CN114917259B - Two-dimensional hydrosilylene-microorganism composite material and preparation method and application thereof - Google Patents
Two-dimensional hydrosilylene-microorganism composite material and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a two-dimensional hydrosilylene-microorganism composite material, which comprises the following components: a microbial carrier and two-dimensional hydrogen bonding silylene nano-sheets loaded on the microbial carrier, wherein the total amount of the two-dimensional hydrogen bonding silylene nano-sheets loaded on each microbial carrier is 10 –6 ‑10 –11 Mu g. The invention also provides a preparation method of the two-dimensional hydrosilylene-microorganism composite material. The invention also provides application of the two-dimensional hydrosilylene-microorganism composite material as an intestinal delivery carrier and a free radical scavenger in preparing medicines for treating inflammatory bowel diseases. The two-dimensional hydrosilylene-microorganism composite material provided by the invention is used in stomachThe microbial agent has excellent stability under the acidic condition, can be effectively coated on the surface of the microorganism for a long time, protects the microorganism from being attacked by gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganism in stomach.
Description
Technical Field
The invention belongs to the technical field of biological nano materials, and particularly relates to a two-dimensional hydrosilylene-microorganism composite material with intestinal microenvironment response characteristics, a preparation method thereof and related applications in the aspects of probiotic intestinal delivery and inflammatory bowel disease treatment.
Background
The human intestinal flora, which is composed of numerous bacteria and other microorganisms, has very complex physiological functions and is closely related to the occurrence and development of a variety of diseases (Science 2018,362,776-780). How to regulate the intestinal flora in a safe and effective way and thus achieve a treatment of diseases is one of the most interesting problems. Probiotic therapy is a research hotspot in the field in recent years. Compared with the traditional treatment method, the probiotic therapy has the advantages of high safety, difficult generation of drug resistance and the like, can assist the reconstruction of intestinal flora steady state, and can play a role in treating various diseases. Among them, there has been a great deal of attention to the treatment of enteritis (Nat. Rev. Gastroenterol. Hepatol.2019,16, 605-616). Probiotic therapy may not only have a beneficial effect on the intestinal epithelium by direct or indirect means, but may also further reproduce in the intestine to improve intestinal flora homeostasis, thus acting as a long-term treatment. However, probiotics still face a lot of difficulties in practical use, among which the most important point is that probiotics are easy to lose activity under the acidic condition of stomach and the impact of enzyme, difficult to reach intestinal tract smoothly for colonization, and exert therapeutic effect.
Therefore, development of a high-efficiency microbial intestinal delivery strategy is needed, which has an effective protection effect on microorganisms in the extreme environment of the stomach, promotes intestinal colonization of probiotics and improves the enteritis treatment effect.
Disclosure of Invention
Aiming at the limitations of the prior art, the invention aims to provide a two-dimensional hydrosilylene-microorganism composite material with intestinal microenvironment response characteristic, a preparation method thereof and related application in the aspects of probiotic intestinal delivery and inflammatory bowel disease treatment, and solves the problems of low stomach survival rate, insufficient intestinal implantation efficiency and limited treatment effect of microorganisms.
In a first aspect, the present invention provides a two-dimensional hydrosilylene-microorganism composite material comprising: a microbial carrier and two-dimensional hydrogen bonding silylene nano-sheets loaded on the microbial carrier, wherein the total amount of the two-dimensional hydrogen bonding silylene nano-sheets loaded on each microbial carrier is 10 –6 -10 –11 μg。
Preferably, the microbial carrier is a fungus or a bacterium, preferably a probiotic, comprising one or more of yeast, trichoderma reesei, escherichia coli, bifidobacterium, lactobacillus, bacillus subtilis, staphylococcus aureus and bacillus proteus, preferably escherichia coli Nissel 1917 subtype.
Preferably, the two-dimensional hydrogen bonding silylene nano-sheet is of a lamellar structure, the transverse dimension is preferably 300-500 nm, and the thickness is preferably 0.5-10nm.
Preferably, the preparation method further comprises a macromolecule surface modifier for promoting the two-dimensional hydrogen bonding of the silylene nano-sheet to be loaded.
Preferably, the macromolecule surface modifier is a positively charged amphiphilic macromolecule, preferably vitamin E-polyethylene glycol-polyethyleneimine.
Preferably, in the vitamin E-polyethylene glycol-polyethyleneimine, the molecular weight of polyethylene glycol is 500-10000, preferably 2000; the molecular weight of the polyethyleneimine is 400-10000, preferably 600.
In a second aspect, the present invention also provides a method for preparing the two-dimensional hydrosilylene-microorganism composite material as described above, comprising the steps of:
step (1), providing two-dimensional hydrogen bonding silylene nano-sheets;
step (2), mixing the two-dimensional hydrogen bonding silylene nano-sheets and a high molecular modifier in a solvent, drying the solvent by using inert gas, dispersing the product in water, and centrifugally cleaning;
and (3) incubating the product obtained in the step (2) with a microbial carrier at a low temperature, centrifuging and washing to obtain the two-dimensional hydrosilylene-microbial composite material.
Preferably, in the step (1), the preparation method of the two-dimensional hydrogen bonding silylene nanomaterial comprises the following steps: will CaSi 2 Immersing the powder into concentrated hydrochloric acid solution, stirring and mixing at the temperature of minus 30 ℃ to minus 10 ℃, centrifuging, removing supernatant, and washing the centrifuged product to obtain the two-dimensional hydrogen bonding silylene nanomaterial.
Preferably, the CaSi 2 The grain diameter of the powder is 5-10 mu m, and the CaSi 2 The concentration of (2) is 5-20mg/mL; stirring and mixing at the temperature of minus 30 ℃ to minus 10 ℃ to form magnetic stirring, wherein the magnetic rotation speed of the magnetic stirring is 500-1000 revolutions per molecule, the time of the magnetic stirring is 3-10 days, and the magnetic stirring and mixing are carried out under the protection of inert gas, and the inert gas is preferably argon; the rotational speed of the centrifugal treatment is 13000 rpm-20000 rpm, and the time is 15-20 minutes; the solution adopted in the washing is absolute ethyl alcohol or deionized water, and the washing times are 3-4 times; the method also comprises the step of ultrasonic crushing the material after washing the centrifugal product in absolute ethyl alcohol, wherein the ultrasonic crushing power is 500-800W, and the ultrasonic crushing time is 8-15h.
Preferably, in the step (2), the solvent is selected from one or more of water, ethanol, acetone and chloroform, preferably absolute ethanol, and the inert gas is nitrogen; the macromolecule modifier is vitamin E-polyethylene glycol-polyethyleneimine, and the mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen bonding silylene nano-sheet is preferably (1-20): 1.
preferably, in step (3)The concentration of the microbial carrier is 10 6 -10 12 The concentration of the two-dimensional hydrogen bonding silylene nano-sheet modified by the macromolecule modifier is 5-200 mug/mL, and the co-incubation temperature is 0-10 ℃, preferably 4 ℃.
In a third aspect, the invention also provides the use of a two-dimensional hydrosilylene-microorganism composite material as described above as an intestinal delivery vehicle and a free radical scavenger for the preparation of a medicament for the treatment of inflammatory bowel disease.
According to the invention, the two-dimensional hydrogen-bonded silylene nano-sheet is loaded on a probiotic microorganism carrier, and the probiotic microorganism carrier wrapped by the two-dimensional hydrogen-bonded silylene nano-sheet can protect probiotic microorganisms from being supplied with gastric acid and enzyme, so that the survival time and the survival rate of the probiotic microorganisms in the stomach are improved. The probiotic microorganisms wrapped by the two-dimensional hydrogen bonding silylene nanosheets have good microenvironment response characteristics, and the two-dimensional hydrogen silylene-microorganism composite material can rapidly degrade and release the internally wrapped probiotic microorganisms after reaching the intestinal tract, so that the metabolism and proliferation of the microorganisms are not influenced, and the intestinal tract field planting efficiency of the microorganisms is greatly improved. In the invention, the two-dimensional hydrosilylene-microorganism composite material also comprises a high molecular surface modifier for promoting the two-dimensional hydrogen bonding silylene nano-sheet to be loaded, so that the loading rate of the Gao Erwei hydrogen bonding silylene nano-sheet loaded on the probiotic microorganism carrier is further improved, and simultaneously, vitamin E-polyethylene glycol-polyethyleneimine is preferably used as the high molecular surface modifier, so that the microbial activity is not reduced in the modification process.
