CN109897789A - Novel Klebsiella or Candida spp and microbiological fuel cell comprising it - Google Patents
Novel Klebsiella or Candida spp and microbiological fuel cell comprising it Download PDFInfo
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- CN109897789A CN109897789A CN201910329033.3A CN201910329033A CN109897789A CN 109897789 A CN109897789 A CN 109897789A CN 201910329033 A CN201910329033 A CN 201910329033A CN 109897789 A CN109897789 A CN 109897789A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- 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
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- 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
- C12N1/14—Fungi; Culture media therefor
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- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
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- 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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- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M8/00—Fuel cells; Manufacture thereof
- H01M8/16—Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/22—Klebsiella
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/72—Candida
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/50—Fuel cells
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P70/00—Climate change mitigation technologies in the production process for final industrial or consumer products
- Y02P70/50—Manufacturing or production processes characterised by the final manufactured product
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Abstract
The present invention relates to a kind of novel Klebsiella IR21 (Klebsiella sp.IR21) or Mycotoruloides (Candida sp.IR11) bacterial strains, the composition for being used for wastewater treatment or purification of waste water comprising the bacterial strain, the method for carrying out wastewater treatment or purification using the composition, microbiological fuel cell comprising the bacterial strain, the method produced electricity using the microbiological fuel cell, the wastewater treatment equipment for having the microbiological fuel cell, and utilize wastewater treatment equipment progress wastewater treatment or the method for purification.Utilize novel Klebsiella IR21 bacterial strain of the invention or Mycotoruloides IR11, it not only can be achieved at the same time to high chemical oxygen demand (COD, Chemical Oxygen Demand) value waste water purification and electricity production, but also can develop it is a kind of have the wastewater treatment equipment of microbiological fuel cell without what external power supply can run wastewater treatment equipment, therefore can be widely applied to more efficient field of waste water treatment.
Description
The application is divisional application, and the applying date of original application is on April 23rd, 2015, application No. is
201580000154.3 entitled " novel Klebsiella or Candida spp and the Microbial fuel comprising it
Battery ".
Technical field
The present invention relates to a kind of novel Klebsiella or Candida spp and comprising its microbiological fuel cell,
More particularly it relates to a kind of novel Klebsiella IR21 (Klebsiella sp.IR21) or Mycotoruloides
IR11 (Candida sp.IR11) bacterial strain, utilizes institute at the composition for being used for wastewater treatment or purification of waste water comprising the bacterial strain
Composition is stated to carry out the method for wastewater treatment or purification, the microbiological fuel cell comprising the bacterial strain, utilize the microorganism
The method of fuel cell electricity production, the wastewater treatment equipment for having the microbiological fuel cell and the utilization wastewater treatment fill
Set the method for carrying out wastewater treatment or purification.
Background technique
Microbiological fuel cell (microbial fuel cell:MFC) be it is a kind of by microorganism be used as catalyst and will make
The device of electric energy is converted to for the chemical energy that the matrix of electron donor has.It specifically refers to following device: including by matrix because micro-
The metabolism of biology and the chemical energy generated during decomposing are converted to the equipment and the recycling electric energy converted out of electric energy
Equipment is finally produced electricity using microorganism from matrix.In general, microbiological fuel cell is by being inoculated with the anode (Anode) of microorganism
Electrode, cathode (cathode) electrode, reaction tank and cation-exchange membrane are constituted, in the reaction tank, with cation exchange
Centered on film and opposite side is arranged in anode electrode and cathode electrode each other.If in the training that reaction tank filling includes matrix
Nutrient solution generates hydrogen ion and electricity as decomposition product then the microorganism for being seeded in anode electrode decomposes the matrix
Son, the hydrogen ion of the generation penetrate cation-exchange membrane and are transmitted to cathode electrode, the electronics of the generation from anode electrode to
Cathode electrode is mobile, finally generates electric current.Described micro- because as long as matrix generates hydrogen ion and electronics under the action of microorganism
Biological fuel cell can produce electric current, as long as that is, sustainably generation is electric so constantly supplying matrix to the reaction tank
Stream.The core of this microbiological fuel cell is to be seeded in the microorganism of anode electrode, and the microorganism divides matrix
The efficiency for solving and generating hydrogen ion and electronics is higher, and the electricity production of microbiological fuel cell is more.
On the other hand, sewage or waste water processing unit generally comprises: sedimentation basin, to stand flowed into sewage or give up
Water makes the solid content to suspend at removing it after fractional precipitation；Aerobic tank is connect with the sedimentation basin, has pure oxygen aeration dress
It sets；And anaerobic pond, it is connect with the aerobic tank, under anaerobic state, passes through microorganism decomposition organic matter.Using the dirt
In the expense that water or wastewater treatment equipment handle contaminant water, pure oxygen aeration and excess sludge handle shared expense highest.
The power consumption of pure oxygen aeration device increases with the size of process object, moreover, if passing through pure oxygen aeration processing pollution
Water then a large amount of aerobic microorganism i.e. excess sludge will be generated, therefore causes excess sludge processing cost to increase.Cause
This, it is believed that the sewage or waste water treatment effeciency of described device is mentioned with the electricity supplied to sewage or waste water processing unit
It is high.
Therefore, it is actively being implemented following research: by the anaerobic pond for having the sewage or waste water processing unit
In conjunction with microbiological fuel cell, described device electricity consumption is reduced.For example, being disclosed in Korean Patent No. 2013-0029530
The power self-support type advanced waste treatment apparatus of a kind of fusion microbiological fuel cell and microorganism electrolysis cell and described in utilizing
The wastewater treatment method of device.But the dress of even this combination sewage or waste water processing unit and microbiological fuel cell
It sets, performance is still driven by used microorganism efficiency, so organic matter is converted to hydrogen there is an urgent need to develop a kind of out
The outstanding microorganism of the efficiency of ion, but up to now, it there is no report to disclose such microorganism.
