CN104225669B - Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof - Google Patents
Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof Download PDFInfo
- Publication number
- CN104225669B CN104225669B CN201410421901.8A CN201410421901A CN104225669B CN 104225669 B CN104225669 B CN 104225669B CN 201410421901 A CN201410421901 A CN 201410421901A CN 104225669 B CN104225669 B CN 104225669B
- Authority
- CN
- China
- Prior art keywords
- preparation
- zein
- film
- water
- bacterial cellulose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W90/00—Enabling technologies or technologies with a potential or indirect contribution to greenhouse gas [GHG] emissions mitigation
- Y02W90/10—Bio-packaging, e.g. packing containers made from renewable resources or bio-plastics
Abstract
The invention discloses biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof.The method first carries out the preparation and purification of bacteria cellulose film; Bacteria cellulose film after purification is put into distilled water and is stirred, and shears, through high pressure microjet homogenizing, obtain nanofiber dispersion liquid with high-speed shearing machine; Zein and slightly water-soluble bioactive substance are dissolved in ethanol-water solution, obtain alcoholic solution; Sodium caseinate is dissolved in water, adds in described alcoholic solution under stirring, obtain mixed liquor; Removing ethanol, centrifugal, obtain the zein nano-particle being rich in bioactive molecule; Nanofiber dispersion liquid be rich in bioactive zein granule and mixed, stir, and carry out supersound process; Vacuum filtration, fast filming; Dry.Composite film material of the present invention has good mechanical performance, biocompatibility, biodegradable, and this film possesses the biological activitys such as regulatable antibacterial, antioxidation.
Description
Technical field
The present invention relates to the preparation method of a kind of high strength bioactive Bacterial cellulose-Compound Film of Zein material, raw material all adopt food stage and can be degradable biomaterial, belong to the association area such as the bio-medical materials such as wound dressing, food industry technology.
Background technology
Bacterial cellulose (BacterialCellulose, BC) is fermented by Pseudomonas such as acetobacter xylinums and produces, and the nanofibre polymer without side chain be made up of β-Isosorbide-5-Nitrae glycosidic bond, just more and more receives the concern of people.Compared with traditional cellulose, Bacterial cellulose has many advantages, as purity is high, there is high mechanical properties, high retentiveness, good biocompatibility, breathable moisture permeability, biodegradable, avirulence, to liquid, there is the characteristics such as very strong absorbability, be therefore widely used in the fields such as bio-medical material, food, papermaking, cosmetics (Biofibre facial mask, eye mask).But owing to limitting by self property, the practical ranges of single BC material is often narrower, limits it and further applies.
In order to give bacteria cellulose material with functions such as specific antibacterial antioxidation, mostly prior art is to adopt direct infusion method Bacterial cellulose to be placed in some bioactive substance solution, rear lyophilizing, or topple over after solution blending, the methods such as oven dry prepare composite film material, these class methods length consuming time, is unfavorable for suitability for industrialized production.And most of bioactive substance, as Polyphenols antimicrobial and antioxidant agent, silver sulfadiazine etc., often there is the problems such as poor solubility, stability is not enough, therefore the antibacterial antioxidation class material actual utility prepared of the method is little.
Summary of the invention
In order to solve above-mentioned the deficiencies in the prior art part, the object of the present invention is to provide the preparation method of a kind of high strength bioactive Bacterial cellulose-Compound Film of Zein material, the method overcome prior art and prepare the defects such as bacterial cellulose composite membrane material material length consuming time, actual utility be low.
Zein (zein) is the main protein in Semen Maydis, water insoluble, dissolve in the alcoholic solution of 60% ~ 95%, this albumen is as natural degradable material, itself has strong-hydrophobicity and does not need to be cross-linked, can not in water violent released load thing, self assembly can be there is during solvent evaporation and formed there is the nano-particle of different shape, therefore, the first-selection of bag carrier material is become.But Bacterial cellulose hydrophilic is extremely strong, how realizes the homogeneous dispersibility of Hydrophilic Nanofibrous in hydrophobic proteins, become key of the present invention.
First the present invention utilizes anti-solvent technology to prepare can the water solublity zein granule of load active factors (natural antibacterial agent/antioxidant), carries out homogeneous compound with the Bacterial cellulose nanofiber through high pressure microjet homogeneous preparation; And adopt the mode of the industrial copy paper of simulation to prepare high strength bacteria cellulose-base active bio composite film material fast first.The present invention adopts nanorize embedding techniques (i.e. the preparation of anti-solvent technology can the granule of load active factors), select suitable bag carrier material, the embedding of these active substances is wherein formed reactive nanoparticles, reach and strengthen its dissolubility and stability, and reaching the object extending the useful effect cycle (controlled release), rear simulation industry papermaking technology prepares composite film material fast.Obtained composite film material has the antibacterial antioxidative biological activity of specificity, the feature such as high strength, environmental protection on the one hand; On the other hand, this film preparation mode Lv color Jie Neng ﹑ rapidly and efficiently, for large-scale industrial production high strength bioactive bacteria cellulose-base composite film material provides New Century Planned Textbook and technical foundation.