The invention has the beneficial effects that:
the two-dimensional hydrosilylene-microorganism composite material and the preparation method thereof provided by the invention have the advantages of mild preparation conditions, simplicity and convenience in operation, and no decrease in microorganism activity in the modification process; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the stomach acidic condition, can be effectively wrapped on the surface of microorganisms for a long time, protects the microorganisms from being attacked by gastric acid and enzymes, and greatly improves the survival time and survival rate of the microorganisms in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good microenvironment response characteristics, can rapidly degrade and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
Drawings
FIG. 1 shows a flow chart for preparing a two-dimensional hydrosilylene-microorganism composite material according to one embodiment of the present invention.
Fig. 2 shows the morphology of two-dimensional hydrosilylene nanoplatelets (SiH) prepared according to example 1 of the present invention and the modified hydrosilylene nanoplatelets with vitamin E-polyethylene glycol-polyethylenimine (sih@tpgs-PEI) under a transmission electron microscope.
FIG. 3 shows a scanning electron microscope image of a two-dimensional hydrosilylene-microorganism composite material prepared according to example 1 of the present invention.
Fig. 4 shows a graph of the uv-vis absorption spectrum in artificial gastric juice of the vitamin E-polyethylene glycol-polyethylenimine modified hydrosilylene nanoplatelets according to example 2 of the present invention over time.
Fig. 5 shows a graph of the uv-vis absorption spectrum of the vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nanoplatelets in artificial intestinal juice according to example 2 of the present invention over time.
Figure 6 shows bacterial survival of two-dimensional hydrosilylene-microorganism composite material in artificial gastric juice according to example 3 of the present invention.
Fig. 7 shows a graph of evaluation of survival time of two-dimensional hydrosilylene-microorganism composite material in artificial gastric juice according to example 4 of the present invention.
Detailed Description
The invention will be further described with reference to the accompanying drawings and the following embodiments, it being understood that the drawings and the following embodiments are only for illustrating the invention, not for limiting the invention.
According to a first aspect of the present invention there is provided a two-dimensional hydrosilylene-microorganism composite material comprising: the microbial carrier and the two-dimensional hydrogen bonding silylene nano sheet loaded on the microbial carrier optionally further comprise a macromolecule surface modifier for promoting the loading of the two-dimensional hydrogen bonding silylene nano sheet.
According to the invention, the microbial carrier is a fungus or a bacterium, and in the embodiment of the invention is a probiotic, including one or more of yeast, trichoderma reesei, escherichia coli, bifidobacterium, lactobacillus, bacillus subtilis, staphylococcus aureus and bacillus proteus, and in the embodiment of the invention is an escherichia coli Nissel 1917 subtype.
Probiotics are fungi or bacteria beneficial to human bodies or animals, and are active microorganisms beneficial to a host by being planted in the human body to change the flora composition of a certain part of the host. By regulating the immune function of host mucous membrane and system or regulating the balance of flora in intestinal tract, the effect of promoting nutrient absorption and maintaining intestinal health is achieved, so that single microorganism or mixed microorganism with definite composition beneficial to health is produced.
However, probiotics still face a lot of difficulties in practical use, among which the most important point is that probiotics are easy to lose activity under the acidic condition of stomach and the impact of enzyme, difficult to reach intestinal tract smoothly for colonization, and exert therapeutic effect. Therefore, the invention also loads the two-dimensional hydrogen bonding silylene nano-sheet on the probiotic microorganism carrier on the basis of the original probiotic microorganism carrier. The probiotic microorganism carrier wrapped by the two-dimensional hydrogen bonding silylene nanosheets can protect probiotic microorganisms from being supplied by gastric acid and enzymes, and can improve the survival time and survival rate of the probiotic microorganisms in the stomach. The probiotic microorganisms wrapped by the two-dimensional hydrogen bonding silylene nanosheets have good microenvironment response characteristics, and the two-dimensional hydrogen silylene-microorganism composite material can rapidly degrade and release the internally wrapped probiotic microorganisms after reaching the intestinal tract, so that the metabolism and proliferation of the microorganisms are not influenced, and the intestinal tract field planting efficiency of the microorganisms is greatly improved.
According to the invention, the two-dimensional hydrogen bonding silylene nano-sheet is of a lamellar structure, the transverse dimension is preferably 300-500 nm, and the thickness is preferably 0.5-10nm.
In order to promote the loading of the two-dimensional hydrogen bonding silylene nano sheet, the invention also optionally comprises a macromolecule surface modifier for modifying the surface of the two-dimensional hydrogen bonding silylene nano sheet. The macromolecule surface modifier is positively charged amphiphilic macromolecule, and vitamin E-polyethylene glycol-polyethyleneimine is adopted in the embodiment of the invention.
The two-dimensional hydrogen-silylene-microorganism composite material also comprises a high molecular surface modifier for modifying the surface of the two-dimensional hydrogen-bonding silylene nano sheet so as to promote the two-dimensional hydrogen-bonding silylene nano sheet to be loaded on a probiotic microorganism carrier, so that the loading rate of the Gao Erwei hydrogen-bonding silylene nano sheet loaded on the probiotic microorganism carrier is further improved, and simultaneously, vitamin E-polyethylene glycol-polyethyleneimine is preferably used as the high molecular surface modifier, so that the microbial activity is not reduced in the modification process.
In a preferred embodiment, in the vitamin E-polyethylene glycol-polyethyleneimine, the polyethylene glycol has a molecular weight of 500 to 10000, such as 2000; the molecular weight of the polyethyleneimine is 400-10000, for example 600.
The invention also provides a preparation method of the two-dimensional hydrosilylene-microorganism composite material, which is mainly divided into three parts, including preparation of a hydrosilylene nano sheet, modification of the hydrosilylene nano sheet and assembly of the modified hydrosilylene nano sheet and microorganisms. The preparation method comprises the following steps:
step 1, preparing a two-dimensional hydrosilylene nano sheet: will CaSi 2 Immersing the powder into concentrated hydrochloric acid solution, magnetically stirring and mixing at the temperature of minus 30 ℃ to minus 10 ℃ under the protection of inert gas, centrifuging, removing supernatant, and then washing the centrifuged product to obtain the two-dimensional hydrogen bonding silylene nanomaterial.
In this step 1, the CaSi 2 The particle size of the powder is 5-10 μm, and the concentration is 5-20mg/mL, for example 10mg/mL; the magnetic stirring magnet has a rotation speed of 500-1000 rpm for 3-10 days, such as 7 days; the inert gas is preferably argon; the rotational speed of the centrifugal treatment is 13000 rpm-20000 rpm, and the time is 15-20 minutes; the solution adopted for washing is absolute ethyl alcohol or deionized water, andthe washing times are 3-4 times.
In the preparation process of the two-dimensional hydrosilylene nano sheet, the method further comprises the step of ultrasonic crushing the material after washing the centrifugal product in absolute ethyl alcohol, wherein the ultrasonic crushing power is 500-800W, and the ultrasonic crushing time is 8-15h.
Step 2, modification of two-dimensional hydrosilylene nano sheets: mixing the two-dimensional hydrogen bonding silylene nano-sheet and a high molecular modifier in a solvent, drying the solvent by using inert gas, dispersing the product in water, and centrifugally cleaning.
In the step 2, the solvent is selected from one or more of water, ethanol, acetone and chloroform, preferably absolute ethanol, and the inert gas is nitrogen; in the embodiment of the invention, the macromolecule modifier is vitamin E-polyethylene glycol-polyethyleneimine, and the mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen bonding silylene nano-sheet is preferably (1-20): 1, for example 10:1.
step 3, assembling the modified two-dimensional hydrosilylene nano-sheet and a probiotic microorganism carrier: and (3) incubating the product obtained in the step (2) with a probiotic microorganism carrier at a low temperature of, for example, 4 ℃, centrifuging and washing to obtain the two-dimensional hydrosilylene-microorganism composite material.
In this step 3, the concentration of the microorganism carrier is 10 6 -10 12 CFU/mL, e.g. 10 8 CFU/mL; the concentration of the two-dimensional hydrogen bonding silylene nano-sheet modified by the macromolecule modifier is 5-200 mug/mL, for example 30 mug/mL.
The invention also provides application of the two-dimensional hydrosilylene-microorganism composite material as an intestinal delivery carrier and a free radical scavenger in preparing medicines for treating inflammatory bowel diseases. According to the invention, the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the stomach acidic condition, can be effectively wrapped on the surface of microorganisms for a long time, protects the microorganisms from being attacked by gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganisms in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good microenvironment response characteristics, can rapidly degrade and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
The present invention will be further illustrated by the following examples. It is also to be understood that the following examples are given solely for the purpose of illustration and are not to be construed as limitations upon the scope of the invention, as many insubstantial modifications and variations are within the scope of the invention as would be apparent to those skilled in the art in light of the foregoing disclosure. The specific process parameters and the like described below are also merely examples of suitable ranges, i.e., one skilled in the art can make a suitable selection from the description herein without necessarily limiting the specific values described below.
Example 1:
preparation of two-dimensional hydrosilylene-microorganism composite material: the preparation steps mainly comprise three parts, namely the preparation of the hydrosilylene nano-sheet, the modification of the hydrosilylene nano-sheet and the assembly of the modified hydrosilylene nano-sheet and probiotic microorganisms. The specific preparation flow is shown in fig. 1, and the following three steps are respectively described:
preparation of a hydrosilylene nanosheet: 1g of calcium silicide is added into 100mL of pre-cooled concentrated hydrochloric acid and stirred vigorously for 7 days under the protection of argon at-20 ℃. After the reaction, the mixture was centrifuged at 12000rpm for 10min, and the precipitate was washed with acetone and absolute ethanol in this order and resuspended in absolute ethanol. And then carrying out ultrasonic treatment for 8 hours by an ice bath probe to obtain the hydrosilylene nano-sheet, wherein the transmission electron microscope result is shown in figure 2.
Modification of the hydrosilylene nanosheets: 1mg of the hydrosilylene nano-sheet and 10mg of vitamin E-polyethylene glycol-polyethyleneimine are dispersed in absolute ethyl alcohol together, and after being dried by nitrogen, the mixture is resuspended by ultrapure water, and the mixture is centrifugally washed for 2 times to obtain the vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nano-sheet, and the transmission electron microscope result is shown in figure 2.
Assembling the modified hydrosilylene nano sheet and probiotic microorganisms: and (3) re-suspending the modified hydrosilylene nanosheets in ultrapure water, and diluting the modified hydrosilylene nanosheets into the hydrosilylene suspension with different concentrations. The probiotic microorganisms were centrifuged at 7000rpm for 5min, washed twice and dispersed in ultrapure water. Mixing the hydrosilylene suspension with the probiotic microorganism suspension, incubating for 20-30min at 4 ℃, and centrifugally washing for 3 times to obtain the two-dimensional hydrosilylene-microorganism composite material. The scanning electron microscope results are shown in fig. 3.
Example 2:
the vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nano-sheet is dispersed in artificial gastric juice and artificial intestinal juice, and the stability and the responsiveness of the vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nano-sheet under different physiological environments are judged by monitoring the change of the ultraviolet-visible absorption spectrum. As can be seen from fig. 4 and 5, the material has good stability in artificial gastric juice (fig. 4), and the ultraviolet-visible absorption spectrum is hardly changed. Whereas in artificial intestinal juice (fig. 5), the uv-visible absorption value decreases rapidly, indicating that the material can degrade rapidly in artificial intestinal juice.
Example 3:
evaluation of bacterial viability of two-dimensional hydrosilylene-microorganism composite in gastric juice. 1mL of probiotics in the logarithmic growth phase is centrifuged and resuspended in 100 mu L of ultrapure water, and mixed with different concentrations of vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nanoplatelets in sterile water, and finally the content of the hydrosilylene is 0, 3, 15 and 30 mu g. And (5) after half an hour of co-incubation, centrifugally cleaning to obtain the two-dimensional hydrosilylene-probiotic composite material. Dispersing the obtained composite material in artificial gastric juice, incubating for 15min, and counting the survival rate by a flat plate coating method. As shown in fig. 6, the unmodified probiotics have no bacteria survival after the probiotics are reacted with gastric juice, and the hydrosilylene-probiotics composite material shows a good probiotic protection effect, so that the survival rate of the probiotics in gastric juice is greatly improved.
Example 4:
evaluation of the survival time of the two-dimensional hydrosilylene-microorganism composite in gastric juice. 1mL of probiotics in the logarithmic growth phase is centrifuged and then resuspended in 100 mu L of ultrapure water, and mixed with vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nanoplatelets in sterile water, and the final content of the hydrosilylene is 30 mu g. And (5) after half an hour of co-incubation, centrifugally cleaning to obtain the two-dimensional hydrosilylene-probiotic composite material. The obtained composite material is dispersed in 1mL of artificial gastric juice, the survival rate of the composite material is counted by a flat plate coating method after 15min, 30min and 60min of incubation, the counting result is shown in figure 7, and the survival rate of probiotics can be greatly improved within 1 hour.
Industrial applicability:
the two-dimensional hydrosilylene-microorganism composite material and the preparation method thereof provided by the invention have the advantages of mild preparation conditions, simplicity and convenience in operation, and no decrease in microorganism activity in the modification process; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the stomach acidic condition, can be effectively wrapped on the surface of microorganisms for a long time, protects the microorganisms from being attacked by gastric acid and enzymes, and greatly improves the survival time and survival rate of the microorganisms in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good microenvironment response characteristics, can rapidly degrade and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
Claims (7)
1. The preparation method of the two-dimensional hydrosilylene-microorganism composite material is characterized by comprising the following steps of:
step (1), providing two-dimensional hydrogen bonding silylene nano-sheets;
step (2), mixing the two-dimensional hydrogen bonding silylene nano-sheets and positively charged amphiphilic polymer modifier in a solvent, drying the solvent by using inert gas, dispersing the product in water, and centrifugally cleaning;
incubating the product obtained in the step (2) with a microbial carrier at a low temperature, centrifuging and washing to obtain the two-dimensional hydrosilylene-microbial composite material;
in the step (2), the positively charged amphiphilic polymer modifier is vitamin E-polyethylene glycol-polyethyleneimine, and the mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen bonding silylene nano-sheet is (1-20): 1, a step of;
in the step (3), the microbial carrier is one or more of saccharomycetes, bifidobacteria, lactobacillus and bacillus subtilis; the concentration of the microbial carrier is 10 6 -10 12 The concentration of the two-dimensional hydrogen bonding silylene nano-sheet modified by the positively charged amphiphilic polymer modifier is 5-200 mug/mL, and the co-incubation temperature is 0-10 ℃.
2. The method of claim 1, wherein in step (1), the method of preparing the two-dimensional hydrogen-bonded silylene nanoplatelets comprises the steps of: will CaSi 2 Immersing the powder into concentrated hydrochloric acid solution, stirring and mixing at-30 ℃ to-10 ℃, centrifuging, removing supernatant, and washing the centrifuged product to obtain the two-dimensional hydrogen bonding silylene nano-sheet.
3. The method of claim 2, wherein the CaSi is 2 The grain diameter of the powder is 5-10 mm, and the CaSi 2 The concentration of (2) is 5-20mg/mL; stirring and mixing at-30 ℃ to-10 ℃ to obtain magnetic stirring, wherein the magnetic rotation speed of the magnetic stirring is 500-1000 revolutions per molecule, the time of the magnetic stirring is 3-10 days, and the magnetic stirring and the mixing are carried out under the protection of inert gas; the rotational speed of the centrifugal treatment is 13000 rpm-20000 rpm, and the time is 15-20 minutes; the solution adopted in the washing is absolute ethyl alcohol or deionized water, and the washing times are 3-4 times; the method also comprises the step of ultrasonic crushing the material after washing the centrifugal product in absolute ethyl alcohol, wherein the ultrasonic crushing power is 500-800W, and the ultrasonic crushing time is 8-15h.
4. The method according to claim 1, wherein in the step (2), the solvent is one or more selected from the group consisting of water, ethanol, acetone and chloroform, and the inert gas is nitrogen.
5. The method according to claim 1, wherein in the vitamin E-polyethylene glycol-polyethyleneimine, the molecular weight of polyethylene glycol is 500 to 10000 and the molecular weight of polyethyleneimine is 400 to 10000.
6. A two-dimensional hydrosilylene-microorganism composite material obtained by the preparation method according to claim 1, comprising: the nano-chip comprises a microbial carrier and a two-dimensional hydrogen bonding silylene nano-chip which is loaded on the microbial carrier and is modified by positively charged amphiphilic polymer modifier.
7. The two-dimensional hydrosilylene-microorganism composite material according to claim 6, wherein the two-dimensional hydrogen bonding silylene nano-sheet is a layered lamellar structure, and has a transverse dimension of 300-500 nm and a thickness of 0.5-10nm.
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