CN105132273A - Culture reactor for multiple bacterial cellulose membranes and application of culture reactor - Google Patents
Culture reactor for multiple bacterial cellulose membranes and application of culture reactor Download PDFInfo
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- CN105132273A CN105132273A CN201510601339.1A CN201510601339A CN105132273A CN 105132273 A CN105132273 A CN 105132273A CN 201510601339 A CN201510601339 A CN 201510601339A CN 105132273 A CN105132273 A CN 105132273A
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- 229920002749 Bacterial cellulose Polymers 0.000 title claims abstract description 33
- 239000005016 bacterial cellulose Substances 0.000 title claims abstract description 33
- 239000012528 membrane Substances 0.000 title abstract description 10
- 229920002678 cellulose Polymers 0.000 claims abstract description 107
- 239000001913 cellulose Substances 0.000 claims abstract description 106
- 239000011229 interlayer Substances 0.000 claims abstract description 30
- 238000000855 fermentation Methods 0.000 claims abstract description 18
- 230000004151 fermentation Effects 0.000 claims abstract description 18
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 17
- 241000894006 Bacteria Species 0.000 claims description 108
- 239000007788 liquid Substances 0.000 claims description 72
- 239000006052 feed supplement Substances 0.000 claims description 20
- 230000003068 static effect Effects 0.000 claims description 17
- 238000002360 preparation method Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 238000000034 method Methods 0.000 abstract description 19
- 238000011169 microbiological contamination Methods 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 3
- 238000009776 industrial production Methods 0.000 abstract 1
- 239000013589 supplement Substances 0.000 abstract 1
- 235000010980 cellulose Nutrition 0.000 description 97
- 239000010408 film Substances 0.000 description 81
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 230000001954 sterilising effect Effects 0.000 description 11
- 238000004659 sterilization and disinfection Methods 0.000 description 10
- 108010025899 gelatin film Proteins 0.000 description 9
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 230000007935 neutral effect Effects 0.000 description 6
- 235000002837 Acetobacter xylinum Nutrition 0.000 description 5
- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 5
- 241001136169 Komagataeibacter xylinus Species 0.000 description 5
- 238000011081 inoculation Methods 0.000 description 5
- 239000010410 layer Substances 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003570 biosynthesizing effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000015071 dressings Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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Abstract
The invention discloses a culture reactor for multiple bacterial cellulose membranes. The culture reactor comprises a main vessel body and a cover body, wherein the vessel opening section of the main vessel body is a semicircular protrusion; an opening is formed in the upper part and the lower part of the side wall of the main vessel body; the upper opening and the lower opening are connected with hoses; the hoses are connected with a peristaltic pump; a stop valve is arranged on the hose that is connected with the lower opening; a porous interlayer is arranged between the upper opening and the lower opening of the main vessel body; the interlayer is fixed on an interlayer bracket; holes are uniformly formed in the interlayer; besides, the peristaltic pump is connected with a material supplement bottle through a hose. With the adoption of the culture reactor for multiple bacterial cellulose membranes and the application of the culture reactor, provided by the invention, multiple cellulose membranes can be obtained in the same reactor, the space is economized, the fermentation liquor is fully utilized, the productivity of bacterial cellulose membranes is improved, and at the same time, grown bacterial cellulose membranes do not need to be taken out, and the risk of microbiological contamination during the operation process is reduced, therefore the culture reactor for multiple bacterial cellulose membranes, provided by the invention, has a broad application prospect in the industrial production of a single membrane.
Description
Technical field
The invention belongs to technical field of microbial fermentation, be specifically related to a kind of bacteria cellulose multi-disc Membrance cuiture reactor and application thereof.
Background technology
Bacteria cellulose, compared with natural cellulose, has character and the advantage of many uniquenesses: (1) bacteria cellulose is a kind of " pure cellulose ", has high chemical purity and high-crystallinity; (2) there is very strong water-holding power; (3) there is higher biological fitness, can directly degrade at nature, free from environmental pollution; (4) Fibre diameter is between 0.01 ~ 0.1 micron, and fiber modulus is more than the several times to ten times of general fiber, and tensile strength is high; (5) during biosynthesizing, there is Modulatory character; (6) matrix widely can be utilized to produce; (7) there is very strong wetting ability, toughness and stability.Above-mentioned characteristic makes it be with a wide range of applications in papermaking, medicine, cosmetic industry, biomedical engineering and foodstuffs industry.