The present invention to achieve these goals, adopts following technical scheme:
The preparation method of biological activity Bacterial cellulose-Compound Film of Zein material, comprises the following steps:
1) preparation of bacteria cellulose film: utilize acetobacter xylinum strain inoculation to prepare bacteria cellulose film in seed culture fluid;
2) purification of bacteria cellulose film
Be soaked in aqueous slkali after bacteria cellulose film clear water is rinsed, boil; Bacteria cellulose film is soaked in distilled water and boils, change water; Repeat to boil 2-5 time for twice; Then by bacteria cellulose film, it is soaked in distilled water, until neutral;
3) preparation of nanofiber dispersion liquid
By step 2) in bacteria cellulose film after the purification that obtains put into distilled water and stir, shear with high-speed shearing machine, then through high pressure microjet homogenizing, obtain nanofiber dispersion liquid;
4) preparation of the zein nano-particle of bioactive molecule is rich in
Zein and slightly water-soluble bioactive substance are dissolved in ethanol-water solution, obtain alcoholic solution; Sodium caseinate is dissolved in water, adds in described alcoholic solution under stirring, obtain mixed liquor; After stirring, remove ethanol by Rotary Evaporators, centrifugal, obtain the zein nano-particle being rich in bioactive molecule; In described mixed liquor, the mass fraction of zein is 1% ~ 5%, and the mass fraction of bioactive substance is 0.1% ~ 1%, and the mass fraction of sodium caseinate is 1% ~ 5%; Described slightly water-soluble bioactive substance is the antibacterial of slightly water-soluble and/or antioxidant;
5) preparation of Bacterial cellulose nanofiber and zein nano-particle mixed liquor
By step 3) in obtained nanofiber dispersion liquid and step 4) the bioactive zein granule that has been rich in prepared mixes, stirs, and carry out supersound process, obtain nanofiber and zein nano-particle mixed liquor; The mass ratio of described nanofiber and zein granule is (1 ~ 10): 1;
6) the biological compounded wet film preparation of Bacterial cellulose
Nanofiber and zein nano-particle mixed liquor are first carried out vacuum outgas, simulates industrial papermaking technology, vacuum filtration, film forming, obtain that there is the biological compounded wet film of bioactive Bacterial cellulose;
7) dry
The biological compounded wet film of bioactive Bacterial cellulose that has obtained in step (6) is carried out freeze-day with constant temperature or hot-pressing drying, obtains biological activity Bacterial cellulose-Compound Film of Zein.
For realizing the object of the invention further, preferably, step 3) in, described distilled water and the mass ratio of Bacterial cellulose are (1 ~ 5): 1.
Step 3) in, the rotating speed of described high-speed shearing machine is 8000 ~ 12000rpm, and shear time is 10 ~ 20min.
Step 3) in, the homogenization pressure of described high pressure microjet is 5000 ~ 20000psi, and homogenization cycles is 1 ~ 20 time.
Step 4) in, in ethanol-water solution, ethanol mass concentration is 60% ~ 95%; Described centrifugal rotating speed is 1000rpm ~ 5000rpm, and the centrifugal time is 10min ~ 30min; Antibacterial and/or the antioxidant of described slightly water-soluble is erythromycin, thymol or resveratrol.
Step 5) in, the time of supersound process is 10min ~ 20min, and ultrasonic power is 100W ~ 200W, and work efficiency is 40% ~ 60%, and operating temperature is 20 DEG C ~ 30 DEG C.
Step 6) in, the time of described vacuum outgas is 20min ~ 60min; Described vacuum filtration selects aperture to be that the cellulose microporosity filter membrane of 0.45-0.65 μm carries out vacuum filtration, fast filming to bacterial fibers and zein granule mixed liquor.
Step 7) in, described freeze-day with constant temperature is 25 ~ 60 DEG C of freeze-day with constant temperature; Described hot-pressing drying is 85 ~ 100 DEG C, carries out drying under 0.1 ~ 0.2MPa.
Described utilize acetobacter xylinum strain inoculation to prepare bacteria cellulose film be get activation in seed culture fluid after acetobacter xylinum strain inoculation in seed culture fluid, shaken cultivation, the mass ratio by 1 ~ 10% is inoculated in fermentation culture, shakes up, quiescent culture, obtains bacteria cellulose film.Fermentation culture composition: 1.8%-2.2%D-glucose, 0.45%-0.6% peptone, 0.45%-0.6% yeast extract, 0.11%-0.115% citric acid, 0.25%-0.30%Na
2hPO
412H
2o, all the other are distilled water, pH3-5; Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C-35 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 1 ~ 10%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
Product of the present invention can be used for the bio-medical materials such as wound dressing, Living Packaging Materials in Foodstuff and beauty and make-up material;
Compared with existing product, the present invention has following beneficial effect:
(1) the present invention utilizes anti-solvent technology to prepare the water solublity zein granule being rich in bioactive substance uniquely, gives Bacterial cellulose-Compound Film of Zein material biological activity and exceptional function.
(2) mode that the present invention proposes to simulate paper industry copy paper first prepares the ideamonger of bacteria cellulose-base composite film material fast, make material preparation method rapidly and efficiently, green safety is pollution-free, for large-scale industrial production provides New Century Planned Textbook and technical foundation.
