CN104560688A - Device and method for culturing hybrid microorganisms - Google Patents
Device and method for culturing hybrid microorganisms Download PDFInfo
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- CN104560688A CN104560688A CN201310509183.5A CN201310509183A CN104560688A CN 104560688 A CN104560688 A CN 104560688A CN 201310509183 A CN201310509183 A CN 201310509183A CN 104560688 A CN104560688 A CN 104560688A
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- interior pipe
- dialysis membrane
- fixing device
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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
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- 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
- C12M23/24—Gas permeable parts
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- 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
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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- 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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention relates to the field of hybrid culturing of microorganisms and specifically relates to a device and a method for culturing hybrid microorganisms. The device comprises a vent plug, an outer barrel, an inner tube, an inner tube holder and a dialysis membrane holder; one end of the outer barrel is closed, while the other end of the outer barrel is open; one end of the inner tube is connected with the dialysis membrane holder; the inner tube is fixed in the outer barrel by use of at least one inner tube holder; a culture medium is added to each of the outer barrel and the inner tube, respectively, and the outer barrel and the inner tube are inoculated with different bacterial strains, respectively; and the port of the outer barrel is sealed by the vent plug. The device is capable of simulating hybrid bacteria culture conditions, and capable of obtaining the pure culture of two target bacterial strains under different culture states in hybrid culturing, and is convenient for studying the mutual relation between the hybrid bacteria. The device is simple to manufacture and convenient to use, and can be widely applied to mutual relation study under various hybrid bacteria conditions.
Description
Technical field
The present invention relates to the field of mixed culture microorganism, being specially a kind of device for cultivating mixing microorganisms and cultural method thereof.
Background technology
Long-term production practice and increasing scientific research show, numerous microbial processes needs the synergy of multiple bacterial strain (or efficient) to complete, not only in fermentation industry field, as fermentation process for vitamin production, food fermentation and wine, vinegar are brewageed, relating to environmental protection and the field of biological energy source of microorganism, also the report having bacterial strain mixed culture to improve conversion and bioaccumulation efficiency more.Along with Hybrid NC machine tool technology is in the widespread use of every field, the mixed bacterium interphase interaction relation of research has become one of study hotspot of domestic and international scientist to improve mixed bacterium conversion (degraded) efficiency.
In the research of hybrid bacterial strain interphase interaction relation, obtaining through mixed culture and aimed strain after interacting with other bacterial strain purebred, is the problem first needing to solve.Design one can ensure can highly imitate mixed culture condition, can obtain the purebred experimental installation of aimed strain fast again, could meet the needs that we study mixed bacterium relation.And such device rarely has report.Similar patent such as Chinese Patent Application No. 201220446864.2 disclosed " use for laboratory anerobe dialysis culturing device " utilizes semi-permeable membranes as dialysis membrane, and be the dialysis culture for independent anaerobion, and this device is unsuitable for studying aerobic bacteria.The dialysis culturing device complex structure that " microbial dialysis incubator " of Chinese Patent Application No. 200920104149.9 is mentioned, cost is higher.Also there is similar patent report abroad, but be all utilize semi-permeable membranes as dialysis membrane, and complex structure, high expensive.Traditional method being used for studying relation between mixed bacterium joins in target microorganism nutrient solution after filtration by the nutrient solution of a kind of microorganism, carries out shake-flask culture.This method is not only difficult to simulation mixed culture condition, and wastes time and energy, and microbiological contamination probability is high.
Summary of the invention
The technical issues that need to address of the present invention are to provide a kind of not only simple but also a kind of device for cultivating mixing microorganisms and cultural method thereof efficiently.
For solving above technical problem, the technical scheme that present method adopts is:
For cultivating a device for mixing microorganisms, comprise porous plug, urceolus, interior pipe, interior pipe fixing device and dialysis membrane fixing device; Urceolus one end is airtight, and the other end is open type, and interior pipe one end connects dialysis membrane fixing device, and interior pipe is fixed in urceolus by pipe fixing device at least one, and urceolus open end is sealed by porous plug.
Outer tube one end is round bottom or flat, described in be connected with dialysis membrane fixing device interior pipe one end with abut bottom urceolus or hung in outer tube by interior pipe fixing device.
Described interior pipe one end is provided with outside screw, is provided with externally threaded interior pipe one end and is threadedly connected with dialysis membrane fixing device.
Described interior pipe is provided with between externally threaded one end and dialysis membrane fixing device and is provided with silicone gasket, dialysis membrane, dialysis membrane fixed network from top to bottom successively.
Described silicone gasket outside diameter is equal with the interior diameter of dialysis membrane fixing device.
Described dialysis membrane is aperture at the cellulose filter membrane of 0.22-0.45 μm.
Described porous plug, urceolus, interior pipe, interior pipe fixing device and dialysis membrane fixing device are thermostable transparent material and make.
Cultural method: add substratum respectively in urceolus and interior pipe, and access different bacterial strains respectively, utilize the pressure reduction between two pipes, bacterial metabolism material in shaking culture in pipe substratum with medium flow to outer tube, pipe in taking out after cultivating for some time, the bacterium in urceolus substratum is the target microorganism of the pure culture obtained.Described urceolus and interior pipe pressure reduction are 1-3cm water column.
