CN101509025A - Method of preparing bacteria cellulose composite material - Google Patents
Method of preparing bacteria cellulose composite material Download PDFInfo
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- CN101509025A CN101509025A CNA2009100199512A CN200910019951A CN101509025A CN 101509025 A CN101509025 A CN 101509025A CN A2009100199512 A CNA2009100199512 A CN A2009100199512A CN 200910019951 A CN200910019951 A CN 200910019951A CN 101509025 A CN101509025 A CN 101509025A
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Abstract
The invention discloses a method for preparing bacterial cellulose composite material, comprising steps as follows: the production medium of bacterial cellulose is prepared and then acetobacter xylinum strain is put in the production medium of bacterial cellulose; sodium alginate, collagen, water-soluble chitin, gelatin or hyaluronic acid polymer material are added into the production medium in step b for cultivating a bacterial cellulose composite membrane, wherein, the adding amount of the polymer material is 1% to 5% of the weight of the production medium of bacterial cellulose; later the bacterial cellulose composite membrane which is prepared from step c is processed and washed by distilled water till being in a neutral state, and then tiled on filter paper and dried for becoming a finished product. The invention directly adds the polymer material to the production medium of bacterial cellulose; in the process of growth, the bacterial cellulose is directly composited with the added polymer material; the preparation process is simple; and the structure of the composite material of bacterial cellulose is uniform.
Description
Technical field
The invention belongs to the Biocomposite material field, particularly relate to the preparation method of bacteria cellulose based composites.
Background technology
As everyone knows, traditional Mierocrystalline cellulose derives from green plants, mainly is cotton and timber.Discover that certain micro-organisms is synthetic cellulose efficiently also, and has very big suitability for industrialized production potentiality.In order to distinguish mutually, the Mierocrystalline cellulose that derives from bacterium is referred to as " bacteria cellulose (bacterial cellulose) " or " micro organism cellulose (microbial cellulose) " with the Mierocrystalline cellulose of plant origin.Bacteria cellulose is found in 1886 by Britain scientist Brown the earliest, when he cultivates acetobacter under the condition of leaving standstill, finds that the gas-liquid surface of substratum forms the gelatinous membrane of one deck white, through chemistry and physical method analysis, determines that its composition is a Mierocrystalline cellulose.
Bacteria cellulose is close with common plant cellulose on chemical constitution and molecular structure, all is by β-1, the 4 glucosidic bond straight chain macromole that is formed by connecting.Compare with common plant cellulose, bacteria cellulose not only resultant velocity is fast, the productive rate height, and have many excellent characteristic, as high purity, high-crystallinity, high-polymerization degree, good physical and mechanical properties and air permeability and good, bacteria cellulose has good biological fitness simultaneously, and is biodegradable at occurring in nature, free from environmental pollution, be considered to the present best fiber of performance in the world, become one of focus of current research.Oneself successfully is applied to various fields such as food, medicine, chemical industry, papermaking, high-end audio equipment, filter membrane permeable membrane present bacteria cellulose.Under the situation exhausted day by day at petroleum resources, that world population increases to become increasingly conspicuous with the limited contradiction of ploughing, bacteria cellulose is being contained infinite commercial opportunities and fine development prospect as a kind of purposes biomaterial very widely.
Summary of the invention
Task of the present invention is to provide a kind of preparation method of bacteria cellulose composite material.
Its technical solution is:
A kind of preparation method of bacteria cellulose composite material may further comprise the steps:
The preparation of a bacteria cellulose production culture medium;
B inserts bacteria cellulose production culture medium with the acetobacter xylinum bacterial classification;
Adding includes but not limited to a certain single thing in sodium alginate, collagen protein, N,O-CMC, gelatin, the hyaluronic acid macromolecular material or the assembly thing of being made up of aforementioned two or more single things in the bacteria cellulose production culture medium of c in step b, turns out bacteria cellulose composite membrane; Above-mentioned sodium alginate, collagen protein, N,O-CMC, gelatin or hyaluronic add-on respectively are 1%～5% of bacteria cellulose production culture medium weight;
D directly or through after other respective handling is washed till neutrality with distilled water with the bacteria cellulose composite membrane that step c produces, and is tiled on the filter paper, and oven dry is finished product.