The training method of existing bacteria cellulose mainly comprises quiescent culture and stirs aerated culture.During quiescent culture, bacteria cellulose goes out to produce layer of gel film on fermented liquid surface, and quiescent culture synthesis bacteria cellulose realizes by single stage method or two step method, and single stage method is direct quiescent culture after referring to culture medium inoculated; And two step method refers to first obtain by aerated culture the bacterial classification that raised growth is good, vigor is high, then carry out quiescent culture.
The cellulose membrane that quiescent culture obtains can sterilizing, porous, flexible, good with skin-friendliness, liquid can be provided to arrive dry matrix, now for making the wound dressingss such as artificial skin, gauze, bandage.In addition, in view of the three-dimensional net structure of its uniqueness, bacteria cellulose has been developed and has been developed into mask base-material.These application requiring bacteria celluloses have certain thickness.For obtaining monolithic bacterial cellulose film, main employing two kinds of methods, a kind of for the thickness of acquisition is formed film more than the bacteria cellulose film section of 2cm, but there are a large amount of tool marks by the bacterial cellulose film that section obtains, and yield rate is low, affect result of use; Another kind is then after fermentation for some time, and regularly take out bacterial cellulose film sheet, bacteria cellulose film prepared by this method yields poorly, and the easy microbiological contamination of operating process.
Summary of the invention
The object of the present invention is to provide a kind of bacteria cellulose multi-disc Membrance cuiture reactor, quiescent culture bacteria cellulose in the reactor, after bacteria cellulose film grows to ideal thickness, lower floor is fermented liquid pump on bacteria cellulose film, continue fermentation, in addition can when carbon source exhaust in the reactor, by the timely feed supplement of pump, fermenting process is controlled flexibly.The output of static cultivation is by S/V ratio (area/volume ratio) impact, and because the surface-area holding the container of nutrient solution is certain, with the increase of gel-film thickness, dissolved oxygen diminishes, and suppresses thalline to produce Mierocrystalline cellulose.The present invention is by this reactor of design, the superposition growth of multi-disc bacteria cellulose film can be realized, improve the output capacity of bacteria cellulose, the bacteria cellulose film grown does not need to take out, below liquid level can be settled down to, reduce the microbiological contamination risk of process, the present invention is that bacteria cellulose monolithic thin film suitability for industrialized production provides the foundation.
The technical scheme realizing the object of the invention is: a kind of bacteria cellulose multi-disc Membrance cuiture reactor, comprise main body container and lid, the height of main body container is according to needing setting, the vessel port section of main body container is semicircle outstanding, the vessel port section of described main body container is the contact part of main body container and lid, to improve reactor and extraneous gaseous interchange.Respectively there is an opening top and the bottom of the sidewall of main body container, and upper shed is used for fermented liquid and pumps into, and lower opening is used for fermented liquid and pumps, and upper and lower opening is by hose connection peristaltic pump, and the flexible pipe connecting lower opening is provided with stopping valve.Be provided with porous interlayer between the upper and lower opening of main body container, interlayer is fixed on interlayer support, and interlayer is uniformly distributed hole, and interlayer is mainly used in evenly trickling down after fermented liquid pumps into fermented liquid surface to reactor.
Further, described peristaltic pump is also by hose connection feed supplement bottle, and flexible pipe is provided with stopping valve, when fermented liquid exhausts in a reservoir, adds fermented liquid by peristaltic pump in container, effectively improves the production efficiency of bacteria cellulose.
The present invention also provides a kind of method utilizing bacteria cellulose multi-disc Membrance cuiture reactor to prepare bacteria cellulose multi-disc film, first bacteria cellulose producing strains is carried out seed expansion cultivation, then be seeded in the bacteria cellulose multi-disc Membrance cuiture reactor containing fermented liquid and ferment, formed after bacterial cellulose film until fluid surface, by the fermentation liquid pump in lower floor's fermented liquid or feed supplement bottle to bacterial cellulose film upper strata, repeat above-mentioned pumping into or feed supplement step, until no longer form bacteria cellulose film, the bacteria cellulose film that final purification obtains.
