CN109295108A - The method for preparing different crystal system calcium carbonate using microbial mineralization - Google Patents
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Abstract
The present invention provides a kind of methods for preparing different crystal system calcium carbonate using microbial mineralization, the bacteria suspension that washing after the completion of culture is resuspended is inoculated into the mixed liquor containing calcium salt and urea with certain proportion, it stands and controls different mineralising reaction conditions and carry out mineralising reaction, deposit is taken out after the reaction was completed to be washed and dried, it can be obtained the carbonic acid calcium product of different crystal forms, wherein the microorganism is urase superior strain Pasteur's sporosarcina.The present invention prepares different crystal system calcium carbonate using microbial mineralization, the microbial resources wherein utilized are abundant, enzymic catalytic reaction is efficient, simple production process, it is low in cost, the calcium carbonate sample with the different crystal forms of different stability and compact structure is obtained by control mineralising reaction condition, the application effect of calcium carbonate is improved, is applied to calcium carbonate widely in many fields.
Description
Technical field
The present invention relates to the preparation methods of calcium carbonate, especially a kind of to prepare different crystal system calcium carbonate using microbial mineralization
Method.
Background technique
Calcium carbonate is the inorganic chemical industry material that a kind of dosage is big, widely used and a kind of typical biomineralization product,
It is widely used in the different field such as rubber, plastics, papermaking, ink, coating, medicine, environmental protection, food.The crystal form packet of calcium carbonate
Calcite, aragonite, vaterite three types are included, wherein calcite structure is most stable, and aragonite and vaterite belong to unstable state, no
There is also certain differences for its compact structure of isomorphous calcium carbonate and pattern, thus the crystal form of calcium carbonate is applied to effect
It has a certain impact.
Currently used calcium carbonate preparation method can be divided into mechanical crushing method and chemical reaction method.Mechanical crushing method is usually
Natural limestone, Bai Zhishi are crushed and obtain calcium carbonate, the calcium carbonate made from this method is powdered whiting, partial size compared with
Greatly, generally micron order.Chemical reaction method obtained is precipitated calcium carbonate, including carbonizatin method, precipitation reaction method and gas diffusion
Method etc..The product size that the former obtains is big, and producing process energy consumption is high, and dust is big, and resource is limited.Latter process's process is more multiple
Miscellaneous, energy consumption is high, and easily causes environmental pollution.
Biomineralization be a kind of microorganism by life activity to form minerals the phenomenon that, utilize microorganism mine
It is the method more at present with development potentiality that change, which prepares calcium carbonate, and the microbial resources utilized in this method are abundant, and its institute
The enzymatic reaction of dependence is single-minded and efficient, environmental-friendly, and calcium carbonate obtained has excellent mineralogical character, has been widely used
In many fields.Urease-producing microorganism is a kind of common biomineralization microorganism, with high efficiency and controllability.At present
It is not described also about the method for preparing different crystal system calcium carbonate using microbial mineralization, therefore, the present invention is intended to provide a kind of
The method for preparing different crystal system calcium carbonate using microbial mineralization.
Calcium carbonate, which is prepared, using microbial mineralization at this stage has become a kind of widely applied technology:
It is mentioned in 1778934 A of patent CN and decomposes substrate urea using urase caused by Bacillus pasteurii, generated
CO3 2-Environment pH is improved simultaneously, in incorporation Ca2+In the case where induction generate calcium carbonate, to obtain carbonic acid calcium product.
Carbonic acid is deposited using urease-producing microorganism Bacillus pasteurii decomposing urea mineralising in 104818719 A of patent CN
Calcium.Microbial inoculum is mixed with nutritive salt first, is in the milk again with certain proportion and calcium salt soln blending after 0.5~1h, this side
Method can effectively improve the yield of early precipitation process object.
It mentions using carbonic anhydrase caused by bacillus cereus in 102121033 A of patent CN respectively in calcium ion
CO is discharged with bicarbonate ion solution system and calcium ion and using ammonium bicarbonate soln2Gas diffusion system chats system
Obtain calcium carbonate.
In the existing invention for preparing calcium carbonate using microbial mineralization, microbe mineralising skill only it has been related to
Art prepares calcium carbonate, or improves its yield for generating calcium carbonate, does not relate to the carbonic acid that different crystal forms are prepared using the technology
Calcium product.Because the calcium carbonate of different crystal forms its stability, compact structure and its pattern have a certain difference, this is also by shadow
It rings and arrives the application effect of calcium carbonate in practical applications.
