CN103897219A - Preparation method of bacterial cellulose/polyacrylamide composite membrane - Google Patents
Preparation method of bacterial cellulose/polyacrylamide composite membrane Download PDFInfo
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- CN103897219A CN103897219A CN201410068805.XA CN201410068805A CN103897219A CN 103897219 A CN103897219 A CN 103897219A CN 201410068805 A CN201410068805 A CN 201410068805A CN 103897219 A CN103897219 A CN 103897219A
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Abstract
The invention discloses a preparation method of a bacterial cellulose/polyacrylamide composite membrane. The preparation method of the bacterial cellulose/polyacrylamide composite membrane comprises the following steps: taking and inoculating 1-2 ring activated gluconacetobacterxylinum CGMCCNo.1.1812 or CGMCCNo.1.2378 into a seed culture medium, carrying out shaking culture to obtain a seed solution; then inoculating the seed solution into a polyacrylamide-containing fermentation culture medium, uniformly mixing the seed solution with the fermentation culture medium, then standing and culturing for 6-14 days at the temperature of 28-32 DEG C, generating a bacterial cellulose/polyacrylamide composite membrane floating on the surface of liquid, and then removing culture medium and impurities in the surface of the bacterial cellulose/polyacrylamide composite membrane and in the bacterial cellulose/polyacrylamide composite membrane, so that the bacterial cellulose/polyacrylamide is obtained. The preparation method of the bacterial cellulose/polyacrylamide composite membrane has the advantages that adoption of toxic chemical reagents like a crosslinking agent can be avoided, the environment is protected and operation is simple.
Description
Technical field
The present invention relates to the preparation field of bacteria cellulose composite membrane, particularly relate to a kind of method that biological composite algorithm is prepared bacteria cellulose/polyacrylamide composite membrane.
Background technology
Bacteria cellulose is that Brown finds first, and its diameter is only 1/10 of people's hairline, is 10-100nm, is a kind of novel nano meter biomaterial.The key distinction of bacteria cellulose and plant cellulose other polysaccharide that are to undope, as xylogen, hemicellulose etc., therefore there is the character of a lot of uniquenesses, as high-crystallinity, high chemical purity, high-tensile, high elastic coefficient, high water holding capacity etc., the aspects such as widespread use and medicine, food, environment.Meanwhile, bacteria cellulose is also a kind of biodegradable, and the nano biological fiber of not degrading in human body.
Bacteria cellulose can be used as water-soluble polymer as the fortifying fibre in (polyacrylamide) composite membrane, this composite membrane is widely used, especially aspect medical science, therefore the correlative study of this new bio nano material is subject to various countries investigator's attention day by day.
Summary of the invention
The object of the invention is in order to solve above-mentioned bacteria cellulose/polyacrylamide composite membrane complicated operation of preparing, introduce the technical problems such as chemical reagent pollutes and a kind of preparation method of bacteria cellulose/polyacrylamide composite membrane is provided.This preparation method is prepared into bacteria cellulose/polyacrylamide composite membrane by adding polyacrylamide in the fermention medium to production bacteria cellulose, it is simple that this preparation method has film-forming process, the features such as cost is low, environmentally friendly have a good application prospect in suitability for industrialized production.