(3) the present invention adopts repeatedly through high pressure microjet homogeneous preparation Bacterial cellulose nanofiber stable uniform dispersion liquid, and adopt anti-solvent technology to prepare the water-solubility protein granule being rich in bioactive substance, achieve the homogeneous dispersion of Bacterial cellulose nanofiber in hydrophobic proteins of strongly hydrophilic.
Accompanying drawing explanation
Fig. 1 is the surface atom power plane graph of embodiment 2, sweep limits 5 microns × 5 microns, scale 500nm;
Fig. 2 is the surface atom power plane graph of comparative example 2, sweep limits 5 microns × 5 microns, scale 500nm;
Fig. 3 is the projection Electronic Speculum figure of zein granule.The form of the zein granule using anti-solvent technology to prepare is shown in figure;
Fig. 4 is the transmission electron microscope Electronic Speculum figure of embodiment 3 Bacterial cellulose nanofiber, the pure molten protein body of Semen Maydis and erythromycin mixed liquor.The homogeneous dispersion of hydrophilic Bacterial cellulose nanofiber in hydrophobin is shown in figure.
Detailed description of the invention
For understanding the present invention better, below in conjunction with meeting with embodiment, the invention will be further described, but embodiments of the present invention are not limited thereto.
Embodiment 1
The preparation of step 1, bacteria cellulose film
Medium component: 2.0%D-glucose, 0.5% peptone, 0.5% yeast extract, 0.115% citric acid, 0.27%Na
2hPO
412H
2o, all the other are distilled water, pH5.0
Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 10%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
The purification of step 2, bacteria cellulose film
Get the bacteria cellulose film of above-mentioned preparation, 3 times are rinsed with clear water, being soaked in concentration is in 0.2mol/LNaOH solution, takes out, then be soaked in distilled water by this film at 90 DEG C after boiling 30min, 30min is boiled at 90 DEG C, change water, repeat 3 times, be then soaked in distilled water, until neutral, obtain the bacteria cellulose film after purification;
The preparation of step 3, Bacterial cellulose nanofiber aqueous solution
The Bacterial cellulose of above-mentioned purification is put into distilled water, the mass ratio of distilled water and Bacterial cellulose is 1:1, after stirring, with high-speed shearing machine high speed shear 20min under 12000rpm, afterwards by high pressure microjet homogenizing 10 times under 20000psi, obtain Bacterial cellulose nanofiber aqueous solution;
Step 4, be rich in the preparation of biological activity zein nano-particle
Accurately take the thymol that 1.25g zein and 0.25g non-modified are crossed, being dissolved in 25mL ethanol contend percentage composition is in the ethanol-water solution of 80%, obtains zein solution; 1.25g sodium caseinate is dissolved in 62.5mL deionized water, it is poured into rapidly in the zein solution in stirring, uses magnetic stirring apparatus fully to stir and makes its mix homogeneously.After this mixed liquor is placed in Rotary Evaporators, removing ethanol, then under 4000rpm centrifugal 10min, obtain being rich in bioactive zein nano-particle;
The preparation of step 5, Bacterial cellulose nanofiber and zein nano-particle mixed liquor
Mix obtaining Bacterial cellulose nanofiber aqueous solution in step 3 with the bioactive protein body that is rich in that step 4 obtains, Bacterial cellulose nanofiber (dry weight) is 5:1 with the mass ratio of protein body, after stirring, this mixed liquor is placed in ultrasonication machine, selection ultrasonic time is 20min, ultrasonic power is 120W, and work efficiency is 50%, and design temperature is 25 DEG C.
Step 6, vacuum filtration, fast filming
Get mixed liquor obtained in step 5, degassed 30min under the vacuum condition of 1.0MPa, selects the cellulose microporosity filter membrane in 0.65um aperture to carry out vacuum filtration to this mixed liquor, simulates industrial papermaking technology, fast filming, obtains having the biological compounded wet film of high-intensity fine fungin of antibacterial activity;
Step 7, drying
The biological compounded wet film of Bacterial cellulose step 6 obtained is at 90 DEG C, and hot-pressing drying under 0.1MPa, namely obtains the high-intensity fine fungin biological composite membrane material with antibacterial activity.
Comparative example 1
The preparation of step 1, bacteria cellulose film
Medium component: 2.0%D-glucose, 0.5% peptone, 0.5% yeast extract, 0.115% citric acid, 0.27%Na
2hPO
412H
2o, all the other are distilled water, pH4.5.
Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 10%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
The purification of step 2, bacteria cellulose film
Get the bacteria cellulose film of above-mentioned preparation, 3 times are rinsed with clear water, be soaked in 0.2mol/LNaOH solution, take out boil 30min at 90 DEG C after, then this film is soaked in distilled water, 30min is boiled at 90 DEG C, change water, repeat 3 times, be then soaked in distilled water, until neutral, obtain the bacteria cellulose film after purification;
The preparation of step 3, Bacterial cellulose nanofiber aqueous solution
The Bacterial cellulose of above-mentioned purification is put into distilled water, the mass ratio of distilled water and Bacterial cellulose is 1:1, after stirring, with high-speed shearing machine high speed shear 20min under 12000rpm, afterwards by high pressure microjet homogenizing 10 times under 20000psi, obtain Bacterial cellulose nanofiber aqueous solution;
The preparation of step 4, Bacterial cellulose nanofiber mixed liquor
Mix obtaining Bacterial cellulose nanofiber aqueous solution in step 3 according to a certain percentage with distilled water, the mass ratio of nanofiber and distilled water is 1:(500 ~ 1000), after stirring, be placed in ultrasonication machine, selection ultrasonic time is 30min, ultrasonic power is 320W, and work efficiency is 45%, and design temperature is 25 DEG C.
Step 5, vacuum filtration, fast filming
Get mixed liquor obtained in step 4, degassed 30min under the vacuum condition of 1.0MPa, selects the cellulose microporosity filter membrane in 0.65um aperture to carry out vacuum filtration to this mixed liquor, simulates industrial papermaking technology, fast filming, obtain high strength bacterial cellulose wet-coating;
Step 6, drying
The biological compounded wet film of Bacterial cellulose step 5 obtained is at 90 DEG C, and hot-pressing drying under 0.1MPa, namely obtains having high-intensity fine fungin biomembrane material.
Obtained composite film material will be tested above carry out the test of mechanical strength, obtain as following table 1 result:
The contrast of table 1 embodiment 1 and comparative example 1
As shown in Table 1, embodiment 1 is compared with comparative example 1, and the Young's modulus of composite film material, hot strength and elongation at break are all significantly improved, and particularly hot strength improves larger.Illustrate compared with original technology, present invention achieves the good combination of hydrophilic Bacterial cellulose and hydrophobicity zein, the composite film material mechanical performance of preparation gets a promotion, overcome prior art and prepare the defects such as protein-based membrane material mechanical strength is not enough, the composite film material simultaneously prepared has the biological activitys such as specific antibacterial antioxidation, can be applicable to the medical field such as active food packaging material or wound adjuvant.
Embodiment 2
The preparation of step 1, bacteria cellulose film
Medium component: 1.8%D-glucose, 0.45% peptone, 0.45% yeast extract, 0.120% citric acid, 0.25%Na
2hPO
412H
2o, all the other are distilled water, pH4.8.
Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 15%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
The purification of step 2, bacteria cellulose film
Get the bacteria cellulose film of above-mentioned preparation, 3 times are rinsed with clear water, be soaked in 0.4mol/LNaOH solution, take out boil 30min at 90 DEG C after, then this film is soaked in distilled water, 30min is boiled at 90 DEG C, change water, repeat 4 times, be then soaked in distilled water, until neutral, obtain the bacteria cellulose film after purification;
The preparation of step 3, Bacterial cellulose nanofiber aqueous solution
The Bacterial cellulose of above-mentioned purification is put into distilled water, the mass ratio of distilled water and Bacterial cellulose is 1:1, after stirring, with high-speed shearing machine high speed shear 20min under 12000rpm, afterwards by high pressure microjet homogenizing 8 times under 20000psi, obtain Bacterial cellulose nanofiber aqueous solution;
Step 4, be rich in the preparation of biological activity zein nano-particle
Accurately take 1.25g zein and 0.125g without the erythromycin of modification, be dissolved in 25mL80% ethanol-water solution; 1.25g sodium caseinate is dissolved in 62.5mL deionized water, it is poured into rapidly in the alcoholic solution in stirring, uses magnetic stirring apparatus fully to stir and makes its mix homogeneously.After this mixed liquor is placed in Rotary Evaporators, removing ethanol, then under 4000rpm centrifugal 10min, obtain the zein nano-particle with antibiotic property;
The preparation of step 5, Bacterial cellulose nanofiber and zein nano-particle mixed liquor
Mix obtaining Bacterial cellulose nanofiber aqueous solution in step 3 with the bioactive protein body that is rich in that step 4 obtains, Bacterial cellulose nanofiber (dry weight) is 5:2 with the mass ratio of protein body, after stirring, this mixed liquor is placed in ultrasonication machine, selection ultrasonic time is 20min, ultrasonic power is 120W, and work efficiency is 40%, and design temperature is 25 DEG C.
Step 6, vacuum filtration, fast filming
Get mixed liquor obtained in step 5, degassed 30min under the vacuum condition of 1.0MPa, selects the cellulose microporosity filter membrane in 0.45um aperture to carry out vacuum filtration to this mixed liquor, simulates industrial papermaking technology, fast filming, obtains having the biological compounded wet film of high-intensity fine fungin of antibacterial activity;
Step 7, drying
The biological compounded wet film of Bacterial cellulose step 6 obtained is at 90 DEG C, and hot-pressing drying under 0.1MPa, namely obtains the high-intensity fine fungin biological composite membrane material with antibacterial activity.
Comparative example 2
The preparation of step 1, bacteria cellulose film
Medium component: 1.8%D-glucose, 0.45% peptone, 0.45% yeast extract, 0.120% citric acid, 0.25%Na
2hPO
412H
2o, all the other are distilled water, pH4.5.
Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 15%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
The purification of step 2, bacteria cellulose film
Get the bacteria cellulose film of above-mentioned preparation, 3 times are rinsed with clear water, be soaked in 0.4mol/LNaOH solution, take out boil 30min at 90 DEG C after, then this film is soaked in distilled water, 30min is boiled at 90 DEG C, change water, repeat 4 times, be then soaked in distilled water, until neutral, obtain the bacteria cellulose film after purification;
The preparation of step 3, Bacterial cellulose nanofiber aqueous solution
The Bacterial cellulose of above-mentioned purification is put into distilled water, the mass ratio of distilled water and Bacterial cellulose is 1:1, after stirring, with high-speed shearing machine high speed shear 20min under 12000rpm, afterwards by high pressure microjet homogenizing 8 times under 20000psi, obtain Bacterial cellulose nanofiber aqueous solution;
Step 4, be rich in the preparation of biological activity zein nano-particle
Accurately take 1.25g zein, be dissolved in 25mL80% ethanol-water solution; 1.25g sodium caseinate is dissolved in 62.5mL deionized water, it is poured into rapidly in the alcoholic solution in stirring, uses magnetic stirring apparatus fully to stir and makes its mix homogeneously.After this mixed liquor is placed in Rotary Evaporators, removing ethanol, then under 4000rpm centrifugal 10min, obtain the zein nano-particle with antibiotic property;
The preparation of step 5, Bacterial cellulose nanofiber and zein nano-particle mixed liquor
Mix obtaining Bacterial cellulose nanofiber aqueous solution in step 3 with the bioactive protein body that is rich in that step 4 obtains, Bacterial cellulose nanofiber (dry weight) is 5:2 with the mass ratio of protein body, after stirring, this mixed liquor is placed in ultrasonication machine, selection ultrasonic time is 20min, ultrasonic power is 120W, and work efficiency is 40%, and design temperature is 25 DEG C.
Step 6, vacuum filtration, fast filming
Get mixed liquor obtained in step 5, degassed 30min under the vacuum condition of 1.0MPa, selects the cellulose microporosity filter membrane in 0.45um aperture to carry out vacuum filtration to this mixed liquor, simulates industrial papermaking technology, fast filming, obtains having the biological compounded wet film of high-intensity fine fungin of antibacterial activity;
Step 7, drying
The biological compounded wet film of Bacterial cellulose step 6 obtained is at 90 DEG C, and hot-pressing drying under 0.1MPa, namely obtains the high-intensity fine fungin biological composite membrane material with antibacterial activity.
Obtained composite film material will be tested above carry out the test of atomic force microscope and mechanical strength, obtain following result:
The contrast of table 2 embodiment 2 and comparative example 2
As shown in Table 2, embodiment 2 is compared with comparative example 2, and the roughness change on film surface is little, also can find out significantly from accompanying drawing 3 and the atomic plane graph of accompanying drawing 4 composite film surface, two kinds of film surface changes are little, and surface is all in graininess, and roughness is similar.And as can be seen from hot strength and Young's modulus data also, the mechanical strength change of two kinds of films is little, illustrates and add active component on the roughness on film surface and mechanical strength impact thereof not quite.Compared with original technology, this patent, while maintaining bacteria cellulose-base composite film material mechanical strength, also imparts composite film material biological activity, has widened its range of application.
Embodiment 3
The preparation of step 1, bacteria cellulose film
Medium component: 2.2%D-glucose, 0.6% peptone, 0.6% yeast extract, 0.110% citric acid, 0.30%Na
2hPO
412H
2o, all the other are distilled water, pH4.0.
Cultural method: inoculation acetobacter xylinum is to seed culture fluid, and at 30 DEG C after shaken cultivation 12h, the ratio inoculation fermentation culture medium of 12%, shakes up, and quiescent culture 5 days, obtains bacteria cellulose film at 30 DEG C.
The purification of step 2, bacteria cellulose film
Get the bacteria cellulose film of above-mentioned preparation, 3 times are rinsed with clear water, be soaked in 0.2mol/LKOH solution, take out boil 30min at 90 DEG C after, then this film is soaked in distilled water, 30min is boiled at 90 DEG C, change water, repeat 5 times, be then soaked in distilled water, until neutral, obtain the bacteria cellulose film after purification;
The preparation of step 3, Bacterial cellulose nanofiber aqueous solution
The Bacterial cellulose of above-mentioned purification is put into distilled water, the mass ratio of distilled water and Bacterial cellulose is 1:1, after stirring, with high-speed shearing machine high speed shear 20min under 12000rpm, afterwards by high pressure microjet homogenizing 9 times under 20000psi, obtain Bacterial cellulose nanofiber aqueous solution;
Step 4, be rich in the preparation of biological activity zein nano-particle
Accurately take 1.25g zein and 0.5g without the erythromycin of modification, be dissolved in 25mL80% ethanol-water solution; 1.25g sodium caseinate is dissolved in 62.5mL deionized water, it is poured into rapidly in the alcoholic solution in stirring, uses magnetic stirring apparatus fully to stir and makes its mix homogeneously.After this mixed liquor is placed in Rotary Evaporators, removing ethanol, then under 4000rpm centrifugal 10min, obtain the zein nano-particle with antibiotic property;
The preparation of step 5, Bacterial cellulose nanofiber and zein nano-particle mixed liquor
Mix obtaining Bacterial cellulose nanofiber aqueous solution in step 3 with the bioactive protein body that is rich in that step 4 obtains, Bacterial cellulose nanofiber (dry weight) is 5:2 with the mass ratio of protein body, after stirring, this mixed liquor is placed in ultrasonication machine, selection ultrasonic time is 20min, ultrasonic power is 120W, and work efficiency is 60%, and design temperature is 25 DEG C.