Summary of the invention
The purpose of the present invention overcomes the shortcomings of the prior art place, according to problem above, provides a kind of utilization micro- life
The method that object mineralising prepares different crystal system calcium carbonate.
The present invention realizes that the technical solution of purpose is as follows:
A method of different crystal system calcium carbonate being prepared using microbial mineralization, the bacterium that washing after the completion of culture is resuspended is hanged
Liquid is inoculated into the mixed liquor containing calcium salt and urea with certain proportion, is stood and is controlled different mineralising reaction conditions progress mineralisings
Reaction takes out deposit after the reaction was completed and is washed and dried, can be obtained the carbonic acid calcium product of different crystal forms；
The microorganism is urase superior strain Pasteur's sporosarcina.
Moreover, Pasteur's sporosarcina culture used medium component are as follows: 19~21g/L of yeast extract, ammonium sulfate 9
The Tris-Hcl of the pH 8.5~9.5 of~11g/L, 0.13mol/L.
Moreover, the activation of Pasteur's sporosarcina and expansion condition of culture are 25~37 DEG C, 150~200r/min is shaken
Bed culture 16~for 24 hours.
Moreover, the cell concentration of bacteria suspension is 7 × 10 using being resuspended after brine thallus 2~3 times5~7
×107A/mL.
Moreover, the volume ratio of bacteria suspension and calcium salt and urea mixed solution is 1:1.
Moreover, controlling mineralising reaction condition as 0.5~1.0mol/L of calcium salt, the ratio of the amount of the substance of calcium salt and urea is 1
~1.5:1,45~50 DEG C of temperature, reaction 20~obtain pure calcite type calcium carbonate for 24 hours；
Control mineralising reaction condition is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:
1,4~8g/L of yeast powder, 30~35 DEG C of temperature, 18~22h of reaction obtains pure vaterite-type calcium carbonate；
Control mineralising reaction condition is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:
1,35~40 DEG C of temperature, 16~20h of reaction obtains the calcium carbonate containing two kinds of crystal forms of calcite and vaterite.
Moreover, deposit is washed with distilled water 2~3 times, dry 2~3h is placed at 60 DEG C.
Moreover, the calcium salt includes inorganic calcium salt calcium chloride, calcium nitrate and organic calcium salt calcium lactate, calcium formate and acetic acid
Calcium.
The present invention compared with prior art the advantages of and good effect it is as follows:
The present invention prepares calcium carbonate using microbial mineralization, and on the one hand overcoming traditional calcium carbonate preparation method, energy consumption is high,
It is complicated for operation, easily cause the difficult points such as environmental pollution, this method is easy to operate, and controllability is strong and calcium carbonate generation efficiency is higher.Separately
On the one hand the invention provides a kind of preparation methods of different crystal system calcium carbonate, can select the application not isomorphous according to actual needs
The carbonic acid calcium product of type, the whole application effect for improving calcium carbonate.
The present invention is compared with existing invention, while preparing calcium carbonate using microbial mineralization, also controls generation carbon
The crystal form of sour calcium makes the calcium carbonate generated because the difference of crystal type is fined classification, is also more convenient for according to different needs
To the carbonic acid calcium product of application different crystal forms.
Detailed description of the invention
Fig. 1 is the XRD spectrum of the calcium carbonate obtained under different condition, and wherein C represents calcite, and V represents vaterite.a,b,
C, d respectively represents the XRD spectrum of the calcium carbonate obtained under the conditions of embodiment 1,2,3,4.
Specific embodiment:
The method of the present invention is described below by specific embodiment.Unless stated otherwise, technology used in the present invention
Means are method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, rather than this hair is limited
Bright range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, it is not carrying on the back
Under the premise of from spirit and scope of the present invention, in these embodiments material component and dosage carry out various changes or change
It is dynamic to also belong to protection scope of the present invention.