Technical scheme of the present invention
A preparation method for bacteria cellulose/polyacrylamide composite membrane, comprising:
(1), the activation of bacterial classification
First 1-2 being encircled to slant strains is forwarded to and in fresh slant medium, controls temperature and be 27-32 ℃ and activate the inclined-plane seed that must activate after 72h;
Described bacterial classification is acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.1812 or acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.2378;
Described slant medium, count by weight percentage, by glucose, fructose or the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, the citric acid of 0.1-0.4%, the calcium carbonate of 2-3%, the agar of 2-4% and the deionized water of surplus composition, adjusting pH is 5-6;
Above-mentioned slant medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), seed liquor is cultivated
Get the inclined-plane seed that 1-2 ring step (1) activated and be seeded in the seed culture medium of 100-250ml, control temperature is 20-32 ℃, and rotating speed is 140-200r/min, cultivates 18-30h, obtains seed liquor;
Described seed culture medium, count by weight percentage, by glucose, fructose or the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, the citric acid of 0.1-0.4% and the deionized water of surplus composition, adjusting pH is 5-6, and above-mentioned seed culture medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 6-10:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, then leave standstill and cultivate 6-14 days in 28-32 ℃, generate bacteria cellulose/polyacrylamide composite membrane and float on liquid level, then remove substratum and impurity in film surface and the film of bacteria cellulose/polyacrylamide composite membrane, obtain bacteria cellulose/polyacrylamide composite membrane;
Described fermention medium, count by weight percentage, by glucose, fructose, the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, the citric acid of 0.1-0.4%, the polyacrylamide of 0.01-0.5% and the deionized water of surplus composition, adjusting pH is 5-6, above-mentioned fermention medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
Substratum and the impurity on the above-mentioned film surface of removing bacteria cellulose/polyacrylamide composite membrane, get bacteria cellulose/polyacrylamide composite membrane that step (3) floats on liquid level, water rinses 5-15 time, to remove substratum and the impurity on film surface of bacteria cellulose/polyacrylamide composite membrane;
Again above-mentioned bacteria cellulose/polyacrylamide composite membrane of removing surperficial substratum and impurity is soaked in alkaline solution, control temperature is 80-100 ℃, boil 30-80min, with thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment the film of bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then stop in the time that the pH of effluent liquid is 7.0-7.2 with distilled water flushing rinsing, obtain bacteria cellulose/polyacrylamide composite membrane:
Described alkaline solution is the NaOH aqueous solution, the KOH aqueous solution or the Na2CO3 aqueous solution that concentration is 0.05-0.4mol/l.
Bacteria cellulose/polyacrylamide composite membrane of preparation method's gained of above-mentioned a kind of bacteria cellulose/polyacrylamide composite membrane, its thickness is 0.05-3mm, water ratio 97.4-98.6%, tensile strength 20MPa-100MPa, elongation at break is 2-4%.
Beneficial effect of the present invention
The preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane of the present invention, have simple to operate, the advantages such as environmentally safe, and because this preparation method is prepared from bacteria cellulose/polyacrylamide composite membrane by adding polyacrylamide in the fermention medium to production bacteria cellulose, therefore there is film-forming process simple, the features such as cost is low, environmentally friendly have a good application prospect in suitability for industrialized production.
The preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane of the present invention; water-soluble polymer polyacrylamide is joined in the fermention medium of bacteria cellulose; owing to adopting biological composite algorithm to obtain bacteria cellulose/polyacrylamide composite membrane; and the preparation of bacteria cellulose/polyacrylamide composite membrane just can be realized in the time that bacteria cellulose is synthetic; simple to operate; avoid using chemical cross-linking agent, protected compared with the conventional method environment.
Tensile strength and the elongation at break of a kind of bacteria cellulose/polyacrylamide composite membrane of the present invention increase compared with bacteria cellulose, are applicable to medical field.
Embodiment
Below by specific embodiment, the present invention is further set forth, but do not limit the present invention.
The milscale that in the present invention embodiment, the thickness instrument model of bacteria cellulose/polyacrylamide composite membrane of gained is 0-25mm, is produced by Shanghai Measuring and Cutting Tools Plant;
The instrument model that water ratio detects is YP1002N electronic balance: produced by balance company of upper Nereid section;
The model of the instrument that tensile strength and elongation at break detect is TA-XTPlus Exponent32 property tester, is produced by Stable Micro System company of Britain;
Various raw materials used in various embodiments of the present invention, as glucose, peptone, yeast powder, citric acid, Sodium phosphate dibasic, fructose, sucrose is analytical pure, is provided by Chemical Reagent Co., Ltd., Sinopharm Group.
Embodiment 1
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 1 ring acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.1812 slant strains to the slant medium in step (1), in constant incubator, control 27 ℃ of temperature and cultivate 72h, the inclined-plane seed that must activate;
Described slant medium, count by weight percentage, by 2% glucose, 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid, 2% calcium carbonate, 2% agar and the deionized water of surplus composition, regulating pH is 5, and above-mentioned slant medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 1 ring step (1) and be linked in the seed culture medium of 100ml, 30 ℃ of shaking culture 18h, shaking speed is 140r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 2% glucose, 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid and surplus deionized water composition, regulating pH is 5, and above-mentioned seed culture medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 6:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 6 days at 28 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 2% glucose, 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid, 0.01% polyacrylamide and the deionized water of surplus composition, regulating pH is 5, above-mentioned fermention medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 5 times, to remove substratum and the impurity on film surface of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the NaOH aqueous solution of 0.05mol/l, 80 ℃ are boiled 80min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then with distilled water flushing until the pH value of effluent liquid be within 7.2 o'clock, stop rinse, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 0.05mm, water ratio are 97.4% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 20MPa, elongation at break are 2%.