Step 6, vacuum filtration, fast filming
Get mixed liquor obtained in step 5, degassed 30min under the vacuum condition of 1.0MPa, selects the cellulose microporosity filter membrane in 0.65um aperture to carry out vacuum filtration to this mixed liquor, simulates industrial papermaking technology, fast filming, obtains having the biological compounded wet film of high-intensity fine fungin of antibacterial activity;
Step 7, drying
The biological compounded wet film of Bacterial cellulose step 6 obtained is at 90 DEG C, and hot-pressing drying under 0.1MPa, namely obtains the high-intensity fine fungin biological composite membrane material with antibacterial activity.
Composite film material obtained is above carried out bacteriostatic test test, obtains result as follows:
The contrast of table 3 embodiment and comparative example's bacteriostatic test
From the contrast of table 3 pair staphylococcus aureus inhibition zone, composite film material prepared by embodiment has larger inhibition zone, comparative example does not then all have inhibition zone, not there is antibiotic property, illustrate that the present invention is after with the addition of active component (antibacterial), really bacterial cellulose composite membrane material material antibacterial activity is imparted, compared with original technology, the present invention has stronger broad spectrum activity, when namely preparing activated protein granule by anti-solvent technology, correspondingly can add dissimilar bioactive ingredients (antibacterial/antioxidation) according to demand, the bacteria cellulose composite membrane correspondingly prepared has the biological activity such as antibacterial/antioxidation accordingly, and there is higher mechanical strength, range of application is more extensive.
Claims (9)
1. the preparation method of biological activity Bacterial cellulose-Compound Film of Zein material, is characterized in that, comprise the following steps:
1) preparation of bacteria cellulose film: utilize acetobacter xylinum strain inoculation to prepare bacteria cellulose film in seed culture fluid;
2) purification of bacteria cellulose film
Be soaked in aqueous slkali after bacteria cellulose film clear water is rinsed, boil; Bacteria cellulose film is soaked in distilled water and boils, change water; Repeat to boil 2-5 time for twice; Then bacteria cellulose film is soaked in distilled water, until neutral;
3) preparation of nanofiber dispersion liquid
By step 2) in bacteria cellulose film after the purification that obtains put into distilled water and stir, shear with high-speed shearing machine, then through high pressure microjet homogenizing, obtain nanofiber dispersion liquid;
4) preparation of the zein nano-particle of bioactive molecule is rich in
Zein and slightly water-soluble bioactive substance are dissolved in ethanol-water solution, obtain alcoholic solution; Sodium caseinate is dissolved in water, adds in described alcoholic solution under stirring, obtain mixed liquor; After stirring, remove ethanol by Rotary Evaporators, centrifugal, obtain the zein nano-particle being rich in bioactive molecule; In described mixed liquor, the mass fraction of zein is 1% ~ 5%, and the mass fraction of bioactive substance is 0.1% ~ 1%, and the mass fraction of sodium caseinate is 1% ~ 5%; Described slightly water-soluble bioactive substance is the antibacterial of slightly water-soluble and/or antioxidant;
5) preparation of Bacterial cellulose nanofiber and zein nano-particle mixed liquor
By step 3) in obtained nanofiber dispersion liquid and step 4) the zein nano-particle being rich in bioactive molecule prepared mixes, stir, and carry out supersound process, obtain nanofiber and zein nano-particle mixed liquor; The mass ratio of described nanofiber and zein granule is (1 ~ 10): 1;
6) the biological compounded wet film preparation of Bacterial cellulose
Nanofiber and zein nano-particle mixed liquor are first carried out vacuum outgas, simulates industrial papermaking technology, vacuum filtration, film forming, obtain that there is the biological compounded wet film of bioactive Bacterial cellulose;
7) dry
The biological compounded wet film of bioactive Bacterial cellulose that has obtained in step (6) is carried out freeze-day with constant temperature or hot-pressing drying, obtains biological activity Bacterial cellulose-Compound Film of Zein.
2. the preparation method according to right 1, is characterized in that, step 3) in, described distilled water and the mass ratio of Bacterial cellulose are (1 ~ 5): 1.
3. the preparation method according to right 1, is characterized in that, step 3) in, the rotating speed of described high-speed shearing machine is 8000 ~ 12000rpm, and shear time is 10 ~ 20min.
4. the preparation method according to right 1, is characterized in that, step 3) in, the homogenization pressure of described high pressure microjet is 5000 ~ 20000psi, and homogenization cycles is 1 ~ 20 time.
