CN103627636A - Airlift type magnetic anion bioreactor - Google Patents

Airlift type magnetic anion bioreactor Download PDF

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Publication number
CN103627636A
CN103627636A CN201310645712.4A CN201310645712A CN103627636A CN 103627636 A CN103627636 A CN 103627636A CN 201310645712 A CN201310645712 A CN 201310645712A CN 103627636 A CN103627636 A CN 103627636A
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air
circulation duct
oxygen
liquid culture
tank
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CN103627636B (en
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董静洲
朱蕾
雷灿
王瑛
艾训儒
郑小江
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Hubei University for Nationalities
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Hubei University for Nationalities
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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
    • C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
    • C12M29/08—Air lift
    • 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
    • C12M29/26—Conditioning fluids entering or exiting the reaction vessel
    • 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
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C12M37/02—Filters
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution

Abstract

The invention discloses an airlift type magnetic anion bioreactor. The airlift type magnetic anion bioreactor comprises a liquid cultivation tank, wherein a tank top cover is arranged at the top of the liquid cultivation tank; the bottom of the liquid cultivation tank is communicated with the top of the liquid cultivation tank through an air-circulating passage; an air pump for circulating air and a temperature controller for heating the temperature in the air-circulating passage are arranged in the air-circulating passage; the air-circulating passage is further sleeved with a magnetic field-high pressure generator. The airlift type magnetic anion bioreactor further comprises an electrolytic oxygen water tank, wherein the output pipeline of the electrolytic positive electrode of the electrolytic oxygen water tank is communicated with the air-circulating passage. The airlift type magnetic anion bioreactor also comprises an oxygen electrode arranged in the liquid cultivation tank and used for detecting the oxygen dissolution rate, as well as a pH meter and a thermometer which are used for detecting the pH value and the temperature respectively. According to the invention, oxygen required by cells, tissues or organisms in a culture solution is supplied by electrolytic oxygen; the anion generation method is adopted to adjust the pH value and the oxidation-reduction potential of the culture solution and neutralize oxidants or free radicals generated in the cell growing process.

