CN112126641B - Shell-silicon dioxide microorganism immobilization carrier and preparation method thereof - Google Patents
Shell-silicon dioxide microorganism immobilization carrier and preparation method thereof Download PDFInfo
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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Abstract
The invention relates to the technical field of microorganism application, in particular to a shell-silicon dioxide microorganism immobilization carrier and a preparation method thereof. The shell-silicon dioxide microorganism immobilization carrier is particles with the diameter of 4-7mm, and the porosity of the immobilization carrier is 40% -55%. The preparation method comprises the following steps of mixing shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, adding cement with a mass fraction of 5-10% as an adhesive, and uniformly mixing to form a mixed raw material; and uniformly mixing the mixed raw materials and water according to the ratio of 10-8: 1, and granulating to obtain the microorganism immobilized carrier particles. The microorganism immobilization carrier has no toxicity to microorganisms, good mechanical strength and low cost; the microorganism fixed by the microorganism fixed carrier has high activity.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to a shell-silicon dioxide microorganism immobilization carrier and a preparation method thereof.
Background
The stichopus japonicus has extremely high nutritional and medicinal values and is one of the aquaculture varieties with the highest economic value in the north of China. In recent years, large-scale industrial culture and offshore culture are rapidly developed, the sediment oxygen consumption rate is increased, the ammonia nitrogen content is remarkably increased, the benthic environment is gradually worsened, the disease problem is increasingly serious, and the environmental benefit of the benthic ecosystem and the economic benefit of stichopus japonicus culture are greatly influenced. Although the antibiotics can inhibit the occurrence of plant diseases and insect pests, the antibiotics also bring secondary pollution to a culture pond and an offshore ecological environment. Beneficial microbial floras become a new research direction for healthy cultivation and restoration of benthic ecological environment and biodiversity, and the beneficial microbial floras are widely researched and applied with the advantages of being green, safe, environment-friendly, inhibiting pathogenic microorganisms, purifying water quality and the like. At present, the application mode of the microbial preparation in the stichopus japonicus culture is mainly a free addition mode or a mixed addition mode with feed, and the main purpose is to adjust the balance of intestinal flora in the growth process of the stichopus japonicus, promote the growth and development of the stichopus japonicus and improve the economic benefit of the stichopus japonicus culture.
The immobilized microorganism technology is started from the 60 th 20 th century, is mainly developed by an immobilized enzyme technology, and the existing microorganism immobilizing technology mainly comprises four methods, namely an adsorption method, an embedding method, a covalent bonding method and a crosslinking method, wherein the adsorption method is low in cost, the immobilization process is simple and easy to operate, and the embedding method and the cell crosslinking method are complex in technology and high in cost. The immobilized microorganism technology is mainly used for degrading organic matters or other pollutants in water and soil, such as denitrification of wastewater, removal of pollutants of petroleum hydrocarbon and heavy metal in soil and the like. Artificial high polymer materials such as polyurethane, polyacrylamide and the like are common carriers, and the microbial toxicity of the artificial high polymer materials needs to be researched for a long time; natural polymer materials such as carrageenan and agarose plants and biological wastes such as straws are also common fixing materials, but the mechanical strength and the antimicrobial decomposition performance need to be further enhanced; the charcoal has high porosity and strong adsorbability, is a common material for fixing microorganisms, and has the defect of high cost. Thus, there is a need to provide a novel environment-friendly microorganism-immobilized carrier with a low price.
Disclosure of Invention
In view of the above, the invention provides a preparation method of a shell-silica microorganism immobilization carrier, thereby solving the problems of poor mechanical strength and high cost of the microorganism immobilization carrier and poor activity of loaded microorganisms.
In order to achieve the above purpose, the present invention mainly provides the following technical solutions:
a shell-silica microorganism immobilization support comprising: the shell-silicon dioxide microorganism fixing carrier is particles with the size of 4-7mm, the porosity of the fixing carrier is 40% -55%, the shell-silicon dioxide microorganism fixing carrier comprises shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, and cement with the mass fraction of 5-10% is used as an adhesive for granulation.
On the other hand, a method for preparing a shell-silica microorganism-immobilized carrier, comprising the steps of,
mixing raw materials: mixing shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, adding cement in a mass fraction of 5-10% as an adhesive, and uniformly mixing to form a mixed raw material;
and (3) granulation: and uniformly mixing the mixed raw materials and water according to the ratio of 10-8: 1, and granulating to obtain the microorganism immobilized carrier particles.