Comprise step among the above-mentioned steps d:
D1 inserts bacteria cellulose composite membrane in certain density calcium chloride solution, certain density acid or the alkaline solution and handles certain hour, after washing, insert again to handle in 80 ℃ of distilled water and be washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
Among the above-mentioned steps c, in bacteria cellulose production culture medium, add sodium alginate or contain the assembly thing of sodium alginate; Among the above-mentioned steps d, it is that 1%～3% calcium chloride solution is handled after after the washing that bacteria cellulose composite membrane is inserted concentration, insert in 80 ℃ of distilled water to handle again and be washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
Among the above-mentioned steps c, in cellulose production culture medium, add water-soluble chitin; Among the above-mentioned steps d, bacteria cellulose composite membrane with after the distillation washing, is inserted to handle in 80 ℃ of distilled water again and was washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
The invention has the beneficial effects as follows:
(1) single things of macromolecular material such as sodium alginate, collagen protein, N,O-CMC, gelatin, hyaluronic acid or assembly thing are added directly in the bacteria cellulose growth medium (fermention medium), bacteria cellulose is directly compound with the macromolecular material that adds in process of growth, preparation process is simple, gained bacteria cellulose composite material even structure.
(2) can prepare the different bacteria cellulose composite material of performance and satisfy actual needs by selecting the macromolecular material of different functions.As add a certain amount of N,O-CMC, can improve bacteria cellulose blend film strength and biocidal property; Add a certain amount of sodium alginate, can improve the absorptivity of physiological saline and the blood and the metal ion of bacteria cellulose blend film, improve the ion exchangeable of bacteria cellulose blend film etc.Add a certain amount of collagen protein, can improve the biocompatibility of bacteria cellulose blend film etc.Widen the Application Areas of bacteria cellulose and functional.
(3) simple, the safety of preparation process.
Embodiment
The preparation method of bacteria cellulose composite material of the present invention comprises following concrete steps:
(1) bacterial classification and substratum are selected
Bacterial classification: acetobacter xylinum (Gluconacetobacter xylinum)
The preparation of slant strains substratum: glucose 100g, yeast powder 10g, lime carbonate 20g, agar 18g, distilled water 1000mL, pH=6.8;
The preparation of seed culture medium: glucose 2%, peptone 0.5%, yeast powder 0.5%, disodium hydrogen phosphate,anhydrous 0.27%, citric acid monohydrate compound 0.115%, sal epsom 057%, pH6.0, autoclave sterilization 20 minutes;
The preparation of bacteria cellulose production culture medium (fermention medium): glucose 2%, peptone 0.5%, yeast powder 0.5%, disodium hydrogen phosphate,anhydrous 0.27%, citric acid monohydrate compound 0.115%, sal epsom 057%, ethanol, pH6.0, autoclave sterilization 20min.
(2) microbial culture
Get the good slant strains of activation and insert seed culture medium, 30 ℃ of shaking culture 24 hours, shaking speed is 150r/min.Insert fermention medium with 8% inoculum size, add macromolecular materials such as a certain amount of sodium alginate, collagen protein, N,O-CMC, gelatin, hyaluronic acid in the substratum, fully vibrate during inoculation, discharging thalline, 30 ℃ of static cultivations of constant temperature 7 days.
(3) extraction of bacteria cellulose composite membrane and processing
Static cultivation is after 7 days, take out bacteria cellulose composite membrane, insert and handle certain hour (kind according to the interpolation macromolecular material is determined) in certain density calcium chloride solution or certain density acid, the alkaline solution, after the distilled water washing, insert in 80 ℃ of distilled water and handled 30 minutes, be washed till neutrality, this moment, composite membrane was creamy white translucent, it is tiled on the filter paper, dries to constant weight in 60～80 ℃.
Prepared bacteria cellulose composite function film can be directly used in medical dressing etc.