5. the preparation method according to right 1, is characterized in that, step 4) in, in ethanol-water solution, ethanol mass concentration is 60% ~ 95%; Described centrifugal rotating speed is 1000rpm ~ 5000rpm, and the centrifugal time is 10min ~ 30min; Antibacterial and/or the antioxidant of described slightly water-soluble is erythromycin, thymol or resveratrol.
6. the preparation method according to right 1, is characterized in that, step 5) in, the time of supersound process is 10min ~ 20min, and ultrasonic power is 100W ~ 200W, and work efficiency is 40% ~ 60%, and operating temperature is 20 DEG C ~ 30 DEG C.
7. the preparation method according to right 1, is characterized in that, step 6) in, the time of described vacuum outgas is 20min ~ 60min; Described vacuum filtration selects aperture to be that the cellulose microporosity filter membrane of 0.45-0.65 μm carries out vacuum filtration, fast filming to bacterial fibers and zein granule mixed liquor.
8. the preparation method according to right 1, is characterized in that, step 7) in, described freeze-day with constant temperature is 25 ~ 60 DEG C of freeze-day with constant temperature; Described hot-pressing drying is 85 ~ 100 DEG C, carries out drying under 0.1 ~ 0.2MPa.
9. the preparation method according to right 1, it is characterized in that, described utilize acetobacter xylinum strain inoculation to prepare bacteria cellulose film be get activation in seed culture fluid after acetobacter xylinum strain inoculation in seed culture fluid, shaken cultivation, mass ratio by 1 ~ 10% is inoculated in fermentation culture, shake up, quiescent culture, obtain bacteria cellulose film.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410421901.8A CN104225669B (en) | 2014-08-25 | 2014-08-25 | Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410421901.8A CN104225669B (en) | 2014-08-25 | 2014-08-25 | Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104225669A CN104225669A (en) | 2014-12-24 |
| CN104225669B true CN104225669B (en) | 2016-04-20 |
Family
ID=52215138
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410421901.8A Active CN104225669B (en) | 2014-08-25 | 2014-08-25 | Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104225669B (en) |
Families Citing this family (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105833738B (en) * | 2016-05-05 | 2019-02-05 | 北京科技大学 | Nano-cellulose/soybean protein composite filter material and preparation method, purposes |
| CN106234914B (en) * | 2016-07-30 | 2019-08-09 | 江南大学 | A kind of preparation method and applications of the bacteriostatic agent containing Herba Origani Vulgaris quintessence oil and resveratrol |
| CN106727428A (en) * | 2016-12-23 | 2017-05-31 | 青岛农业大学 | A kind of rutin zeins casein sodium composite nanoparticle and preparation method thereof |
| CN106983871A (en) * | 2017-03-31 | 2017-07-28 | 中国农业大学 | A kind of preparation method of large biological molecule compound stable state quercetagetin |
| CN107569718A (en) * | 2017-08-30 | 2018-01-12 | 哈尔滨工业大学 | A kind of preparation method of 3D printing PLA and bacteria cellulose compound support frame material |
| CN107880567A (en) * | 2017-11-24 | 2018-04-06 | 四川农业大学 | A kind of wheat-bran fiber element/gluten protein composite membrane and preparation method thereof |
| CN109771693A (en) * | 2018-04-13 | 2019-05-21 | 中国人民解放军第五医院 | A kind of preparation method for the new injectable spontaneous coagulation cmposite artificial bone carrying rhBMP_2 microballoon |
| CN108456966B (en) * | 2018-06-14 | 2021-06-01 | 汕头市富莱美服饰实业有限公司 | Preparation method of environment-friendly soft tencel fabric |
| CN109180990A (en) * | 2018-07-23 | 2019-01-11 | 德清顾舒家华高分子材料有限公司 | A kind of preparation method of composite aerogel sponge |
| CN109369961B (en) * | 2018-10-16 | 2020-11-20 | 河南工程学院 | Polypeptide-enhanced nanocellulose-based thin film material and preparation method thereof |
| CN111117000A (en) * | 2018-10-31 | 2020-05-08 | 西北农林科技大学 | Bacterial cellulose as performance modifier of biodegradable film |
| CN109289079B (en) * | 2018-11-08 | 2021-09-17 | 广州润虹医药科技股份有限公司 | Medical dressing for inhibiting scars and preparation method and application thereof |
| CN109776872A (en) * | 2018-12-04 | 2019-05-21 | 厦门大学 | A kind of water-resistant degradable thin-film material and preparation method thereof |
| CN115141409A (en) * | 2021-03-30 | 2022-10-04 | 海南光宇生物科技有限公司 | Bacterial cellulose and plant protein composite material and preparation method and application thereof |
| CN113016933B (en) * | 2021-04-26 | 2021-12-07 | 江南大学 | Preparation method of antioxidant fiber additive for vegetable protein meat |
| CN115584657B (en) * | 2021-07-06 | 2024-04-12 | 中国科学院理化技术研究所 | Hydrophobic slurry and preparation method and application thereof |
| CN114588127A (en) * | 2022-03-04 | 2022-06-07 | 辽宁大学 | Modified zein nano drug delivery system wrapped by outer membrane vesicles of bacteria and preparation method and application thereof |
| CN114534527B (en) * | 2022-04-18 | 2023-07-04 | 重庆文理学院 | Membrane filtration assembly |
