CN114314807A - Method for rapidly domesticating and enriching electrogenic bacteria - Google Patents
Method for rapidly domesticating and enriching electrogenic bacteria Download PDFInfo
- Publication number
- CN114314807A CN114314807A CN202111535858.4A CN202111535858A CN114314807A CN 114314807 A CN114314807 A CN 114314807A CN 202111535858 A CN202111535858 A CN 202111535858A CN 114314807 A CN114314807 A CN 114314807A
- Authority
- CN
- China
- Prior art keywords
- biochar
- chamber
- electrogenic bacteria
- enriching
- anode
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 29
- 244000005700 microbiome Species 0.000 claims abstract description 20
- 230000000813 microbial effect Effects 0.000 claims abstract description 18
- 239000002245 particle Substances 0.000 claims abstract description 18
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 9
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims abstract description 9
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 9
- 239000001632 sodium acetate Substances 0.000 claims abstract description 9
- 235000017281 sodium acetate Nutrition 0.000 claims abstract description 9
- JXKPEJDQGNYQSM-UHFFFAOYSA-M sodium propionate Chemical compound [Na+].CCC([O-])=O JXKPEJDQGNYQSM-UHFFFAOYSA-M 0.000 claims abstract description 8
- 239000004324 sodium propionate Substances 0.000 claims abstract description 8
- 235000010334 sodium propionate Nutrition 0.000 claims abstract description 8
- 229960003212 sodium propionate Drugs 0.000 claims abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 7
- -1 potassium ferricyanide Chemical compound 0.000 claims abstract description 6
- 230000000694 effects Effects 0.000 claims abstract description 4
- 239000007772 electrode material Substances 0.000 claims abstract description 4
- 239000003575 carbonaceous material Substances 0.000 claims abstract description 3
- 238000012544 monitoring process Methods 0.000 claims abstract description 3
- 239000010802 sludge Substances 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 8
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims description 8
- 239000000919 ceramic Substances 0.000 claims description 5
- 239000002023 wood Substances 0.000 claims description 5
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 4
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 claims description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 4
- 238000003760 magnetic stirring Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 claims description 3
- 238000005341 cation exchange Methods 0.000 claims description 3
- 229910001447 ferric ion Inorganic materials 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 210000002966 serum Anatomy 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 claims description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 claims description 2
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 claims description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 2
- 238000005273 aeration Methods 0.000 claims description 2
- 235000019270 ammonium chloride Nutrition 0.000 claims description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 2
- 239000004327 boric acid Substances 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims description 2
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 claims description 2
- BNBLBRISEAQIHU-UHFFFAOYSA-N disodium dioxido(dioxo)manganese Chemical compound [Na+].[Na+].[O-][Mn]([O-])(=O)=O BNBLBRISEAQIHU-UHFFFAOYSA-N 0.000 claims description 2
- 239000011790 ferrous sulphate Substances 0.000 claims description 2
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 2
- 239000003112 inhibitor Substances 0.000 claims description 2
- 238000005342 ion exchange Methods 0.000 claims description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 2
- 239000011565 manganese chloride Substances 0.000 claims description 2
- 235000002867 manganese chloride Nutrition 0.000 claims description 2
- 229940099607 manganese chloride Drugs 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 239000001103 potassium chloride Substances 0.000 claims description 2
- 235000011164 potassium chloride Nutrition 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 238000000197 pyrolysis Methods 0.000 claims description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 2
- 235000011152 sodium sulphate Nutrition 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- UEUXEKPTXMALOB-UHFFFAOYSA-J tetrasodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O UEUXEKPTXMALOB-UHFFFAOYSA-J 0.000 claims description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 2
- 229960001763 zinc sulfate Drugs 0.000 claims description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 2
- WKVSEJXEIVEFJT-UHFFFAOYSA-N bromo ethanesulfonate Chemical compound CCS(=O)(=O)OBr WKVSEJXEIVEFJT-UHFFFAOYSA-N 0.000 claims 1
- 230000000696 methanogenic effect Effects 0.000 claims 1
- 239000011259 mixed solution Substances 0.000 claims 1
- DAKAQNVUSAGTRS-UHFFFAOYSA-M sodium;1-bromoethanesulfonate Chemical compound [Na+].CC(Br)S([O-])(=O)=O DAKAQNVUSAGTRS-UHFFFAOYSA-M 0.000 claims 1
- 239000000243 solution Substances 0.000 claims 1
- 230000005611 electricity Effects 0.000 abstract description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 8
- 239000000370 acceptor Substances 0.000 description 5
- 238000012136 culture method Methods 0.000 description 4
- 235000019260 propionic acid Nutrition 0.000 description 4