Summary of the invention
[problems to be solved by the invention]
In this background, the present inventor spares no effort to study that a kind of that organic matter is converted to hydrionic efficiency is outstanding
And the bacterial strain outstanding to the applicability of microbiological fuel cell, finally confirm novel Klebsiella or Candida spp
There is outstanding applicability to microbiological fuel cell, and then complete the present invention.
[means solved the problems, such as]
One of the objects of the present invention is to provide a kind of novel Klebsiella IR21 (Klebsiella sp.IR21)
Or Mycotoruloides IR11 (Candida sp.IR11) bacterial strain.
Another object of the present invention is to provide a kind of for wastewater treatment or the composition of purification of waste water, and it includes described
Klebsiella IR21 or Mycotoruloides IR11 bacterial strain.
Another object of the present invention is to provide the composition described in a kind of utilize for wastewater treatment or purification of waste water, into
Row wastewater treatment or the method for purification.
Another object of the present invention is to provide a kind of microbiological fuel cell, it includes the Klebsiella IR21
Or Mycotoruloides IR11 bacterial strain.
Another object of the present invention is to provide a kind of method produced electricity using the microbiological fuel cell.
Another object of the present invention is to provide a kind of wastewater treatment equipment, has the microbiological fuel cell.
Another object of the present invention is to provide a kind of to carry out wastewater treatment or purification using the wastewater treatment equipment
Method.
[The effect of invention]
Using novel Klebsiella IR21 bacterial strain of the invention or Mycotoruloides IR11, not only can be achieved at the same time to height
The purification and electricity production of the waste water of COD value, but also can develop and a kind of can run wastewater treatment equipment without external power supply
Have the wastewater treatment equipment of microbiological fuel cell, therefore can be widely applied to more efficient field of waste water treatment.
Detailed description of the invention
Fig. 1 a is to indicate that COD and pH value are fired with the microorganism comprising using the Klebsiella IR21 bacterial strain of anaerobic sludge
Expect the chart of the runing time variation of battery.
Fig. 1 b is to indicate voltage with the Microbial fuel electricity comprising using the Klebsiella IR21 bacterial strain of anaerobic sludge
The chart of the runing time variation in pond.
Fig. 2 a is to indicate COD and pH value with the Microbial fuel electricity comprising using the Mycotoruloides IR11 bacterial strain of anaerobic sludge
The chart of the runing time variation in pond.
Fig. 2 b is to indicate voltage with the microbiological fuel cell comprising using the Mycotoruloides IR11 bacterial strain of anaerobic sludge
The chart of runing time variation.
Fig. 3 a is to indicate COD and pH value with the Microbial fuel electricity comprising Klebsiella IR21 bacterial strain and artificial substratum
The chart of the runing time variation in pond.
Fig. 3 b is to indicate voltage with the microbiological fuel cell comprising Klebsiella IR21 bacterial strain and artificial substratum
The chart of runing time variation.
Fig. 4 a is to indicate COD and pH value with micro- life comprising the waste water in Klebsiella IR21 bacterial strain and hydrogen production process
The chart of the runing time variation of object fuel cell.
Fig. 4 b is to indicate that voltage is fired with the microorganism comprising the waste water in Klebsiella IR21 bacterial strain and hydrogen production process
Expect the chart of the runing time variation of battery.
Fig. 5 a is to indicate COD and pH value with the microbiological fuel cell comprising Mycotoruloides IR11 bacterial strain and artificial substratum
The chart of runing time variation.
Fig. 5 b is to indicate voltage with the operation comprising Mycotoruloides IR11 bacterial strain and the microbiological fuel cell of artificial substratum
The chart of time change.
Fig. 6 a is to indicate COD and pH value with micro- life comprising the waste water in Mycotoruloides IR11 bacterial strain and biogas production process
The chart of the runing time variation of object fuel cell.
Fig. 6 b is to indicate that voltage is fired with the microorganism comprising the waste water in Mycotoruloides IR11 bacterial strain and biogas production process
Expect the chart of the runing time variation of battery.
Fig. 7 is to being inoculated with the MFC reaction tank of Klebsiella bacterial strain and be not inoculated with the MFC of Klebsiella bacterial strain
The chart that Klebsiella bacterial strain amount in the biomembrane of reaction tank is compared.
Fig. 8 is to being inoculated with the MFC reaction tank of Candida spp and be not inoculated with the MFC reaction tank of Candida spp
The chart that Candida spp amount in biomembrane is compared.
Specific embodiment
In order to find the bacterial strain suitable for microbiological fuel cell, the present inventor makes micro- life that operation is inoculated with anaerobic sludge
Object fuel cell from the biological UF membrane for being formed in anode electrode and identifies novel Klebsiella (Klebsiella later
Sp.) bacterial strain, that is, Klebsiella IR21 bacterial strain or Mycotoruloides (Candida sp.) bacterial strain, that is, Mycotoruloides IR11.Separately
Outside, the performance of the microbiological fuel cell comprising the Klebsiella IR21 is assessed, as a result confirming will have
In the case where thering is waste water in the hydrogen production process of high-grade COD value to be used as matrix, electric current also can produce, to including the beads
The performance of the microbiological fuel cell of Pseudomonas IR11 is assessed, and is as a result confirmed to the voltage of microbiological fuel cell and is generated
Time needed for until stabilization shortens, and can produce the voltage of relatively high grade, thus the Klebsiella IR21 or
Mycotoruloides IR11 bacterial strain can be used not only as the core element of microbiological fuel cell, but also can combine wastewater treatment
Central role is played in terms of device and microbiological fuel cell.