The advantage that the present invention has:
The present invention adopts thermostable transparent material to make, and after apparatus of the present invention sterilising treatment, in urceolus and interior pipe, adds substratum respectively, makes the substratum liquid level of interior pipe higher than outer tube.After inner and outer pipe accesses different bacterial strains respectively, owing to there is pressure reduction between two pipes, when shaking culture, the bacterial metabolism material in interior pipe substratum can with medium flow to outer tube, and thalline can not pass through.Pipe in careful taking-up after cultivation for some time, and the bacterium in urceolus substratum is the target microorganism of the pure culture obtained, and this microorganism has been subject to the effect of interior pipe microorganism or has affected.The target microorganism of the pure culture obtained, namely can be applicable to the comparative analysis carrying out various biological method and technology, finally reaches the object of understanding two bacterium mutual relationship and effect.
Device of the present invention, not only can altitude simulation Hybrid NC machine tool condition, and can obtain the pure growth of each aimed strain under different cultivation conditions in mixed culture at short notice, between research mixed bacterium, mutual relationship is provided convenience.Apparatus of the present invention make simple, easy to use, can be widely used in the interaction relationship research under various mixed bacterium condition.
Accompanying drawing explanation
Fig. 1 is the structural representation of this experimental installation;
Fig. 2 is the dialysis membrane fixing device schematic diagram of this experimental installation;
Wherein: 1, porous plug, 2, outer tube, 3, interior pipe, 4, interior pipe fixing device, 5, dialysis membrane fixing device, 6, silicone gasket, 7, dialysis membrane, 8, dialysis membrane fixed network.
Embodiment
Embodiment 1
For cultivating the device of mixing microorganisms, comprise porous plug, urceolus, interior pipe, interior pipe fixing device and dialysis membrane fixing device; Urceolus one end airtight (can be round bottom or flat bottom blind end), the other end is open type, and interior pipe one end connects dialysis membrane fixing device, and interior pipe is fixed in urceolus by pipe fixing device at least one, and urceolus port is sealed by porous plug.Described interior pipe one end is provided with outside screw, is provided with externally threaded interior pipe one end and is threadedly connected with dialysis membrane fixing device.Described interior pipe is provided with between externally threaded one end and dialysis membrane fixing device and is provided with silicone gasket, dialysis membrane, dialysis membrane fixed network from top to bottom successively.Described silicone gasket outside diameter is equal with dialysis membrane fixing device interior diameter.Said apparatus kraft paper is wrapped and puts into Autoclave sterilizing, stand-by.
Wherein, ventilative plug can be plug or gauze sealing; Urceolus material is glass, external diameter 35.00mm, internal diameter 30.00mm, long 200.00mm; Interior tubing matter is glass, external diameter 15.00mm, internal diameter 10mm, long 120.00mm; Interior pipe fixing device material is high-temperature resistance plastice, and there is breather hole external diameter 35.00mm, internal diameter 30.00mm, centre; Dialysis membrane fixing device material is high-temperature resistance plastice nut, external diameter 20.00mm, internal diameter 18.00mm, mesopore internal diameter 10.00mm, long 20.00mm; Mesoporous silica gel packing ring material is silica gel, external diameter 18.00mm, mesopore internal diameter 10.00mm; Dialysis membrane is the high temperature resistant filter membranes such as material fibers element filter membrane, 0.22 μm-0.45 μm, aperture diameter 18.00mm; Dialysis membrane fixed network material is stainless steel, diameter 18.00mm.
The assembling of device:
First the internal diameter size of dialysis membrane by interior pipe filter membrane stationary installation 5 is cut, and get wet wetting after, dialysis membrane fixed network 8, filter membrane 7, silicone gasket 6 are put into filter membrane stationary installation 5 successively.Interior pipe is carefully rotated and puts film stationary installation 5 into, can add after screwing suitable water with detect whether completely closed.Be placed in by interior pipe fixing device 4 on interior pipe 3, then carefully put into urceolus 2 by installing in interior pipe fixing device to manage, interior pipe top is to outer pipe bottom or hang in urceolus.Utilize ventilative plug 1 by outer duct occlusion, wrap with kraft paper and put into Autoclave sterilizing.
Application examples
Existing a kind of aerobic microbiological A bacterium, it can transformation substance a be b, but the own poor growth of A bacterium and conversion capability is low.The effect that A bacterium just can must obtain with another kind of aerobic microbiological B bacterium co-cultivation.B bacterium itself can be good at growth.Being so which type of material A bacterium provides to study B bacterium, causing the metabolism and growth change of A bacterium and A bacterium when itself there occurs which change after the impact of the B bacterium be subject to, will obtain A bacterium affect by B bacterium after the A bacterium of pure culture.In gnotobasis, respectively to the substratum adding suitable A bacterium and B bacterium in interior pipe 3 and urceolus 2 respectively, add B bacterium in pipe wherein, urceolus adds a small amount of A bacterium, be stoppered ventilative plug, tiltedly put after for some time is cultivated in concussion according to the training method of existing A bacterium and B bacterium and measure corresponding index.After having cultivated in gnotobasis in urceolus nutrient solution coating or microscopy to guarantee to obtain pure A bacterium.