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241001135750 Geobacter Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000006399 behavior Effects 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000027756 respiratory electron transport chain Effects 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 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 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GRUMFCSJBLIRMQ-UHFFFAOYSA-N bromoethane;sodium Chemical compound [Na].CCBr GRUMFCSJBLIRMQ-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000002910 solid waste Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for rapidly domesticating and enriching electricigens, which comprises the following steps; step 1: constructing a double-chamber microbial electrochemical system, using common suspended anaerobic fermentation microorganisms as seed mud microorganisms, using sodium acetate or sodium propionate as an electron donor to be added into an anode chamber, using potassium ferricyanide as an electron acceptor to be added into a cathode chamber, using conductive carbon material carbon felts as electrode materials to be respectively placed into the anode chamber and the cathode chamber, and domesticating and enriching electricity-producing bacteria; step 2: the method comprises the steps of adding suspended biochar particles into an anode chamber to accelerate the enrichment speed of electrogenic bacteria on the surface of an anode, and evaluating the promotion effect of biochar particle adding on rapid domestication and enrichment of electrogenic bacteria by monitoring the electrogenesis rate, the coulombic efficiency and the population structure of a biomembrane on the surface of a carbon felt of a biochar adding system and a biochar-free adding system. The invention can efficiently and quickly use the common anaerobic microorganism with low cost as the seed mud to realize domestication and enrichment of the electrogenic bacteria.
Description
Technical Field
The invention belongs to the technical field of sewage treatment, and particularly relates to a method for rapidly domesticating and enriching electrogenic bacteria.
Background
The extracellular electron transfer behavior of the microorganism has important application value in the technical fields of environmental engineering such as water environment treatment, pollutant degradation, soil remediation and the like. However, how to rapidly acclimatize and enrich the electricity-generating bacteria with the ability of transferring electrons from the outside of the cell by using common microorganisms is one of the technical bottlenecks in the field of research and application of electron transfer behaviors from the outside of the cell. Therefore, the development of a method for rapidly domesticating and enriching the electrogenic bacteria by using the common anaerobic microorganisms has important significance.
At present, methods for acclimatizing and culturing electrogenic bacteria mainly comprise a culture medium acclimating culture method and a microbial electrochemical system culture method. The culture medium domestication culture method is to domesticate and culture electrogenic bacteria by utilizing a microorganism purification technology and using ferric iron and the like as electron donors, perform colony culture by utilizing an LB culture medium, and purify and separate the bacteria. The method has the disadvantages that the colony culture and the purification separation of the method are carried out in a sterile anaerobic incubator, the operation difficulty is high, and the cost is high; secondly, the microbial electrochemical system culture method is to build a double-chamber microbial electrochemical system, take glucose, sodium acetate and other small molecular organic matters as electron donors, and enrich and culture the electrogenesis bacteria on the surface of the anode through a periodic acclimation process, and has the defects of long culture time period and low efficiency.
Disclosure of Invention
In order to overcome the technical problems, the invention aims to provide a method for quickly domesticating and enriching electrogenic bacteria, which can efficiently and quickly domesticate and enrich electrogenic bacteria by taking common anaerobic microorganisms with low cost as seed mud.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for rapidly domesticating enriched electrogenic bacteria comprises the following steps;
step 1: constructing a double-chamber microbial electrochemical system, taking 50ml of suspended sludge taken from a complete mixed anaerobic fermentation system as sludge microorganisms, taking 150ml of sodium acetate or sodium propionate as an electron donor to be fed into an anode chamber, taking potassium ferricyanide as an electron acceptor to be fed into a cathode chamber, taking conductive carbon material carbon felts as electrode materials to be respectively placed in the anode chamber and the cathode chamber, and domesticating and enriching electrogenic bacteria;
step 2: the method comprises the steps of adding biochar particles into an anode chamber, ensuring that the biochar particles are uniformly suspended in a device through magnetic stirring at 200 revolutions per minute so as to increase the contact area of biochar and suspended microorganisms and accelerate the enrichment speed of electrogenic bacteria on the surface of an anode, and evaluating the promotion effect of biochar particle addition on rapid domestication and enrichment of electrogenic bacteria by monitoring the electrogenesis rate, coulombic efficiency and the population structure of a biomembrane on the surface of a carbon felt of a biochar adding system and a biochar-free adding system.