| CN115232367B (en) * | 2022-07-11 | 2024-03-22 | 华南理工大学 | Nanocellulose-based antibacterial anti-oxidation hydrophobic film and preparation method and application thereof |
| CN116473872B (en) * | 2023-05-26 | 2023-11-07 | 微赛生物技术(广州)有限公司 | Biological fiber mask substrate and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102786702A (en) * | 2012-07-18 | 2012-11-21 | 华南理工大学 | Preparation method of controlled release antibacterial film |
| CN102827388A (en) * | 2012-08-29 | 2012-12-19 | 华南理工大学 | Hydrophobic controlled-release antibacterial film and preparation method thereof |
| CN103357060A (en) * | 2013-07-23 | 2013-10-23 | 海南光宇生物科技有限公司 | Method for preparing bacterial cellulose composite fish collagen wound dressing |
| CN103893820A (en) * | 2013-11-06 | 2014-07-02 | 浙江经贸职业技术学院 | Silk fibroin composite biomaterial scaffold and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| PL212003B1 (en) * | 2003-07-03 | 2012-07-31 | Politechnika Łodzka | Method for obtaining bacterial cellulose, method for bacteria immobilization, method for obtaining immobilized biocatalysts, application of bacterial cellulose, method for modification of cellulose membranes |
-
2014
- 2014-08-25 CN CN201410421901.8A patent/CN104225669B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102786702A (en) * | 2012-07-18 | 2012-11-21 | 华南理工大学 | Preparation method of controlled release antibacterial film |
| CN102827388A (en) * | 2012-08-29 | 2012-12-19 | 华南理工大学 | Hydrophobic controlled-release antibacterial film and preparation method thereof |
| CN103357060A (en) * | 2013-07-23 | 2013-10-23 | 海南光宇生物科技有限公司 | Method for preparing bacterial cellulose composite fish collagen wound dressing |
| CN103893820A (en) * | 2013-11-06 | 2014-07-02 | 浙江经贸职业技术学院 | Silk fibroin composite biomaterial scaffold and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104225669A (en) | 2014-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104225669B (en) | Biological activity Bacterial cellulose-Compound Film of Zein and preparation method thereof | |
| CN103866487B (en) | The preparation method of a kind of nano micro crystal cellulose/chitosan/polyvinyl alcohol composite nanometer film | |
| CN107556495B (en) | A kind of preparation method of functional nano cellulose composite aerogel | |
| CN113354853B (en) | Biodegradable high-barrier-property antibacterial composite membrane and preparation method thereof | |
| CN105646951B (en) | The preparation method of functional solvable edible composite membrane | |
| CN101372536B (en) | Preparation of bacteria cellulose food fresh keeping membrane | |
| CN106220896B (en) | It is a kind of flexible with high-moisture Cellulose/Chitosan base plural gel, its corresponding composite membrane and application | |
| CN106928908A (en) | A kind of new aeroge multifunctional material and preparation method thereof | |
| CN103691005B (en) | A kind of micro--Na fibrous tissue engineering rack and preparation method thereof | |
| CN103361885A (en) | Preparation method of antibacterial silk fibroin fibrous membrane | |
| CN104558677A (en) | Preparation method of nano cellulose/chitosan composite foam | |
| CN106676753A (en) | Antibacterial nano fiber and preparation method thereof | |
| TWI398276B (en) | Method for producing bacterial cellulose with controlled pore size and its use in wound dressing | |
| CN109233231A (en) | A method of lignin/polylactic acid laminated film is prepared using Pickering lotion | |
| CN104947320B (en) | A kind of bio-medical energy-saving and temperature-regulating tunica fibrosa and preparation method thereof | |
| CN105536032A (en) | Preparation method of nanofiber antimicrobial dressing blended from nano-silver containing sericin and polyoxyethylene | |
| CN102321261B (en) | The preparation method of a kind of anti-virus bacteria cellulose protective material and goods thereof | |
| CN103882624A (en) | Preparation method for water-resistant nano microcrystalline cellulose/polyvinyl alcohol/polyacrylic acid/silicon dioxide composite nanofiber membrane | |
| CN104225670B (en) | The preparation method of the hydrophobic Bacterial cellulose-Compound Film of Zein of controllability | |
| CN104153120A (en) | Antibacterial medical dressing film carrying nano-silver and cellulose nanocrystalline hybrid materials and preparation method thereof | |
| CN103194856A (en) | Preparation method of chitosan/sericin composite nanometer fibers with antibiosis and wound healing effects | |
| CN107519540A (en) | A kind of implantable fibroin/bacteria cellulose/graphene composite conductive film of high tensile flexible printing opacity | |
| CN107286546B (en) | A kind of preparation method of Biodegradable high-molecular film | |
| CN112877384A (en) | Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof | |
| Gu et al. | Natural lignin nanoparticles: a promising nano-crosslinker for constructing fluorescent photoswitchable supramolecular hydrogels |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |