CN110872386A - Method for preparing amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization - Google Patents
Method for preparing amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization Download PDFInfo
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Abstract
The invention discloses a method for preparing an amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization. The method comprises the steps of inoculating acetobacter xylinum seed liquid into a fermentation culture medium containing 1.0-2.0 g/L of sodium silicate, carrying out static culture, and repeatedly washing and freezing and thawing after the culture is finished to obtain the amorphous silicon dioxide/bacterial cellulose composite material. According to the invention, the amorphous silicon dioxide/bacterial cellulose composite material is prepared by a biological in-situ mineralization method, glucose is metabolized into acid substances by acetobacter xylinum in the fermentation process, and silicate is converted into amorphous silicon dioxide with low solubility by the acid substances along with the fermentation process by controlling the concentration of sodium silicate in a fermentation medium and is deposited on the generated bacterial cellulose, so that the mineralization of organic polymers and inorganic substances is realized, and the prepared amorphous silicon dioxide/bacterial cellulose composite material has good mechanical properties.
Description
Technical Field
The invention belongs to the technical field of biological materials, and relates to a method for preparing an amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization.
Background
The Bacterial Cellulose (BC) has many unique physical and chemical properties, such as high crystallinity, high water holding capacity, superfine three-dimensional nanofiber structure, high tensile strength, elastic modulus and the like, and simultaneously has good biocompatibility and degradability, thereby having wide application prospects in medicines, papermaking, biomedicine and foods.
The microorganism in-situ mineralization method for preparing the organic-inorganic composite tissue engineering material has remarkable advantages in material preparation, organically integrates two separate crystallization processes into one step, and has incomparable physical and chemical properties compared with the composite material prepared by chemical biomimetic mineralization, such as uniform assembly of organic and inorganic components, strong binding force, excellent interface characteristics and the like.
In the literature (Barud H S, et al. bacterial cellulose-silicon organic-inorganic fibers [ J ]. Journal of Sol-Gel Science and Technology,2008,46(3): 363-. However, in the method, the bacterial cellulose and the inorganic mineral are prepared by two separate steps of crystallization, so that the prepared composite material has weak binding force of each part, poor mechanical property and insufficient organic-inorganic interface characteristics.
Disclosure of Invention
The invention aims to provide a method for preparing an amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization.
The technical scheme for realizing the purpose of the invention is as follows:
the method for preparing the amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization comprises the following steps:
inoculating the acetobacter xylinum seed solution into a fermentation culture medium containing sodium silicate, statically culturing, and repeatedly washing and freezing and thawing after the culture is finished to obtain the amorphous silicon dioxide/bacterial cellulose composite material, wherein the concentration of the sodium silicate is 1.0-2.0 g/L.
Preferably, the inoculation amount of the acetobacter xylinum seed liquid is 8% -12%.
Preferably, the fermentation medium is an HS medium, the pH is 6.0, the static culture temperature is 30 ℃, and the culture time is 5-8 days.
Preferably, the repeated washing is to repeatedly wash the amorphous silicon dioxide/bacterial cellulose composite material with water for 8-10 times until the amorphous silicon dioxide/bacterial cellulose composite material is neutral.
Preferably, the repeated freeze thawing is to cool the composite material after water washing to-20 ℃, then rapidly melt the composite material at room temperature, and the number of times is 5-7.
Compared with the prior art, the invention has the following advantages:
according to the invention, the amorphous silicon dioxide/bacterial cellulose composite material is prepared by a biological in-situ mineralization method, glucose is metabolized into acid substances by acetobacter xylinum in the fermentation process, and silicate is converted into amorphous silicon dioxide with low solubility by the acid substances along with the fermentation process by controlling the concentration of sodium silicate in a fermentation medium and is deposited on the generated bacterial cellulose, so that the mineralization of organic polymers and inorganic substances is realized, and the prepared amorphous silicon dioxide/bacterial cellulose composite material has good mechanical properties.
Drawings
FIG. 1 is a flow chart of the preparation of amorphous silica/bacterial cellulose composite material.
FIG. 2 is a scanning electron microscope comparison image of the amorphous silica/bacterial cellulose composite material obtained in example 1 and bacterial cellulose.
FIG. 3 is an XPS comparison of the amorphous silica/bacterial cellulose composite obtained in example 1 with bacterial cellulose.