And 2, in the step 2, the source of the biochar particles is apple wood chips, the preparation method of the biochar is limited-oxygen pyrolysis, the apple wood chips are placed in a ceramic crucible, are compacted by a ceramic pestle, are covered, are placed in a common muffle furnace, the temperature rise amplitude is controlled to be 12 +/-1 ℃/min, are kept for 2 hours after the temperature reaches 500 ℃, and are taken out for later use after the temperature of the muffle furnace is restored to the room temperature.
In the step 2, the particle size of the biochar particles is 2-5mm, and the adding concentration is 15 g/L.
In the step 1, the double-chamber microbial electrochemical system is made of glass, the reaction volumes of the anode chamber and the cathode chamber are both 200 ml, the double chambers are separated by a cation exchange membrane, ion exchange is facilitated, and the anode and the cathode are connected by a titanium wire with the diameter of 1 mm.
The operating temperature of the double-chamber microbial electrochemical system in the step 1 is room temperature (26 +/-2 ℃).
In the step 1, the anode chamber and the cathode chamber of the two-chamber microbial electrochemical system are aerated for 8 minutes by nitrogen with the purity of 99.9 percent at the aeration intensity of 1L/minute before the reaction is carried out.
The mixing mode of the double-chamber microbial electrochemical system in the step 1 is magnetic stirring, and the stirring speed is 200 revolutions per minute.
In the step 1, the adding concentration of the sodium acetate or the sodium propionate is 1500mg/L, and the adding mode is one-time adding.
In the step 1, the microorganism seed sludge comes from a completely mixed type medium-temperature anaerobic fermentation system and is used for domestication and enrichment of electrogenic bacteria, the sludge is taken out and then is placed in an anaerobic serum bottle for culture, the purpose is to exhaust organic matters in sludge mixed liquor, the daily methane production amount is less than 5 mL/day, the sludge can be used, and the concentration of the sludge added into an anode chamber is 2.3g volatile solids/L.
In the step 1, 50mM/L of bromoethane sodium sulfonate (BES) is added into each anode chamber to serve as a methanogenesis inhibitor, so that methanogenesis caused by sodium propionate or sodium acetate is avoided through methanogenesis by methanogens in anaerobic seed sludge.
The anode culture solution for domesticating enriched electrogenic bacteria in the anode chamber in the step 1 comprises the following components: 500mg/L of ammonium chloride, 200mg/L of monopotassium phosphate, 40mg/L of sodium sulfate, 50mg/L of potassium chloride, 0.5mg/L of aluminum chloride, 10mg/L of calcium chloride, 70mg/L of magnesium chloride, 0.8mg/L of manganese chloride, 1.2mg/L of cobalt chloride, 0.5mg/L of nickel chloride, 3mg/L of EDTA-sodium, 3.2mg/L of ferrous sulfate, 1.1mg/L of copper chloride, 0.1mg/L of sodium manganate, 3.2mg/L of zinc sulfate and 0.2mg/L of boric acid.
And (3) adding potassium ferricyanide in the cathode chamber in the step (1) with the addition concentration of 50mM/L by using ferric ions as an electron acceptor.
The invention has the beneficial effects that:
the invention can realize the rapid domestication and enrichment of functional electrogenic bacteria;
compared with the traditional method, the method uses the biochar green material prepared from solid waste, is environment-friendly and has low cost;
the method is simple to operate and easy to popularize and use.
Drawings
FIG. 1 is a schematic view of the electrochemical experimental apparatus for microorganisms of the present invention.
FIG. 2 is a schematic diagram showing the variation of current density among the groups in the reaction process of the present invention.
FIG. 3 is a schematic diagram showing the degradation of acetic acid and propionic acid and the change of coulomb efficiency of the system during the reaction process of the present invention.
FIG. 4 is a schematic diagram showing the enrichment abundance of the electrogenic bacteria on the carbon felt.