Description

A kind of air lift type magnetic negative ion bio-reactor
Technical field
The invention belongs to the biochemical culture technique of Life Science Laboratory field, more specifically relate to a kind of air lift type magnetic negative ion bio-reactor, the liquid culture research of applicable autotrophic cell, heterotrophic cell and animal and plant cells or tissue.
Background technology
Biochemical engineering is one of life science new and high technology direction.Medicine, food and even tissue or the organ of by biochemical engineering means, producing human needs are one of life science development trends.In the fundamental research and industrial applications of biochemical reaction, cell cultures is the most frequently used a kind of means of studying wild-type cell and the synthetic object product of engineering cell.Air lift type liquid culture is gentle owing to stirring, and is easy to control and is widely used.Yet current aerobic microbiological and animal and plant cells air lift type training method are mainly to adopt outside air oxygen supply.This certainly will reduce greatly because topsoil causes liquid culture efficiency, and for example airborne sulfurous gas exceeds standard and will cause the decline or poisoning rapidly of bacterial classification or cell, and object product output sharply reduces.And airborne oxygen ratio fixes, therefore traditional air lift type liquid culture tank is unfavorable for regulating liquid culture research needs and the need of production of oxygen ratio.
This device adopts the defect of fed oxygen from air except solving at present traditional air lift type liquid culture tank, also emphasis has solved in cell cultivation process, to adopt controlled oxygen concentration to carry out the technical problem of oxygen supply and do not need to add alkaline salt and just can control the problem of pH value, and can maintain the reduction-state of liquid medium, to passing into the gas molecule of fermented liquid, magnetize, can delay cell, tissue aging and improve cell, organizational vitality.
This device carries out the aerobic cultivation of microorganism and animal and plant cells and tissue except being used for, can also or produce for the research of anaerobism liquid culture, no matter in aerobic liquid culture or anaerobism liquid culture, can both directly regulate pH value and redox potential, and do not need by adding alkaline salt, the method can greatly improve the vigor of cell and extend meta-bolites production time and output.
Summary of the invention
The object of the invention is to be the problems referred to above for existing air lift type liquid culture tank existence, a kind of air lift type magnetic negative ion bio-reactor is provided, the liquid culture research of applicable autotrophic cell, heterotrophic cell and animal vegetable tissue or organ and production, can also be for aquatic animals and plants cultivation etc.
To achieve these goals, the present invention is by the following technical solutions:
A kind of air lift type magnetic negative ion bio-reactor, comprise liquid culture tank, the top of liquid culture tank is provided with top ends of cans, the bottom of liquid culture tank is communicated with the top of liquid culture tank by air circulation duct, in air circulation duct, be provided with for the air pump of recirculated air with for heating the temperature regulator of temperature in air circulation duct, in air circulation duct, be also arranged with magnetic field-high pressure producer, also comprise electrolysis oxygen tank, the output channel of the electrolysis positive electrode of electrolysis oxygen tank is communicated with air circulation duct, also comprise the oxygen electrode for detection of dissolved oxygen rate that is arranged in liquid culture tank and for detection of pH meter and the thermometer of pH value and temperature.
The high voltage electrode of magnetic field-high pressure producer as above is deep in air circulation duct in indenting mode.
On top ends of cans as above, be provided with stopple coupon and feed-pipe, on stopple coupon, be provided with sample cock, on feed-pipe, be provided with air filter, in described air circulation duct, be provided with air filter, the part that the bottom of liquid culture tank is communicated with air circulation duct is provided with bottom valve.
Air filter in temperature regulator as above, magnetic field-high pressure producer, air pump and air circulation duct is arranged in order setting from the top of liquid culture tank to bottom, and the air filter in temperature regulator, magnetic field-high pressure producer, air pump and air circulation duct is all higher than the liquid level in liquid culture tank, and the output channel of the electrolysis positive electrode of electrolysis oxygen tank is communicated with the part between magnetic field-high pressure producer with temperature regulator in air circulation duct.
Compared with prior art, this device has the following advantages and effect in the present invention:
1, by electrolysis oxygen, supply cell, tissue or organism in nutrient solution required oxygen, oxygen concn is adjustable, can by oxygen concn, regulate easily the metabolism and growth of cell, tissue or organism in nutrient solution, the object that reaches research or produce;
2, air lift type stirs air cycle used and adopts internal recycle gas, the disadvantageous effect of having avoided topsoil (as sulfurous gas) to bring the cell in nutrient solution, tissue or organism;
3, no matter in aerobic cultivation or anaerobism, cultivate, can adopt the mode of generating negative ions to regulate pH value and the redox potential of nutrient solution, and do not need to add acid-base modifier, and whole process can adjust pH and redox potential, the link that has solved the problem that in traditional zymotic process, restive pH value changes and avoided adding acid-alkali accommodation reagent;
4, no matter in aerobic cultivation or anaerobism, cultivate, can adopt in the mode of generating negative ions and the oxygenant or the free radical that in cell growth process, produce, thereby improve vigor and meta-bolites production time and the output of cell;
5, in internal recycle gas, contain a large amount of gases such as vapour molecule, by high-strength magnetic field, these gases especially vapour molecule are magnetized, again these magnetized molecules are passed in fermentor tank, can improve significantly Growth of Cells vigor and increment and promote metabolism.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention.
In figure: 1-electrolysis oxygen tank, 2-magnetic field-high pressure producer, 3-electrolysis positive electrode, 4-electrolysis negative potential, 5-oxygen electrode, 6-power-supply controller of electric, 7-temperature regulator, 8-feed-pipe, 9-air pump, 10-bottom valve, 11-liquid culture tank, 12-air circulation duct, 13-pH takes into account thermometer, 14-thief hole and valve thereof, 15-top ends of cans, 16-air filter.
Embodiment
Below in conjunction with accompanying drawing, the present invention is described in further detail:
Implement 1:
As shown in Figure 1, a kind of air lift type magnetic negative ion bio-reactor, comprise electrolysis oxygen tank 1, be arranged on the magnetic field-high pressure producer 2 on the oxygen path on electrolysis oxygen tank 1, the air filter 16 in temperature regulator 7, feed-pipe 8 and air pump 9, the pH meter being connected with controller and thermometer 13, thief hole and the valve 14 thereof of electrolysis positive electrode 3 and electrolysis negative potential 4, oxygen electrode 5 and power-supply controller of electric 6, liquid culture tank 11 and bottom valve 10 thereof, air circulation duct 12 and inside thereof, top ends of cans 15, air circulation duct 12.
