sizes were deposited.
The efficient expression and stability of carbonic anhydrase have been achieved, which can quickly deposit calcium carbonate of different crystal sizes. It is suitable for environments such as water and concrete, with low cost, high yield and stable performance.
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Figure CN120796397A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the field of microbial deposition of carbonate minerals, and particularly relates to a method for microbial deposition of carbonate minerals with different crystal size. BACKGROUND
[0002] There are more than 100 kinds of carbonate minerals known at present. Carbonate minerals are divided into different mineral types according to the cation components, such as calcium ions which can form three isomeric crystalline forms of calcite, aragonite and vaterite. Microbial deposition of carbonate minerals is a common phenomenon in nature and has become one of the research hotspots in geology microbiology. The action mode is that microorganisms produce carbonic anhydrase (CA) to catalyze the reversible reaction between CO2 and HCO3 - , thus promoting carbonate mineralization. Therefore, carbonic anhydrase is also one of the key enzymes for carbonate mineralization.
[0003] Carbonic anhydrase mineralization bacteria are a kind of mineralization microorganisms that can produce carbonic anhydrase (CA) and can efficiently catalyze the hydration reaction of CO2 in the air. Carbonic anhydrase is a metal enzyme with zinc as the active center, which widely exists in metabolic diversity of prokaryotes in animal, plant, archaea and bacteria, and is one of the fastest enzymes in terms of reaction rate. In nature, the hydration reaction rate of CO2 is quite slow, and the conversion number is only 1.3×10 -1 / s. When carbonic anhydrase catalyzes the above reaction, the conversion number of CO2 hydration is significantly improved to 1.4×10 7 / s, which is about 10 7 times the rate in nature. Carbonic anhydrase can catalyze the mutual transformation reaction between CO2 and HCO3 - , and each carbonic anhydrase molecule can catalyze 1.4×10 6 CO2 molecules to combine with water to form H2CO3 in 1s. Microorganisms accelerate CO2 hydration by generating carbonic anhydrase, rapidly generate a large amount of HCO3 － , and form a supersaturated solution with Ca 2+ in the environment to produce calcium carbonate precipitation.
[0004] Due to the problems of low enzyme activity, low expression amount, poor stability and easy to be affected by environmental conditions in natural bacteria, the use of genetic engineering technology to construct recombinant bacteria to improve the expression amount of enzyme can effectively solve this problem. The engineering bacteria are artificially constructed strains by introducing target into bacteria through biological technology or genetic engineering means in order to enhance the required characteristics. The rational design is based on the known protein structure and function mechanism, and the mutation site is designed by using the mechanism, and the obtained mutant has the expected function. The use of genetic engineering technology to construct CA heterologous expression recombinant bacteria can improve the expression amount of enzyme, realize the rapid and large-scale production of CA, and construct the engineering bacteria with the "superpower" of fixing CO2, which becomes an effective way to realize large-scale industrialization. SUMMARY
[0005] The technical problem to be solved by the present application is to provide a method for depositing carbonate minerals of different crystal size by controllable microorganisms, which is used for depositing calcium carbonate of different crystal size.
[0006] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:
[0007] A method for depositing carbonate minerals of different crystal size by controllable microorganisms, comprising the following steps:
[0008] (1) Construction of engineering bacteria: introducing a plasmid containing a CA gene protein sequence, a long hinge and a His tag into a chassis bacteria to construct engineering bacteria for depositing carbonate minerals of different crystal size;
[0009] (2) Preparation of engineering bacteria liquid: culturing the engineering bacteria obtained in step (1) in a culture medium to obtain an engineering bacteria liquid;
[0010] (3) Carbonate mineral deposition: applying the engineering bacteria liquid obtained in step (2) to deposit carbonate minerals of different crystal size, and depositing carbonate minerals of different crystal size by adjusting the reaction conditions.