The present invention provides a kind of method for preparing different crystal system calcium carbonate using microbial mineralization.First will in the method
Microorganism carries out activation culture, then will washing is resuspended after the completion of culture bacteria suspension be inoculated into certain proportion containing calcium salt and
In the mixed liquor of urea, different mineralising reaction conditions progress mineralising reactions are stood and controlled, take out deposit after the reaction was completed
It is washed and is dried, can be obtained the carbonic acid calcium product of different crystal forms；The invention is mainly characterized by using microbial mineralization
Calcium carbonate is prepared, while the carbonic acid calcium product of different crystal forms being made by control mineralising reaction condition.Microbial mineralization prepares carbon
The microbial resources of sour calcium application are abundant, and enzymatic reaction is single-minded and efficient, and entire reaction controllability is strong.Different crystal forms obtained
Carbonic acid calcium product can preferably meet application demand.
In the present invention, urase catalyzing urea caused by Pasteur's sporosarcina hydrolyzes, and can mention for calcium carbonate generation
For CO3 2-, while environment pH is improved, introducing Ca2+Afterwards, can calcium carbonate be induced to generate under the action of microorganism, it is entire to react
Process is as follows:
NH2COOH+H2O→NH3+H2CO3
It is involved in the present invention to a kind of method for preparing different crystal system calcium carbonate using microbial mineralization, specifically include with
Lower step:
(1) picking Pasteur sporosarcina single colonie is seeded in the sterilized culture medium of 20mL in superclean bench
Activation culture is carried out, then the bacterium solution that activation is completed is forwarded in sterilized fresh culture and is expanded culture, is activated
It is 25~37 DEG C, 150~200r/min with the condition for expanding culture, shaking table culture 16~for 24 hours.
(2) bacteria suspension that washing after the completion of culture is resuspended is inoculated into the mixed liquor containing calcium salt and urea with certain proportion
In, different mineralising reaction conditions progress mineralising reactions are stood and controlled, deposit is taken out after the reaction was completed and is washed and dried
It is dry, it can be obtained the carbonic acid calcium product of different crystal forms.Wherein using being resuspended after brine thallus 2~3 times, bacterium is outstanding
The cell concentration of liquid is 7 × 105~7 × 107The volume ratio of a/mL, bacteria suspension and calcium salt and urea mixed solution is 1:1.Control mine
Change reaction condition is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:1, temperature 45~50
DEG C, reaction 20~obtain pure calcite type calcium carbonate for 24 hours；Control mineralising reaction condition be 0.5~1.0mol/L of calcium salt, calcium salt with
The ratio of the amount of the substance of urea is 1~1.5:1,4~8g/L of yeast powder, and 30~35 DEG C of temperature, 18~22h of reaction obtains pure ball graupel
Stone-type calcium carbonate；Control mineralising reaction condition is 0.5~1.0mol/L of calcium salt, the ratio of the amount of the substance of calcium salt and urea is 1~
1.5:1,35~40 DEG C of temperature, 16~20h of reaction obtains the calcium carbonate containing two kinds of crystal forms of calcite and vaterite.The precipitating of acquisition
Substance is washed with distilled water 2~3 times, is placed at 60 DEG C and is dried 2~3h to get the carbonic acid calcium product of different crystal forms.
The urase producing bacterial strain that the present invention can be used is as follows:
1, Pasteur's sporosarcina ATCC 11859
2, Pasteur's sporosarcina MTCC 1761
3, bacillus megaterium SS3
4, bacillus licheniformis ATCC 9789
5, Bacillus sphaericus NZRM 4381
6, spherical lysine bacillus ATCC 4525
7, bacillus subtilis ATCC 6633
The specific effect of the above method is illustrated by following 4 embodiments.
Embodiment 1
A method of different crystal system calcium carbonate being prepared using microbial mineralization, steps are as follows:
(1) it prepares Pasteur's sporosarcina culture used medium: weighing Tris 15.75g and be dissolved in 1L distilled water,
With 50% salt acid for adjusting pH, it is configured to the Tris-Hcl buffer of pH 9.0, then weighs yeast extract 20g, ammonium sulfate
10g is dissolved in 1LTris-Hcl buffer.The fluid nutrient medium of preparation is sub-packed in conical flask, after 121 DEG C of sterilizing 20min
It takes out spare.
(2) anhydrous calcium chloride 83.3g is weighed, urea 45.0g is dissolved in 1L distilled water, is uniformly mixed, in the mixed solution
The concentration of calcium chloride is about 0.75mol/L, and the ratio of the amount of the substance of calcium chloride and urea is about 1:1.
(3) picking Pasteur sporosarcina single colonie is seeded to the sterilized culture medium of 20mL in superclean bench
In, 30 DEG C are placed in, shaking table culture 16h under the conditions of 180r/min, the bacterium solution for then taking 1mL activation to complete is forwarded to 100mL and has gone out
In the fresh culture of bacterium, 30 DEG C are placed in, shaking table culture 18h under the conditions of 180r/min.