As the embodiment to reach the purpose, the present invention provides a kind of novel Klebsiella
(Klebsiella sp.) bacterial strain, that is, Klebsiella IR21 (Klebsiella sp.IR21) bacterial strain (KCTC 12571BP),
Or novel Mycotoruloides (Candida sp.) bacterial strain, that is, Mycotoruloides IR11 (Candida sp.IR11) bacterial strain (KCTC
12570BP)。
Term " Klebsiella IR21 (Klebsiella sp.IR21) bacterial strain " in the present invention refers to from anaerobism
Sludge in the past from unreported Klebsiella (Klebsiella sp.) bacterial strain being disclosed, and is referred to April 3 in 2014
Day is saved into the Korea Institute of Bioengineering Biological Resource Center as one of international saving mechanism in budapest treaty
(South Korea's Culture Collection, Korean Collection for Type Culture) and it is endowed preservation number
The bacterial strain of KCTC12571BP.
Term " Mycotoruloides IR11 (Candida sp.IR11) bacterial strain " in the present invention refers to from anaerobic sludge, with
Toward from unreported Mycotoruloides (Candida sp.) bacterial strain being disclosed, and refers to and be saved on April 3rd, 2014 as cloth
The Korea Institute of Bioengineering Biological Resource Center of one of international saving mechanism in Da Peisi treaty (protect by South Korea's type species
Hiding center, Korean Collection for Type Culture) and it is endowed the bacterial strain for saving number KCTC 12570BP.
In the present invention, in the case where microbiological fuel cell includes the Klebsiella IR21 bacterial strain, even if
Use the waste water of high-grade COD value as matrix used in microbiological fuel cell, also can reach the effect for generating electric current,
In the case where microbiological fuel cell includes the Mycotoruloides IR11 bacterial strain, the electricity of microbiological fuel cell generation can be shortened
Pressure tends towards stability the required time, and can increase voltage class, therefore can be used as the microorganism provided in the present invention combustion
Expect the component part of battery.
According to one embodiment of present invention, in order to screen the bacterium for microbiological fuel cell for being present in anaerobic sludge
Strain runs microbiological fuel cell using the anaerobic sludge, from the anode electrode for being formed in the microbiological fuel cell
Biological membrane choosing is used for the bacterial strain of microbiological fuel cell and is identified, as a result, isolating novel Klebsiella
(Klebsiella sp.) bacterial strain or Mycotoruloides (Candida sp.) bacterial strain, are respectively designated as " citric acid for the bacterial strain
Pseudomonas IR21 (Klebsiella sp.IR21) " or " Mycotoruloides IR11 (Candida sp.IR11) ", and April 3 in 2014
Day the bacterial strain is saved into raw as the Korea Institute of Bioengineering of one of international saving mechanism in budapest treaty
Object resource center (South Korea's Culture Collection, Korean Collection for Type Culture) is simultaneously assigned respectively
It gives and saves number KCTC 12571BP and KCTC 12570BP.On the other hand, to the identified Klebsiella IR21
Or the effect that Mycotoruloides IR11 plays microbiological fuel cell confirms, as a result confirms the bacterial strain to artificial base
Organic matter included in the waste water in waste water or biogas production process in glucose, hydrogen production process included in matter carries out
It decomposes, generates hydrogen ion and electronics and the hydrogen ion and electronics of transferring the generation, thus can drive microbiological fuel cell.It removes
Except this, further acknowledges the microbiological fuel cell comprising the Klebsiella IR21 and do not include micro- life of the bacterial strain
The case where object fuel cell is compared, and the waste water in the hydrogen production process for using artificial substratum or high-grade COD value is as matrix
Under, remain to generate electric current (Fig. 3 a, 3b, 4a and 4b), the microbiological fuel cell comprising the Mycotoruloides IR11 with do not include
The microbiological fuel cell of the bacterial strain is compared, use the waste water in artificial substratum or biogas production process as the feelings of matrix
Under condition, also can be shortened to voltage generate tend towards stability until needed for time, generate relatively high grade voltage (Fig. 5 a, 5b,
6a and 6b).
Therefore, it is known that Klebsiella IR21 bacterial strain or Mycotoruloides IR11 bacterial strain provided in the present invention not only can be used
Make the core element of microbiological fuel cell, but also device and the Microbial fuel electricity to wastewater treatment can combined
Central role is played in terms of pond.
As another embodiment to reach the purpose, the present invention provides a kind of comprising the Klebsiella
IR21 bacterial strain or Mycotoruloides IR11 bacterial strain for wastewater treatment or purification of waste water composition and using the composition into
Row wastewater treatment or the method for purification.
The Klebsiella IR21 bacterial strain is originated from anaerobic sludge, to organic included in the waste water in hydrogen production process
Object is decomposed, and is generated hydrogen ion and electronics and the hydrogen ion and electronics of transferring the generation, thus can be driven Microbial fuel
Battery, therefore the bacterial strain can be used for organic matter included in anaerobic digestion waste water.
The Mycotoruloides IR11 bacterial strain is originated from anaerobic sludge, to organic included in the waste water in biogas production process
Object is decomposed and generates hydrogen ion and electronics, thus can drive microbiological fuel cell, therefore the bacterial strain can be used for anaerobism
Organic matter included in Decomposition Wastewater.
It is provided in the present invention to be used for waste water other than the Klebsiella IR21 or Mycotoruloides IR11 bacterial strain
The composition of processing or purification of waste water also may include other bacterial strains that can promote wastewater treatment or purification.Its that also may include
There is no particular restriction for his bacterial strain, but the bacterial strain of organic matter included in preferably decomposable waste water, construction inhibit spoilage organisms raw
The bacterial strain etc. of elongate member；The saccharomyces (Saccharomyces sp.) of organic matter included in more preferably decomposable waste water
Bacterial strain, bacillus (Bacillus sp.) bacterial strain reduce pH value and build the intestines ball for inhibiting spoilage organisms growth conditions
Pseudomonas (Enterococcus sp.) bacterial strain etc.；It most preferably can alone or in combination include saccharomyces cerevisiae (Saccharomyces
Cerevisiae), hay bacillus (Bacillus subtilis), enterococcus faecium (Enterococcus faecium), excrement intestines ball
Bacterium (Enterococcus faecalis) etc..
In addition, including by the composition pair using the method that the composition of the invention carries out wastewater treatment or purification
The step of waste water is handled, and reacts it under anaerobic.
At this point, the composition of the invention because of the reaction can divide carbohydrate component included in waste water
Solution, there is no particular restriction relative to the treating capacity of the waste water for composition provided in the present invention, as long as can reach wastewater treatment
Or purification effect, but preferably according to 1,000 ton of waste water using the composition 50 to the amount of 200kg/day at
Reason；More preferably handled according to 1,000 tons of waste water using the amount of composition 100 to the 150kg/day；Most preferably press
It is handled according to 1,000 tons of waste water using the amount of the composition 120kg/day.
As the another embodiment to reach the purpose, the present invention provides a kind of comprising the Klebsiella
The method of the microbiological fuel cell and utilization microbiological fuel cell electricity production of IR21 or Mycotoruloides IR11 bacterial strain.
Term " microbiological fuel cell " in the present invention refers to following device: comprising by matrix due to the metabolism of microorganism
The chemical energy generated during decomposition is converted to the equipment of electric energy and the equipment of the recycling electric energy converted out, final benefit
It is produced electricity with microorganism from matrix.In general, microbiological fuel cell includes: anode electrode is inoculated with microorganism；Cathode electrode；
Culture solution, it includes can provide the culture environment of microorganism by the matrix of the microorganism decomposition；Electric wire connects the electricity
Pole；Reaction tank has the inflow entrance and outflux of described two electrodes, the culture solution；And cation-exchange membrane, setting
In the reaction tank, it is located between two electrodes；In the reaction tank, centered on cation-exchange membrane and anode electrode and
Opposite side is arranged in cathode electrode each other, and the culture liquid flow inlet and outflux that the reaction tank has, which are located at, is provided with anode
The position of electrode.When the inflow entrance by the reaction tank fills the culture solution comprising matrix to reaction tank, it is seeded in sun
The microorganism of pole electrode decomposes matrix included in the culture solution, as decomposition product, generates hydrogen ion and electricity
Son, the hydrogen ion of the generation penetrates cation-exchange membrane and is transmitted to cathode electrode, while electronics is electric from anode electrode to cathode
Ghandler motion is dynamic, finally generates electric current.Because as long as matrix generates hydrogen ion and electronics, the microorganism combustion under the action of microorganism
Material battery can produce electric current, as long as so the inflow entrance and outflux that have by the reaction tank are continually provided base
Matter sustainably generates electric current.
In the present invention, the microbiological fuel cell includes: (a) anode electrode is inoculated with provided in the present invention
Klebsiella IR21 or Mycotoruloides IR11 bacterial strain；(b) cathode electrode；(c) culture solution, it includes can be by the microorganism
The matrix of decomposition provides the culture environment of microorganism；(d) electric wire connects the electrode；(e) reaction tank has described two
The inflow entrance and outflux of a electrode, the culture solution；And (f) cation-exchange membrane, setting are located at two in the reaction tank
Between a electrode.At this point, there is no particular restriction for the anode electrode, as long as Klebsiella IR21 or Mycotoruloides can be inoculated with
IR11 bacterial strain, but the anode electrode manufactured by materials such as platinum, stainless (steel) wire, carbon felt, carbon papers preferably can be used；The yin
There is no particular restriction for pole electrode, but the cathode electrode manufactured by materials such as carbon felt, carbon paper, carbon cloths preferably can be used or use empty
Gas cathode (air-cathode)；There is no particular restriction for the matrix, as long as can be by Klebsiella IR21 or Mycotoruloides
IR11 bacterial strain decomposes and generates hydrogen ion and electronics, but the mixture of glucose and acetate preferably can be used, through pre-
The waste water in the mixture (1: 1, w/w) or hydrogen production process of glucose and acetate more preferably can be used in the waste water etc. of processing
Or the waste water in biogas production process；There is no particular restriction for the culture solution, as long as can provide Klebsiella IR21 or thought
The culture environment of pearl Pseudomonas IR11 bacterial strain and the hydrogen ion and electronics for transmitting generation, but preferably can be used comprising carbon source and
Electron acceptor, and the buffer solution of pH neutral can be kept, the more preferably usable glucose comprising as carbon source and conduct
The Fe of electron acceptor4O21P6Or C6H5FeO7, and the phosphate buffer that pH value is 7.0, most preferably can be used includes conduct
The Fe of the glucose of the 20mM of carbon source and the 50mM as electron acceptor4O21P6Or C6H5FeO7, and the 100mM that pH value is 7.0
Phosphate buffer.
In addition, being comprised the following steps using the method that the microbiological fuel cell produces electricity: (a) passing through Microbial fuel electricity
The inflow entrance and outflux that the reaction tank in pond has, to being inoculated with Klebsiella IR21 or Mycotoruloides IR11 bacterial strain
Anode electrode continuously feeds the step of including the culture solution of matrix；And (b) it is seeded in the Klebsiella of the anode electrode
Belong to IR21 or Mycotoruloides IR11 bacterial strain to decompose the matrix, generate hydrogen ion and electronics, by the hydrogen ion and
Electronics generates electric current.At this point, the conditions such as used matrix, culture solution are identical as condition as described above.
As the another embodiment to reach the purpose, the present invention, which provides one kind, to be had comprising the citric acid
The wastewater treatment equipment and the utilization wastewater treatment of the microbiological fuel cell of Pseudomonas IR21 or Mycotoruloides IR11 bacterial strain fill
The method set while carrying out purification of waste water and electricity production.