[0011] Preferably, in step (1), the construction of engineering bacteria is as follows:
[0012] (a) Synthesis of plasmid: enzyme cutting site Nde I + CA gene protein sequence + long hinge + His tag + stop codon + enzyme cutting site Xho I gene sequence;
[0013] (b) Selection of vector: selecting a vector matching the enzyme cutting site in step (a), and the vector is suitable for the protein expression system of the chassis bacteria;
[0014] (c) Strain transformation: the plasmid is added to the competent cells of the chassis strain and mixed, first in ice water bath for 20-50 min, then in 42℃ water bath for 60 s, and finally quickly placed in ice water bath for 1-5 min to obtain the transformation solution;
[0015] (d) Strain screening culture: the transformation solution obtained in step (c) is spread on the culture medium plate containing the corresponding antibiotic to the carrier resistance, or mixed with fresh liquid culture medium and culture medium containing the corresponding antibiotic to the carrier resistance, and then poured into the plate, and after 1-3 h culture at 37℃, the plate is inverted and cultured in the 37℃ incubator overnight;
[0016] (e) Engineering bacteria amplification: single colonies on the plate are selected and cultured in the culture medium to obtain the engineering bacteria.
[0017] Preferably, in step (b), the pET-21a vector is selected, and the resistance is ampicillin (Amp) or kanamycin (KnaR), and more preferably Amp.
[0018] Preferably, in step (c), the ice water bath time is 30 min, then 42℃ water bath for 60 s, and finally quickly placed in ice water bath for 2 min.
[0019] Preferably, in step (c), the culture medium is a mixture of one or more of LB medium, TB medium or NA medium.
[0020] Preferably, in step (c), the antibiotic is ampicillin (Amp) or kanamycin (KnaR).
[0021] Preferably, in step (d), the culture medium is a mixture of one or more of LB medium, TB medium or NA medium.
[0022] More preferably, in steps (d) and (e), the culture medium is LB medium.
[0023] Preferably, in step (d), the antibiotic is ampicillin (Amp) or kanamycin (KnaR).
[0024] Preferably, in step (d), the 37℃ culture is 1 h.
[0025] Preferably, in step (1), the chassis strain is one or more of Escherichia coli BL21, Bacillus subtilis or yeast. More preferably, the chassis strain is Escherichia coli BL21.
[0026] Preferably, in step (2), the obtained engineered bacteria in step (1) are mixed into a culture medium, and cultured at 20-50°C for 6-24 hours to obtain an engineered bacteria solution; wherein the culture medium is a mixture of one or more of LB medium, TB medium or NA medium. More preferably, the culture medium is LB medium, the culture temperature of the engineered bacteria is 37°C, and the culture time is 12 hours.
[0027] Preferably, in step (2), the culture medium contains a water-soluble Ca 2+ compound.
[0028] Preferably, the water-soluble Ca 2+ compound is one or more of CaCl2, CaSO4, Ca(CH3COO)2, Ca3(C6H5O7)2. More preferably, the water-soluble Ca 2+ compound is CaCl2.
[0029] Preferably, in step (2), the concentration of Ca 2+ in the culture medium is 10-100 mM.
[0030] Preferably, in step (3), the precipitated carbonate mineral includes one or more of precipitated calcium carbonate mineral or magnesium carbonate mineral. More preferably, the carbonate mineral is calcium carbonate mineral.
[0031] Preferably, in step (3), the reaction conditions include reaction temperature, pH value, and reaction time.
[0032] Preferably, in step (3), the reaction conditions are 20°C, pH 5-6, and 1-3 days, and the precipitated calcium carbonate is nanometer calcite. More preferably, the pH is 5.5.
[0033] Preferably, in step (3), the reaction conditions are 20°C, pH 7-8, and 1-3 days, and the precipitated calcium carbonate is vaterite. More preferably, the pH is 7.
[0034] Preferably, in step (3), the reaction conditions are 35°C, pH 5-6, and 1-3 days, and the precipitated calcium carbonate is calcite. More preferably, the pH is 5.5.
[0035] Preferably, in step (3), the reaction conditions are 35°C, pH 7-8, and 1-3 days, and the precipitated calcium carbonate is calcite or vaterite. More preferably, the pH is 7.
[0036] Preferably, in step (3), the precipitation process is used in combination with a bio-based carrier.
[0037] Preferably, the bio-based carrier is a bio-based nanomaterial.
[0038] Preferably, in step (3), the engineered bacteria solution is applied to water.
