CN110343602A - The pure bacterium Geobacter microorganism belonging to genus electro-chemical systems that can simply start in aerobic environment - Google Patents
The pure bacterium Geobacter microorganism belonging to genus electro-chemical systems that can simply start in aerobic environment Download PDFInfo
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- CN110343602A CN110343602A CN201910580223.2A CN201910580223A CN110343602A CN 110343602 A CN110343602 A CN 110343602A CN 201910580223 A CN201910580223 A CN 201910580223A CN 110343602 A CN110343602 A CN 110343602A
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- geobacter
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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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
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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
- 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
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Abstract
The present invention relates to microbial culture techniques, it is desirable to provide a kind of pure bacterium Geobacter microorganism belonging to genus electro-chemical systems that can simply start in aerobic environment.The system is to form the electrode material of carbon electrode material or modification amorphous carbon as the carrier of electro-chemical activity biomembrane, and using the Geobacter microorganism belonging to genus of pure culture as inoculation source, culture solution is configured by PBS solution, matrix and aerobic nutrient chemical；The microorganism electrochemical system is run under aerobic conditions, and head space partial pressure of oxygen is 0 < PO2≤ 21%；The matrix of the culture solution is acetic acid, ethyl alcohol or glucose；Aerobic nutrient chemical is the mixture of one or more of trivalent iron salt, fumarate, sexavalence chromic salts, pentavalent vanadic salts or sexavalence uranium salt.The present invention more meets that practical application request, highly-safe, universality is wide, easy to operate for the first time using the method for the aerobic nutrient chemical aerobic culture Geobacter microorganism belonging to genus in electro-chemical systems is added.
Description
Technical field
The present invention relates to microbial culture techniques, in particular to a kind of to start pure bacterium Geobacter category in aerobic environment
The method of microorganism electrochemical system.This method can be applied to soil remediation, heavy metal recovery, sewage treatment etc., belong to micro- life
Object, bioelectrochemistry field.
Background technique
Valuable substance is recycled while wastewater treatment has important meaning to alleviating energy crisis and problem of environmental pollution
Justice meets the great demand of country.Microorganism electrochemical system (MESs) utilizes the electro-chemical activity microorganism oxygen adhered on anode
Change the organic matter in waste water, receives the electronics of anode generation using cathode, reach and not only handle waste water but also recycle biomass energy, it can
Realize wastewater treatment process from energy consumption to production capacity revolutionary change, it is considered to be the hope of the following sustainable wastewater treatment.
The microorganism that Geobacter belongs to is that one kind has compared with the chemically active microorganism of forceful electric power, in the anode biology of MESs
Usually have in film and occupies an leading position.Study reason of the electrochemical properties for MESs of the Geobacter microorganism belonging to genus of pure culture
It is all of great significance by research and practical application.However, since the Geobacter microorganism belonged to is a kind of micro- life of strictly anaerobic
Object, it is extremely sensitive to oxygen, it is difficult to anode biomembrane is grown and formed in aerobic situation.Therefore, pure bacterium Geobacter belongs to micro-
The culture of biology must carry out under conditions of strictly anaerobic, higher to the operation level requirement of anaerobism equipment and personnel, operation
Very complicated, success rate are low.
In order to reduce operation difficulty, the culture success ratio of pure bacterium Geobacter microorganism belonging to genus is improved, usually in the medium
A certain amount of hydrogen reduction agent such as cysteine, vulcanized sodium etc. is added to reduce oxygen to the shadow of pure bacterium Geobacter microorganism belonging to genus
It rings.However, these hydrogen reduction agent are based on sulfur-containing compound, on the one hand, excessive hydrogen reduction agent may be to pure bacterium Geobacter
The microorganism of category generates toxic side effect, influences the growth of microorganism；On the other hand, these sulfur-containing compounds are formed after being oxidized
Sulphur simple substance is readily adsorbed in electrode surface, influences pure bacterium Geobacter microorganism belonging to genus in the growth and absorption of electrode surface.Cause
This, need to find in aerobic environment it is highly-safe, universality is wide, the culture side of Geobacter microorganism belonging to genus easy to operate
Method.
Summary of the invention
The technical problem to be solved by the present invention is to overcome deficiency in the prior art, provide one kind and open in aerobic environment
The method for moving pure bacterium Geobacter microorganism belonging to genus electro-chemical systems.