Embodiment 2
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 1 ring acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.1812 slant strains to the slant medium in step (1), in constant incubator, cultivate 72h for 32 ℃, as the inclined-plane seed having activated;
Described slant medium, count by weight percentage, by 4% glucose, 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 3% calcium carbonate, 4% agar and the deionized water of surplus composition, adjusting pH is 5-6, and above-mentioned slant medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 250ml, 32 ℃ of shaking culture 30h, shaking speed is 200r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 4% glucose, 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid and the deionized water of surplus composition, regulating pH is 6, and above-mentioned seed culture medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 10:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 14 days at 32 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 4% glucose, 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 0.5% polyacrylamide and the deionized water of surplus composition, regulating pH is 6, above-mentioned fermention medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 15 times, to remove film surface medium and the impurity of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the KOH aqueous solution of 0.4mol/l, 100 ℃ are boiled 80min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then be within 7.2 o'clock, to stop rinsing by distilled water flushing to the pH value of effluent liquid, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 3mm, water ratio are 98% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 90MPa, elongation at break are 3.2%.
Embodiment 3
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 2 ring acetobacter xylinums (Gluconacetobacter xylinum) CGMCC No.1.1812 slant strains to the slant medium in step (1), in constant incubator, cultivate 72h, the inclined-plane seed that must activate for 30 ℃;
Described slant medium, count by weight percentage, by 3% glucose, 0.7% yeast powder, 0.6% Tryptones, 0.3% Na2HPO4,0.2% citric acid, 3% calcium carbonate, 3% agar and the deionized water of surplus composition, regulating pH is 6, and above-mentioned slant medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 100ml, 30 ℃ of shaking culture 24h, shaking speed is 160r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 3% glucose, 0.8% yeast powder, 0.9% Tryptones, 0.4% Na2HPO4, the citric acid of 0.3 % and the deionized water of surplus composition, regulating pH is 6, and above-mentioned seed culture medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 10:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 7 days at 30 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 3% glucose, 0.6% yeast powder, 0.6% Tryptones, 0.3% Na2HPO4,0.2% citric acid, 0.2% polyacrylamide and the deionized water of surplus composition, regulating pH is 6, above-mentioned fermention medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 10 times, to remove film surface medium and the impurity of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the Na2CO3 aqueous solution of 0.1mol/l, 60 ℃ are boiled 60min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then stop in the time that the pH value of effluent liquid is 7.0-7.2 with distilled water flushing rinsing, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 2.6mm, water ratio are 98.7% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 79MPa, elongation at break are 2.7%.
Embodiment 4
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 2 ring acetobacter xylinums (Gluconacetobacter xylinum) CGMCC No.1.1812 slant strains to the slant medium in step (1), in constant incubator, cultivate 72h, the inclined-plane seed that must activate for 28 ℃;
Described slant medium, count by weight percentage, by 2% fructose, 0.5% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 3% calcium carbonate, 4% agar and the deionized water of surplus composition, regulating pH is 6, and above-mentioned slant medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 250ml, 32 ℃ of shaking culture 28h, shaking speed is 180r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 2% fructose, 1% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid and the deionized water of surplus composition, regulating pH is 6, and above-mentioned seed culture medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 8:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 10 days at 30 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 4% sucrose, 1% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.3% citric acid, 0.3% polyacrylamide and the deionized water of surplus composition, regulating pH is 6, above-mentioned fermention medium is sterilizing 20min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 5-15 time, to remove film surface medium and the impurity of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the Na2CO3 aqueous solution of 0.1mol/l, 100 ℃ are boiled 30min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then be within 7.2 o'clock, to stop rinsing by distilled water flushing to the pH value of effluent liquid, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 1.6mm, water ratio are 97.3% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 40MPa, elongation at break are 4%.