(4) with being resuspended after brine thallus 2 times, the cell concentration of bacteria suspension is 7 × 106A/mL, by bacterium
Suspension is that 1:1 is mixed with volume ratio with calcium salt and urea mixed solution, is statically placed at 45 DEG C and reacts 22h.Deposit is taken out to steam
Distilled water is washed 3 times, is taken out after being placed at 60 DEG C dry 2h.
Embodiment 2
A method of different crystal system calcium carbonate being prepared using microbial mineralization, steps are as follows:
(3) it prepares Pasteur's sporosarcina culture used medium: weighing Tris 15.75g and be dissolved in 1L distilled water,
With 50% salt acid for adjusting pH, it is configured to the Tris-Hcl buffer of pH 9.0, then weighs yeast extract 20g, ammonium sulfate
10g is dissolved in 1LTris-Hcl buffer.The fluid nutrient medium of preparation is sub-packed in conical flask, after 121 DEG C of sterilizing 20min
It takes out spare.
(4) anhydrous calcium chloride 83.3g is weighed, urea 45.0g is dissolved in 1L distilled water, is uniformly mixed, in the mixed solution
The concentration of calcium chloride is about 0.75mol/L, and the ratio of the amount of the substance of calcium chloride and urea is about 1:1.
(3) picking Pasteur sporosarcina single colonie is seeded to the sterilized culture medium of 20mL in superclean bench
In, 30 DEG C are placed in, shaking table culture 16h under the conditions of 180r/min, the bacterium solution for then taking 1mL activation to complete is forwarded to 100mL and has gone out
In the fresh culture of bacterium, 30 DEG C are placed in, shaking table culture 18h under the conditions of 180r/min.
(4) with being resuspended after brine thallus 2 times, the cell concentration of bacteria suspension is 7 × 106A/mL, by bacterium
Suspension is that 1:1 is mixed, while yeast powder 6g/L being added thereto with volume ratio with calcium salt and urea mixed solution, is statically placed at 30 DEG C
React 20h.It takes out deposit to be washed with distilled water 3 times, be taken out after being placed at 60 DEG C dry 2h.
Embodiment 3
A method of different crystal system calcium carbonate being prepared using microbial mineralization, steps are as follows:
(1) it prepares Pasteur's sporosarcina culture used medium: weighing Tris 15.75g and be dissolved in 1L distilled water,
With 50% salt acid for adjusting pH, it is configured to the Tris-Hcl buffer of pH 9.0, then weighs yeast extract 20g, ammonium sulfate
10g is dissolved in 1LTris-Hcl buffer.The fluid nutrient medium of preparation is sub-packed in conical flask, after 121 DEG C of sterilizing 20min
It takes out spare.
(2) anhydrous calcium chloride 83.3g is weighed, urea 45.0g is dissolved in 1L distilled water, is uniformly mixed, in the mixed solution
The concentration of calcium chloride is about 0.75mol/L, and the ratio of the amount of the substance of calcium chloride and urea is about 1:1.
(3) picking Pasteur sporosarcina single colonie is seeded to the sterilized culture medium of 20mL in superclean bench
In, 30 DEG C are placed in, shaking table culture 16h under the conditions of 180r/min, the bacterium solution for then taking 1mL activation to complete is forwarded to 100mL and has gone out
In the fresh culture of bacterium, 30 DEG C are placed in, shaking table culture 18h under the conditions of 180r/min.
(4) with being resuspended after brine thallus 2 times, the cell concentration of bacteria suspension is 7 × 106A/mL, by bacterium
Suspension is that 1:1 is mixed with volume ratio with calcium salt and urea mixed solution, is statically placed at 35 DEG C and reacts 16h.Deposit is taken out to steam
Distilled water is washed 3 times, is taken out after being placed at 60 DEG C dry 2h.
Embodiment 4
A method of different crystal system calcium carbonate being prepared using microbial mineralization, steps are as follows:
(1) it prepares Pasteur's sporosarcina culture used medium: weighing Tris 15.75g and be dissolved in 1L distilled water,
With 50% salt acid for adjusting pH, it is configured to the Tris-Hcl buffer of pH 9.0, then weighs yeast extract 20g, ammonium sulfate
10g is dissolved in 1LTris-Hcl buffer.The fluid nutrient medium of preparation is sub-packed in conical flask, after 121 DEG C of sterilizing 20min
It takes out spare.
