CN204981845U - Reactor is cultivateed to bacteria cellulose multi -disc membrane - Google Patents
Reactor is cultivateed to bacteria cellulose multi -disc membrane Download PDFInfo
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- CN204981845U CN204981845U CN201520728452.1U CN201520728452U CN204981845U CN 204981845 U CN204981845 U CN 204981845U CN 201520728452 U CN201520728452 U CN 201520728452U CN 204981845 U CN204981845 U CN 204981845U
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Abstract
The utility model discloses a reactor is cultivateed to bacteria cellulose multi -disc membrane, including main part container and lid, the container mouth section of main part container is semi -circular outstanding, and the upper and lower parts of the lateral wall of main part container respectively has an opening, goes up under shed coupling hose, and the hose connection peristaltic pump is equipped with the stop valve on the hose of connection under shed, is equipped with porous interlayer between the last under shed of main part container, and the interlayer is fixed on the interlayer support, interlayer evenly distributed hole. In addition, peristaltic pump accessible hose connection feed supplement bottle still. The utility model discloses a multi -disc cellulose membrane can be realized in same reactor obtaining by the reactor, and the zymotic fluid of having practiced thrift space and make full use of has improved bacterium cellulose membrane's productivity ratio, and simultaneously, the bacterium cellulose membrane who has grown need not to take out, has reduced operation process's microbiological contamination risk. The utility model discloses a reactor is cultivateed to bacteria cellulose multi -disc membrane has wide application prospect in the industrial production of monolithic film.
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 (2)
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.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132273A (en) * | 2015-09-18 | 2015-12-09 | 南京荣之盛生物科技有限公司 | Culture reactor for multiple bacterial cellulose membranes and application of culture reactor |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132273A (en) * | 2015-09-18 | 2015-12-09 | 南京荣之盛生物科技有限公司 | Culture reactor for multiple bacterial cellulose membranes and application of culture reactor |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160120 Termination date: 20180918 |