In order to solve the above technical problems, solution of the invention is:
A kind of pure bacterium Geobacter microorganism belonging to genus electro-chemical systems that can simply start in aerobic environment are provided, including
Anode, cathode and electro-chemical activity biomembrane；The microorganism electrochemical system is amorphous to form carbon electrode material or modification
Carrier of the electrode material of carbon as electro-chemical activity biomembrane, using the Geobacter microorganism belonging to genus of pure culture as inoculation source, training
Nutrient solution is configured by PBS solution, matrix and aerobic nutrient chemical；The microorganism electrochemical system is run under aerobic conditions,
Head space partial pressure of oxygen is 0 < PO2≤ 21%；
The matrix of the culture solution is acetic acid, ethyl alcohol or glucose；Aerobic nutrient chemical be trivalent iron salt, fumarate,
The mixture of one or more of sexavalence chromic salts, pentavalent vanadic salts or sexavalence uranium salt.
In the present invention, in the culture solution, the proportion relation of PBS solution, matrix and aerobic nutrient chemical are as follows: 14mL PBS is molten
Liquid: 1.5g/L sodium acetate: the aerobic nutrient chemical of 20mM.
In the present invention, 2.45g/L NaH is contained in the PBS solution2PO4·H2O, 4.58g/L Na2HPO4, 0.31g/L
NH4Cl, 0.13g/L KCl, surplus are water.
In the present invention, the molding carbon material is any in graphite flake, carbon cloth, graphite rod, carbon paper, carbon felt or carbon fiber
It is a kind of.
In the present invention, the electrode material of the modification amorphous carbon, which refers to, has wrapped up graphene, carbon nanohorn, carbon nanotube
Or the metal electrode material or molding carbon electrode material of graphite particle.
It is in microorganism electrochemical system invention further provides the operation method of the microorganism electrochemical system
Operational process in implement any one following control means: between the anode and cathode apply 0.3~1.0V voltage, or control
Anode potential is -0.48~0.4V (vs Ag/AgCl ,+0.202V), or control external resistance value is 10~10000 Ω, or
Control anode current is 0.1~15A/m2。
Inventive principle description:
The present invention moves pure bacterium Geobacter microorganism belonging to genus electrification by adding aerobic nutrient chemical, realization in aerobic environment
System.Compared to the Geobacter microorganism belonging to genus cultural method of tradition addition hydrogen reduction agent, the aerobic culture that the present invention adds
Agent is one of available electron acceptor of Geobacter microorganism belonging to genus, and on the one hand aerobic nutrient chemical is dissolved oxygen in vivo
Competitive electron acceptor, added in culture solution aerobic nutrient chemical can inhibit microorganism transmitting electronics to oxygen molecule, thus
The generation of active oxygen species is reduced, toxic action of the oxygen to Geobacter microorganism belonging to genus is reduced；On the other hand, aerobic culture
Agent is good reducing agent by the reduzate after micro-reduction, can be anti-with the dissolved oxygen in culture solution, active oxygen species
It answers, the growth for Geobacter microorganism belonging to genus provides anaerobic environment.The cultural method is highly-safe, universality is wide, operation letter
Just.
Compared with prior art, the medicine have the advantages that
(1) reported pure bacterium Geobacter microorganism belonging to genus electro-chemical systems start method, and limitation is big, complicated for operation.
The present invention uses the method for adding aerobic nutrient chemical aerobic culture Geobacter microorganism belonging to genus in electro-chemical systems for the first time.Phase
Than previous cultural method, this method can be in 0 < PO2Geobacter microorganism belonging to genus is cultivated under the conditions of≤21% partial pressure of oxygen, more
Meet practical application request.
(2) cultural method is highly-safe, universality is wide, easy to operate.
Detailed description of the invention
Fig. 1 is the start-up course of pure bacterium Geobacter microorganism belonging to genus electro-chemical systems in embodiment 1.Wherein, ironic citrate
Belong to MESs to add the pure bacterium Geobacter of aerobic nutrient chemical starting.As reference pair ratio, Control is traditional conventional method
The pure bacterium Geobacter of starting belongs to MESs.
Specific embodiment
Below in conjunction with drawings and concrete examples, the invention will be further described, and its object is to preferably explain the present invention
Rather than limitation of the present invention.
