CN117568247B - Preparation method and application of metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst - Google Patents
Preparation method and application of metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst Download PDFInfo
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- 241000588724 Escherichia coli Species 0.000 title claims abstract description 65
- 239000010411 electrocatalyst Substances 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 13
- 239000008103 glucose Substances 0.000 claims abstract description 13
- 230000000813 microbial effect Effects 0.000 claims abstract description 9
- 239000000446 fuel Substances 0.000 claims abstract description 8
- 239000002244 precipitate Substances 0.000 claims abstract description 8
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 15
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 10
- 229940033123 tannic acid Drugs 0.000 claims description 10
- 229920002258 tannic acid Polymers 0.000 claims description 10
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- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 claims description 7
- 235000015523 tannic acid Nutrition 0.000 claims description 7
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 6
- 229910052742 iron Inorganic materials 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 5
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 4
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- 230000008569 process Effects 0.000 claims description 3
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 claims description 2
- VEQPNABPJHWNSG-UHFFFAOYSA-N Nickel(2+) Chemical compound [Ni+2] VEQPNABPJHWNSG-UHFFFAOYSA-N 0.000 claims description 2
- 229910052782 aluminium Inorganic materials 0.000 claims description 2
- PFTAWBLQPZVEMU-UHFFFAOYSA-N catechin Chemical compound OC1CC2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UHFFFAOYSA-N 0.000 claims description 2
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 claims description 2
- 235000005487 catechin Nutrition 0.000 claims description 2
- 229950001002 cianidanol Drugs 0.000 claims description 2
- 229910001429 cobalt ion Inorganic materials 0.000 claims description 2
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 claims description 2
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- 238000009630 liquid culture Methods 0.000 claims description 2
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- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 16
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- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 2
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- 235000002867 manganese chloride Nutrition 0.000 description 2
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- 239000002609 medium Substances 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 238000011056 performance test Methods 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
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- 239000002028 Biomass Substances 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- 229920000557 Nafion® Polymers 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
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- 229910021386 carbon form Inorganic materials 0.000 description 1
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical group OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 1
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- 229910010272 inorganic material Inorganic materials 0.000 description 1
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- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
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- 230000035899 viability Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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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
- H01M4/00—Electrodes
- H01M4/86—Inert electrodes with catalytic activity, e.g. for fuel cells
- H01M4/90—Selection of catalytic material
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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
- C12R2001/185—Escherichia
- C12R2001/19—Escherichia coli
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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
- 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
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Abstract
The invention discloses a preparation method and application of a metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst, wherein escherichia coli culture solution is centrifuged to obtain bacterial precipitate, and the bacterial precipitate is centrifugally washed by PBS buffer solution, and then suspended in PBS solution of plant polyphenol to obtain bacterial solution; adding a PBS solution of metal ions into the bacterial solution, centrifugally separating, and pouring supernatant to obtain the metal-polyphenol nano complex coated engineering escherichia coli electrocatalyst. When the electrocatalyst prepared by the invention is used as a cathode oxygen reduction catalyst of a microbial fuel cell, the electrocatalyst shows excellent electrocatalytic and organic pollutant degradation performances in domestic wastewater, and the full-cell power output and the glucose-containing model degradation capability respectively reach the maximum power density of 213 mu W cm −2 And the degradation amount of glucose was 13.5mM in 140 hours.
Description
Technical Field
The invention belongs to the technical field of microbial fuel cell air cathode biological materials, and particularly relates to a preparation method and application of a metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst.
Background
The large amount of sewage discharged from agriculture, city and industry poses a serious threat to water and public health, and an innovative and sustainable sewage treatment method is to convert pollutants into valuable products. Because sewage is rich in carbohydrates, bioelectrochemical processes based on microorganisms that utilize carbohydrates as a carbon source are an environmentally friendly renewable way of degrading organic pollutants.