The mode of the electrolysis positive electrode 3 of electrolysis oxygen tank 1 by brine electrolysis discharges oxygen and enters magnetic field-high pressure producer 2, the high voltage electrode that is positioned at the magnetic field-high pressure producer 2 in air circulation duct 12 is deep into pipeline in indenting mode, described magnetic field-high pressure producer 2 is high direct voltage, high pressure range is at 2000-20000 volt, enters air circulation duct 12 again enter liquid culture tank 11 again through the gas of magnetic field-high pressure producer 2; Described 2 pairs of oxygen that pass through of magnetic field-high pressure producer and carbonic acid gas ionize and produce negative oxygen ion; The oxygen electrode 5 that is positioned at liquid culture tank 11 is connected with power-supply controller of electric 6.This power-supply controller of electric 6 can be according to switch or the current/voltage that scope control electrolysis positive electrode 3 is set of oxygen electrode 5, and the dissolved oxygen rate scope that described oxygen electrode 5 is measured is at 0-30.0mg/L,
The gases such as 2 pairs of oxygen that pass through of magnetic field-high pressure producer, carbonic acid gas and water molecules gas ionize and produce negative ion.When anaerobically fermenting, in internal recycle gas with a large amount of vapour molecules, nitrogen molecule and carbon dioxide molecule, nitrogen, carbonic acid gas and vapour molecule under the promotion of high-pressure pump through magnetic field-high pressure producer in, the gas molecules such as carbonic acid gas are with negative charge to form active negative ion under the catalysis of water molecules, this negative ion can improve the pH value of fermented liquid after entering fermented liquid on the one hand, on the other hand can in and the free radical, the reduction-state that maintains fermented liquid that in cell growth process, produce, improve cell viability; When aerobic fermentation, the oxygen molecule passing into becomes the main negative ion of internal recycle gas, plays as above beneficial effect.
Magnetic field-high pressure producer 2 is with high-intensity magnetic field, to through gas molecule especially water molecules after magnetizing, pass into again in fermentor tank, can improve fermentor tank inner cell vigor, promote Growth and metabolism.Magneticstrength is 1000-7000mT(milli tesla), described magnetic field can be that additional permanent magnet produces, and can be also that electro-magnet energising produces.
The pH meter and the thermometer 13 that are positioned at liquid culture tank 11 are connected with power-supply controller of electric 6.This power-supply controller of electric 6 can carry out the work of controlling magnetic field-high pressure producer 2 according to the redox potential data of liquid culture pot liquid nutrient solution, according to the work of the variation control heater of fermented liquid temperature.
Embodiment 2: the application example in aerobic cell cultures.
To the north of Chinese caterpillar fungus hypha fermentation be example:
Step 1, first according to the optimal temperature set temperature controller of Cordyceps militaris growth, temperature range in liquid culture tank 11 is controlled at 15-16 degree Celsius, by recycle gas, regulate the temperature in liquid culture tank 11, according to conventional aseptic technique, aseptic culture medium is injected in tank body again, the composition of aseptic culture fluid comprises deionized water, yeast extract paste, glucose, potassium primary phosphate and zinc sulfate, quality ratio is: 1500(g): 5(g): 50(g): 0.3(g): 0.3(g), under aseptic condition, access Cordyceps militaris bacterial classification (Cordyceps militatirs).
Step 2, open air pump 9, the gas in air circulation duct 12 starts circulation, the fermented liquid in liquid culture tank 11 is stirred, to power-supply controller of electric 6 arrange dissolved oxygen rate scope in 16-18mg/L and pH value scope at 6.0-7.5.
Step 3, when the dissolved oxygen rate of fermented liquid is during lower than 16mg/L, power-supply controller of electric 6 is opened electrolysis power automatically, electrolysis positive electrode 3 produces oxygen and enters the airborne oxygen concentration of circulating line replenishment cycles, until the dissolved oxygen rate in fermented liquid reaches after 18mg/L, electrolysis power is closed automatically; PH value in fermented liquid is when setting 6.0, temperature regulator 7 is opened the power supply of magnetic field-high pressure producer 2 automatically, magnetic field-high pressure producer 2 produces 2500 volts of voltages, recycle gas is carried out to high pressure ionization and produce negative ion, a large amount of negative ions enters fermented liquid and causes pH value to raise, when reaching 7.5, the power supply of magnetic field-high pressure producer 2 is closed automatically.So automatically the dissolved oxygen rate of controlled fermentation liquid is stabilized in 16-18mg/L and pH value all-the-time stable at 6.0-7.5, and does not need the artificial adjusting reagent that adds.
Magnetic field-high pressure producer 2 is except with high voltage electrode, and also with high intensity electromagnetic field, magneticstrength is arranged on 7000mT(milli tesla), this magnetic field is that electro-magnet passes into the high-strength magnetic field that civil power produces.Magnetize the gases such as internal recycle water molecules that pass through in this magnetic field, and these magnetized negative ions or magnetization molecule enter the vigor that can improve cell after fermented liquid, promote metabolism and growth.
Adopt as above method to cultivate Cordyceps militaris bacterial classification, with ordinary method fermentation in contrast, result is as follows:
Figure BDA0000429441900000051
Embodiment 2: the application example in anaerobic cell is cultivated.
Take intestinal bacteria cultivation as example,
Step 1, first according to the optimal temperature set temperature controller of Escherichia coli Growth, temperature in liquid culture tank 11 is controlled at 36-37 degree Celsius, according to conventional aseptic technique, in tank body, inject LB aseptic culture medium again, under aseptic condition, access intestinal bacteria bacterial classification (TOP10).Open air pump, gas in circulating line starts circulation, fermented liquid in tank body is stirred, the electrolysis power of closing controller carries out anaerobically fermenting, controller (6) is arranged to pH value scope at 6.0-7.0, pH value in fermented liquid is lower than 6.0 time, controller is opened the power supply of magnetic field-high pressure producer automatically, recycle gas is carried out to high pressure ionization and produce negative ion, a large amount of negative ions enters fermented liquid and causes pH value to raise, when reaching 7.0, the power supply of magnetic field-high pressure producer 2 is closed automatically.So automatic controlled fermentation liquid pH value.
Adopt as above method to cultivate intestinal bacteria, compare with cellar culture method (the gas for stirring not being carried out to high pressure ionization), colibacillary increment is more than doubled, and the bacterial classification paracme has postponed more than 2 weeks.
Specific embodiment described herein is only to the explanation for example of the present invention's spirit.Those skilled in the art can make various modifications or supplement or adopt similar mode to substitute described specific embodiment, but can't depart from spirit of the present invention or surmount the defined scope of appended claims.

