CN103451252B - Chrysanthemum shape bacteria cellulose and its fermentation preparation - Google Patents
Chrysanthemum shape bacteria cellulose and its fermentation preparation Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 145
- 229920002678 cellulose Polymers 0.000 title claims abstract description 140
- 239000001913 cellulose Substances 0.000 title claims abstract description 139
- 241000723353 Chrysanthemum Species 0.000 title claims abstract description 91
- 235000007516 Chrysanthemum Nutrition 0.000 title claims abstract description 91
- 238000000855 fermentation Methods 0.000 title claims abstract description 55
- 230000004151 fermentation Effects 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 238000005457 optimization Methods 0.000 claims abstract description 16
- 230000002093 peripheral effect Effects 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- 239000012153 distilled water Substances 0.000 claims description 15
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 230000010355 oscillation Effects 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 241000032681 Gluconacetobacter Species 0.000 claims description 5
- 241000589216 Komagataeibacter hansenii Species 0.000 claims description 5
- 229920001410 Microfiber Polymers 0.000 claims description 4
- 239000003658 microfiber Substances 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 239000004310 lactic acid Substances 0.000 claims description 3
- 235000014655 lactic acid Nutrition 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 238000004804 winding Methods 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 2
- 238000000280 densification Methods 0.000 abstract description 3
- 238000000034 method Methods 0.000 abstract description 2
- 235000010980 cellulose Nutrition 0.000 description 125
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 32
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 30
- 238000004088 simulation Methods 0.000 description 25
- 229930014669 anthocyanidin Natural products 0.000 description 23
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 23
- 235000008758 anthocyanidins Nutrition 0.000 description 23
- 238000002474 experimental method Methods 0.000 description 23
- 229930003944 flavone Natural products 0.000 description 23
- 150000002213 flavones Chemical class 0.000 description 23
- 235000011949 flavones Nutrition 0.000 description 23
- 235000003095 Vaccinium corymbosum Nutrition 0.000 description 22
- 235000017537 Vaccinium myrtillus Nutrition 0.000 description 22
- 235000021014 blueberries Nutrition 0.000 description 22
- 240000000851 Vaccinium corymbosum Species 0.000 description 21
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 14
- 229930003268 Vitamin C Natural products 0.000 description 14
- 210000004051 gastric juice Anatomy 0.000 description 14
- 239000000243 solution Substances 0.000 description 14
- 235000019154 vitamin C Nutrition 0.000 description 14
- 239000011718 vitamin C Substances 0.000 description 14
- 230000000968 intestinal effect Effects 0.000 description 13
- 239000002609 medium Substances 0.000 description 12
- 238000010521 absorption reaction Methods 0.000 description 8
- 238000010586 diagram Methods 0.000 description 8
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- 239000000835 fiber Substances 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 6
- 235000016709 nutrition Nutrition 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 150000007946 flavonol Chemical class 0.000 description 4
- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Natural products O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 description 4
- 235000011957 flavonols Nutrition 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000013268 sustained release Methods 0.000 description 3
- 239000012730 sustained-release form Substances 0.000 description 3
- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 239000011609 ammonium molybdate Substances 0.000 description 2
- 235000018660 ammonium molybdate Nutrition 0.000 description 2
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 2
- 229940010552 ammonium molybdate Drugs 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
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- 239000006071 cream Substances 0.000 description 2
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- 235000015097 nutrients Nutrition 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
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- 238000002791 soaking Methods 0.000 description 2
- 239000000052 vinegar Substances 0.000 description 2
- 235000021419 vinegar Nutrition 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 241000589220 Acetobacter Species 0.000 description 1
- 229920002749 Bacterial cellulose Polymers 0.000 description 1
- 244000283207 Indigofera tinctoria Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Chrysanthemum shape bacteria cellulose and its fermentation preparation.It is related to a kind of bacteria cellulose and its fermentation preparation.It provides a kind of chrysanthemum shape bacteria cellulose for being shaped differently than existing spherical bacteria cellulose, and there is provided the method that fermentation prepares chrysanthemum shape bacteria cellulose.The shape of chrysanthemum shape bacteria cellulose is in chrysanthemum shape, and peripheral thorn is more and long, and milky is glossy, homogeneous densification.Preparation method：First, seed fermentation liquid is prepared；2nd, the seed fermentation liquid of step one shaking table culture is seeded in optimization fermentation medium, then ferments, that is, obtain chrysanthemum shape bacteria cellulose.The present invention is used for bacteria cellulose and produces preparation field.