In the preparation method of the shell-silicon dioxide microorganism immobilization carrier, the shell powder is scallop shell powder which is obtained by grinding scallop shells into 60-40 meshes;
the silica particles are medium-fine-grained silica particles of 60-40 meshes;
in the granulating step, a disk granulator is used for granulating to form spherical or sphere-like granules with the size of 4-7mm of the microorganism immobilized carrier granules.
The preparation method of the shell-silicon dioxide microorganism immobilization carrier also comprises the following steps,
modification step: and (3) placing the microorganism fixed carrier particles obtained in the granulation step into a modification solution, reacting for 10-20 min, taking out the microorganism fixed carrier particles, washing, and naturally drying to obtain the modified microorganism fixed carrier particles.
In the preparation method of the shell-silica microorganism immobilization carrier, the modified solution is a citric acid solution, the citric acid solution contains 0.005-0.01g of citric acid per 100mL of water, and the reaction equation is as follows:
2C6H8O7+3CaCO3＝(C6H5O7)2Ca3+3CO2↑+3H2O。
in the preparation method of the shell-silicon dioxide microorganism immobilization carrier, 60-80 g of microorganism immobilization carrier particles are added into 100mL of citric acid solution for modification;
and soaking and washing the modified microorganism fixed carrier particles by using seawater, naturally drying, and then placing into a refrigerator at 4 ℃ for storage.
The preparation method of the shell-silicon dioxide microorganism immobilization carrier also comprises the following microorganism loading steps:
preparing a microorganism fixed carrier particle nutrient solution, transferring the microorganism to be loaded into the microorganism fixed carrier particle nutrient solution, and performing shake culture at 25-30 ℃ for 12-24 h to obtain the microorganism fixed carrier particle with fixed bacteria.
In the above method for preparing a shell-silica microorganism immobilization carrier, the method for preparing the microorganism immobilization carrier particle nutrient solution comprises the following steps: adding 50-70 g of microorganism immobilization carrier particles into every 100mL of liquid culture medium, sterilizing at 121 ℃ for 15-30 min, and cooling to room temperature.
In the above method for preparing a shell-silica microorganism immobilization carrier, the type of the microorganism to be loaded is bacillus or photosynthetic bacteria.
In the preparation method of the shell-silicon dioxide microorganism immobilization carrier, the preparation method of the microorganism to be loaded is to take 200mL of fermented bacterial liquid to be loaded, and the concentration of the bacteria is 3 multiplied by 1010And (5) centrifuging and concentrating the mixture at the speed of 3000r/min by using a CFU/mL, and removing supernatant fluid to obtain the microorganism to be loaded.
In the preparation method of the shell-silicon dioxide microorganism immobilization carrier, the microorganism immobilization carrier particles immobilized with bacteria are frozen and stored at the temperature of-20 ℃ after being frozen and dried.
By the technical scheme, the preparation method of the shell-silicon dioxide microorganism immobilization carrier and the preparation method thereof provided by the invention at least have the following advantages:
1) the raw materials of the microorganism fixing carrier are shell, silicon dioxide and cement, so that the microorganism fixing carrier is non-toxic to microorganisms and low in cost;
2) the microorganism immobilization carrier can improve the mechanical strength of the immobilization carrier by increasing the cement content;
3) the microorganism fixed by the microorganism fixed carrier has high activity.
The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical solutions of the present invention more clearly understood and to implement the technical solutions according to the contents of the description, the following detailed description of the preferred embodiments of the present invention is given with reference to the accompanying drawings.
Drawings
FIG. 1 is a linear plot of the effect of different cement additions on immobilization carrier fracture rate as provided by an example of the present invention;
FIG. 2 is a flowchart of a method for preparing a microorganism-immobilized carrier according to an embodiment of the present invention.
Detailed Description
To further illustrate the technical means and effects of the present invention for achieving the predetermined objects, the following detailed description will be given to the specific implementation, structure, characteristics and effects of the method for preparing the shell-silica microorganism immobilization carrier according to the present invention with reference to the preferred embodiments. In the following description, different "one embodiment" or "an embodiment" refers to not necessarily the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
Examples of microorganism-immobilizing carriers
The embodiment discloses a shell-silicon dioxide microorganism immobilization carrier, which comprises: the shell-silicon dioxide microorganism fixing carrier is particles with the diameter of 4-7mm, comprises shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, and is granulated by taking cement with the mass fraction of 5-10% as an adhesive.
The shell-silicon dioxide microorganism fixing carrier is used for fixing microorganisms, such as photosynthetic bacteria such as rhodopseudomonas palustris and beneficial microorganism flora such as bacillus, wherein the photosynthetic bacteria decompose and utilize organic matters in a water body in an environment with light and oxygen deficiency, the oxygen consumption of the organic matters is reduced, the bacillus can decompose harmful substances such as the organic matters and nitrite in the water body, the eutrophication level of the water body is reduced, the micro-ecological imbalance is adjusted, meanwhile, the balance of flora in intestinal tracts of stichopus japonicus can be maintained, the propagation of harmful bacteria is inhibited, and the immunity of the stichopus japonicus is enhanced.
The shell-silicon dioxide microorganism immobilization carrier is prepared into particles with diameters of 4-7mm, so that the microorganism immobilization carrier has high mechanical strength, and a large attachment area is provided for microorganisms. The diameter of the microorganism-immobilized carrier particles is of such a size that it does not affect the implementation of the present solution. The microorganism-immobilized carrier particles may be spherical, cubic, rugby-ball or other irregular shapes.
The proportion of the content of the shell powder to the content of the silicon dioxide in the microorganism immobilization carrier influences the number of microorganisms immobilized by the carrier. The cement content in the microorganism immobilization carrier affects the mechanical strength of the immobilization carrier itself.
In order to make the mechanical hardness of the microorganism immobilization carrier meet the requirement and detect the mechanical hardness under the influence of the addition amount of cement, the embodiment provides a mechanical hardness detection method. The preparation method of the shell-silica microorganism immobilization carrier is shown in example 1, wherein the addition amounts of cement are 3, 4, 5, 6, 7, 8, 9 and 10g, respectively. The detection method of the mechanical hardness comprises the following steps:
randomly taking 50 fixed carriers, and fixedly placing the fixed carriers in a net bag at the bottom of a culture pond;
the fixed carrier is impacted by flowing water with the flow velocity of 0.67m/s, and the continuous impact is carried out for 7 days;
the mechanical strength of the cement is measured by counting the breakage rate, and the result is shown in figure 1, wherein when the addition amount of the cement is 5% -10%, the better mechanical strength can be maintained.
The microorganism immobilization carrier provided by the embodiment can be applied to the apostichopus japonicus breeding benthic environment modifier.
Preparation method example 1
As shown in fig. 2, this embodiment discloses a method for preparing a shell-silica microorganism immobilization carrier, comprising the following steps: cleaning scallop shells, drying in an oven, and grinding into scallop shell powder of 40 meshes;
taking 80g of the scallop shell powder, and uniformly mixing with 20g of medium-fine silicon dioxide;
and (3) putting the uniformly mixed scallop shell-silicon dioxide into a granulator, adding 5g of cement powder, and spraying a proper amount of water to prepare scallop shell-silicon dioxide carrier particles with the particle size of 5 mm.
Mixing raw materials: mixing shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, wherein the influence on the number of fixed microorganisms is not obvious in the ratio range, and the magnitude order is not changed;
adding cement with the mass fraction of 5-10% as an adhesive, and uniformly mixing to form a mixed raw material;
and (3) granulation: and uniformly mixing the mixed raw materials and water according to the proportion of 10:1, and granulating to obtain the microorganism immobilized carrier particles.
In specific implementation, the scallop shells are ground into scallop shell powder of 60-40 meshes; the silica particles are medium-fine-grained silica particles of 60 to 40 mesh. In the embodiment, the scallop shells are selected as one of the raw materials of the fixed carrier because the porosity of the scallop shells is rich, and the scallop culture quantity is large and easy to obtain. Of course, as a variant embodiment, other seashells may be used as the material of the immobilization carrier instead of scallop shells.
In the preparation of the microorganism immobilization carrier provided in this embodiment, the citric acid modified scallop shell-silica microorganism carrier is a microorganism immobilization carrier material obtained by cleaning scallop shells, pulverizing the cleaned scallop shells into powder, mixing the powder with medium-fine silica particles, granulating with cement No. 300 as an adhesive, modifying with citric acid to increase particle pores, soaking in seawater, and washing. The invention also provides a preparation method of the citric acid modified scallop shell-silicon dioxide microbial carrier, and the use method of the immobilized particles is that the immobilized particles are used as a carrier material for immobilized microorganisms.