Contain 2.45g/L NaH in PBS solution used in following embodiment2PO4·H2O, 4.58g/L Na2HPO4, 0.31g/L
NH4Cl, 0.13g/L KCl, surplus are water.Except especially expressing, agents useful for same is commercial products in embodiment.
Geobacter microorganism belonging to genus purchase used in the present invention is in Germany Microbiological Culture Collection Center (DSMZ).This
Field technical staff can be obtained by accordingly disclosing approach.Because belonging to the prior art, therefore the present invention repeats no more.
Specific embodiment 1
Using graphite flake as carrier, trivalent iron salt culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2High temperature pyrolysis graphite plate successively uses P400 sand paper, P800 sand paper and 0.05 μm of aluminium powder polishing
Extremely smooth, in 70% ethanol water after ultrasound 10min, deionized water cleans drying as electro-chemical activity biomembrane
Carrier.Using Geobacter sulfurreducens PCA as inoculation source, 14mL sterilizing PBS solution is added, 1.5g/L's
The culture solution that the ironic citrate of sodium acetate and 20mM are configured to, metathesis reactor headspace gas make oxygen partial pressure 1%.Using straight
Galvanic electricity source is outside battery both ends plus 0.7V voltage can start Geobacter microorganism belonging to genus.It is compared as reference, conventional method
It is distinguished as that 14mL sterilizing PBS solution, the culture solution that the sodium acetate of 1.5g/L is configured to is added.
Using graphite flake as carrier, Geobacter belongs to the characterization of biomembrane starting: acquiring external resistance two using data collecting instrument
The voltage at end, as a result as shown in Figure 1.Under 0.7V applied voltage, after the ironic citrate of 20mM is added, MESs can be in 200h
Quick start, after more renewing culture solution, it is 5A/m that reactor, which reaches peak current density,2.As referring to comparing, lemon is not added
The reference group of lemon acid iron cannot start after running more than 700h.It is surveyed using the microorganism that Sanger method obtains culture
Sequence, the results are shown in Table 1, shows that the pure bacterium Geobacter cultivated belongs to the microbe species in MESs.
The sequencing gained 16s segment of table 1 corresponds to microbial species
Specific embodiment 2
Using graphite rod as carrier, fumarate culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2Graphite rod be successively polishing to P400 sand paper, P800 sand paper and 0.05 μm of aluminium powder it is smooth,
In 70% ethanol water after ultrasound 10min, deionized water cleans carrier of the drying as electro-chemical activity biomembrane.It adopts
It uses Geobacter metallireducens as inoculation source, 14mL sterilizing PBS solution, the ethyl alcohol and 20mM of 1.5g/L is added
The culture solution that is configured to of fumaric acid sodium, metathesis reactor headspace gas makes oxygen partial pressure 5%.Using DC power supply in electricity
The additional 0.3V voltage in pond both ends can start Geobacter microorganism belonging to genus.
Specific embodiment 3
Using carbon cloth as carrier, sodium dichromate culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2Carbon cloth after ultrasound 10min, impregnates 1 day, deionized water in 70% ethanol water
It cleans, handles 1h in 370 DEG C, the carrier as electro-chemical activity biomembrane.Using Geobacter metallireducens
As inoculation source, 14mL sterilizing PBS solution, the ethyl alcohol of 1.5g/L and the culture solution of 20mM sodium dichromate being configured to is added, sets
Changing reactor top air body makes oxygen partial pressure 21%.Add 1V voltage that can start outside battery both ends using DC power supply
Geobacter microorganism belonging to genus.
Specific embodiment 4
Using carbon paper as carrier, pentavalent vanadic salts culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2Carbon paper after ultrasound 10min, impregnates 1 day, deionized water in 70% ethanol water
It cleans, handles 1h in 370 DEG C, the carrier as electro-chemical activity biomembrane.Using Geobacter sulfurreducens
14mL sterilizing PBS solution, the culture that the sodium acetate of 1.5g/L and the positive vanadate of 20mM are configured to is added as inoculation source in PCA
Liquid, metathesis reactor headspace gas make oxygen partial pressure 5%.Control anode potential is that -0.48V can start Geobacter category
Microorganism.