A preparation method for bacteria cellulose multi-disc film, concrete steps are as follows:
Step 1, carries out seed expansion cultivation by bacteria cellulose producing strains;
Step 2, preparation fermented liquid, then inoculates seed solution, in bacteria cellulose multi-disc Membrance cuiture reactor, carries out static cultivation;
Step 3, is formed after bacterial cellulose film until fluid surface, by the fermentation liquid pump in lower floor's fermented liquid or feed supplement bottle to the upper strata of bacterial cellulose film;
Step 4, repeating step 3, until no longer formation bacteria cellulose film or liquid level reach porous interlayer;
Step 5, the bacteria cellulose film that purifying obtains.
Further, the preparation method of a kind of bacteria cellulose multi-disc film provided by the invention, concrete steps are as follows:
Step 1, carries out seed expansion cultivation by bacteria cellulose producing strains;
Step 2, preparation fermented liquid, then inoculates seed solution, in bacteria cellulose multi-disc Membrance cuiture reactor, carries out static cultivation;
Step 3, is formed after bacterial cellulose film until fluid surface, the upper strata of liquid pump to bacterial cellulose film of lower floor being fermented;
Step 4, repeating step 3, until no longer form bacteria cellulose film;
Step 5, by the fermentation liquid pump in feed supplement bottle to the upper strata of bacterial cellulose film, continues static cultivation;
Step 6, repeating step 5, until liquid level reaches porous interlayer;
Step 7, the bacteria cellulose film that purifying obtains.
Compared with prior art, beneficial effect of the present invention is: reactor of the present invention can realize obtaining more cellulosic film in same reactor, saves space and takes full advantage of fermented liquid, improve the productivity of bacteria cellulose film.Meanwhile, the bacteria cellulose film grown, without the need to taking out, reduces the microbiological contamination risk of operating process.
Accompanying drawing explanation
Fig. 1 is the structural representation of bacteria cellulose multi-disc Membrance cuiture reactor of the present invention.
Fig. 2 is the structural representation being connected with the bacteria cellulose multi-disc Membrance cuiture reactor of feed supplement bottle of the present invention.
Fig. 3 is the longitudinal section of main body container.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail.
In accompanying drawing 1, accompanying drawing 2 and accompanying drawing 3,1 is sidewall upper shed, and 2 is sidewall lower opening, and 3 is flexible pipe, and 4 is stopping valve, and 5 is peristaltic pump, and 6 is interlayer support, and 7 is porous interlayer, and 8 is vessel port section, and 9 is lid, and 10 is main body container, and 11 is feed supplement bottle.
With reference to accompanying drawing 1, accompanying drawing 2 and accompanying drawing 3, a kind of bacteria cellulose multi-disc Membrance cuiture reactor, comprise main body container (10) and lid (9), the height of main body container is according to needing setting, the vessel port section (8) of main body container is semicircle outstanding, the vessel port section of described main body container is the contact part of main body container and lid, the semicircle outstanding gaseous interchange that can improve reactor and the external world.Respectively there is an opening top and the bottom of the sidewall of main body container, upper shed (1) pumps into for fermented liquid, lower opening (2) pumps for fermented liquid, upper and lower opening connects peristaltic pump (5) by flexible pipe (3), the flexible pipe connecting lower opening is provided with stopping valve (4), porous interlayer (7) is provided with between the upper and lower opening of main body container, interlayer (7) is fixed on interlayer support (6), interlayer is uniformly distributed hole, interlayer is mainly used in evenly trickling down after fermented liquid pumps into fermented liquid surface to reactor, further, peristaltic pump (5) also connects feed supplement bottle (11) by flexible pipe (3), flexible pipe (3) is provided with stopping valve (4), when fermented liquid exhausts in a reservoir, in container, fermented liquid is added by peristaltic pump.
The application process of bacteria cellulose multi-disc Membrance cuiture reactor of the present invention is as follows: first bacteria cellulose producing strains is carried out seed expansion cultivation, then be seeded in the bacteria cellulose multi-disc Membrance cuiture reactor containing fermented liquid and ferment, formed after bacterial cellulose film until fluid surface, lower floor is fermented liquid pump to bacterial cellulose film upper strata, repeat above-mentionedly to pump into step, until no longer form bacteria cellulose film, the bacteria cellulose film that final purification obtains, or after no longer forming bacteria cellulose film, by the fermentation liquid pump in feed supplement bottle to the upper strata of bacterial cellulose film, repeat feed supplement step, until liquid level reaches porous interlayer, the bacteria cellulose film that final purification obtains.