(2) anhydrous calcium chloride 83.3g is weighed, urea 45.0g is dissolved in 1L distilled water, is uniformly mixed, in the mixed solution
The concentration of calcium chloride is about 0.75mol/L, and the ratio of the amount of the substance of calcium chloride and urea is about 1:1.
(3) picking Pasteur sporosarcina single colonie is seeded to the sterilized culture medium of 20mL in superclean bench
In, 30 DEG C are placed in, shaking table culture 16h under the conditions of 180r/min, the bacterium solution for then taking 1mL activation to complete is forwarded to 100mL and has gone out
In the fresh culture of bacterium, 30 DEG C are placed in, shaking table culture 18h under the conditions of 180r/min.
(4) with being resuspended after brine thallus 2 times, the cell concentration of bacteria suspension is 7 × 106A/mL, by bacterium
Suspension is that 1:1 is mixed with volume ratio with calcium salt and urea mixed solution, is statically placed at 40 DEG C and reacts 20h.Deposit is taken out to steam
Distilled water is washed 3 times, is taken out after being placed at 60 DEG C dry 2h.
Table 1 is the crystal form content analysis result for the calcium carbonate sample that 4 embodiments obtain.
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | |
| Calcite (%) | 100 | 0 | 18% | 59% |
| Vaterite (%) | 0 | 100 | 82% | 41% |
Claims (8)
1. a kind of method for preparing different crystal system calcium carbonate using microbial mineralization, it is characterised in that: will be washed after the completion of culture
The bacteria suspension of resuspension is inoculated into the mixed liquor containing calcium salt and urea with certain proportion, is stood and is controlled different mineralising reaction items
Part carries out mineralising reaction, takes out deposit after the reaction was completed and is washed and dried, can be obtained the calcium carbonate of different crystal forms
Product；The bacteria suspension is urase superior strain Pasteur sporosarcina bacteria suspension.
2. the method according to claim 1 for preparing different crystal system calcium carbonate using microbial mineralization, it is characterised in that: bar
Family name's sporosarcina culture used medium component are as follows: 19~21g/L of yeast extract, 9~11g/L of ammonium sulfate,
The Tris-Hcl of the pH 8.5~9.5 of 0.13mol/L.
3. a kind of method for preparing different crystal system calcium carbonate using microbial mineralization according to claim 1, feature exist
In: Pasteur's sporosarcina activation and to expand condition of culture be 25~37 DEG C, 150~200r/min, shaking table culture 16~
24h。
4. the method according to claim 1 for preparing different crystal system calcium carbonate using microbial mineralization, it is characterised in that: bacterium
Suspension is, using being resuspended after brine thallus 2~3 times, the cell concentration of bacteria suspension is 7 × 105~7 × 107A/
mL。
5. the method according to claim 1 or 4 for preparing different crystal system calcium carbonate using microbial mineralization, feature exist
In: the volume ratio of bacteria suspension and calcium salt and urea mixed solution is 1:1.
6. the method according to claim 1 for preparing different crystal system calcium carbonate using microbial mineralization, it is characterised in that: control
Mineralising reaction condition processed is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:1, temperature 45~
50 DEG C, reaction 20~obtain pure calcite type calcium carbonate for 24 hours；
Control mineralising reaction condition is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:1, ferment
4~8g/L of female powder, 30~35 DEG C of temperature, 18~22h of reaction obtains pure vaterite-type calcium carbonate；
Control mineralising reaction condition is 0.5~1.0mol/L of calcium salt, and the ratio of the amount of the substance of calcium salt and urea is 1~1.5:1, temperature
35~40 DEG C of degree, 16~20h of reaction obtain the calcium carbonate mixture containing two kinds of crystal forms of calcite and vaterite.
7. a kind of method for preparing different crystal system calcium carbonate using microbial mineralization according to claim 1, feature exist
In: deposit is washed with distilled water 2~3 times, is placed at 60 DEG C dry 2~3h.
8. a kind of method for preparing different crystal system calcium carbonate using microbial mineralization according to claim 1, feature exist
In: the calcium salt includes inorganic calcium salt calcium chloride, calcium nitrate and organic calcium salt calcium lactate, calcium formate and calcium acetate.
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