Specific embodiment 5
Using carbon felt as carrier, sodium diuranate culture Geobacter microorganism belonging to genus is added
It is 1.5cm by surface area2Carbon felt after ultrasound 10min, is impregnated 1 day, deionized water in 70% ethanol water
It cleans, 1h is handled in 370 DEG C, as the carrier of electro-chemical activity biomembrane, using Geobacter sulfurreducens
14mL sterilizing PBS solution, the culture that the sodium acetate of 1.5g/L and the sodium diuranate of 20mM are configured to is added as inoculation source in PCA
Liquid, metathesis reactor headspace gas make oxygen partial pressure 5%.Control anode potential can start Geobacter for -0.3V and belong to micro-
Biology.
Specific embodiment 6
Using surface modification graphene metal as carrier, trivalent iron salt culture Geobacter microorganism belonging to genus is added.
Carrier using the stainless steel substrates of surface modification graphene as electro-chemical activity biomembrane.Using Geobacter
14mL sterilizing PBS solution, the sodium acetate of 1.5g/L and the citric acid of 20mM is added as inoculation source in sulfurreducens PCA
The culture solution that iron is configured to, metathesis reactor headspace gas make oxygen partial pressure 5%.Control anode potential is that 0.4V can start
Geobacter microorganism belonging to genus.
Specific embodiment 7
Using the molding carbon material of surface modification carbon nanohorn as carrier, addition trivalent iron salt culture Geobacter belongs to micro- life
Object.
Carrier using the graphite flake of surface modification carbon nanohorn as electro-chemical activity biomembrane.Using Geobacter
14mL sterilizing PBS solution, the sodium acetate of 1.5g/L and the citric acid of 20mM is added as inoculation source in sulfurreducens PCA
The culture solution that iron is configured to, metathesis reactor headspace gas make oxygen partial pressure 5%.Control external resistance value is 10 Ω
Start Geobacter microorganism belonging to genus.
Specific embodiment 8
Using the molding carbon material of surface modification graphite particle as carrier, addition trivalent iron salt culture Geobacter belongs to micro- life
Object.
Carrier using the graphite flake of surface modification carbon nanohorn as electro-chemical activity biomembrane.Using Geobacter
14mL sterilizing PBS solution, the sodium acetate of 1.5g/L and the citric acid of 20mM is added as inoculation source in sulfurreducens PCA
The culture solution that iron is configured to, metathesis reactor headspace gas make oxygen partial pressure 5%.Control external resistance value is that 1000 Ω are
Geobacter microorganism belonging to genus can be started.
Specific embodiment 9
Using the molding carbon material of surface modification graphite particle as carrier, addition trivalent iron salt culture Geobacter belongs to micro- life
Object.
Carrier using the graphite flake of surface modification carbon nanohorn as electro-chemical activity biomembrane.Using Geobacter
14mL sterilizing PBS solution, the sodium acetate of 1.5g/L and the citric acid of 20mM is added as inoculation source in sulfurreducens PCA
The culture solution that iron is configured to, metathesis reactor headspace gas make oxygen partial pressure 5%.Control external resistance value is 10000 Ω
Geobacter microorganism belonging to genus can be started.
Specific embodiment 10
Using the molding carbon material of surface modification graphite particle as carrier, addition trivalent iron salt culture Geobacter belongs to micro- life
Object.
Carrier using the graphite flake of surface modification carbon nanohorn as electro-chemical activity biomembrane.Using Geobacter
14mL sterilizing PBS solution, the sodium acetate of 1.5g/L and the citric acid of 20mM is added as inoculation source in sulfurreducens PCA
The culture solution that iron is configured to, metathesis reactor headspace gas make oxygen partial pressure 5%.Control anode current is 0.1A/m2?
Start Geobacter microorganism belonging to genus.
Specific embodiment 11
Using graphite flake as carrier, trivalent iron salt culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2High temperature pyrolysis graphite plate successively uses P400 sand paper, P800 sand paper and 0.05 μm of aluminium powder polishing
Extremely smooth, in 70% ethanol water after ultrasound 10min, deionized water cleans drying as electro-chemical activity biomembrane
Carrier.Using Geobacter sulfurreducens PCA as inoculation source, 14mL sterilizing PBS solution is added, 1.5g/L's
The culture solution that the ironic citrate of sodium acetate and 20mM are configured to, metathesis reactor headspace gas make oxygen partial pressure 1%.Control sun
Electrode current is 1A/m2Geobacter microorganism belonging to genus can be started.