Microbial Fuel Cells (MFCs) are a promising technology option with dual efficiency of power generation and pollutant degradation, since biodegradation of sugar-containing domestic wastewater not only requires no external energy input, but also can convert other environmentally-unsafe organic substances into biomass fuels in low-carbon form for power generation.
Patent document publication No. CN 111604094A discloses a mixed iron oxide nanomaterial of escherichia coli, a biomimetic mineralization method and application thereof, wherein the biomimetic mineralization method comprises the following steps: culturing escherichia coli, collecting escherichia coli thalli, and washing to obtain escherichia coli wet thalli; suspending the coliform wet cell in Fe 3+ And (3) carrying out biomimetic mineralization synthesis reaction in the solution, sampling and centrifuging at different time, washing and drying to obtain the escherichia coli mixed iron oxide nano material. The material has good dispersibility and stability, so that the material has high organic pollutant rapid degradation rate and electrocatalytic hydrogen evolution efficiency, and can be well applied to the fields of photocatalytic degradation of organic pollutants and electrocatalytic hydrogen evolution. However, the technical solution of the patent document has the following disadvantages: in terms of the preparation method, the patent document adopts the post-treatment of centrifugal drying and high-temperature pyrolysis (400-700 ℃) to biomimetic mineralized escherichia coli, and the preparation process is complex; on the synthesized product, the patent document finally obtains the iron oxide inorganic nano material which has no life activity and noIs beneficial to green sustainable development and the like.
The air cathode catalyst of the MFC is mainly inorganic materials and oxidoreductase at present, however, the preparation process of the air cathode catalyst is complex, the air cathode catalyst cannot be regenerated and can not be used for generating electricity and degrading pollutants, and compared with the microbial whole-cell electrocatalyst provided by the invention, the air cathode catalyst has the advantages of low cost, high stability, simple culture, capability of reproducing and regenerating electricity, capability of degrading organic pollutants and the like, and is particularly important for operation under complex environments such as sewage and the like. The catalytic activity of microbial cells can be improved by a plurality of nano materials, such as the electric conductivity of cell membranes is enhanced by cell surface engineering and membrane proteins are structurally modified, so that the nano hybrid whole cells constructed by the method are expected to obtain excellent electrocatalytic activity and stability. Currently, there is no report on this aspect.
Disclosure of Invention
The invention solves the technical problem of providing a preparation method of a metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst with simple and rapid synthesis and mild reaction conditions and application of the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst in preparation of an air cathode of a microbial fuel cell.
The invention adopts the following technical proposal to solve the technical problems, and the preparation method of the metal-polyphenol nano complex coated engineering escherichia coli electrocatalyst is characterized by comprising the following specific steps:
step S1: centrifuging 50-200mL of a biosafety escherichia coli culture solution to obtain bacterial precipitate, centrifuging and washing for multiple times by using PBS buffer solution, and suspending the finally obtained bacteria in 0.5-6 mg mL −1 PBS solution of plant polyphenolSwirling for 1min to obtain bacterial solution, wherein the plant polyphenol is one or more of tannic acid, gallic acid, catechin or tannic acid;
step S2: 5-80 mg mL −1 Adding a PBS solution of metal ions into the bacterial solution obtained in the step S1, swirling for 1min to obtain a blue-black bacterial solution, centrifuging, pouring supernatant, and removing metal-polyphenol nano-complexes which do not react with bacteria to finally obtain the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst, wherein the metal ions are one or more of iron ions, cobalt ions, nickel ions, manganese ions, aluminum ions or copper ions.
Further defined, in the step S2, the pH of the blue-black bacterial solution is 4 to 8.
Further defined, the volume ratio of the PBS solution of the metal ions to the PBS solution of the plant polyphenol in step S2 is 1:1.