Below in conjunction with embodiment the present invention is made further explanation:
Embodiment 1
Choosing of bacterial classification: acetobacter xylinum
The preparation of slant strains substratum: glucose 100g, yeast powder 10g, lime carbonate 20g, agar 18g, distilled water 1000mL, pH=6.8;
The preparation of seed culture medium: glucose 2%, peptone 0.5%, yeast powder 0.5%, disodium hydrogen phosphate,anhydrous 0.27%, citric acid monohydrate compound 0.115%, sal epsom 057%, pH6.0, autoclave sterilization 20min;
The preparation of fermention medium: glucose 2%, peptone 0.5%, yeast powder 0.5%, disodium hydrogen phosphate,anhydrous 0.27%, citric acid monohydrate compound 0.115%, sal epsom 057%, ethanol, pH6.0, autoclave sterilization 20 minutes.
Get the good slant strains of activation and insert seed culture medium, 30 ℃ of shaking culture 24 hours, shaking speed is 150r/min.Insert fermention medium with 8% inoculum size, in fermention medium, add the sodium alginate that accounts for this fermention medium weight 1%, 2%, 4% or 5%, fully vibrate during inoculation, discharging thalline, 30 ℃ of static cultivations of constant temperature 7 days.
Take out bacteria cellulose-sodium alginate composite membrane, insert concentration and be in 1%, 2% or 3% the calcium chloride solution and handled 1 minute, after the distilled water washing, insert in 80 ℃ of distilled water and handled 30 minutes, be washed till neutrality, it is tiled on the filter paper, dry to constant weight in 60 ℃, 70 ℃ or 80 ℃.
The absorptivity of bacteria cellulose-calcium alginate composite membrane is measured as follows:
Take by weighing a certain amount of bacteria cellulose-calcium alginate composite membrane sample, at room temperature put into distilled water, physiological saline and artificial plasm (the imitation blood sodium ion that British Pharmacopoeia is stipulated and the concentration of calcium ion respectively, the concentration of sodium ion is 142mmol/L among the artificial plasm, the concentration of calcium ion is 2.5mmol/L) in fully wetting, take out after 1 hour, have in the centrifuge shield of aperture draining 2～3min under state of nature bottom packing into.Centrifuge shield is put into whizzer centrifuge dehydration 15min again, taken out centrifuge shield, the water of bottom is poured out, behind the centrifuge dehydration 45min, take out the film sample again, G weighs rapidly
0Membrane sample is put into baking oven, and oven temperature is at 105 ℃～110 ℃, and the time is 1 hour.Take out sample, the G that in moisture eliminator, weighs behind the cooling 30min, absorptivity is represented with liquid absorption N:
Adopt formula (1) to calculate the liquid absorption of bacteria cellulose-calcium alginate composite membrane.Utilize thickness tester working sample thickness, utilize universal testing machine to measure breaking tenacity, measurement result sees Table 1.
The absorptivity of table 1 bacteria cellulose-sodium alginate film
Embodiment 2
Preparation process and method be substantially with embodiment 1, only is selected at 1%, 3%, 4% or 5% the N,O-CMC that is incorporated as this fermention medium weight in the fermention medium, fully vibrates 30 ℃ of static cultivations of constant temperature 7 days during inoculation.
Resulting bacteria cellulose-N,O-CMC composite membrane is inserted in 80 ℃ of distilled water and was handled 30 minutes after the distilled water washing, is washed till neutrality, and it is tiled on the filter paper, dries to constant weight in 60 ℃, 70 ℃ or 80 ℃.
Bacteria cellulose-N,O-CMC composite membrane the results are shown in Table 2 by U.S. AATCC Test Method 100 standard test bacteriostasis rates.
The bacteriostasis rate of table 2 bacteria cellulose-N,O-CMC film
Embodiment 3
Preparation process and method are substantially with embodiment 1, only be selected at and add the assembly thing that contains sodium alginate in the substratum, wherein sodium alginate is 1%, 3%, 4% or 5% of a fermention medium weight, N,O-CMC is 3% or 1% of a fermention medium weight, collagen protein is 2% or 1% of a fermention medium weight, fully vibrate 30 ℃ of static cultivations of constant temperature 7 days during inoculation.
Resulting bacteria cellulose composite membrane was handled 1 minute in 1%, 2% or 3% calcium chloride solution, after the distilled water washing, inserted in 80 ℃ of distilled water and handled 30 minutes, was washed till neutrality, and it is tiled on the filter paper, dried to constant weight in 60 ℃, 70 ℃ or 80 ℃.