Utilize bacteria cellulose multi-disc Membrance cuiture reactor of the present invention to prepare the method for multi-disc bacteria cellulose film, concrete steps are as follows:
Step 1, be 25-32 DEG C by acetobacter xylinum in temperature, shaking speed is carry out seed expansion cultivation under 90-250rpm condition, cultivates 6-72h;
Step 2, preparation fermented liquid, after high pressure steam sterilization, is cooled to room temperature;
Step 3, after bacteria cellulose multi-disc Membrance cuiture reactor high pressure steam sterilization, adds fermented liquid, inoculation 2-20% (v/v) seed liquor, static cultivation;
Step 4, after static cultivation 12-36 hour, fermented liquid surface forms the thin bacterial cellulose gel film of one deck, by peristaltic pump by the fermentation liquid pump in lower floor's fermented liquid or feed supplement bottle to the surface of bacteria cellulose film, continue quiescent culture;
Step 5, repeating step 4, until no longer formation bacteria cellulose film or liquid level reach porous interlayer;
Step 6, film step 5 obtained is immersed in the H of NaOH, the 1-10 ‰ (v/v) containing massfraction 1-10 ‰ (w/v)
2o
2in solution, at 70-100 DEG C of Water Under bath 0.5-3.0 hour, remove residual bacterium and fermented liquid;
Step 7, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Further, the above-mentioned bacteria cellulose multi-disc Membrance cuiture reactor of the present invention that utilizes prepares the method for multi-disc bacteria cellulose film, through step 5 until after no longer forming bacteria cellulose film, carry out following steps:
Step 6, by the fermented liquid in feed supplement bottle by peristaltic pump to the surface of bacterial cellulose film, continue quiescent culture;
Step 7, repeating step 6, until liquid level reaches porous interlayer;
Step 8, film step 7 obtained is immersed in containing massfraction the H of NaOH, the 1-10 ‰ (v/v) being 1-10 ‰ (w/v)
2o
2in solution, at 70-100 DEG C of Water Under bath 0.5-3.0 hour, remove residual bacterium and fermented liquid;
Step 9, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Comparative example
Step 1, acetobacter xylinum, temperature 30 DEG C, carries out seed expansion cultivation under shaking speed 160rpm condition, cultivates 24h;
Step 2, fermented liquid is prepared, and high pressure steam sterilization, is cooled to room temperature;
Step 3, reactor high pressure steam sterilization, with 8% inoculum size inoculation, carries out static cultivation in conventional tray;
Step 4, static fermentation, after 7 days, forms the bacterial cellulose gel film of thick layer on fermented liquid surface;
Step 5, film step 4 obtained is immersed in the H of NaOH containing massfraction 3 ‰ (w/v), 3 ‰ (v/v)
2o
2in solution, bathe 2 hours at 80 DEG C of Water Unders, remove residual bacterium and fermented liquid;
Step 6, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Comparative example reactor is the reactor of identical material, but only has the main body container part of reactor involved in the present invention, container side wall without upper and lower opening, without feeding-system, without interlayer and interlayer support.
The bacteria cellulose film that comparative example obtains is monolithic membrane, and thickness is 11mm.
Embodiment 1
By reference to the accompanying drawings 1, utilize bacteria cellulose multi-disc Membrance cuiture reactor of the present invention, adopt and the lower floor's fermentation liquid pump in container to the method on bacteria cellulose film upper strata is prepared bacteria cellulose multi-disc film, concrete steps are as follows:
Step 1, acetobacter xylinum, temperature 30 DEG C, carries out seed expansion cultivation under shaking speed 160rpm condition, cultivates 24h;
Step 2, fermented liquid is prepared, and high pressure steam sterilization, is cooled to room temperature;
Step 3, reactor high pressure steam sterilization, with 8% inoculum size inoculation, carries out static cultivation in bacteria cellulose multi-disc Membrance cuiture reactor;
Step 4, after static fermentation 24h, forms the thin bacterial cellulose gel film of one deck on fermented liquid surface, by gel-film lower floor fermented liquid by peristaltic pump to the surface of bacteria cellulose film, continue quiescent culture;
Step 5, repeating step 4, until no longer form bacteria cellulose film, takes out all bacteria cellulose films in reactor;
Step 6, film step 5 obtained is immersed in the H of NaOH containing massfraction 3 ‰ (w/v), 3 ‰ (v/v)
2o
2in solution, bathe 2 hours at 80 DEG C of Water Unders, remove residual bacterium and fermented liquid;
Step 7, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Embodiment 1 obtains 11 layers of bacterial cellulose film, and mean thickness is 5mm.