Preparation method example 2
In order to further increase the attachment area of microorganisms, this example modified the microorganism-immobilized carrier particles prepared in preparation method example 1. The modification step specifically comprises the following steps:
and (2) placing the microorganism fixed carrier particles obtained in the granulation step into a modified solution, reacting for 10min-20min, specifically, adding 70g of the microorganism fixed carrier particles into 100mL of the modified solution, and fully mixing the microorganism fixed carrier and the modified solution in a stirring or vibrating manner, so that holes formed on the microorganism fixed carrier are more uniform. And then placing the mixed solution of the microorganism fixed carrier and the modified solution in an oscillator with the rotating speed of 75-100r/min for reaction, taking out the microorganism fixed carrier particles, washing and naturally drying to obtain the modified microorganism fixed carrier particles.
In the embodiment, the microorganism immobilization carrier particles are placed in seawater to be soaked and washed for 1h, so that the seawater environment can be simulated, other impurities can be washed away by using seawater, and a more appropriate microenvironment is provided for bacteria. Taking out after soaking, and naturally drying for use.
The modified solution is citric acid solution, and the citric acid solution contains 0.005-0.01g of citric acid per 100mL of water. The citric acid content is within the threshold range, the higher the content is, the stronger the acidity is, the more violent the reaction with the shell is, the less the shell content is remained in the shell-dioxide mixed material, and the number of the microorganisms capable of being loaded is relatively reduced.
And (3) storing the modified microorganism immobilization carrier particles in a refrigerator at 4 ℃.
Preparation method example 3
In order to realize the improvement of the stichopus japonicus culture benthic environment by the microorganism immobilization carrier, the microorganism immobilization carrier is required to be loaded with microorganisms, and specifically, the microorganism loading step comprises the following steps: fermenting the microorganism to be loaded to the required concentration, concentrating the microorganism fermentation liquor, preparing the fixed carrier particle nutrient solution, transferring the concentrated microorganism to the microorganism fixed carrier, and obtaining the carrier particle with the fixed bacteria.
In the embodiment, the specific operation steps for preparing the microorganism immobilization carrier particle nutrient solution are that 40g of modified scallop shell-silicon dioxide carrier particles are put into 60mL of LB liquid culture medium, sterilized at 121 ℃ for 20min, and naturally cooled.
In this embodiment, the type of the microorganism to be loaded is bacillus. The preparation method of the microorganism to be loaded comprises taking the fermented microorganism to be loaded with a concentration of 3 × 1010200mL of CFU/mL bacterial liquid and 3000 r-And (5) centrifuging and concentrating for min, and removing supernatant to obtain the microorganism (bacillus concentrating) to be loaded. Transferring the concentrated bacillus into a cooled scallop shell-silicon dioxide nutrient solution, and performing shake culture at 25 ℃ for 24 hours; freeze-drying the microorganism immobilized carrier particles with the bacteria immobilized thereon, and refrigerating and storing at-20 deg.C
The preparation method of the microorganism immobilized carrier particle nutrient solution comprises the following steps: adding 50-70 g of microorganism immobilization carrier particles into every 100mL of liquid culture medium, sterilizing at 121 ℃ for 15-30 min, and cooling to room temperature.
Preparation method example 4
This example differs from preparation method example 3 in that the microorganism to be supported is a mixed bacterium of Rhodospirillum Molisch and Bacillus subtilis, which is subjected to amplification fermentation to 3X 10 with LB liquid medium10CFU/mL of bacterial liquid.
In this example, the step of concentrating the microorganism to be loaded is as follows: and (4) taking 200mL of fermented bacterial liquid, carrying out centrifugal concentration at 3000r/min, and removing supernatant to obtain the concentrated microbial liquid to be loaded.
In the embodiment, the specific operation steps for preparing the microorganism immobilization carrier particle nutrient solution are that 40g of modified scallop shell-silicon dioxide carrier particles are put into 60mL of LB liquid culture medium, sterilized at 121 ℃ for 20min, and naturally cooled.
In the example, the microorganism immobilization method comprises the steps of transferring the bacillus concentration into a cooled scallop shell-silicon dioxide nutrient solution, and performing shake culture at 25 ℃ for 24 hours; freeze-drying the microorganism immobilized carrier particles with the fixed bacteria, and refrigerating and storing at-20 ℃.