As described above, Klebsiella IR21 provided in the present invention or Mycotoruloides IR11 bacterial strain can pass through microorganism
Fuel cell is produced electricity to being decomposed through organic matter included in the pretreated waste water of wastewater treatment equipment, so if
The microbiological fuel cell is set on wastewater treatment equipment, electricity needed for supplying wastewater treatment equipment with this, then described useless
Even if water treatment facilities can also independently carry out wastewater treatment or purification without external power supply.
In the wastewater treatment equipment for having the microbiological fuel cell, it should remove to the maximum extent and be supplied to microorganism
Meeting in the pretreated waste water of fuel cell inhibits the active ingredient of microbiological fuel cell, therefore can be used by common
The wastewater treatment equipment sedimentation basin, the aerobic tank that have etc. comprising organic matter but to remove most of ingredient other than organic matter
Mode carry out pre-processing resulting waste water.Be inoculated on the anode electrode of the microbiological fuel cell can to organic matter into
The Klebsiella IR21 or Mycotoruloides IR11 bacterial strain of row anaerobic digestion, therefore the microbiological fuel cell can be applied flexibly
Make anaerobic pond, when multiple small-sized microbiological fuel cells are arranged in order to improve the efficiency of fuel cell generation of the microbiological fuel cell
When, the sum total of these small-sized microbiological fuel cells and the capacity of anaerobic pond are close, it is not necessary that anaerobic pond is especially arranged, therefore also have
There is the advantages of can simplifying the wastewater treatment equipment structure.
In the present invention, having the wastewater treatment equipment of the microbiological fuel cell, there is no particular restriction, but preferably
It may include sedimentation basin, aerobic tank and microbiological fuel cell, also may include waste water inflow entrance, waste water discharge on this basis
Mouth, electrical storage device etc..
On the other hand, purification of waste water is carried out while in the present invention and the method for electricity production comprises the following steps: (a) making waste water
It is purified after being flowed into the wastewater treatment equipment for having the microbiological fuel cell, it is useless to obtain the pretreatment containing organic matter
The step of water；And the step of pretreated waste water that above-mentioned steps obtain is produced electricity (b) is fed through to microbiological fuel cell.
Implementation form
Hereinafter, the present invention will be described in more detail by embodiment.But these embodiments are only to schematic
Ground illustrates the present invention, and it's not limited to that for the scope of the present invention.
Embodiment 1: the screening and identification of the bacterial strain for microbiological fuel cell
In order to screen the bacterial strain for microbiological fuel cell being present in anaerobic sludge, transported using the anaerobic sludge
After row microbiological fuel cell, the bacterial strain of microbiological fuel cell is used for from the biological membrane choosing being formed on anode (Anode)
And it is identified.
Specifically, the microbiological fuel cell utilizes Single chamber systems (Anode: as 450
Carry out pre-processing for 30 minutes the carbon felt (9 × 14cm) of resulting anode electrode at DEG C；Air yin as cathode electrode
Pole (air-cathode, ODE75, MEET, Korea)；Proton exchange membrane:Nafion 117 (Dupont,
USA)；Reactor:7 × 15 × 10cm (L × H × W) per single chamber (1-L volume)；Voltage
Monitoring system:digital multimeter (GL220, Japan)；Line:titanium wire (Sigma,
USA)), wherein the anode electrode that the microbiological fuel cell is included is connected with cathode electrode outside platinum electrode and 100 Ω
Resistance.In addition, as the culture solution for microbiological fuel cell, using including 100mM phosphate buffer
(NH4Cl0.31g/l；KCl0.13g/l；Na2HPO4-12H2O21.838g/l；And NaH2PO4-2H2) and conduct O6.084g/l
Glucose and acetate mixture (1: 1, the w: culture solution w) that the COD of matrix is about 1,500mg/L.
To the microbiological fuel cell filling 1l described in culture solution, and as Initial Inoculum be inoculated with anaerobic sludge after,
Using multi meter (GL220_820-APS, GRAPHTEC, Japan) every 10 minutes observation primary voltages, operation about 80
Its (Fig. 1 a, 1b, 2a and 2b).
Fig. 1 a is to indicate that COD and pH value are fired with the microorganism comprising using the Klebsiella IR21 bacterial strain of anaerobic sludge
Expect the chart of the runing time variation of battery, Fig. 2 a is to indicate COD and pH value with the Mycotoruloides comprising using anaerobic sludge
The chart of the runing time variation of the microbiological fuel cell of IR11 bacterial strain.Fig. 1 b is to indicate voltage with comprising using anaerobic sludge
Klebsiella IR21 bacterial strain microbiological fuel cell runing time variation chart, Fig. 2 b is to indicate voltage with packet
The chart of the runing time variation of microbiological fuel cell containing the Mycotoruloides IR11 bacterial strain for using anaerobic sludge.
As shown in Fig. 1 a and Fig. 1 b, confirm comprising Klebsiella IR21 bacterial strain or Mycotoruloides IR11 bacterium
In the microbiological fuel cell of strain, voltage generates unstable, generation 200mV or less voltage.
As shown in Fig. 2 a and Fig. 2 b, confirm comprising Klebsiella IR21 bacterial strain or Mycotoruloides IR11 bacterium
In the microbiological fuel cell of strain, the required time is about 40 days or more until voltage is generated and tended towards stability, and is produced after stabilization
Raw 200mV voltage below.
After the operation for terminating the microbiological fuel cell, recycling is formed with the anode of biomembrane (biofilm)
(Anode), therefrom separate and identify aimed strain.