Embodiment 2
By reference to the accompanying drawings 2, utilize bacteria cellulose multi-disc Membrance cuiture reactor of the present invention, adopt and the fermentation liquid pump in feed supplement bottle to the method on bacteria cellulose film upper strata is prepared bacteria cellulose multi-disc film, concrete steps are as follows:
Step 1, acetobacter xylinum, temperature 29 DEG C, carries out seed expansion cultivation under shaking speed 150rpm condition, cultivates 30h;
Step 2, fermented liquid is prepared, and high pressure steam sterilization, is cooled to room temperature;
Step 3, reactor high pressure steam sterilization, with 8% inoculum size inoculation, carries out static cultivation in bacteria cellulose multi-disc Membrance cuiture reactor;
Step 4, after static fermentation 20h, forms the thin bacterial cellulose gel film of one deck on fermented liquid surface, by fermented liquid in feed supplement bottle by peristaltic pump to the surface of bacteria cellulose film, continue quiescent culture;
Step 5, repeating step 4, until liquid level reaches porous interlayer, takes out all bacteria cellulose films in reactor;
Step 6, film step 5 obtained is immersed in the H of NaOH containing massfraction 4 ‰ (w/v), 4 ‰ (v/v)
2o
2in solution, bathe 1 hour at 85 DEG C of Water Unders, remove residual bacterium and fermented liquid;
Step 7, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Embodiment 2 obtains 16 layers of bacterial cellulose film, and mean thickness is 4mm.
Embodiment 3
By reference to the accompanying drawings 2, utilize bacteria cellulose multi-disc Membrance cuiture reactor of the present invention, adopt and first liquid pump is fermented to bacteria cellulose film upper strata in the lower floor in container, until no longer form bacteria cellulose film, adopt the fermentation liquid pump in feed supplement bottle to the method on bacteria cellulose film upper strata to prepare bacteria cellulose multi-disc film again, concrete steps are as follows:
Step 1, acetobacter xylinum, temperature 30 DEG C, carries out seed expansion cultivation under shaking speed 180rpm condition, cultivates 20h;
Step 2, fermented liquid is prepared, and high pressure steam sterilization, is cooled to room temperature;
Step 3, reactor high pressure steam sterilization, with 10% inoculum size inoculation, carries out static cultivation in bacteria cellulose multi-disc Membrance cuiture reactor;
Step 4, after static fermentation 18h, forms the thin bacterial cellulose gel film of one deck on fermented liquid surface, by gel-film lower floor fermented liquid by peristaltic pump to the surface of bacteria cellulose film, continue quiescent culture;
Step 5, repeating step 4, until no longer form bacteria cellulose film;
Step 6, by fermented liquid in feed supplement bottle by peristaltic pump to the surface of bacteria cellulose film, continue quiescent culture;
Step 7, repeating step 6, until liquid level reaches porous interlayer, takes out all bacteria cellulose films in reactor;
Step 8, film step 7 obtained is immersed in the H of NaOH containing massfraction 3 ‰ (w/v), 4 ‰ (v/v)
2o
2in solution, bathe 1 hour at 90 DEG C of Water Unders, remove residual bacterium and fermented liquid;
Step 9, extremely neutral with the deionized water rinsing bacteria cellulose film of flowing, obtain pure bacteria cellulose film.
Embodiment 3 obtains 19 layers of bacterial cellulose film, and mean thickness is 4mm.
The foregoing is only detailed description of the invention, but the present invention is not limited to above-mentioned embodiment.It should be pointed out that for those skilled in the art, allly do not departing under the present invention conceives prerequisite, any amendment made, equivalent replacement, improvement etc., all should be included in protection scope of the present invention.