Method of use embodiment
The embodiment provides a use method of a citric acid modified scallop shell-silicon dioxide carrier immobilized microorganism, and particularly the use method comprises the following step of applying a mixture of Rhodospirillum Rhodospirillum Molisch and Bacillus subtilis immobilized by a citric acid modified scallop shell-silicon dioxide material to sediment bottom mud in a stichopus japonicus culture pond for an experiment with a period of 2 weeks.
Selecting three culture ponds (80cm multiplied by 40cm) with the same model number, namely No. 1, No. 2 and No. 3;
paving sediment in ocean with the thickness of 5cm in a culture pond, adding seawater, aerating, acclimatizing for 1d, and discharging the seawater;
no matter is added on the upper part of sediment bottom mud of the No. 1 culture pond, and only 5g of stichopus japonicus culture feed is added as a control; adding free Rhodospirillum Rhodospirillum Molisch and Bacillus subtilis mixed bacteria into a No. 2 culture pond, wherein the mixed bacteria are concentrated mixed bacteria obtained by concentrating 16L mixed bacteria; 3200g of shell-silicon dioxide carriers with fixed bacteria are paved in a No. 3 culture pond with the thickness of 3cm, and are placed on the upper part of the bottom sediment to be added with 5g of stichopus japonicus culture feed;
adding seawater, aerating, acclimating and adapting for 2 d;
5 young stichopus japonicus seedlings with the mass of 15-20g are placed in the culture pond for culture for 2 weeks.
Ammonia nitrogen, nitrate nitrogen and BOD of culture pond sediments in the experimental process5The parameters are used as indexes for measuring the action of the immobilization carrier. Wherein ammonia nitrogen and nitrate nitrogen are determined by potassium chloride solution extraction-spectrophotometry (HJ 634-5The content of BOD in the water body at the water-sediment interface5Substitution (HJ05-2009), the formula for the daily growth rate (SGR) of stichopus japonicus is as follows:
SGR＝[ln(Wt/W0)]/T×100％]
wherein WtRepresents the mass (g) of the stichopus japonicus selenka when weighed; w0Represents the initial mass (g) of the stichopus japonicus selenka.
TABLE 1 indexes of relevant parameters of sediment and Stichopus japonicus
| Group of | Ammonia nitrogen (mg/L) | Nitrate nitrogen (mg/L) | BOD5(mg/L) | Stichopus japonicus SGR (g) |
| No. 1 pool | 1.4a | 0.02a | 11a | 2.21a |
| No. 2 pool | 1.0a | 0.01b | 8b | 3.11b |
| No. 3 pool | 0.5b | 0.006c | 5c | 2.98b |
Remarking: lower case letters in the table represent significance of difference.
Ammonia nitrogen: only bacillus and photosynthetic bacteria are added, the degradation effect on ammonia nitrogen in sediments is not obvious, the ammonia nitrogen of the bacillus and photosynthetic bacteria fixed by citric acid modified scallop shell-silicon dioxide material is obviously reduced, and the microbial cell activity of the microbial fixed carrier is increased;
nitrate nitrogen and BOD5: free bacillus, light and bacteria, and can also reduce nitrate nitrogen and BOD significantly5But adding citric acid modified scallop shell-silicon dioxide material fixed bacillus and photosynthetic bacteria,nitrate nitrogen and BOD in the sediment5The content reduction is more obvious;
daily growth rate (SGR) of stichopus japonicus: free bacillus, light and bacteria, and bacillus and photosynthetic bacteria fixed by citric acid modified scallop shell-silicon dioxide material can obviously improve the SGR of stichopus japonicus.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and any simple modification, equivalent change and modification made to the above embodiment according to the technical spirit of the present invention are still within the scope of the technical solution of the present invention.
Claims (8)
1. A shell-silica microorganism immobilization carrier, comprising:
the shell-silicon dioxide microorganism fixing carrier is particles with the size of 4-7mm, the porosity of the fixing carrier is 40% -55%, the shell-silicon dioxide microorganism fixing carrier comprises shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, and cement with the mass fraction of 5-10% is used as an adhesive for granulation;
placing the microorganism fixed carrier particles obtained after granulation into a modification solution, reacting for 10-20 min, taking out the microorganism fixed carrier particles, washing, and naturally drying to obtain modified microorganism fixed carrier particles; the modified solution is citric acid solution, the citric acid solution contains 0.005-0.01g of citric acid per 100mL of water, and the reaction equation is as follows: 2C6H8O7+3CaCO3＝(C6H5O7)2Ca3+3CO2↑+3H2O; 60-80 g of microorganism immobilization carrier particles are added into 100mL of citric acid solution for modification.