Specifically, in 100mM phosphate buffer (NH4Cl0.31g/l；KCl0.13g/l；Na2HPO4-
12H2O21.838g/l；And NaH2PO4-2H2O6.084g/l, pH value 7.0) in the Fe comprising 20mM glucose and 50mM4O21P6
Or C6H5FeO720ml inoculated and cultured liquid in, the anode (Anode) for being formed with the biomembrane is inoculated with, by nitrogen
In the state of saturation, with 35 DEG C of stir cultures.If the color of inoculated and cultured liquid becomes buff and turbidity increases, execute
4 times to this addition 10 times of volumes new inoculated and cultured liquid be diluted the step of and be diluted to 104Volume again, by the dilution
Culture be applied in the inoculated and cultured liquid comprising agarose solid medium on, in 35 DEG C of anaerobic reaction condition
It is cultivated under (nitrogen saturation conditions), to form bacterium colony.Each thallus is obtained from the bacterium colony of the formation, and is obtained described
The thallus taken is applied to NucleoSpin Soil Kit (Macherey-Nagel, Germany) and obtains the deoxidation of each thallus
Ribonucleic acid (DNA, Deoxyribonucleic Acid) is carried out using the DNA of the acquisition as template using following primer
PCR, to obtain amplified production.At this point, using 2720Thermal cycler (Applied Biosystems, USA) conduct
PCR instrument, PCR condition are as follows: 95 DEG C, 4 minutes；(95 DEG C, 30 seconds of 30cycle；55 DEG C, 30 seconds；And 72 DEG C, 30 seconds)；And 72 DEG C,
5 minutes.
Forward primer: 341f 5'-CCT ACG GGA GGC AGC AG-3'(sequence number 1)
Reverse primer: 907r 5'-CCC CGT CAA TTC ATT TGA GTT T-3'(sequence number 2)
Forward primer ITS1:5'-TCC GTA GGT GAA CCT GCG G-3'(sequence number 3)
Reverse primer ITS4:5'-TCC TCC GCT TAT TGA TAT GC-3'(sequence number 4)
The base sequence (MACROGEN, Korea) for determining the amplified production of the acquisition carries out determining base sequence
Analyze (NCBI Blast, www.ncbi.nim.nih.gov/), and to the thallus for each bacterium colony for generating the amplified production into
Row identification, therefrom screens novel strain.As a result, confirming one in the identified thallus to belong to fungi
(Fungi) (boundary), sac fungus (Ascomycota) (door), saccharomycete (Saccharomycotina) (subphylum), saccharomycete
(Saccharomycetales) the novel Klebsiella of (mesh), saccharomycete (Saccharomycetaceae) (section)
(Klebsiella sp.) or Mycotoruloides (Candida sp.) bacterial strain, and be " Klebsiella by the Strain Designation
IR21 (Klebsiella sp.IR21) " or " Mycotoruloides IR11 (Candida sp.IR11) ", on April 3rd, 2014 by institute
It states bacterial strain and is saved into Korea Institute of Bioengineering living resources as one of international saving mechanism in budapest treaty
Center (South Korea's Culture Collection, Korean Collection for Type Culture) is simultaneously endowed preservation number
KCTC 12571BP and KCTC 12570BP.
It knows under anaerobic state, the Klebsiella IR21 or Mycotoruloides IR11 bacterial strain can also use glucose
Make carbon source.
Embodiment 2: the assessment of Klebsiella IR21 bacterial strain or the microbiological fuel cell of Mycotoruloides IR11 is utilized
Operation includes the micro- of Klebsiella IR21 or Mycotoruloides IR11 bacterial strain identified in the embodiment 1
Biological fuel cell assesses its result.
Embodiment 2-1: microorganism culture environment and voltage change
Specifically, in the bacterium colony obtained by the embodiment 1, from being equivalent to Klebsiella IR21 or beads
The bacterium colony of Pseudomonas IR11 bacterial strain obtains thallus and is cultivated after being inoculated into inoculated and cultured liquid and obtain culture.
After mixing the culture and anaerobic sludge, the mixture is used as initial inoculation source, uses work as matrix
The glucose and acetate mixture (1: 1, w/w) or COD that COD for artificial substratum is about 1,500mg/L are about 15,
In addition to this waste water in the hydrogen production process of 000mg/L by method identical with the embodiment 1, runs Microbial fuel
Battery (Fig. 3 a, 3b, 4a and 4b).
In addition, mixing the culture and anaerobic sludge, the mixture is used as initial inoculation source, is used as matrix
The glucose and acetate mixture (1: 1, w/w) or COD that COD as artificial substratum is about 500mg/L are about 500mg/L
Biogas production process in waste water, in addition to this, by with 1 same procedure of embodiment, run microbiological fuel cell
(Fig. 5 a, 5b, 6a and 6b).
Firstly, Fig. 3 a and Fig. 5 a are to indicate COD and pH value with comprising Klebsiella IR21 bacterial strain or Mycotoruloides
The microbiological fuel cell of IR11 bacterial strain and artificial substratum runing time variation chart, Fig. 3 b and Fig. 5 b be indicate voltage with
When the operation of the microbiological fuel cell comprising Klebsiella IR21 bacterial strain or Mycotoruloides IR11 bacterial strain and artificial substratum
Between the chart that changes.
As shown in described Fig. 3 a, 3b, 5a and 5b, the voltage generated in the microbiological fuel cell is confirmed from the beginning
With regard to stabilization, and generate 200mV or more voltage.
Secondly, Fig. 4 a is to indicate COD and pH value with comprising the waste water in Klebsiella IR21 bacterial strain and hydrogen production process
Microbiological fuel cell runing time variation chart, Fig. 4 b is to indicate voltage with comprising Klebsiella IR21 bacterial strain
The chart changed with the runing time of the microbiological fuel cell of the waste water in hydrogen production process.
As shown in Fig. 4 a and 4b, although the voltage generation for confirming microbiological fuel cell is unstable, using
In the case that waste water in hydrogen production process with high-grade COD is as matrix, remain to generate electric current.
Again, Fig. 6 a is to indicate COD and pH value with comprising the waste water in Mycotoruloides IR11 bacterial strain and biogas production process
Microbiological fuel cell runing time variation chart, Fig. 6 b is to indicate voltage with comprising Mycotoruloides IR11 bacterial strain and natural pond
The chart of the runing time variation of the microbiological fuel cell of waste water in gas production process.