Specific embodiment 12
Using graphite flake as carrier, trivalent iron salt culture Geobacter microorganism belonging to genus is added.
It is 1.5cm by surface area2High temperature pyrolysis graphite plate successively uses P400 sand paper, P800 sand paper and 0.05 μm of aluminium powder polishing
Extremely smooth, in 70% ethanol water after ultrasound 10min, deionized water cleans drying as electro-chemical activity biomembrane
Carrier.Using Geobacter sulfurreducens PCA as inoculation source, 14mL sterilizing PBS solution is added, 1.5g/L's
The culture solution that the ironic citrate of sodium acetate and 20mM are configured to, metathesis reactor headspace gas make oxygen partial pressure 1%.Control sun
Electrode current is 15A/m2Geobacter microorganism belonging to genus can be started.
Claims (6)
1. a kind of pure bacterium Geobacter microorganism belonging to genus electro-chemical systems that can simply start in aerobic environment, including anode, yin
Pole and electro-chemical activity biomembrane；It is characterized in that, the microorganism electrochemical system is to form carbon electrode material or modification nothing
Carrier of the electrode material of setting carbon as electro-chemical activity biomembrane, is inoculation with the Geobacter microorganism belonging to genus of pure culture
Source, culture solution are configured by PBS solution, matrix and aerobic nutrient chemical；The microorganism electrochemical system is transported under aerobic conditions
Row, head space partial pressure of oxygen are 0 < PO2≤ 21%；
The matrix of the culture solution is acetic acid, ethyl alcohol or glucose；Aerobic nutrient chemical is trivalent iron salt, fumarate, sexavalence
The mixture of one or more of chromic salts, pentavalent vanadic salts or sexavalence uranium salt.
2. microorganism electrochemical system according to claim 1, which is characterized in that in the culture solution, PBS solution, base
The proportion relation of matter and aerobic nutrient chemical are as follows: 14mL PBS solution: 1.5g/L sodium acetate: the aerobic nutrient chemical of 20mM.
3. microorganism electrochemical system according to claim 1, which is characterized in that contain 2.45g/L in the PBS solution
NaH2PO4·H2O, 4.58g/L Na2HPO4, 0.31g/L NH4Cl, 0.13g/L KCl, surplus are water.
4. microorganism electrochemical system according to claim 1, which is characterized in that the molding carbon material be graphite flake,
Any one in carbon cloth, graphite rod, carbon paper, carbon felt or carbon fiber.
5. microorganism electrochemical system according to claim 1, which is characterized in that the electrode material of the modification amorphous carbon
Material refers to the metal electrode material for having wrapped up graphene, carbon nanohorn, carbon nanotube or graphite particle or molding carbon electrode material.
6. the operation method of microorganism electrochemical system described in claim 1, which is characterized in that be in microorganism electrochemical system
Operational process in implement any one following control means: between the anode and cathode apply 0.3~1.0V voltage, or control
Anode potential is -0.48~0.4V (vs Ag/AgCl ,+0.202V), or control external resistance value is 10~10000 Ω, or
Control anode current is 0.1~15A/m2。
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107342428A (en) * | 2017-06-08 | 2017-11-10 | 中国科学技术大学 | A kind of method of the extracellular electron transmission of enhancement microbiological in microorganism electrochemical system |
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- 2019-06-28 CN CN201910580223.2A patent/CN110343602A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107342428A (en) * | 2017-06-08 | 2017-11-10 | 中国科学技术大学 | A kind of method of the extracellular electron transmission of enhancement microbiological in microorganism electrochemical system |
Non-Patent Citations (4)
| Title |
|---|
| LIN ET AL.: "Geobacter sulfurreducens Can Grow with Oxygen as a Terminal Electron Acceptor", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| YANG ET AL.: "Sensitivity to Oxygen in Microbial Electrochemical Systems Biofilms", 《ISCIENCE》 * |
| 曹效鑫等: "阳极电势对Geobacter sulfurreducens产电性能的影响", 《高等学校化学学报》 * |
| 赵昕宇等: "典型胞外呼吸细菌的胞内电子转移机制研究进展", 《生态学报》 * |
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