Description
Technical field
The present invention relates to a kind of bacteria cellulose and its fermentation preparation.
Background technology
Bacteria cellulose (Bacterial Cellulose, abbreviation BC) is a kind of bacterial polysaccharides, is had with animal and plant fiber element
By similar structure and constitute, and with the incomparable high-purity of animal and plant fiber element, high polymerization degree, high-crystallinity, high parent
Unique property such as aqueous, hyperfine network structure and high biocompatibility.Because bacteria cellulose has biosynthesis can
Control, structure modifiability, do not need the operational advantage such as pre-treatment and purification, therefore, in food industry, biomedicine, make
It is widely applied in terms of paper, acoustics equipment and oil exploitation.
The content of the invention
The chrysanthemum shape bacteria cellulose of existing spherical bacteria cellulose is shaped differently than the invention provides a kind of, and is provided
The method that fermentation prepares chrysanthemum shape bacteria cellulose.
The shape of chrysanthemum shape bacteria cellulose is in chrysanthemum shape, and peripheral thorn is more and long, and milky is glossy, homogeneous
It is fine and close.
Above-mentioned chrysanthemum shape bacteria cellulose ferments preparation according to the following steps：
First, by Han Shi gluconacetobacters HDM1-3（Gluconacetobacter hansenii HDM1-3）Streak inoculation
In bacteria cellulose solid medium, then the constant temperature quiescent culture 48h under the conditions of 28 DEG C is inoculated with oese picking lawn
In 100ml seed fermentation liquid, remix it is uniform after be placed in the shaking table that temperature is 28 DEG C, revolution is 130r/min and cultivate 24h；
2nd, the seed fermentation liquid of step one shaking table culture is seeded to by the inoculum concentration of optimization fermented liquid fraction 7%
In 75ml optimization fermentation medium, then temperature be 28 DEG C, rotating speed be oscillation and fermentation 48h under conditions of 150r/min, i.e.,
Obtain chrysanthemum shape bacteria cellulose；
Wherein, in step one bacteria cellulose solid medium by mass percentage by 5% glucose, 0.5% yeast
Cream, 0.5% peptone, 0.1% citric acid, 0.2% Na2HPO4·H2O, 0.1% K2HPO4, 0.025% MgSO4·
7H2O, 2% agar and surplus distilled water composition, pH value is 5.8；Seed fermentation liquid is by mass percentage by 2% in step one
Sucrose, 0.3% yeast extract, 0.5% peptone, 0.2% potassium dihydrogen phosphate, the distillation of 0.015% magnesium sulfate and surplus
Water is constituted, and pH value is 5.8；Step 2 optimizes glucose, 0.34g beef extract, 0.19g of the every 100ml of fermentation medium by 5g
Yeast extract, 0.22g disodium hydrogen phosphate, 0.46g dipotassium hydrogen phosphate, 0.2g lactic acid, 0.025g magnesium sulfate, 1ml
Ethanol and the distilled water of surplus composition, pH value is 6.5.
Not only shape is novel for chrysanthemum shape bacteria cellulose of the present invention, good appearance and with more excellent adsorptivity,
Viscosity, adhesivity and chewiness, elasticity, glutinous poly- property and resilience are also better than spherical bacteria cellulose.
Modulatory character and shape-plastic of the present invention based on bacteria cellulose biosynthesis, by adjusting fermentation process,
Biogenic reworking is carried out, is produced a kind of suitable for food, medicine trade, the high chrysanthemum shape bacterial fibers of good appearance, added value
Element.Chrysanthemum shape bacterial fibers have substantial amounts of " petal ", while increasing attractive in appearance, increase the table of chrysanthemum shape bacteria cellulose
Area, therefore chrysanthemum shape bacteria cellulose has stronger adsorption capacity and good sustained release performance.Long and many furcella thing energy
Increase the surface area of chrysanthemum shape bacteria cellulose, increase its ability and releasability for adsorbing nutriment.If being added into indigo plant
In certain kind of berries fruit juice, nutrition adsorption capacity is added, and because of its preferable sustained release performance, extends its residence time in enteral,
Reach that nutrition continues the purpose slowly supplied.