As shown in Fig. 6 a and 6b, needed for confirming until the voltage of microbiological fuel cell is generated and tended towards stability
Time is about 20 days, and 200mV or more voltage is generated after stabilization.
In addition, confirming by comparing Fig. 3 a and 3b and Fig. 4 a and Fig. 4 b and using artificial discrete phase ratio, by hydrogen manufacturing
When waste water in the process is used as matrix, voltage generation is difficult to stablize.But significantly larger than artificial base can will be had by confirming to have
Waste water in the hydrogen production process of the COD value of matter generates the ability of electric current as matrix, it is believed that its reason is the supply
Klebsiella IR21 bacterial strain.
Similarly, it by comparing Fig. 5 a and Fig. 5 b and Fig. 6 a and Fig. 6 b, confirms and uses artificial discrete phase ratio, it will
When waste water in biogas production process is used as matrix, certain time, the voltage of generation are needed until voltage generation tends towards stability
Grade it is low.But by comparing the result of Fig. 6 a and Fig. 6 b and Fig. 2 a and Fig. 2 b's as a result, confirm with using artificial
Matrix (Fig. 2 a and Fig. 2 b) is compared, and when using waste water (Fig. 2 a and Fig. 2 b) in biogas production process, not only voltage generation tends to
The stable time shortens, and the grade of voltage generated is relatively high, it is believed that its reason is the provided beads
Pseudomonas IR11 bacterial strain.
Embodiment 2-2: coulombic efficiency (coulombic efficiency) assessment
Coulombic efficiency refers to that the electronics for aoxidizing the microorganism in microbiological fuel cell to organic matter and generating turns
Be changed to the efficiency of electric energy, to coulombic efficiency whether be inoculated with Candida spp of the invention and change and assess.
That is, other than being inoculated with or not being inoculated with Candida spp of the invention, according to identical with the embodiment 2-1
Condition runs microbiological fuel cell, calculates coulombic efficiency, (table 1) is compared to it.
[table 1]
As shown in the table 1, it is not inoculated with Candida spp of the invention if confirmed, in the matrix flowed into
About 14% is converted to electric energy, and if being inoculated with Candida spp of the invention, about 22% conversion in the matrix flowed into
For electric energy.Even if coulombic efficiency is also due to microbe inoculation that is, using identical matrix in identical microbiological fuel cell
It obviously increases.
It is thus identified that may originate to Klebsiella IR21 bacterial strain provided in the present invention or Mycotoruloides IR11 bacterial strain
To carry out the anaerobic pond of wastewater treatment and apply in wastewater treatment processing procedure, also acts as and improve microbiological fuel cell electricity production
The bacterial strain of efficiency, the bacterial strain can be used not only as the core element of microbiological fuel cell, but also can combine waste water
Central role is played in terms of processing unit and microbiological fuel cell.
Embodiment 3: quantitative analysis is carried out to Klebsiella or Candida spp in MFC reaction tank
In two kinds of MFC reaction tanks (inoculation or the MFC for not being inoculated with Klebsiella or Candida spp), to Cray
Primary Bordetella or Candida spp concentration carry out quantitative analysis, it is confirmed that the Klebsiella of inoculation or Mycotoruloides bacterium
Strain survival participates in wastewater treatment and electricity production in MFC reaction tank.
Embodiment 3-1: biomembrane sampling
It is acquired from the MFC reaction tank and nonvaccinated MFC reaction tank for being inoculated with Klebsiella or Candida spp
The biomembrane being formed on anode (anode).In addition to strain inoculated, makes and run identically as the reaction tank of inoculation and do not connect
The MFC reaction tank of kind Klebsiella bacterial strain.
Embodiment 3-2: quantitative PCR (quantitative real time PCR)
After the biomembrane of acquisition extraction RNA, passes through Omniscript RT Kit (Qiagen Inc., USA) and synthesize
cDNA.Using synthesized cDNA as template, based on the base sequence information of Klebsiella pneumoniae for separating identification, design two
Kind specific primer group, carries out quantitative PCR.Specific primer uses Kleb_F1 (5'-GGCAGGCTGGAGTCTTGTAG-3') (sequence
And Kleb_R1 (5'-GCCACTCCTCAAGGGAACAA-3') (sequence number 6), Kleb_F2 (5'- number 5)
GGCAGGCTGGAGTCTTGTAG-3') (sequence number 7) and Kleb_R2 (5'-AAGCCACTCCTCAAGGGAAC-3') (sequence number
8).PCR carries out following steps: 4 minutes denaturations are carried out at 95 DEG C；It carries out being denaturalized for 30 seconds at 95 DEG C；At 57 DEG C into
Annealing (annealing) in row 30 seconds；It carries out extending for 30 seconds at 72 DEG C；Final extension in 30 seconds is carried out at 82 DEG C；Finally 82
Carried out at DEG C 30 seconds it is final extend, using 7300Real-time PCR system (Applied Biosystems, USA) altogether into
Row 35 times circulations.
Similarly, from the biomembrane of acquisition extract RNA after, by Omniscript RT Kit (Qiagen Inc.,
USA cDNA) is synthesized.Using synthesized cDNA as template, using with the gene (SAPT and ERG gene) of Mycotoruloides is target
Two kinds of specific primer groups carry out quantitative PCR.Specific primer uses SAPT3-F (5'-ACTTGGATTTCCAGCGAAGA-3') (sequence
Row number 9) and SAPT3-R (5'-AGCCCTTCCAATGCCTAAAT-3') (sequence number 10), ERG11-F (5'-
ATGGCTATTGTTGTACTGC-3') (sequence number 11) and ERG11-R (5'-GCATTGTAAATGAATTCGTG-3') (sequence number
12).PCR carries out following steps: 3 minutes denaturations are carried out at 95 DEG C；It carries out being denaturalized for 45 seconds at 95 DEG C；At 55 DEG C into
Annealing (annealing) in row 15 seconds；It carries out extending for 30 seconds at 72 DEG C；Final extension in 30 seconds is finally carried out at 82 DEG C, is utilized
7300Real-time PCR system (Applied Biosystems, USA) carries out 35 circulations altogether.