2. A preparation method of a shell-silicon dioxide microorganism immobilization carrier is characterized by comprising the following steps,
mixing raw materials: mixing shell powder and silicon dioxide particles in a mass ratio of 4-2: 1, adding cement in a mass fraction of 5-10% as an adhesive, and uniformly mixing to form a mixed raw material;
and (3) granulation: uniformly mixing the mixed raw materials and water according to the ratio of 10-8: 1, and granulating to form microorganism immobilized carrier particles;
modification step: placing the microorganism fixed carrier particles obtained in the granulation step into a modification solution, reacting for 10-20 min, taking out the microorganism fixed carrier particles, washing, and naturally drying to obtain modified microorganism fixed carrier particles;
the modified solution is citric acid solution, the citric acid solution contains 0.005-0.01g of citric acid per 100mL of water, and the reaction equation is as follows:
2C6H8O7+3CaCO3＝(C6H5O7)2Ca3+3CO2↑+3H2O；
60-80 g of microorganism immobilization carrier particles are added into 100mL of citric acid solution for modification.
3. The method for preparing a shell-silica microorganism-immobilized carrier according to claim 2, wherein the silica microorganism-immobilized carrier is a silica-immobilized carrier,
the shell powder is obtained by grinding scallop shell into 60-40 mesh powder;
the silica particles are medium-fine-grained silica particles of 60-40 meshes;
in the granulating step, a disk granulator is used for granulating to form spherical or sphere-like granules with the size of 4-7mm of the microorganism immobilized carrier granules.
4. The method for preparing a shell-silica microorganism-immobilized carrier according to claim 2, wherein the silica microorganism-immobilized carrier is a silica-immobilized carrier,
and soaking and washing the modified microorganism fixed carrier particles by using seawater, naturally drying, and then placing into a refrigerator at 4 ℃ for storage.
5. The method for preparing a shell-silica microorganism-immobilized carrier according to any one of claims 2 to 4, further comprising a microorganism-loading step of:
preparing a microorganism fixed carrier particle nutrient solution, transferring the microorganism to be loaded into the microorganism fixed carrier particle nutrient solution, and performing shake culture at 25-30 ℃ for 12-24 h to obtain the microorganism fixed carrier particle with fixed bacteria.
6. The method for preparing a shell-silica microorganism-immobilized carrier according to claim 5, wherein the silica microorganism-immobilized carrier is a silica-immobilized carrier,
the preparation method of the microorganism immobilized carrier particle nutrient solution comprises the following steps: adding 50-70 g of microorganism immobilization carrier particles into every 100mL of liquid culture medium, sterilizing at 121 ℃ for 15-30 min, and cooling to room temperature.
7. The method for preparing a shell-silica microorganism-immobilized carrier according to claim 5, wherein the silica microorganism-immobilized carrier is a silica-immobilized carrier,
the type of the microorganism to be loaded is bacillus or photosynthetic bacteria.
8. The method for preparing a shell-silica microorganism-immobilized carrier according to claim 5, wherein the silica microorganism-immobilized carrier is a silica-immobilized carrier,
the preparation method of the microorganism to be loaded comprises the steps of taking 200mL of fermented bacterial liquid to be loaded, wherein the bacterial concentration is 3 multiplied by 1010Centrifuging and concentrating the CFU/mL solution at 3000r/min, and removing supernatant to obtain the microorganism to be loaded;
freeze-drying the microorganism immobilized carrier particles with the fixed bacteria, and refrigerating and storing at-20 ℃.
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| CN106135080A (en) * | 2016-06-26 | 2016-11-23 | 仇颖莹 | A kind of preparation method of shell powder artificial reef material |
| CN106745806A (en) * | 2016-12-20 | 2017-05-31 | 大连赛姆生物工程技术有限公司 | A kind of apostichopus japonicus culture microbial water-purifying agent and preparation method thereof |
| CN108584974B (en) * | 2018-08-01 | 2021-09-24 | 中科亿诚(珠海)环保技术有限公司 | Artificial zeolite using shell and construction waste as raw material and its preparation method |
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