When the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst is used for electrocatalytic ORR, hydrophobic phenolic hydroxyl groups of plant polyphenol enable the metal-polyphenol nano-complex to be deeply embedded into the hydrophobic interior of a phospholipid bilayer of a cell membrane through hydrophobic interaction, electrostatic interaction, intermolecular interaction such as hydrogen bond and the like, and the precise positioning of the iron-tannic acid nano-complex effectively regulates and controls the coordination conformation and electronic structure of an active center of a catalytic active membrane protein cytochrome c (Cyt c) and promotes the O of the active center 2 The adsorption and activation capacity of the bacillus coli and the electrocatalytic ORR performance of the bacillus coli are obviously improved.
The application of the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst in preparing an air cathode of a microbial fuel cell is characterized by comprising the following specific processes: the carbon cloth electrode attached with the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst is used as a cathode, a catholyte is glucose solution of 20mM, a carbon cloth electrode coated with commercial Pt/C is used as an anode, an anolyte is glucose solution of 1M, the two chambers are separated by a proton exchange membrane, an MFC is placed in an incubator of 30-37 ℃, 2000 ohm resistors are externally connected, oxygen is introduced into the catholyte, and nitrogen is introduced into the anolyte.
Further defined, the specific preparation process of the cathode comprises the following steps: suspension of metal-polyphenol nano-complex coated engineered E.coli electrocatalyst in LB liquid Medium (10 g L) −1 Tryptone, 5g L −1 Yeast extract and 5g L −1 Sodium chloride), and placing the carbon cloth electrode in a liquid culture medium for co-incubation with bacteria to finally obtain the carbon cloth electrode, namely a cathode, attached with the metal-polyphenol nano-complex coating engineering escherichia coli electrocatalyst biological membrane.
Further defined, the co-incubation time is 24-48 hours.
The invention relates to an application of a metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst in preparing a working electrode of a three-electrode half-cell, wherein a platinum sheet and a saturated silver/silver chloride electrode in the three-electrode half-cell are respectively used as a counter electrode and a reference electrode.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the preparation method is simple, quick and efficient, mild in reaction condition, low in cost and environment-friendly, good in biocompatibility of plant-derived polyphenol, almost free of influence on viability, self-replication and self-repair capability of cells, and wide in application potential.
2. The invention utilizes the biological activity and coordination of plant polyphenol, and the coordination effect of catechol groups and iron ions in the molecular structure effectively constructs a double-coordination metal-polyphenol nano complex assembly material at proper PH.
3. The invention utilizes the permeability of phenolic hydrophobic groups and iron ions in the molecular structure of polyphenol, reasonably embeds the metal-polyphenol nano-complex synthesized in the escherichia coli cells into the cell membranes of the escherichia coli, effectively promotes the intermolecular interaction between the metal-polyphenol nano-complex and the catalytic activity membrane protein cytochrome c (Cyt c), controllably adjusts the conformation and coordination structure of the Cyt c, and the electronic structure and the para-O of heme catalytic activity center 2 Is used for adsorption and activation.
4. The metal-polyphenol nano complex coated engineering escherichia coli electrocatalyst provided by the invention not only has excellent electrocatalytic ORR activity in half cells, but also has good biological power generation capacity and glucose-containing sewage degradation capacity when being used as a catalytic material of an MFC air cathode to be assembled into a full cell.