Embodiment 5
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 1 ring acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.1812 slant strains to the slant medium in step (1), in constant incubator, cultivate 72h, the inclined-plane seed that must activate for 30 ℃;
Described slant medium, count by weight percentage, by 4% sucrose, 0.8% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.2% citric acid, 2% calcium carbonate, 2% agar and the deionized water of surplus composition, regulating pH is 6, and above-mentioned slant medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 1-2 ring step (1) and be linked in the seed culture medium of 250ml, 30 ℃ of shaking culture 26h, shaking speed is 180r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 3% sucrose, 0.9% yeast powder, 0.9% Tryptones, 0.4% Na2HPO4,0.1% citric acid and the deionized water of surplus composition, regulating pH is 5, and above-mentioned seed culture medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 8:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 14 days at 30 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 2% sucrose, 0.5% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.3% citric acid, 0.4% polyacrylamide and the deionized water of surplus composition, regulating pH is 5, above-mentioned fermention medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 10 times, to remove film surface medium and the impurity of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the NaOH aqueous solution of 0.4mol/l, 100 ℃ are boiled 80min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then be within 7.2 o'clock, to stop rinsing by distilled water flushing to the pH value of effluent liquid, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 2.8mm, water ratio are 97.8% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 80MPa, elongation at break are 3.6%.
Embodiment 6
A preparation method for bacteria cellulose/polyacrylamide composite membrane, specifically comprises the steps:
(1), the activation of bacterial classification
Get 1 ring acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.2378 slant strains to the slant medium in step (1), in constant incubator, cultivate 72h, the inclined-plane seed that must activate for 30 ℃;
Described slant medium, count by weight percentage, by 4% sucrose, 0.6% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.2% citric acid, 2% calcium carbonate, 2% agar and the deionized water of surplus composition, regulating pH is 6, and above-mentioned slant medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get the inclined-plane seed having activated in 1-2 ring step (1) and be linked in the seed culture medium of 250ml, 30 ℃ of shaking culture 30h, shaking speed is 200r/min, obtains seed liquor;
Described seed culture medium, count by weight percentage, by 3% sucrose, 0.9% yeast powder, 0.9% Tryptones, 0.4% Na2HPO4,0.1% citric acid and the deionized water of surplus composition, regulating pH is 5, and above-mentioned seed culture medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 8:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, and then leave standstill and cultivate 14 days at 30 ℃, generate bacteria cellulose/polyacrylamide composite membrane crude product and float on liquid level;
Described fermention medium, count by weight percentage, by 2% sucrose, 0.5% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.3% citric acid, 0.4% polyacrylamide and the deionized water of surplus composition, regulating pH is 5, above-mentioned fermention medium is sterilizing 25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000;
By bacteria cellulose/polyacrylamide composite membrane crude product of above-mentioned generation, water rinses 10 times, to remove film surface medium and the impurity of bacteria cellulose/polyacrylamide composite membrane, again bacteria cellulose/polyacrylamide composite membrane is soaked in the NaOH aqueous solution of 0.4mol/l, 100 ℃ are boiled 80min, thalline and residual substratum in the film of removal bacteria cellulose/polyacrylamide composite membrane, at this moment bacteria cellulose/polyacrylamide composite membrane is creamy white translucent, then be within 7.2 o'clock, to stop rinsing by distilled water flushing to the pH value of effluent liquid, obtain bacteria cellulose/polyacrylamide composite membrane.
After testing, its thickness is that 2.7mm, water ratio are 98.6% to bacteria cellulose/polyacrylamide composite membrane of above-mentioned gained, tensile strength 86MPa, elongation at break are 3.3%.
Above embodiment is only not used in and limits the scope of the invention for the present invention is described.In addition, after having read the content of the present invention's instruction, those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally.