As shown in fig. 7, when being quantified using two kinds of primers of Kleb_F1 and Kleb_F2, from inoculation Klebsiella
The bacterial strain amount of Klebsiella in the biomembrane of the MFC reaction tank acquisition of bacterial strain is than acquiring from nonvaccinated reaction tank
Biomembrane has more 2 times or more (Fig. 7).This shows that Klebsiella bacterial strain stablizes survival in MFC reaction tank, facilitates waste water
Processing (removal COD), while helping to improve efficiency of fuel cell generation.
In addition, as shown in figure 8, when being quantified using two kinds of primers of SAPT3 and ERG11, from inoculation Mycotoruloides bacterium
The bacterial strain amount of Mycotoruloides in the biomembrane of the MFC reaction tank acquisition of strain is than the biomembrane that acquires from nonvaccinated reaction tank
It is higher by 3 times or more (Fig. 8).This shows that Candida spp stablizes survival in MFC reaction tank, helps to improve efficiency of fuel cell generation.
In the description, personnel commonsense in the technical field of the invention can be fully understood by being omitted
The detailed record of the content of deduction can not depart from technology of the invention other than concrete example documented in this specification
Various modifications are realized in the range of thought or the necessary composition of change.Therefore, the present invention can by with illustrate example in specification
The different mode of the mode shown is implemented, and commonsense personnel should can fully understand the thing in the technical field of the invention
?.
Sequence table
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Claims (14)
1. a kind of Mycotoruloides (Candida) IR11 bacterial strain (KCTC 12570BP).
2. a kind of for wastewater treatment or the composition of purification, it includes Mycotoruloides IR11 bacterium according to claim 1
Strain.
3. composition according to claim 2, also comprising being selected from saccharomyces (Saccharomyces) bacterial strain, bacillus
Belong to (Bacillus) bacterial strain, enterococcus spp (Enterococcus) bacterial strain, and combinations thereof.
4. a kind of wastewater treatment or purification method, including by the composition as claimed in claim 3 for being used for wastewater treatment or purification
It is handled in waste water, then reacts gains under anaerobic.
5. a kind of microbiological fuel cell, it includes:
(a) anode electrode is inoculated with Mycotoruloides IR11 bacterial strain；
(b) cathode electrode；
(c) culture medium provides the culture environment of microorganism it includes by the matrix of the microorganism decomposition；
(d) electric wire connects described two electrodes；
(e) reaction tank, wherein providing the outflux of described two electrodes, the inflow entrance of the culture solution and the culture solution；And
(f) cation-exchange membrane is arranged in the reaction tank, between described two electrodes.
6. microbiological fuel cell according to claim 5, wherein the anode electrode, which is used, is selected from platinum, stainless (steel) wire, carbon
Felt, carbon paper, and combinations thereof material manufacture.
7. microbiological fuel cell according to claim 5, wherein the cathode electrode use selected from carbon felt, carbon paper, carbon cloth,
And combinations thereof material manufacture.
8. microbiological fuel cell according to claim 5, wherein the matrix is decomposed by Mycotoruloides IR11 bacterial strain
And generate the substance of hydrogen ion and electronics.
9. microbiological fuel cell according to claim 5, wherein the culture solution be comprising carbon source and electron acceptor simultaneously
It is able to maintain the buffer solution of neutral pH.
10. a kind of electricity production method, comprising:
(a) inflow entrance and outflux having by the reaction tank of microbiological fuel cell according to claim 5, to
The anode electrode for being inoculated with Mycotoruloides IR11 bacterial strain continuously feeds the culture solution comprising matrix；And
(b) hydrogen is generated and the Mycotoruloides IR11 bacterial strain being inoculated into anode electrode decomposes the matrix of the supply
Then ion and electronics generate electric current by the hydrogen ion and electronics.
11. a kind of wastewater treatment equipment, it includes microbiological fuel cells according to claim 5, sedimentation basin and aerobic
Pond.
12. wastewater treatment equipment according to claim 11 can carry out wastewater treatment and electricity production simultaneously.
13. wastewater treatment equipment according to claim 11 also includes selected from following one kind: waste water inflow entrance gives up
Water export, electrical storage device, and combinations thereof.
14. a kind of method for carrying out wastewater treatment and electricity production simultaneously, comprising:
(a) so that waste water is flowed into wastewater treatment equipment according to claim 11 to purify the waste water, and contained
There is the pretreated waste water of organic matter；And
(b) thus obtained pretreated waste water is supplied to produce electricity to microbiological fuel cell.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
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| KR10-2014-0049399 | 2014-04-24 | ||
| KR1020140049397A KR101686706B1 (en) | 2014-04-24 | 2014-04-24 | Novel Candida sp. and microbial fuel cell comprising the same |
| KR10-2014-0049397 | 2014-04-24 | ||
| KR1020140049399A KR101726479B1 (en) | 2014-04-24 | 2014-04-24 | Novel Klebsiella sp. and microbial fuel cell comprising the same |
| PCT/KR2015/004055 WO2015163708A1 (en) | 2014-04-24 | 2015-04-23 | Novel klebsiella sp. or candida sp. strain, and microbial fuel cell containing same |
| CN201580000154.3A CN105765056B (en) | 2014-04-24 | 2015-04-23 | Klebsiella or Candida strains and microbial fuel cells comprising same |
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| CN105765056B (en) | 2020-11-24 |
| CN109897789B (en) | 2023-02-17 |
| WO2015163708A1 (en) | 2015-10-29 |
| CN105765056A (en) | 2016-07-13 |
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