5. The technical scheme disclosed by the invention has the following substantial differences compared with the patent document of publication number CN 111604094A: (1) In the preparation method, the patent document adopts the post-treatment of centrifugal drying and high-temperature pyrolysis (400-700 ℃) on the biomimetic mineralized escherichia coli, the preparation process is complicated, and the application research is directly carried out after the nano-complex is used for coating the engineering escherichia coli; (2) On a synthetic product, the iron oxide inorganic nano material finally obtained by the patent document has no bioactivity and is unfavorable for green sustainable development, and the nano complex coating engineering escherichia coli prepared by the method is a living bacterial biological material, has bioactivity, is green and renewable, and can be self-repaired and propagated; (3) On the raw materials, except Fe 3+ In addition, the invention uses the biocompatible organic molecule of polyphenol, and can fully interact with intracellular catalytic active protein under the condition of not damaging the biological activity of cells; (4) In terms of reaction mechanism, the patent literature is biomimetic mineralization to generate iron oxide, and the coordination reaction of iron and polyphenol is utilized to generate a complex; (5) In terms of catalytic mechanism, the catalytic active substance of the patent in the document is an iron oxide nano material, and the catalytic active substance of the invention is catalytic active protein in living escherichia coli; (6) In application, the escherichia coli mixed iron oxide nano-material obtained by centrifugal drying in the patent literature is used for photocatalytic degradation of organic pollutants, the escherichia coli mixed iron oxide material obtained by high-temperature pyrolysis is used for electrocatalytic hydrogen evolution, and the obtained metal-polyphenol nano-complex coats an engineering escherichia coli as an ORR electrocatalyst of an MFC cathode, so that the double-effect functions of biological power generation and pollutant degradation can be simultaneously exerted.
Drawings
FIG. 1 is an SEM (a) and an ultra-thin section TEM (b) image of the catalyst prepared in example 1;
FIG. 2 is an SEM (a) and an ultra-thin section TEM (b) image of the catalyst prepared in example 2;
FIG. 3 is an SEM (a) and an ultra-thin section TEM (b) image of the catalyst prepared in example 3;
FIG. 4 is a Raman spectrum of the catalyst prepared in example 1;
FIG. 5 is a CV (a) and LSV (b) diagram of ORR of the catalyst prepared in example 1;
FIG. 6 is a graph of polarization curve versus power density for the catalyst prepared in example 1 as an MFC air cathode material;
FIG. 7 is a graph showing the glucose consumption of the catalyst prepared in example 1 as an MFC air cathode material.
Detailed Description
The above-described matters of the present invention will be described in further detail by way of examples, but it should not be construed that the scope of the above-described subject matter of the present invention is limited to the following examples, and all techniques realized based on the above-described matters of the present invention are within the scope of the present invention.
Example 1
Step S1: centrifuging 100mL of the biosafety Escherichia coli culture solution to obtain bacterial precipitate, centrifuging and washing twice with PBS buffer solution, and suspending the bacterial precipitate in 5mL of 2mg mL −1 In PBS solution of vegetable polyphenol tannic acid with pH=5, swirling for 1min to obtain bacterial solution;
step S2: 5mL of 40mg mL −1 Rapidly pouring the PBS solution of ferric chloride with the pH value of=5 into the bacterial solution obtained in the step S1, swirling for 1min to obtain a blue-black bacterial solution, centrifuging, pouring supernatant, removing metal-polyphenol nano-complex which does not react with bacteria, and finally obtaining the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst.
Example 2
Step S1: centrifuging 50mL of the biosafety Escherichia coli culture solution to obtain bacterial precipitate, centrifuging and washing twice with PBS buffer solution, and suspending the bacterial precipitate in 5mL of 1mg mL −1 Plants with ph=5In PBS solution of polyphenol tannic acid, swirling for 1min to obtain bacterial solution;
step S2: 5mL of 40mg mL −1 Rapidly pouring the PBS solution of manganese chloride with the pH value of=5 into the bacterial solution obtained in the step S1, swirling for 1min to obtain a blue-black bacterial solution, centrifuging, pouring supernatant, removing metal-polyphenol nano-complex which does not react with bacteria, and finally obtaining the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst.
Example 3
Step S1: centrifuging 200mL of a biosafety escherichia coli culture solution to obtain bacterial sediment, centrifuging and washing twice by using PBS buffer solution, and suspending the bacterial sediment obtained finally in 10mL of 6mg mL −1 In PBS solution of plant polyphenol gallic acid with pH=7, swirling for 1min to obtain bacterial solution;
step S2: 10mL 60mg mL −1 Rapidly pouring the PBS solution of manganese chloride with the pH value of 7 into the bacterial solution obtained in the step S1, swirling for 1min to obtain a blue-black bacterial solution, centrifuging, pouring supernatant, removing metal-polyphenol nano-complex which does not react with bacteria, and finally obtaining the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst.
The metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared in examples 1-3 is characterized by SEM and ultrathin section TEM, a in fig. 1 is an SEM image of the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared in example 1, the prepared engineered escherichia coli is coated with a large number of nano-particles, and b in fig. 1 is an ultrathin section TEM image of the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared in example 1, and the prepared engineered escherichia coli coated nano-particles are deeply embedded in cell membranes. SEM and ultrathin section TEM characterization of the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared in the embodiment 2-3 are respectively shown in the fig. 2-3, and the morphology is similar to that of the embodiment 1. FIG. 4 is a Raman spectrum diagram of the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared in example 1, and the characteristic peak of the single iron-tannic acid nano-complex is consistent with the characteristic peak of the iron-tannic acid nano-complex engineered escherichia coli, which shows that metal iron ions and tannic acid are successfully coordinated on the cell surface to obtain the nano-complex.
Example 4
The application of the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst in preparing the air cathode biological material of the microbial fuel cell comprises the following steps: the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst obtained in examples 1-4 is suspended in LB liquid medium (10 g L) −1 Tryptone, 5g L −1 Yeast extract and 5g L −1 Sodium chloride), and placing a carbon cloth electrode with the length of 2cm multiplied by 2cm in a culture medium to incubate with bacteria for 36 hours, and finally obtaining the carbon cloth electrode attached with the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst biological membrane.
Half cell electrochemical performance test: ORR performance tests were performed on a CHI 760E electrochemical workstation (Shanghai Chenhua, china) using a typical three-electrode set-up with the carbon cloth electrode (1 cm x 1 cm) prepared as described above as the working electrode, and a platinum sheet and saturated silver/silver chloride electrode as the counter and reference electrodes, respectively. In the reaction from 22mM KH 2 PO 4 、42mM Na 2 HPO 4 、85.5mM NaCl、1.0mM MgSO 4 And 0.1mM CaCl 2 Electrochemical measurements were performed in 80mL buffer (ph=7) of composition. At O 2 After saturation, at 50mV s −1 Is measured at a scanning rate of 5mV s −1 A Linear Sweep Voltammetric (LSV) curve test was performed at the scan rate of (c).
MFC full cell test: the internal working volume was 50mL separated by proton exchange membrane (nafion 211, duPont, U.S.) using a classical H-type dual chamber MFC. Carbon cloth (2 cm. Times.2 cm) was used as the cathode electrode and the anode electrode base electrode. The catholyte was a 20mM glucose solution, purged with oxygen for 30min, and dissolved nitrogen was removed. The anolyte was a 1M glucose solution. The carbon cloth electrode prepared above is used as a biological cathode, the carbon cloth electrode coated with commercial Pt/C is used as an anode, the anode catalyst is 40wt% Pt/C,the loading was 2mg cm −2 Assembled MFC voltage measurements were evaluated on the CHI 760E electrochemical workstation (Shanghai Chenhua, china). When the MFC voltage reaches a steady state, the polarization and power density curve is obtained by changing the external resistance (10-300000Ω). Output performance of the MFC under long term operation was tested with a Keithley 2700 data acquisition system at an external resistance of 1000 Ω. MFC was run at 37 ℃ and repeated 3 times.
FIG. 5 is a graph showing the electrocatalytic ORR performance of the metal-polyphenol nano-complex coated engineered E.coli electrocatalyst prepared in example 1, showing that the prepared catalyst has excellent electrocatalytic activity and a maximum current density of 3.1mA cm −2 The initial potential was 0.66V. FIGS. 6 and 7 are respectively a polarization curve-power density diagram and a glucose consumption diagram of the metal-polyphenol nano-complex coated engineered E.coli electrocatalyst prepared in example 1 as an MFC air cathode material. As can be seen from the figure, the maximum power density of the MFC assembled with the prepared electrocatalyst was 213 μw cm −2 The degradation amount of glucose in 140 hours is 13.5mM, which shows that the metal-polyphenol nano complex coating engineering escherichia coli electrocatalyst simultaneously shows excellent electrogenesis performance and glucose-containing wastewater degradation capability.