Claims (4)

1. an air lift type magnetic negative ion bio-reactor, comprise liquid culture tank (11), it is characterized in that, the top of liquid culture tank (11) is provided with top ends of cans (15), the bottom of liquid culture tank (11) is communicated with the top of liquid culture tank (11) by air circulation duct (12), in air circulation duct (12), be provided with air pump (9) for recirculated air and for heating the temperature regulator (7) of temperature in air circulation duct, in air circulation duct (12), be also arranged with magnetic field-high pressure producer (2), also comprise electrolysis oxygen tank (1), the output channel of the electrolysis positive electrode of electrolysis oxygen tank (1) is communicated with air circulation duct (12), also comprise the oxygen electrode for detection of dissolved oxygen rate (5) that is arranged in liquid culture tank (11) and for detection of pH meter and the thermometer (13) of pH value and temperature.
2. a kind of air lift type magnetic negative ion bio-reactor according to claim 1, is characterized in that, the high voltage electrode of described magnetic field-high pressure producer (2) is deep in air circulation duct (12) in indenting mode.
3. a kind of air lift type magnetic negative ion bio-reactor according to claim 2, it is characterized in that, on described top ends of cans (15), be provided with stopple coupon and feed-pipe (8), on stopple coupon, be provided with sample cock, on feed-pipe, be provided with air filter, described air circulation duct is provided with air filter in (12), and the part that the bottom of liquid culture tank (11) is communicated with air circulation duct (12) is provided with bottom valve (10).
4. a kind of air lift type magnetic negative ion bio-reactor according to claim 3, it is characterized in that, described temperature regulator (7), magnetic field-high pressure producer (2), air filter in air pump (9) and air circulation duct (12) is arranged in order setting from the top of liquid culture tank (11) to bottom, and temperature regulator (7), magnetic field-high pressure producer (2), air filter in air pump (9) and air circulation duct (12) is all higher than the liquid level in liquid culture tank (11), the output channel of the electrolysis positive electrode of electrolysis oxygen tank (1) is communicated with the part between magnetic field-high pressure producer (2) with the upper temperature regulator (7) of air circulation duct (12).
CN201310645712.4A 2013-12-04 2013-12-04 Airlift type magnetic anion bioreactor Expired - Fee Related CN103627636B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106434293A (en) * 2016-09-20 2017-02-22 成都测迪森生物科技有限公司 Rotary type biological fermentation container

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1153114A (en) * 1967-02-06 1969-05-21 Ajinomoto Kk Fermentation processes and apparatus for Carrying Out the same
US20070218540A1 (en) * 2004-05-26 2007-09-20 Serge Guiot Bioelectrolytic Methanogenic/Methanotrophic Coupling for Bioremediation of Ground Water
CN102576892A (en) * 2009-08-07 2012-07-11 西安大略大学 Bio-fuel cell system
CN203593760U (en) * 2013-12-04 2014-05-14 湖北民族学院 Air-lift magnetic anion bio-reactor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1153114A (en) * 1967-02-06 1969-05-21 Ajinomoto Kk Fermentation processes and apparatus for Carrying Out the same
US20070218540A1 (en) * 2004-05-26 2007-09-20 Serge Guiot Bioelectrolytic Methanogenic/Methanotrophic Coupling for Bioremediation of Ground Water
CN102576892A (en) * 2009-08-07 2012-07-11 西安大略大学 Bio-fuel cell system
CN203593760U (en) * 2013-12-04 2014-05-14 湖北民族学院 Air-lift magnetic anion bio-reactor

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106434293A (en) * 2016-09-20 2017-02-22 成都测迪森生物科技有限公司 Rotary type biological fermentation container

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