Detailed Description
The present invention will be described in further detail with reference to examples.
The system of the invention comprises the following construction and operation steps:
(1) preparing biochar particles: apple wood chips with the plane size of 1-3 square centimeters are placed in a ceramic crucible of 200 milliliters, the volume of the crucible is filled, and gaps are not left as much as possible, so that an oxygen-deficient environment is created. Covering the crucible with a cover, placing the crucible in a muffle furnace, setting the heating rate of the muffle furnace to be 12 +/-1 ℃/min, keeping the temperature for 2 hours at 500 ℃, then automatically closing the crucible, and removing the crucible after the muffle furnace is cooled to the room temperature to obtain the biochar. Sieving the biochar with stainless steel sieve until the particle diameter is 2-5mm, storing in a plastic package bag, and placing in a dry and dark place for later use.
(2) And (3) source and pre-culture of the microbial seed sludge: 500 mL of anaerobic sludge is taken from a stably operated medium-temperature anaerobic fermentation system, is pre-cultured in an anaerobic fermentation serum bottle for 10 to 15 days, and is stored in an anaerobic environment for later use after the daily methane production of the anaerobic sludge is lower than 5 mL/day.
(3) Construction of a two-chamber microbial electrochemical system: the two-chamber microbial electrochemical system is shown in fig. 1, and is made of glass, and the working volume is 200 ml. The upper part of the glass bottle is sealed by a rubber plug and a plastic bottle cap. Two round holes are distributed on the side surface of each bottle, and the rubber plug and the aluminum cover are used for sealing in the experimental process. The joint of the anode chamber and the cathode chamber is separated by a cation exchange membrane, and is fixed and sealed by a water stop belt and a stainless steel water stop clamp. The electrode material used in the system is conductive carbon felt with areas of 6cm respectively2. The anode and the cathode were connected by a titanium wire having a diameter of 1 mm.
(4) The two-chamber microbial electrochemical system is set up in groups: according to the existence of biochar particle addition and the type of an electron donor, an acetic acid-biochar addition group (Ac-BC), an acetic acid-control group (Ac-CT), a propionic acid-biochar addition group (Pr-BC) and a propionic acid-control group (Pr-CT) are arranged together.
(5) Starting and operating the electrochemical system of the double-chamber microorganism: 50mL of anaerobic sludge with the concentration of 2.3g/L and 150mL of microorganism culture solution are added into an anode chamber of the system, sodium acetate with the concentration of 1500mg/L is added into two groups of systems taking acetic acid as an electron donor, and sodium propionate with the concentration of 1500mg/L is added into two groups of systems taking propionic acid as an electron donor. Adding biochar particles into the anode chamber of each group with biochar, wherein the concentration is 15 g/L. Potassium ferricyanide is uniformly added into the cathode chamber, ferric ions are used as electron acceptors, and the adding concentration is 50 mM/L.
(6) And (3) measuring system operation indexes: in the operation process of the system, 2 ml of mixed liquid is taken from the anode chamber by using a 5ml plastic needle tube at regular intervals, a filter membrane with the aperture of 0.45 micron is used for filtering, a gas chromatograph (flame ionization detector) is used for measuring the concentration of acetic acid or propionic acid, a resistor with the diameter of 10 ohms is connected in series in an external circuit, the voltage at two ends of the resistor is measured in real time by a data acquisition unit and a computer, and the current intensity of the system is calculated by using the ohm's law. And (4) taking down the microorganism sample on the surface of the carbon felt for high-throughput sequencing analysis after the reaction is finished.
(7) The analysis method of the system operation result comprises the following steps: the electricity generation efficiency of each group of microorganisms consuming the electron donor was calculated using coulomb's law. According to the high-throughput sequencing result of the microorganism, the enrichment condition of typical electrogenic bacteria among all groups is quantified from the family and genus levels, and the important effect of biochar addition on the rapid domestication and enrichment of the electrogenic bacteria is analyzed.