While the basic principles, principal features and advantages of the present invention have been described in the foregoing examples, it will be appreciated by those skilled in the art that the present invention is not limited by the foregoing examples, but is merely illustrative of the principles of the invention, and various changes and modifications can be made without departing from the scope of the invention, which is defined by the appended claims.
Claims (8)
1. The preparation method of the metal-polyphenol nano complex coated engineering escherichia coli electrocatalyst is characterized by comprising the following specific steps of:
step S1: centrifuging 50-200mL of a biosafety escherichia coli culture solution to obtain bacterial precipitate, centrifuging and washing for multiple times by using PBS buffer solution, and suspending the finally obtained bacteria in 0.5-6 mg mL −1 PBS (Poly Butylene succinate) solution of plant polyphenolSwirling for 1min in the liquid to obtain bacterial solution, wherein the plant polyphenol is one or more of tannic acid, gallic acid, catechin or tannic acid;
step S2: 5-80 mg mL −1 Adding a PBS solution of metal ions into the bacterial solution obtained in the step S1, swirling for 1min to obtain a blue-black bacterial solution, centrifuging, pouring supernatant, and removing metal-polyphenol nano-complexes which do not react with bacteria to finally obtain the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst, wherein the metal ions are one or more of iron ions, cobalt ions, nickel ions, manganese ions, aluminum ions or copper ions.
2. The method for preparing the metal-polyphenol nano complex coated engineered escherichia coli electrocatalyst according to claim 1, wherein the method comprises the following steps of: and (2) the pH value of the blue-black bacterial solution in the step (S2) is 4-8.
3. The method for preparing the metal-polyphenol nano complex coated engineered escherichia coli electrocatalyst according to claim 1, wherein the method comprises the following steps of: the volume ratio of the PBS solution of the metal ions to the PBS solution of the plant polyphenol in the step S2 is 1:1.
4. The application of the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst prepared by the method according to any one of claims 1-3 in preparation of an air cathode of a microbial fuel cell.
5. The application according to claim 4, wherein the specific process is: the carbon cloth electrode attached with the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst is used as a cathode, a catholyte is glucose solution of 20mM, a carbon cloth electrode coated with commercial Pt/C is used as an anode, an anolyte is glucose solution of 1M, the two chambers are separated by a proton exchange membrane, an MFC is placed in an incubator of 30-37 ℃, 2000 ohm resistors are externally connected, oxygen is introduced into the catholyte, and nitrogen is introduced into the anolyte.
6. The use according to claim 5, characterized in that the specific preparation process of the cathode is: suspending the metal-polyphenol nano complex coated engineering escherichia coli electrocatalyst in an LB liquid culture medium, wherein the composition of the LB liquid culture medium is 10g L −1 Tryptone, 5g L −1 Yeast extract and 5g L −1 And (3) sodium chloride, and placing the carbon cloth electrode in a liquid culture medium for co-incubation with bacteria to finally obtain the carbon cloth electrode, namely a cathode, attached with the metal-polyphenol nano-complex coated engineering escherichia coli electrocatalyst biomembrane.
7. The use according to claim 6, characterized in that: the co-incubation time is 24-48 h.
8. The application of the metal-polyphenol nano-complex coated engineered escherichia coli electrocatalyst prepared by the method according to any one of claims 1-3 in preparation of a three-electrode half-cell working electrode, wherein a platinum sheet and a saturated silver/silver chloride electrode are respectively used as a counter electrode and a reference electrode.
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