The measuring result of material b:
The purebred A bacterium through the impact of B bacterium is obtained after cultivating in apparatus of the present invention, and not by B fungi pollution.
Claims (8)
1. for cultivating a device for mixing microorganisms, it is characterized in that: device comprises porous plug, urceolus, interior pipe, interior pipe fixing device and dialysis membrane fixing device; Urceolus one end is airtight, and the other end is open type, and interior pipe one end connects dialysis membrane fixing device, and interior pipe is fixed in urceolus by pipe fixing device at least one, and urceolus open end is sealed by porous plug.
2. by the device for cultivating mixing microorganisms according to claim 1, it is characterized in that: described interior pipe one end is provided with outside screw, being provided with externally threaded interior pipe one end and being threadedly connected with dialysis membrane fixing device.
3. by the device for cultivating mixing microorganisms according to claim 2, it is characterized in that: described interior pipe is provided with between externally threaded one end and dialysis membrane fixing device and is provided with silicone gasket, dialysis membrane, dialysis membrane fixed network from top to bottom successively.
4. by the device for cultivating mixing microorganisms according to claim 3, it is characterized in that: described silicone gasket outside diameter is equal with the interior diameter of dialysis membrane fixing device.
5. by the device for cultivating mixing microorganisms according to claim 2, it is characterized in that: described dialysis membrane is aperture at the cellulose filter membrane of 0.22-0.45 μm.
6. by the device for cultivating mixing microorganisms according to claim 1, it is characterized in that: described porous plug, urceolus, interior pipe, interior pipe fixing device and dialysis membrane fixing device are thermostable transparent material and make.
7. the cultural method utilizing cultivation mixing microorganisms device according to claim 1, it is characterized in that: in urceolus and interior pipe, add substratum respectively, and access different bacterial strains respectively, utilize the pressure reduction between two pipes, bacterial metabolism material in shaking culture in pipe substratum with medium flow to outer tube, pipe in taking out after cultivating for some time, the bacterium in urceolus substratum is the target microorganism of the pure culture obtained.
8., by cultural method according to claim 7, it is characterized in that: described urceolus and interior pipe pressure reduction are 1-3cm water column.
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| CN201310509183.5A CN104560688B (en) | 2013-10-23 | 2013-10-23 | A kind of for cultivating device and the cultural method thereof of mixed microorganism |
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| CN201310509183.5A CN104560688B (en) | 2013-10-23 | 2013-10-23 | A kind of for cultivating device and the cultural method thereof of mixed microorganism |
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| CN104560688B CN104560688B (en) | 2016-05-04 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950458A (en) * | 2016-05-27 | 2016-09-21 | 哈尔滨师范大学 | Mixed microbe culturing device |
| CN106148164A (en) * | 2016-06-30 | 2016-11-23 | 南京师范大学 | A kind of dialysis apparatus and comprise the microcosm experiment device of this dialysis apparatus |
| CN108238360A (en) * | 2018-01-12 | 2018-07-03 | 长春理工大学 | A kind of steam sterilizing Packaging Bottle |
| CN111139174A (en) * | 2020-01-15 | 2020-05-12 | 辽东学院 | Mixed microorganism culture device |
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| CN203112832U (en) * | 2012-11-02 | 2013-08-07 | 中华人民共和国上海出入境检验检疫局 | Multifunctional test tube for culturing bacteria |
| CN203559051U (en) * | 2013-10-23 | 2014-04-23 | 中国科学院沈阳应用生态研究所 | Device for cultivating mixed microbes |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2570288Y (en) * | 2002-09-30 | 2003-09-03 | 周晋 | Disposable semipermeabile connection type cell separating combined culture bottle |
| CN201325964Y (en) * | 2008-12-18 | 2009-10-14 | 崔生发 | Open circulation type tubular microorganism dialysis cultivation system |
| CN201459137U (en) * | 2009-08-19 | 2010-05-12 | 林涛 | Microbial dialysis incubator |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950458A (en) * | 2016-05-27 | 2016-09-21 | 哈尔滨师范大学 | Mixed microbe culturing device |
| CN106148164A (en) * | 2016-06-30 | 2016-11-23 | 南京师范大学 | A kind of dialysis apparatus and comprise the microcosm experiment device of this dialysis apparatus |
| CN106148164B (en) * | 2016-06-30 | 2018-09-07 | 南京师范大学 | A kind of dialysis apparatus and the microcosm experiment device comprising the dialysis apparatus |
| CN108238360A (en) * | 2018-01-12 | 2018-07-03 | 长春理工大学 | A kind of steam sterilizing Packaging Bottle |
| CN111139174A (en) * | 2020-01-15 | 2020-05-12 | 辽东学院 | Mixed microorganism culture device |
| CN111139174B (en) * | 2020-01-15 | 2023-08-29 | 辽东学院 | Mixed microorganism culture apparatus |
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