Claims (4)
1. a bacteria cellulose multi-disc Membrance cuiture reactor, comprise main body container and lid, it is characterized in that, the vessel port section of described main body container is semicircle outstanding, respectively there is an opening top and the bottom of the sidewall of described main body container, upper and lower opening is by hose connection peristaltic pump, and the flexible pipe connecting lower opening is provided with stopping valve; Be provided with porous interlayer between the upper and lower opening of main body container, described porous interlayer is fixed on interlayer support.
2. cultivation reactor according to claim 1, is characterized in that, described peristaltic pump is by hose connection feed supplement bottle, and described flexible pipe is provided with stopping valve.
3. a preparation method for bacteria cellulose multi-disc film, is characterized in that, concrete steps are as follows:
Step 1, carries out seed expansion cultivation by bacteria cellulose producing strains;
Step 2, preparation fermented liquid, then inoculates seed solution, in reactor as claimed in claim 1 or 2, carries out static cultivation;
Step 3, is formed after bacterial cellulose film until fluid surface, by the fermentation liquid pump in lower floor's fermented liquid or feed supplement bottle to the upper strata of bacterial cellulose film;
Step 4, repeating step 3, until no longer formation bacteria cellulose film or liquid level reach porous interlayer;
Step 5, the bacteria cellulose film that purifying obtains.
4. the preparation method of bacteria cellulose multi-disc film according to claim 3, it is characterized in that, concrete steps are as follows:
Step 1, carries out seed expansion cultivation by bacteria cellulose producing strains;
Step 2, preparation fermented liquid, then inoculates seed solution, in reactor as claimed in claim 2, carries out static cultivation;
Step 3, is formed after bacterial cellulose film until fluid surface, the upper strata of liquid pump to bacterial cellulose film of lower floor being fermented;
Step 4, repeating step 3, until no longer form bacteria cellulose film;
Step 5, by the fermentation liquid pump in feed supplement bottle to the upper strata of bacterial cellulose film, continues static cultivation;
Step 6, repeating step 5, until liquid level reaches porous interlayer;
Step 7, the bacteria cellulose film that purifying obtains.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510601339.1A CN105132273A (en) | 2015-09-18 | 2015-09-18 | Culture reactor for multiple bacterial cellulose membranes and application of culture reactor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510601339.1A CN105132273A (en) | 2015-09-18 | 2015-09-18 | Culture reactor for multiple bacterial cellulose membranes and application of culture reactor |
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| Publication Number | Publication Date |
|---|---|
| CN105132273A true CN105132273A (en) | 2015-12-09 |
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|---|---|---|---|
| CN201510601339.1A Pending CN105132273A (en) | 2015-09-18 | 2015-09-18 | Culture reactor for multiple bacterial cellulose membranes and application of culture reactor |
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| CN (1) | CN105132273A (en) |
Cited By (5)
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| CN108273130A (en) * | 2018-02-27 | 2018-07-13 | 华东交通大学 | A kind of three-D micro-nano fibrous composite scaffold and preparation method |
| CN108624486A (en) * | 2018-07-05 | 2018-10-09 | 福建师范大学 | A kind of film forming fermentation tank and its application method |
| CN112323494A (en) * | 2020-11-06 | 2021-02-05 | 南京林业大学 | A kind of efficient preparation method of bacterial cellulose/non-woven fiber composite mask base cloth |
| WO2021138837A1 (en) * | 2020-01-08 | 2021-07-15 | 钟春燕 | Device and method for using dynamic fermentation to prepare bacterial cellulose composite material having core-shell structure |
| TWI871706B (en) * | 2023-07-21 | 2025-02-01 | 張瑞能 | Fungal culture device |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108273130A (en) * | 2018-02-27 | 2018-07-13 | 华东交通大学 | A kind of three-D micro-nano fibrous composite scaffold and preparation method |
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| CN112323494A (en) * | 2020-11-06 | 2021-02-05 | 南京林业大学 | A kind of efficient preparation method of bacterial cellulose/non-woven fiber composite mask base cloth |
| CN112323494B (en) * | 2020-11-06 | 2022-06-03 | 南京林业大学 | A kind of efficient preparation method of bacterial cellulose/non-woven fiber composite mask base cloth |
| TWI871706B (en) * | 2023-07-21 | 2025-02-01 | 張瑞能 | Fungal culture device |
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Application publication date: 20151209 |