Fig. 4 is an XRD comparison graph of the amorphous silica/bacterial cellulose composite obtained in example 2 with bacterial cellulose.
Fig. 5 is a graph comparing the tensile strength of the amorphous silica/bacterial cellulose composite obtained in example 2 with that of bacterial cellulose.
Table 1 is a table of the effect of different concentrations of sodium silicate on the tensile strength of amorphous silica/bacterial cellulose composites.
Detailed Description
The invention is further elucidated with reference to the drawings and the embodiments.
In the following examples, the materials and reagents used are as follows;
the Acetobacter xylinum is Acetobacter gluconicum (Komagataibactericina) RSZ 01.
Seed culture medium: 2g of glucose, 0.3g of peptone, 0.225g of yeast extract powder, 0.1g of monopotassium phosphate, 0.04g of magnesium sulfate, 0.6g of ammonium sulfate, 0.04g of sodium carboxymethylcellulose, pH 6.0 and 100mL of water.
HS fermentation medium: 2g of glucose, 0.5g of peptone, 0.5g of yeast extract powder, 0.12g of citric acid, 0.27g of disodium hydrogen phosphate and 100mL of water.
Example 1
Preparing a seed culture medium, sterilizing at 121 ℃ for 20min, inoculating the activated slant strain into the sterilized and cooled seed culture medium, and culturing at 30 ℃ for 24h at 150r/min for later use. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 1g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min. The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 8%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 6 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 6 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
Example 2
Preparing a seed culture medium, sterilizing at 121 ℃ for 20min, inoculating the activated slant strain into the sterilized and cooled seed culture medium, and culturing at 30 ℃ for 24h at 150r/min for later use. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 2g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min. The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 8%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 5 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 7 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
Comparative example 1
Preparing a seed culture medium, sterilizing at 121 ℃ for 20min, inoculating the activated slant strain into the sterilized and cooled seed culture medium, and culturing at 30 ℃ for 24h at 150r/min for later use. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 0.1g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min. The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 10%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 8 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 5 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
Comparative example 2
Preparing seed culture medium, sterilizing at 121 deg.C for 20min, inoculating activated slant strain into sterilized and cooled seed culture medium at 30 deg.CAnd culturing for 24 hours under the condition of 150r/min for standby. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 0.2g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min. The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 10%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 8 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 5 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
Comparative example 3
Preparing a seed culture medium, sterilizing at 121 ℃ for 20min, inoculating the activated slant strain into the sterilized and cooled seed culture medium, and culturing at 30 ℃ for 24h at 150r/min for later use. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 0.5g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min. The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 10%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 7 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 6 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
Comparative example 4
Preparing a seed culture medium, sterilizing at 121 ℃ for 20min, inoculating the activated slant strain into the sterilized and cooled seed culture medium, and culturing at 30 ℃ for 24h at 150r/min for later use. Preparing HS fermentation medium, adding Na2SiO3·9H2O, to a concentration of 5.0g/L, 0.05mol/L hydrochloric acid solution was prepared, and sterilized at 121 ℃ for 20 min.The pH of the mixture of silicate and fermentation broth was adjusted to 6.0 with hydrochloric acid solution. Inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 10%, gently shaking to uniformly disperse the seed solution in the culture medium, and culturing in a constant-temperature incubator at 30 ℃ for 5 days to obtain the amorphous silicon dioxide/bacterial cellulose compound. And repeatedly washing the compound obtained by the culture with deionized water until the compound is neutral. And after the cleaning is finished, cooling the compound to-20 ℃, then rapidly melting the compound at room temperature, and repeatedly freezing and thawing for 6 times to finally obtain the amorphous silicon dioxide/bacterial cellulose biological composite material.
FIG. 1 is a flow chart of the preparation of amorphous silica/bacterial cellulose composite material.
As shown in FIG. 2, comparing SEM images of the bacterial cellulose (a) and the amorphous silica/bacterial cellulose composite material (b), it can be found that the amorphous silica/bacterial cellulose composite material has greatly changed morphology and enlarged size, which indicates that SiO is formed2Compounding is achieved with BC.
As shown in FIG. 3, the XPS comparison of the amorphous silica/bacterial cellulose biocomposite and the bacterial cellulose shows that BC/SiO2BC/SiO compared to the BC curve with Si2s at 152.7eV and Si2p at 102.4eV2Compared with BC curve, the peak signal of O1s is stronger, and the oxygen-carbon ratio is obviously increased, which indicates that SiO2Complexing with BC occurs.