Milky gloss and the smooth chrysanthemum shape bacteria cellulose of tissue surface, illustrate the structure of chrysanthemum shape bacteria cellulose
Surface is more uniform, and preferably, fermentation condition is suitable, is conducive to the absorption of nutriment for elasticity.
Han Shi gluconacetobacters HDM1-3 (Gluconacetobacter hansenii HDM1-3) is Han Shi glucose vinegar bars
Bacterium（Gluconacetobacter hansenii）, belong to glucose acetobacter (Gluconacetobacter)；In being preserved in
State's Type Tissue Collection, preservation address is Wuhan City, and Wuhan University, preservation date is on December 5th, 2010, preserving number
For CCTCC NO：M2010332.
Brief description of the drawings
Fig. 1 is the observation figure of the chrysanthemum shape bacteria cellulose of embodiment one；
Fig. 2 is the chrysanthemum shape bacteria cellulose scanning electron microscopic observation figure of embodiment two；
Fig. 3 is chrysanthemum shape bacteria cellulose vitamin under artificial simulation gastric juices environment in the release experiment of embodiment two
C releasing curve diagrams.
Fig. 4 is that chrysanthemum shape bacteria cellulose flavones is released under artificial simulation gastric juices environment in the release experiment of embodiment two
Put curve map.
Fig. 5 is chrysanthemum shape bacteria cellulose anthocyanidin under artificial simulation gastric juices environment in the release experiment of embodiment two
Releasing curve diagram.
Fig. 6 is spherical bacteria cellulose vitamin C under artificial simulation gastric juices environment in the release experiment of embodiment two
Releasing curve diagram.
Fig. 7 is that spherical bacteria cellulose flavones discharges under artificial simulation gastric juices environment in the release experiment of embodiment two
Curve map.
Fig. 8 is that spherical bacteria cellulose anthocyanidin is released under artificial simulation gastric juices environment in the release experiment of embodiment two
Put curve map.
Fig. 9 is chrysanthemum shape bacteria cellulose vitamin under artificial simulation intestinal juice environment in the release experiment of embodiment two
C releasing curve diagrams.
Figure 10 is chrysanthemum shape bacteria cellulose flavones under artificial simulation intestinal juice environment in the release experiment of embodiment two
Releasing curve diagram.
Figure 11 is chrysanthemum shape bacteria cellulose cyanine under artificial simulation intestinal juice environment in the release experiment of embodiment two
Plain releasing curve diagram.
Figure 12 is spherical bacteria cellulose vitamin C under artificial simulation intestinal juice environment in the release experiment of embodiment two
Releasing curve diagram.
Figure 13 is that spherical bacteria cellulose flavones is released under artificial simulation intestinal juice environment in the release experiment of embodiment two
Put curve map.
Figure 14 is spherical bacteria cellulose anthocyanidin under artificial simulation intestinal juice environment in the release experiment of embodiment two
Releasing curve diagram.
Figure 15 is that chrysanthemum shape bacteria cellulose vitamin C discharges under distillation water environment in the release experiment of embodiment two
Curve map.
Figure 16 is that the release of chrysanthemum shape bacteria cellulose flavones is bent under distillation water environment in the release experiment of embodiment two
Line chart.
Figure 17 is that chrysanthemum shape bacteria cellulose anthocyanidin discharges under distillation water environment in the release experiment of embodiment two
Curve map.
Figure 18 is that the release of spherical bacteria cellulose vitamin C is bent under distillation water environment in the release experiment of embodiment two
Line chart.
Figure 19 is spherical bacteria cellulose flavones release profiles under distillation water environment in the release experiment of embodiment two
Figure.