[0039] Beneficial effects: The method of the application explores strains capable of producing carbonic anhydrase from nature, and artificially constructs the engineering bacteria for depositing calcium carbonate with different crystal sizes, which has strong controllability, high mineralization rate and stable performance.
[0040] The engineering bacteria liquid in the application is suitable for depositing carbonate minerals in water, concrete and other environmental conditions, and can deposit carbonate minerals with different crystal sizes after regulation. The method of the application can be used for depositing carbonate minerals with different crystal sizes, has low cost, high carbonate mineral yield, stable performance and strong controllability. BRIEF DESCRIPTION OF DRAWINGS
[0041] Figure 1 It is the electrophoretogram for detecting the morphological characteristics and proteins of the engineering bacteria in the examples;
[0042] Figure 2 It is the morphology and characterization of the engineering bacteria depositing nanoscale calcite under time, temperature and pH value in the examples; wherein: A, B: SEM images of nanoscale calcite; C: TEM image of nanoscale calcite; D: nanoscale calcite lattice stripe image; E: nanoscale calcite EDS image; F: nanoscale calcite XRD image;
[0043] Figure 3 It is the type and characterization of the engineering bacteria depositing calcium carbonate with different crystal sizes under time, temperature and pH value in the examples; wherein: A: calcite B: vaterite C: calcite and vaterite D: XRD image. DETAILED DESCRIPTION
[0044] The application will be further described below in conjunction with the examples. Those skilled in the art will be able to implement the application based on these descriptions. In addition, the examples of the application involved in the following description are generally only a part of the examples of the application, not all examples. Therefore, based on the examples in the application, all other examples obtained by those skilled in the art without creative labor shall fall within the scope of protection of the application.
[0045] A controllable method for depositing carbonate minerals with different crystal sizes by microorganisms, comprising the following steps:
[0046] (1) Construction of engineering bacteria: introducing a plasmid containing CA gene protein sequence, long hinge and His tag into a chassis bacteria to construct engineering bacteria for depositing carbonate minerals with different crystal sizes;
[0047] In some preferred embodiments of the application, the construction step of the engineering bacteria is as follows:
[0048] (a) Synthesis of plasmid: enzyme cutting site Nde I + CA gene protein sequence + long hinge + His tag + stop codon + enzyme cutting site Xho I gene sequence;
[0049] (b) Selection of vector: select a vector matching the enzyme cutting site in step (a), which is suitable for the protein expression system of the chassis; in some preferred embodiments of the present application, the pET-21a vector is selected, with resistance to ampicillin (Amp) or kanamycin (KnaR), more preferably Amp;
[0050] (c) Strain transformation: add the plasmid to the competent cells of the chassis and mix, first in ice water bath for 20-50 min, then in 42℃ water bath for 60 s, and finally quickly put in ice water bath for 1-5 min to obtain the transformation liquid; in some preferred embodiments of the present application, the ice water bath time is 30 min, then in 42℃ water bath for 60 s, and finally quickly put in ice water bath for 2 min;
[0051] In some preferred embodiments of the present application, the chassis is one or more of Escherichia coli BL21, Bacillus subtilis or yeast. More preferably, the chassis is Escherichia coli BL21.
[0052] (d) Strain screening culture: take the transformation liquid obtained in step (c) and spread on a culture medium plate containing an antibiotic corresponding to the resistance of the vector, or add fresh liquid medium and mix with a culture medium containing an antibiotic corresponding to the resistance of the vector, then pour the plate, and after 1-3 h of culture at 37℃, the plate is inverted and cultured overnight in a 37℃ incubator; wherein the culture medium is a mixture of one or more of LB medium, TB medium or NA medium, preferably LB medium; the antibiotic is ampicillin (Amp) or kanamycin (KnaR);
[0053] (e) Engineering bacteria amplification: select a single colony on the plate and culture in a culture medium to obtain engineering bacteria, wherein the culture medium is a mixture of one or more of LB medium, TB medium or NA medium, preferably LB medium.
[0054] (2) Engineering bacteria liquid preparation: mix the engineering bacteria obtained in step (1) into a culture medium, and culture at 20-50℃ for 6-24 hours to obtain an engineering bacteria liquid; wherein the culture medium is a mixture of one or more of LB medium, TB medium or NA medium. More preferably, the culture medium is LB medium, the engineering bacteria culture temperature is 37℃, and the culture time is 12 hours.
[0055] In some preferred embodiments of the present application, the culture medium is added with a water-soluble Ca 2+ containing compound. Preferably, the Ca 2 +The compound is one or more of CaCl2, CaSO4, Ca(CH3COO)2, Ca3(C6H5O7)2. More preferably, the Ca-containing 2+ The compound is CaCl2.
[0056] In some preferred embodiments of the present application, the concentration of Ca 2+ is 10-100 mM.
[0057] (3) Carbonate mineral deposition: the engineered bacteria solution obtained in step (2) is applied to water, soil, concrete or steel slag environment, and by adjusting the reaction conditions, different crystal size carbonate minerals are deposited.
[0058] In some preferred embodiments of the present application, the reaction conditions include reaction temperature, pH value, reaction time, and one or more of the reaction conditions are adjusted.
[0059] In some preferred embodiments of the present application, the reaction conditions are 20℃, pH 5-6, 1-3 days, and the deposited calcium carbonate is nanometer calcite.
[0060] In some preferred embodiments of the present application, the reaction conditions are 20℃, pH 7-8, 1-3 days, and the deposited calcium carbonate is vaterite.
[0061] In some preferred embodiments of the present application, the reaction conditions are 35℃, pH 5-6, 1-3 days, and the deposited calcium carbonate is calcite.
[0062] In some preferred embodiments of the present application, the reaction conditions are 35℃, pH 7-8, 1-3 days, and the deposited calcium carbonate is calcite, vaterite.
[0063] In some preferred embodiments of the present application, the deposition process is used in combination with a bio-based carrier, and preferably the bio-based carrier is a bio-based nanomaterial.
[0064] In some preferred embodiments of the present application, the engineered bacteria solution is applied to water.
[0065] The present application is further described below in conjunction with examples.
[0066] Example 1: Construction of engineered bacteria
[0067] The present application is implemented by using E. coli (BL21) as a chassis bacteria for experiments to verify the technical solutions of the present application. The engineered bacteria use E. coli (BL21) as a chassis bacteria, and the construction method includes the following steps:
[0068] (1) Synthesis of plasmid: insert gene main components: enzyme cutting site Nde I + CA gene protein sequence + long hinge + His tag + stop codon + enzyme cutting site Xho I.
[0069] (2) Strain transformation: ①Use of vector: pET-21a; resistance: Amp, take 100 μL competent (BL21), thaw on ice; ②Centrifuge the centrifuge tube containing the freeze-dried plasmid in the palm centrifuge for 2 min, add 100 μL of sterile water, then add 5 μL of ligation product or 0.5-2 μL of plasmid, gently mix, and place in ice water bath for 30 min, then gently mix 3-5 times; 42°C water bath for 60 s, quickly place in ice water bath for 2 min; add 900 μL of fresh LB medium, seal and culture for 1-3 h (37°C, 15000 rpm). ③Take 10-100 μL of bacterial solution and add the corresponding antibiotic (Amp / KnaR) to the LB plate (add one thousandth of the antibiotic and shake well to pour the plate), or inoculate 2-4 mL of LB medium containing the corresponding antibiotic (10 mL centrifuge tube), and incubate at 37°C overnight.
[0070] (3) Determination of engineering bacteria In LB medium (40 mL), TB medium (3 L), TPTG mother liquor, pick single colony to LB medium (+ Amp) for culture; 37°C, 170 rpm, turbid culture, observe whether the bacteria grow; preserve bacteria, take 1-2 μL of bacterial solution, PCR verification, take 200 μL of bacterial solution for sequencing verification. The detection results show that the engineering bacteria are successfully constructed, such as Figure 1 .
[0071] Example 2: Time, temperature and pH value control engineering bacteria to deposit calcium carbonate as nanoscale calcite
[0072] The engineering bacteria were inoculated into the LB medium containing 60 mM CaCl2 at an amount of 1% and cultured at 20°C and pH 5.5 for 1 d or 3 d, and the precipitate was collected by centrifugation, washed with deionized water and ethanol for 3 times respectively, and dried. After washing and drying the obtained precipitate, the mineral composition thereof was determined by X-ray diffractometer (XRD, Dmax-B type, Rigaku Corporation, Japan). The determination conditions: Cu target, Kα, tube pressure 35 kV, tube flow 20 mA, 2° / min, step 0.01°, determination range 5°-60°. The morphology of the precipitate was observed and the element composition was determined by Sirion field emission scanning electron microscope (S-3400) with element analysis.
[0073] The detection results show that the precipitated calcium carbonate is nanoscale calcite Figure 2 , Table 1).
[0074] Example 3: Time, temperature and pH value control engineering bacteria to deposit calcium carbonate as micron-scale calcite
[0075] The engineered bacteria were inoculated into a LB medium containing 60 mM CaCl2 at an amount of 1% and cultured at 35°C and pH 5.5 for 1 to 3 days. The precipitate was collected by centrifugation, washed with deionized water and ethanol three times, respectively, and dried. After washing and drying the obtained precipitate, the mineral composition thereof was measured using an X-ray diffractometer (XRD, Dmax-B, Rigaku Corporation, Japan). Measurement conditions: Cu target, Kα, tube voltage 35 kV, tube current 20 mA, 2° / min, step width 0.01°, measurement range 5° to 60°. The morphology of the precipitate was observed and the elemental composition thereof was measured using a Sirion field emission scanning electron microscope (S-3400) with elemental analysis.
[0076] The results of the measurement indicated that the precipitated calcium carbonate was a micrometer-sized calcite (CaCO3) (Table 1, Example 1). Figure 3
[0077] Example 4: Time, temperature, and pH control of deposition of calcium carbonate by engineered bacteria as micrometer-sized calcite
[0078] The engineered bacteria were inoculated into a LB medium containing 60 mM CaCl2 at an amount of 1% and cultured at 35°C and pH 7 for 1 day. The precipitate was collected by centrifugation, washed with deionized water and ethanol three times, respectively, and dried. After washing and drying the obtained precipitate, the mineral composition thereof was measured using an X-ray diffractometer (XRD, Dmax-B, Rigaku Corporation, Japan). Measurement conditions: Cu target, Kα, tube voltage 35 kV, tube current 20 mA, 2° / min, step width 0.01°, measurement range 5° to 60°. The morphology of the precipitate was observed and the elemental composition thereof was measured using a Sirion field emission scanning electron microscope (S-3400) with elemental analysis.
[0079] The results of the measurement indicated that the precipitated calcium carbonate was a micrometer-sized calcite (CaCO3) (Table 1, Example 1). Figure 3
[0080] Example 5: Time, temperature, and pH control of deposition of calcium carbonate by engineered bacteria as micrometer-sized vaterite
[0081] The engineered bacteria were inoculated into a LB medium containing 60 mM CaCl2 at an amount of 1% and cultured at 20°C and pH 7 for 1 to 3 days. The precipitate was collected by centrifugation, washed with deionized water and ethanol three times, respectively, and dried. After washing and drying the obtained precipitate, the mineral composition thereof was measured using an X-ray diffractometer (XRD, Dmax-B, Rigaku Corporation, Japan). Measurement conditions: Cu target, Kα, tube voltage 35 kV, tube current 20 mA, 2° / min, step width 0.01°, measurement range 5° to 60°. The morphology of the precipitate was observed and the elemental composition thereof was measured using a Sirion field emission scanning electron microscope (S-3400) with elemental analysis.
[0082] The results of the detection show that the precipitated calcium carbonate is micron-sized calcite (CaCO3) and micron-sized vaterite (CaCO3). Figure 3 Table 1).
[0083] Example 6: Time, temperature and pH value control the engineered bacteria to deposit calcium carbonate as micron-sized calcite and micron-sized vaterite
[0084] The engineered bacteria were inoculated into the LB medium containing 60 mM CaCl2 at an amount of 1%, and cultured at 35°C and pH 7 for 3 days. The precipitate was collected by centrifugation, washed with deionized water and ethanol for 3 times, respectively, and dried. After the obtained precipitate was washed and dried, its mineral composition was determined by X-ray diffractometer (XRD, Dmax-B type, Rigaku Corporation, Japan). The determination conditions were as follows: Cu target, Kα, tube pressure 35 kV, tube flow 20 mA, 2° / min, step 0.01°, determination range 5°-60°. The morphology of the precipitate was observed and its elemental composition was determined by Sirion field emission scanning electron microscope (S-3400) with elemental analysis.
[0085] The results of the detection show that the precipitated calcium carbonate is micron-sized calcite (CaCO3) and micron-sized vaterite (CaCO3). Figure 3 Table 1).
[0086] Table 1: Influence of time, temperature and pH value on the mineralization of the engineered bacteria
[0087]
[0088] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.
Claims
1. A method for controllable microbial deposition of carbonate minerals of different crystal sizes, characterized by: The steps include: (1) Construction of engineered bacteria: A plasmid containing the CA gene protein sequence, a long hinge, and a His tag was introduced into the chassis bacteria to construct engineered bacteria for depositing carbonate minerals of different crystal sizes; (2) Preparation of engineered bacterial liquid: culturing the engineered bacteria obtained in step (1) in a culture medium to obtain an engineered bacterial liquid; (3) Carbonate mineral deposition: The engineered bacterial solution obtained in step (2) is applied to the deposition of carbonate minerals with different crystal sizes, and by adjusting the reaction conditions, carbonate minerals with different crystal sizes are deposited.
2. The method according to claim 1, wherein: In step (1), the construction steps of the engineered bacteria are as follows: (a) Synthetic plasmid: Nde I restriction enzyme cleavage site + CA gene protein sequence + long hinge + His tag + stop codon + Xho I restriction enzyme cleavage site gene sequence; (b) selecting a vector: selecting a vector that matches the restriction enzyme cleavage site in step (a), wherein the vector is suitable for the protein expression system of the bottom plate bacteria; (c) Transformation: Add the plasmid to the competent cells of the bottom plate bacteria and mix well. Incubate in an ice-water bath for 20-50 minutes, then in a 42°C water bath for 60 seconds, and finally in an ice-water bath for 1-5 minutes to obtain the transformation solution. (d) Strain screening and culture: The transformation solution obtained in step (c) is spread on a culture medium plate containing an antibiotic corresponding to the vector resistance, or fresh culture medium and a culture medium containing an antibiotic corresponding to the vector resistance are added, mixed, and then poured onto the plate. After incubation at 37°C for 1-3 hours, the plate is inverted and cultured in a 37°C incubator overnight. (e) Proliferation of engineered bacteria: Select a single colony on the plate and culture it in a culture medium to obtain the engineered bacteria.
3. The method according to claim 1 or 2, characterized in that: In step (1), the substrate bacteria are one or more of Escherichia coli BL21, Bacillus subtilis or yeast.
4. The method according to claim 1, wherein: In step (2), the engineered bacteria obtained in step (1) are mixed into a culture medium and cultured at 20-50° C. for 6-24 hours to obtain an engineered bacterial solution; wherein the culture medium is a mixture of one or more of LB culture medium, TB culture medium or NA culture medium.
5. The method according to claim 1 or 5, characterized in that: In step (2), water-soluble Ca-containing 2+ Compounds, in which the culture medium contains Ca 2+ The concentration is 10-100mM.
6. The method according to claims 1-6, characterized in that: In step (3), the deposited carbonate minerals include deposited calcium carbonate and magnesium carbonate minerals.
7. The method according to claim 1, wherein: In step (3), the reaction conditions include reaction temperature, pH value, and reaction time.
8. The method according to claim 1, wherein: In step (3), The reaction conditions are 20°C, pH 5-6, and 1-3 days, and the deposited calcium carbonate is nano-calcite; The reaction conditions are 20°C, pH 7-8, and 1-3 days, and the precipitated calcium carbonate is vaterite; The reaction conditions are 35°C, pH 5-6, and 1-3 days, and the precipitated calcium carbonate is calcite; The reaction conditions are 35° C., pH 7-8, and 1-3 days, and the precipitated calcium carbonate is calcite and vaterite.
9. The method according to claim 1, wherein: In step (3), the engineering bacteria liquid is applied to water, desert, beach, rock, soil, concrete or steel slag environment.
10. The method according to claim 1, wherein: In step (3), the deposition process is used in conjunction with a bio-based carrier, which is a bio-based nanomaterial.