Claims (8)
1. a preparation method for bacteria cellulose/polyacrylamide composite membrane, is characterized in that specifically comprising the steps:
(1), first 1-2 being encircled to slant strains is forwarded to and in fresh slant medium, controls temperature and be 27-32 ℃ and activate the inclined-plane seed that must activate after 72h;
Described slant strains is acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.1812 or acetobacter xylinum (Gluconacetobacter xylinum) CGMCC No.1.2378;
Described slant medium, count by weight percentage, by glucose, fructose or the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, 0.1-0.4% citric acid, the calcium carbonate of 2-3%, the agar of 2-4% and the deionized water of surplus composition, adjusting pH is 5-6;
Above-mentioned slant medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(2), the cultivation of seed liquor
Get 1-2 and encircle the inclined-plane seed having activated and be seeded in the seed culture medium of 100-250ml, control temperature is 20-32 ℃, and rotating speed is that 140-200r/min cultivates 18-30h, obtains seed liquor;
Described seed culture medium, count by weight percentage, by glucose, fructose or the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, the citric acid of 0.1-0.4% and the deionized water of surplus composition, adjusting pH is 5-6, and above-mentioned seed culture medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
(3), calculate by volume, it is seed liquor: the ratio that fermention medium is 6-10:100, the seed liquor of step (2) gained is seeded in fermention medium, fully vibration mixes seed liquor and fermention medium, then leave standstill and cultivate 6-14 days in 28-32 ℃, generate bacteria cellulose/polyacrylamide composite membrane and float on liquid level, then remove substratum and impurity in film surface and the film of bacteria cellulose/polyacrylamide composite membrane, obtain bacteria cellulose/polyacrylamide composite membrane;
Described fermention medium, count by weight percentage, by glucose, fructose, the sucrose of 2-4%, the yeast powder of 0.5-1%, the Tryptones of 0.5-1%, the Na2HPO4 of 0.1-0.5%, the citric acid of 0.1-0.4%, the polyacrylamide of 0.01-0.5% and the deionized water of surplus composition, adjusting pH is 5-6, above-mentioned fermention medium is sterilizing 20-25min at the temperature of 121 ℃, for subsequent use after being cooled to 30 ℃;
The molecular weight of described polyacrylamide is 100000-5000000.
2. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that:
In step (1), control 27 ℃ of temperature and cultivate 72h; Described slant medium, counts by weight percentage, by 2% glucose, and 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid, 2% calcium carbonate, 2% agar and the deionized water of surplus composition, regulating pH is 5;
In step (2), get the inclined-plane seed having activated in 1 ring step (1) and be linked in the seed culture medium of 100ml, 30 ℃ of shaking culture 18h, shaking speed is 140r/min; Described seed culture medium, counts by weight percentage, by 2% glucose, and 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid and surplus deionized water composition, regulating pH is 5;
In step (3), calculate by volume i.e. seed liquor: the ratio that fermention medium is 6:100, the seed liquor of step (2) gained is seeded in fermention medium, leave standstill and cultivate 6 days at 28 ℃; Described fermention medium, counts by weight percentage, by 2% glucose, and 0.5% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid, 0.01% polyacrylamide and the deionized water of surplus composition, regulating pH is 5.
3. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that:
It is 32 ℃ of cultivation 72h that step (1) is controlled temperature; Described slant medium, counts by weight percentage, by 4% glucose, and 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 3% calcium carbonate, 4% agar and the deionized water of surplus composition, adjusting pH is 5-6;
In step (2), get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 250ml, 32 ℃ of shaking culture 30h, shaking speed is 200r/min; Described seed culture medium, counts by weight percentage, by 4% glucose, and 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid and the deionized water of surplus composition, regulating pH is 6;
In step (3), calculate by volume i.e. seed liquor: the ratio that fermention medium is 10:100, the seed liquor of step (2) gained is seeded in fermention medium, leave standstill and cultivate 14 days at 32 ℃; Described fermention medium, counts by weight percentage, by 4% glucose, and 1% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 0.5% polyacrylamide and the deionized water of surplus composition, regulating pH is 6.
4. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that:
In step (1), controlling temperature is 30 ℃ of cultivation 72h; Described slant medium, counts by weight percentage, by 3% glucose, and 0.7% yeast powder, 0.6% Tryptones, 0.3% Na2HPO4,0.2% citric acid, 3% calcium carbonate, 3% agar and the deionized water of surplus composition, regulating pH is 6;
In step (2), get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 100ml, 30 ℃ of shaking culture 24h, shaking speed is 160r/min; Described seed culture medium, counts by weight percentage, by 3% glucose, and 0.8% yeast powder, 0.9% Tryptones, 0.4% Na2HPO4, the citric acid of 0.3 % and the deionized water of surplus composition, regulating pH is 6;
In step (3), calculate by volume i.e. seed liquor: the ratio that fermention medium is 10:100, the seed liquor of step (2) gained is seeded in fermention medium, leave standstill and cultivate 7 days at 30 ℃; Described fermention medium, counts by weight percentage, by 3% glucose, and 0.6% yeast powder, 0.6% Tryptones, 0.3% Na2HPO4,0.2% citric acid, 0.2% polyacrylamide and the deionized water of surplus composition, regulating pH is 6.
5. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that:
In step (1), controlling temperature is 28 ℃ of cultivation 72h; Described slant medium, counts by weight percentage, by 2% fructose, and 0.5% yeast powder, 1% Tryptones, 0.5% Na2HPO4,0.4% citric acid, 3% calcium carbonate, 4% agar and the deionized water of surplus composition, regulating pH is 6;
In step (2), get the inclined-plane seed having activated in 2 ring steps (1) and be linked in the seed culture medium of 250ml, 32 ℃ of shaking culture 28h, shaking speed is 180r/min; Described seed culture medium, counts by weight percentage, by 2% fructose, and 1% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.1% citric acid and the deionized water of surplus composition, regulating pH is 6;
In step (3), calculate by volume i.e. seed liquor: the ratio that fermention medium is 8:100, the seed liquor of step (2) gained is seeded in fermention medium, leave standstill and cultivate 10 days at 30 ℃; Described fermention medium, counts by weight percentage, by 4% sucrose, and 1% yeast powder, 0.5% Tryptones, 0.1% Na2HPO4,0.3% citric acid, 0.3% polyacrylamide and the deionized water of surplus composition, regulating pH is 6.
6. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that:
In step (1), controlling temperature is 30 ℃ of cultivation 72h; Described slant medium, counts by weight percentage, by 4% sucrose, and 0.6% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.2% citric acid, 2% calcium carbonate, 2% agar and the deionized water of surplus composition, regulating pH is 6;
In step (2), get the inclined-plane seed having activated in 1-2 ring step (1) and be linked in the seed culture medium of 250ml, 30 ℃ of shaking culture 30h, shaking speed is 200r/min; Described seed culture medium, counts by weight percentage, by 3% sucrose, and 0.9% yeast powder, 0.9% Tryptones, 0.4% Na2HPO4,0.1% citric acid and the deionized water of surplus composition, regulating pH is 5;
In step (3), calculate by volume i.e. seed liquor: the ratio that fermention medium is 8:100, the seed liquor of step (2) gained is seeded in fermention medium, leave standstill and cultivate 14 days at 30 ℃; Described fermention medium, counts by weight percentage, by 2% sucrose, and 0.5% yeast powder, 1% Tryptones, 0.1% Na2HPO4,0.3% citric acid, 0.4% polyacrylamide and the deionized water of surplus composition, regulating pH is 5.
7. the preparation method of a kind of bacteria cellulose/polyacrylamide composite membrane as claimed in claim 1, is characterized in that in step (3):
Substratum and the impurity on the described film surface of removing bacteria cellulose/polyacrylamide composite membrane, get step (3) and float on bacteria cellulose/polyacrylamide composite membrane of liquid level, and water rinses 5-15 time;
Substratum and impurity in the described film of removing bacteria cellulose/polyacrylamide composite membrane film, be about to remove the substratum on bacteria cellulose/polyacrylamide composite membrane film surface and bacteria cellulose/polyacrylamide composite membrane of impurity is soaked in alkaline solution, control temperature is 80-100 ℃ and boils 30-80min, then stop in the time that the pH of effluent liquid is 7.0-7.2 with distilled water flushing rinsing, obtain bacteria cellulose/polyacrylamide composite membrane:
Described alkaline solution is the NaOH aqueous solution, the KOH aqueous solution or the Na2CO3 aqueous solution that concentration is 0.05-0.4mol/l.
8. bacteria cellulose/polyacrylamide composite membrane of the preparation method's gained as described in claim 1-7, is characterized in that its thickness is 0.05-3mm, water ratio 97.4-98.6%, and tensile strength 20MPa-90MPa, elongation at break is 2-4%.
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