Figure 20 is that the release of spherical bacteria cellulose anthocyanidin is bent under distillation water environment in the release experiment of embodiment two
Line chart.
Embodiment
Technical solution of the present invention is not limited to act embodiment set forth below, in addition between each embodiment
Any combination.
Embodiment one：The shape of present embodiment chrysanthemum shape bacteria cellulose is in chrysanthemum shape, and peripheral thorn is more
And it is long, milky is glossy, homogeneous densification.
Present embodiment chrysanthemum shape bacteria cellulose is as shown in figure 1, present embodiment chrysanthemum shape bacteria cellulose periphery thorn
Shape thing is more sturdy.
Embodiment two：Present embodiment chrysanthemum shape bacteria cellulose ferments preparation according to the following steps：
First, by Han Shi gluconacetobacters HDM1-3（Gluconacetobacter hansenii HDM1-3）Streak inoculation
In bacteria cellulose solid medium, then the constant temperature quiescent culture 48h under the conditions of 28 DEG C is inoculated with oese picking lawn
In 100ml seed fermentation liquid, remix it is uniform after be placed in the shaking table that temperature is 28 DEG C, revolution is 130r/min and cultivate 24h；
2nd, the seed fermentation liquid of step one shaking table culture is seeded to by the inoculum concentration of optimization fermented liquid fraction 7%
In 75ml optimization fermentation medium, then temperature be 28 DEG C, rotating speed be oscillation and fermentation 48h under conditions of 150r/min, i.e.,
Obtain chrysanthemum shape bacteria cellulose；
Wherein, in step one bacteria cellulose solid medium by mass percentage by 5% glucose, 0.5% yeast
Cream, 0.5% peptone, 0.1% citric acid, 0.2% Na2HPO4·H2O, 0.1% K2HPO4, 0.025% MgSO4·
7H2O, 2% agar and surplus distilled water composition, pH value is 5.8；Seed fermentation liquid is by mass percentage by 2% in step one
Sucrose, 0.3% yeast extract, 0.5% peptone, 0.2% potassium dihydrogen phosphate, the distillation of 0.015% magnesium sulfate and surplus
Water is constituted, and pH value is 5.8；Step 2 optimizes glucose, 0.34g beef extract, 0.19g of the every 100ml of fermentation medium by 5g
Yeast extract, 0.22g disodium hydrogen phosphate, 0.46g dipotassium hydrogen phosphate, 0.2g lactic acid, 0.025g magnesium sulfate, 1ml
Ethanol and the distilled water of surplus composition, pH value is 6.5.
ESEM（SEM）The surface texture featur of present embodiment chrysanthemum shape bacteria cellulose is observed, dry chrysanthemum is taken
Flower-shaped bacteria cellulose 1cm2Left and right, is adhesive on testboard with two-sided, directly in metalling platinum on ion sputtering instrument, then
It is as shown in Figure 2 that its microstructure is observed in the secure execution mode (sem.The filament of present embodiment chrysanthemum shape bacteria cellulose is very very thin（<
0.1μm）, and ultra-microfibers mutually wind, and constitute ultra-microfibers net, while existing, silk ribbon stratiform is overlapping, silk ribbon is more uniform,
And with pore space structure.The microstructure of chrysanthemum shape bacteria cellulose determines its peculiar property, such as has very strong
Hydrophily, good water penetration and adsorptivity etc., therefore chrysanthemum shape bacteria cellulose has and is more widely applied space.
By with spherical bacteria cellulose it was found that, the mycelia of chrysanthemum shape bacteria cellulose is thinner, and mycelia winding nothing
Rule；And the mycelia of spherical bacteria cellulose is thicker, and mycelia winding is orderly.
Texture TPA is detected：
Using instrumental test hygrometric state chrysanthemum shape bacteria cellulose and spherical bacteria cellulose.Popped one's head in using P/5, set and survey
Preceding speed is 2.00mm/s, and speed is 5.00mm/s after survey, and measuring distance is 70%, and trigger force is 0.1N, and the residence time is 10s.
Each sample retest 5 times, location parameter：Viscosity, elasticity, glutinous poly- property, gumminess, chewiness and recovery.Testing result is such as
Shown in table 1.
Table 1
| Title | Chrysanthemum shape bacteria cellulose | Spherical bacteria cellulose |
| Viscosity | -178.3968±7.00 | -141.5784±24.10 |
| Elasticity | 0.9702±0.01 | 0.9508±0.03 |
| Stick poly- property | 0.652±0.03 | 0.6392±0.08 |
| Gumminess | 469.9096±21.01 | 321.76±63.86 |
| Chewiness | 455.9746±22.69 | 307.1314±68.15 |
| Resilience | 0.0926±0.02 | 0.0742±0.02 |
Texture testing result shows, viscosity, gumminess and the chewiness of chrysanthemum shape bacteria cellulose are significantly better than spherical thin
Fungin, elasticity, glutinous poly- property and resilience are also better than spherical bacteria cellulose.
Adsorption experiment：
The chrysanthemum shape bacteria cellulose that spherical bacteria cellulose and present embodiment are obtained carries out nutrition in blueberry juice
Composition anthocyanidin, flavones and ascorbic adsorption experiment.
Experimental procedure：First, mucus and the bacterium of chrysanthemum shape bacteria cellulose and spherical bacteria cellulose surface residual are gone in washing
Body, is immersed in 0.1mol/L NaOH solution, boiling water boiling 1h, then carries out neutralisation treatment 1h with glacial acetic acid, repeatedly soaking flushing
Remove, untill the close neutrality of pH value, chrysanthemum shape bacteria cellulose and spherical bacteria cellulose are immersed in sterile completely to tart flavour
It is standby in water, using preceding by chrysanthemum shape bacteria cellulose and spherical bacteria cellulose drying process.
2nd, blueberry is put into after being dried in 60 DEG C of baking ovens and smashed, added 100mL distilled water, be filtrated to get blueberry juice.
3rd, the spherical bacteria cellulose and chrysanthemum of 0.8g step one drying process are separately added into two parts of 10mL blueberry juices
Shape bacteria cellulose, immersion 24h makes spherical and chrysanthemum shape bacteria cellulose fully adsorb blueberry juice, after outwell unnecessary blueberry juice.
Weigh the spherical and chrysanthemum shape bacteria cellulose after absorption.By the anthocyanidin of making, flavones and ascorbic standard curve,
Bacteria cellulose absorption anthocyanidin, flavones and ascorbic content for calculating spherical and chrysanthemum shape.
1. the standard curve of anthocyanidin：The blueberry juice for being put into chrysanthemum shape bacteria cellulose and spherical bacteria cellulose is distinguished
It is placed in 50mL triangular flasks, adds 0.1mol/L HCl solution 10mL, rim of a cup is covered tightly with pan paper to prevent moisture evaporation, is placed in 32
In DEG C incubator, impregnate 4 hours.The solution after dipping is taken to survey absorbance under wavelength 530nm with optical path 1cm cuvette, with
0.1mol/L Hcl are blank control, with optical path 1cm cuvette.Calculate the relative amount of anthocyanidin：Content=10 of anthocyanidin
×A×B（A：The absorbance measured, B：Extension rate）.
2. the standard curve of flavones：Accurately flavonol standards product 20mg/L is weighed, plus methanol dissolves and is settled to 100mL.
Flavonol standards solution is made.It is accurate draw flavonol standards solution 1.00,2.00,3.00,4.00,5.00,6.00mL in
In 50mL scale colorimetric cylinders, plus ethanol to cumulative volume is 15mL, sequentially adds aluminum nitrate solution 1mL, liquor kalii acetici 1mL, shakes
Even standing 1h.With cuvette at 414nm, using 30% ethanol solution as blank control, absorbance is determined, flavonol standards are formulated
Curve.
The blueberry juice 1mL into bacteria cellulose is picked and placeed, ethanol is added and is settled to 15mL, add aluminum nitrate solution 1mL, vinegar
Sour potassium solution 1mL, places 1h, determines absorbance, calculates flavones alcohol content.
3. ascorbic standard curve：The accurate vitamin C 0.05g for weighing 60 DEG C of vacuum drying 2h, with oxalic acid-EDTA
Solution constant volume makes concentration of standard solution reach 1mg/mL in 500mL volumetric flasks；Standard vitamin c solution is made.Draw respectively
0.4th, 0.6 it is molten that, 0.8,1.0,1.2,1.4mL standard vitamin c solution adds oxalic acid-EDTA in 50mL volumetric flask, afterwards
Liquid, makes cumulative volume reach 1000mL.1.00mL metaphosphoric acid-acetum and 5% sulfuric acid 2.0mL is added, addition is shaken up
4.00mL ammonium molybdate, using distilled water as blank control, surveys absorbance under 705nm, formulates standard curve.
Blueberry juice addition metaphosphoric acid-acetum and 5% sulfuric acid 2mL that 50mL is put into bacteria cellulose are drawn, is shaken
It is even, 4mL ammonium molybdate solution is added, absorbance is surveyed after being settled to 50mL, 15min with distilled water, calculates Vitamin C content.
Experimental result finds chrysanthemum shape and spherical bacteria cellulose being put into 10mL blueberry juices, chrysanthemum shape bacteria cellulose
Absorption anthocyanidin, flavones and ascorbic effect are substantially better than spherical bacteria cellulose.The chrysanthemum shape bacterial fibers that 0.8g is dried
Element absorption anthocyanidin：13.6209mg, spherical bacteria cellulose absorption anthocyanidin：4.5386mg；Chrysanthemum shape bacteria cellulose is adsorbed
Flavones：3.6094mg, spherical bacteria cellulose absorption flavones：1.2027mg；Chrysanthemum shape bacteria cellulose adsorbs vitamin C：
1.8782mg, the vitamin C of spherical bacteria cellulose absorption：0.6258mg.
Release experiment：
The chrysanthemum shape bacteria cellulose that spherical bacteria cellulose and present embodiment are obtained is immersed in blueberry juice
48h, is then placed in artificial simulation gastric juices, manual simulation's gastro-intestinal Fluid and distilled water, and blueberries are discharged to two kinds of bacteria celluloses
Anthocyanidin, flavones and ascorbic ability are tested in juice.
Experimental procedure：First, it will clean and remove the chrysanthemum shape bacteria cellulose of the identical weight of surface moisture and spherical thin
Fungin is put into the blueberry juice that soaking at room temperature in blueberry juice draws phase homogenous quantities.
2nd, by chrysanthemum shape and spherical bacteria cellulose, it is separately immersed in the artificial simulation gastric juices of isodose, detection 0h,
1h, 2h, 3h, 4h chrysanthemum shape bacteria cellulose and spherical bacteria cellulose discharge the situation of blueberry nutrient material.
3rd, by chrysanthemum shape and spherical bacteria cellulose, it is separately immersed in the artificial simulation gastric juices of isodose after 4h, then divide
Not Jia Ru isodose artificial simulation intestinal juice, respectively detect after 0h, 1h, 2h chrysanthemum shape bacteria cellulose and spherical bacteria fiber
The situation of element release blueberry nutrient material.
4th, by chrysanthemum shape and spherical bacteria cellulose, be separately immersed in the distilled water of isodose, respectively detect 0h, 1h,
2h, 3h, 4h chrysanthemum shape and spherical bacteria cellulose discharge the situation of nutriment in distilled water.
In artificial simulation gastric juices, dimension of the chrysanthemum shape bacteria cellulose in 0h, 1h, 2h, 3h and 4h discharge blueberry juice
Raw element C, flavones and anthocyanidin situation can be seen that the release of the vitamin C, flavones and anthocyanidin of chrysanthemum shape bacteria cellulose
Amount, increase is discharged at 0~1, and release starts reduction during 1~2h, and release during 3h reaches maximum, and presentation is that " M " shape is bent
Line（As seen in figures 3-5）.
In artificial simulation gastric juices, dimension life of the spherical bacteria cellulose in 0h, 1h, 2h, 3h and 4h discharge blueberry juice
Plain C, flavones and anthocyanidin can be seen that in 0~4h that burst size substantially persistently rises（As can be seen from figures 6 to 8）.
It can be seen that the rate of release of chrysanthemum shape bacteria cellulose and spherical bacteria fiber have according to burst size curve map
Very big difference, is calculated by anthocyanidin, flavones and vitamin C standard curve and found, the totality of chrysanthemum shape bacteria cellulose is released
High-volume it is more than spherical bacteria cellulose.Effect of the chrysanthemum shape bacteria cellulose with sustained release in gastric juice, the big advantage of burst size,
It is particularly suitable as the carrier that shaped form in gastric juice discharges medicine.
In artificial simulation intestinal juice, chrysanthemum shape bacteria cellulose 0h, 1h and 2h discharge blueberry juice in vitamin C,
Flavones and anthocyanidin situation as shown in Fig. 9~11, in artificial simulation gastric juices chrysanthemum shape bacteria cellulose not by nutrition into
Divide all release, remaining nutritional ingredient is discharged in artificial simulation intestinal juice with " A " shape curve, start to increase in 0~1h releases
Greatly, release reaches maximum during 1h, and 1~2h releases start reduction.
Vitamin C, flavones of the spherical bacteria cellulose in 0h, 1h and 2h discharge blueberry juice in artificial simulation intestinal juice
With anthocyanidin during 0~2h releases, burst size is constantly increasing, and rate of release is relatively slow as shown in Figure 12~14, it is impossible to
Reach and effectively discharge in short time, cause the waste of nutriment.
Release of the chrysanthemum shape bacteria cellulose in artificial simulation intestinal juice reaches saturation point in 1h, with release time
Increase, burst size is constantly reduced；Its burst size and rate of release are higher than spherical bacteria cellulose, are particularly suitable for medicine in intestinal juice and release
The medicine slowly declined after high-volume reaching a certain height.
In distilled water, vitamin C, flavones of the chrysanthemum shape bacteria cellulose in 0h, 1h, 2h and 3h discharge blueberry juice
And anthocyanidin.It is release constantly increase in 0~1h, in 1h, release reaches maximum, and 1~3h releases start straight line and declined, finally
It is released to zero（As seen in figs. 15-17）.Spherical bacteria cellulose 0h, 1h, 2h and 3h discharge blueberry juice in vitamin C,
Flavones and anthocyanidin are in the slow reduction of 0~3h releases（As shown in Figure 18~20）.Found by calculating, chrysanthemum shape bacteria cellulose
Be higher than spherical bacteria cellulose on total volume, rate of release apparently higher than spherical bacteria cellulose, but in total volume and
Effect in rate of release all not in simulation stomach, intestinal juice is good.
The rate of release of chrysanthemum shape bacteria cellulose is higher than spherical bacteria cellulose in distilled water, and in 1h, chrysanthemum
The burst size of flower-shaped bacteria cellulose is significantly increased higher than initial content.
Release test proves that the burst size of chrysanthemum shape bacteria cellulose is higher than the content of spherical bacteria cellulose.
Embodiment three：The difference of present embodiment and embodiment two is：
Step 2: the seed fermentation liquid of step one shaking table culture is inoculated with by the inoculum concentration of optimization fermented liquid fraction 7%
Into 75ml optimization fermentation medium, then temperature be 28 DEG C, rotating speed be oscillation and fermentation respectively under conditions of 150r/min
24h, 48h and 72h.Other steps and parameter are identical with embodiment two.
The fermentation results of bacteria cellulose are as shown in table 2 under the conditions of present embodiment different fermentations.
Table 2
Embodiment four：The difference of present embodiment and embodiment two is：
Step 2: the seed fermentation liquid of step one shaking table culture is distinguished by the inoculum concentration of optimization fermented liquid fraction 7%
It is seeded in 75ml, 100ml and 125ml optimization fermentation medium, then in the bar that temperature is 28 DEG C, rotating speed is 150r/min
Oscillation and fermentation 48h under part.Other steps and parameter are identical with embodiment two.
The fermentation results of bacteria cellulose are as shown in table 3 under the conditions of present embodiment different fermentations.
Table 3
Embodiment five：The difference of present embodiment and embodiment two is：
Step 2: the seed fermentation liquid of step one shaking table culture being connect by optimization fermented liquid fraction 4%, 7% and 10%
Kind of amount is seeded in 75ml optimization fermentation medium respectively, is then 28 DEG C, under conditions of rotating speed is 150r/min in temperature
Oscillation and fermentation 48h.Other steps and parameter are identical with embodiment two.
The fermentation results of bacteria cellulose are as shown in table 4 under the conditions of present embodiment different fermentations.
Table 4
Embodiment six：The difference of present embodiment and embodiment two is：
Step 2: the seed fermentation liquid of step one shaking table culture is distinguished by the inoculum concentration of optimization fermented liquid fraction 7%
Be seeded in 75ml optimization fermentation medium, then temperature be 28 DEG C, rotating speed be respectively 130r/min, 150r/min and
Oscillation and fermentation 48h under conditions of 180r/min.Other steps and parameter are identical with embodiment two.
The fermentation results of bacteria cellulose are as shown in table 5 under the conditions of present embodiment different fermentations.
Table 5
Embodiment seven：The difference of present embodiment and embodiment two is：Optimize fermentation in step 2
Amount of alcohol added is respectively 1ml, 1.5ml and 2ml in the every 100ml of culture medium.Other steps and parameter are identical with embodiment two.
The fermentation results of bacteria cellulose are as shown in table 6 under the conditions of present embodiment different fermentations.
Table 6
The formation of chrysanthemum shape bacteria cellulose of the present invention need to select specific microbial material, fermentation culture conditions and culture
Based component.The peripheral thorn of chrysanthemum shape bacteria cellulose of the present invention is more, long and sturdy, and milky is glossy, homogeneous densification.
Claims (1)
1. the fermentation preparation of chrysanthemum shape bacteria cellulose, it is characterised in that chrysanthemum shape bacteria cellulose ferments according to the following steps
Prepare：
First, by Han Shi gluconacetobacters HDM1-3 (Gluconacetobacter hansenii HDM1-3) streak inoculation is in thin
In fungin solid medium, then the constant temperature quiescent culture 48h under the conditions of 28 DEG C is inoculated in oese picking lawn
In 100ml seed fermentation liquid, remix it is uniform after be placed in the shaking table that temperature is 28 DEG C, revolution is 130r/min and cultivate 24h；
2nd, the seed fermentation liquid of step one shaking table culture is seeded to 75ml by the inoculum concentration of optimization fermented liquid fraction 7%
Optimization fermentation medium in, then temperature be 28 DEG C, rotating speed be oscillation and fermentation 48h under conditions of 150r/min, that is, obtain
Chrysanthemum shape bacteria cellulose；
Wherein, in step one bacteria cellulose solid medium by mass percentage by 5% glucose, 0.5% yeast extract,
0.5% peptone, 0.1% citric acid, 0.2% Na2HPO4·H2O, 0.1% K2HPO4, 0.025% MgSO4·
7H2O, 2% agar and surplus distilled water composition, pH value is 5.8；In step one seed fermentation liquid by mass percentage by
2% sucrose, 0.3% yeast extract, 0.5% peptone, 0.2% potassium dihydrogen phosphate, 0.015% magnesium sulfate and surplus
Distilled water composition, pH value is 5.8；Step 2 optimization fermentation medium per 100ml by 5g glucose, 0.34g beef extract,
0.19g yeast extract, 0.22g disodium hydrogen phosphate, 0.46g dipotassium hydrogen phosphate, 0.2g lactic acid, 0.025g magnesium sulfate,
1ml ethanol and the distilled water composition of surplus, pH value is 6.5；
The shape of the chrysanthemum shape bacteria cellulose is in chrysanthemum shape, and peripheral thorn is more and long, and milky is glossy, homogeneous to cause
It is close；The filament of chrysanthemum shape bacteria cellulose is very very thin<0.1 μm, ultra-microfibers are mutually wound, and mycelia winding is random,
Ultra-microfibers net is constituted, silk ribbon stratiform is overlapping, silk ribbon is more uniform while existing, and with pore space structure.
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