As shown in fig. 4, the two curves are XRD comparison graphs of the amorphous silica/bacterial cellulose biocomposite and bacterial cellulose, respectively, and it can be found that both curves generate diffraction peaks around 14.7 °, 16.8 ° and 22.7 °, corresponding to (1-10), (110) and (020) crystal planes of BC, respectively. And BC/SiO2Compared with the appearance of SiO in the BC curve2Bao Peak, also evidence SiO2Complexing with BC occurs.
As shown in fig. 5, the two curves are respectively a comparison graph of the tensile strength of the amorphous silica/bacterial cellulose biocomposite and the tensile strength of the bacterial cellulose, and it can be found that the amorphous silica/bacterial cellulose biocomposite has stronger tensile strength and better toughness.
TABLE 1 different concentrations of sodium silicate vs. BC/SiO2Effect of composite tensile Strength
| Sample (I) | Tensile Strength (MPa) | Elongation (%) |
| BC | 4.12 | 7.27 |
| BC/SiO2-0.1 | 3.46 | 4.18 |
| BC/SiO2-0.2 | 3.48 | 6.46 |
| BC/SiO2-0.5 | 3.58 | 7.82 |
| BC/SiO2-1.0 | 5.61 | 7.99 |
| BC/SiO2-2.0 | 6.46 | 8.01 |
| BC/SiO2-5.0 | 3.49 | 4.23 |
As shown in Table 1, the amorphous silica/bacterial cellulose composite prepared by sodium silicate with different concentrations has great difference in tensile strength, so that the preferable concentration range of sodium silicate is 1.0 g/L-2.0 g/L. When the concentration of the sodium silicate is 0.1 g/L-0.5 g/L, the silica generated under the acidic condition is filled between the bacterial cellulose microfibers, so that the formation of a part of hydrogen bonds is hindered, but the dispersed silica particles cannot form larger tensile strength, so that the composite material has lower tensile strength than the bacterial cellulose. And as the concentration of the sodium silicate is increased, the content of the generated silica is increased, at the moment, the bacterial cellulose is used as a supporting material, and the formed silica net structure enables the composite material to have higher tensile strength. But as the sodium silicate concentration continues to rise to 5g/L, the composite again becomes less tensile, likely due to failure of the cellulose microfibers to assemble into ribbons at high silica concentrations.
Claims (5)
1. The method for preparing the amorphous silicon dioxide/bacterial cellulose composite material by in-situ mineralization is characterized by comprising the following steps:
inoculating the acetobacter xylinum seed solution into a fermentation culture medium containing sodium silicate, statically culturing, and repeatedly washing and freezing and thawing after the culture is finished to obtain the amorphous silicon dioxide/bacterial cellulose composite material, wherein the concentration of the sodium silicate is 1.0-2.0 g/L.
2. The method according to claim 1, wherein the inoculation amount of the acetobacter xylinum seed solution is 8% to 12%.
3. The method according to claim 1, wherein the fermentation medium is HS medium, pH is 6.0, the temperature of static culture is 30 ℃, and the culture time is 5-8 days.
4. The method according to claim 1, wherein the repeated washing is that the amorphous silica/bacterial cellulose composite material is washed with water repeatedly for 8-10 times until the amorphous silica/bacterial cellulose composite material is neutral.
5. The method according to claim 1, wherein the repeated freeze-thawing is to cool the composite material after the water washing to-20 ℃, then rapidly melt the composite material at room temperature, and the number of times is 5-7.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114317370A (en) * | 2022-01-11 | 2022-04-12 | 浙江大学 | Preparation method of bionic silicon mineralized bacteria and application of bionic silicon mineralized bacteria in treatment of phenol wastewater |
| WO2023172782A3 (en) * | 2022-01-07 | 2024-04-04 | Northwestern University | Biosynthesis of hierarchical metal organic framework-bacterial cellulose composites |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023172782A3 (en) * | 2022-01-07 | 2024-04-04 | Northwestern University | Biosynthesis of hierarchical metal organic framework-bacterial cellulose composites |
| CN114317370A (en) * | 2022-01-11 | 2022-04-12 | 浙江大学 | Preparation method of bionic silicon mineralized bacteria and application of bionic silicon mineralized bacteria in treatment of phenol wastewater |
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