Embodiment 4
Preparation process and method be substantially with embodiment 1, only is selected at 1%, 3%, 4% or 5% the gelatin that is incorporated as this fermention medium weight in the fermention medium, fully vibrates 30 ℃ of static cultivations of constant temperature 7 days during inoculation.
Resulting bacteria cellulose-gelatin-compounded film is inserted in 80 ℃ of distilled water and was handled 30 minutes after the distilled water washing, is washed till neutrality, and it is tiled on the filter paper, dries to constant weight in 60 ℃, 70 ℃ or 80 ℃.
Embodiment 5
Preparation process and method be substantially with embodiment 1, only is selected at 1%, 3%, 4% or 5% the collagen protein that is incorporated as this fermention medium weight in the fermention medium, fully vibrates 30 ℃ of static cultivations of constant temperature 7 days during inoculation.
Resulting bacteria cellulose-N,O-CMC composite membrane is inserted in 80 ℃ of distilled water and was handled 30 minutes after the distilled water washing, is washed till neutrality, and it is tiled on the filter paper, dries to constant weight in 60 ℃, 70 ℃ or 80 ℃.
Embodiment 6
Preparation process and method be substantially with embodiment 1, only is selected at 1%, 3%, 4% or 5% the hyaluronic acid that is incorporated as this fermention medium weight in the fermention medium, fully vibrates 30 ℃ of static cultivations of constant temperature 7 days during inoculation.
Resulting bacteria cellulose-N,O-CMC composite membrane is inserted in 80 ℃ of distilled water and was handled 30 minutes after the distilled water washing, is washed till neutrality, and it is tiled on the filter paper, dries to constant weight in 60 ℃, 70 ℃ or 80 ℃.
Need to prove at this: the variation that those skilled in the art make under the instruction of above-mentioned description apparently, the variation of different assembly each other etc. such as cited each add-on of macromolecular material of the present invention, and other suitable macromolecular materials of introducing apparently etc., should fall into appended claims of the present invention institute restricted portion equally.
Claims (4)
1, a kind of preparation method of bacteria cellulose composite material is characterized in that may further comprise the steps:
The preparation of a bacteria cellulose production culture medium;
B inserts bacteria cellulose production culture medium with the acetobacter xylinum bacterial classification;
Adding includes but not limited to a certain single thing in sodium alginate, collagen protein, N,O-CMC, gelatin, the hyaluronic acid macromolecular material or the assembly thing of being made up of aforementioned two or more single things in the bacteria cellulose production culture medium of c in step b, turns out bacteria cellulose composite membrane; Above-mentioned sodium alginate, collagen protein, N,O-CMC, gelatin or hyaluronic add-on respectively are 1%～5% of bacteria cellulose production culture medium weight;
D directly or through after other respective handling is washed till neutrality with distilled water with the bacteria cellulose composite membrane that step c produces, and is tiled on the filter paper, and oven dry is finished product.
2, the preparation method of bacteria cellulose composite material according to claim 1 is characterized in that: comprise step in the described steps d:
D1 inserts bacteria cellulose composite membrane in certain density calcium chloride solution, certain density acid or the alkaline solution and handles certain hour, after washing, insert again to handle in 80 ℃ of distilled water and be washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
3, the preparation method of bacteria cellulose composite material according to claim 1 is characterized in that: among the described step c, add sodium alginate or contain the assembly thing of sodium alginate in bacteria cellulose production culture medium; In the described steps d, it is that 1%～3% calcium chloride solution is handled after after the washing that bacteria cellulose composite membrane is inserted concentration, insert in 80 ℃ of distilled water to handle again and be washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
4, the preparation method of bacteria cellulose composite material according to claim 1 is characterized in that: among the described step c, add water-soluble chitin in cellulose production culture medium; In the described steps d, bacteria cellulose composite membrane with after the distillation washing, is inserted to handle in 80 ℃ of distilled water again and was washed till neutrality in 30 minutes, the bacteria cellulose composite membrane that obtains is creamy white translucent, dries to constant weight in 60～80 ℃.
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| CN102210884A (en) * | 2010-04-07 | 2011-10-12 | 南京理工大学 | Preparation method of antibacterial dressing for promoting wound healing |
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