(8) Analyzing the system operation result: fig. 1 shows the current intensity variation for each group. As can be seen from FIG. 2(a), the bulk current density of the Ac-BC group was significantly higher than that of the Ac-CT group when acetic acid was used as the electron acceptor, and the Ac-BC group reached a maximum current density of 1.6A/m on day 2 of operation2In contrast to the Ac-CT group, the current density in the control group reached a maximum of 0.5A/m at day 42As can be seen from FIG. 2(b), when propionic acid is used as the electron acceptor, the overall current density of the Pr-BC group is significantly higher than that of the Pr-CT group, which reaches a maximum current density of 2.3A/m at day 3 of operation2And the time lag of the Pr-CT group reaching the peak value of the current is 1.3A/m276 lower than the Pr-BC group.9 percent. The coulombic efficiency calculation results in fig. 3 also show that the coulombic efficiency of the system is improved as a whole by adding the biochar. Fig. 4 shows the abundance of geobacter (kojic level) typical in the biofilm on the surface of each group of electrodes after the reaction is finished, and the relative abundance of geobacter in the biochar adding group is found to be 26.7-31.7%, which is much higher than that in the biochar-free adding group by 3.4-4.6%. Although the difference of the electron donor influences the Geobactiraceae enrichment species on the genus level, the addition of the biochar can obviously promote the enrichment of typical electrogenic bacteria on the surface of the electrode.
Claims (10)
1. A method for rapidly domesticating and enriching electrogenic bacteria is characterized by comprising the following steps;
step 1: constructing a double-chamber microbial electrochemical system, taking 50ml of suspended sludge taken from a complete mixed anaerobic fermentation system as sludge microorganisms, taking 150ml of sodium acetate or sodium propionate as an electron donor to be fed into an anode chamber, taking potassium ferricyanide as an electron acceptor to be fed into a cathode chamber, taking conductive carbon material carbon felts as electrode materials to be respectively placed in the anode chamber and the cathode chamber, and domesticating and enriching electrogenic bacteria;
step 2: the method comprises the steps of adding biochar particles into an anode chamber, ensuring that the biochar particles are uniformly suspended in a device through magnetic stirring at 200 revolutions per minute so as to increase the contact area of biochar and suspended microorganisms and accelerate the enrichment speed of electrogenic bacteria on the surface of an anode, and evaluating the promotion effect of biochar particle addition on rapid domestication and enrichment of electrogenic bacteria by monitoring the electrogenesis rate, coulombic efficiency and the population structure of a biomembrane on the surface of a carbon felt of a biochar adding system and a biochar-free adding system.
2. The method for rapid domestication and enrichment of electrogenic bacteria as claimed in claim 1, wherein in step 2, the biochar particles are derived from apple wood chips, the biochar is prepared by limited-oxygen pyrolysis, the apple wood chips are placed in a ceramic crucible, compacted by a ceramic pestle, covered by a cover, placed in a common muffle furnace, the temperature rise is controlled to be 12 ± 1 ℃/min, the muffle furnace is kept for 2h after the temperature reaches 500 ℃, and the biochar is taken out for standby after the muffle furnace is returned to room temperature.
3. The method for rapidly acclimatizing and enriching the electrogenic bacteria according to claim 1, wherein the particle size of the biochar particles in the step 2 is 2-5mm, and the adding concentration is 15 g/L.
4. The method for rapidly domesticating enriched electrogenic bacteria according to claim 1, wherein in the step 1, the two-chamber microbial electrochemical system is made of glass, the reaction volumes of the anode chamber and the cathode chamber are both 200 ml, the two chambers are separated by a cation exchange membrane to facilitate ion exchange, and the anode and the cathode are connected by a titanium wire with the diameter of 1 mm.
5. The method for rapidly acclimatizing and enriching the electrogenic bacteria according to claim 1, wherein the operating temperature of the two-chamber microbial electrochemical system in the step 1 is room temperature (26 ± 2 ℃);
in the step 1, the anode chamber and the cathode chamber of the two-chamber microbial electrochemical system are aerated for 8 minutes by nitrogen with the purity of 99.9 percent at the aeration intensity of 1L/minute before the reaction is carried out.
6. The method for rapid domestication and enrichment of electrogenic bacteria as claimed in claim 1, wherein the mixing manner of the two-chamber microbial electrochemical system in step 1 is magnetic stirring, and the stirring is 200 rpm;
in the step 1, the adding concentration of the sodium acetate or the sodium propionate is 1500mg/L, and the adding mode is one-time adding.
7. The method for rapidly acclimatizing and enriching the electrogenic bacteria according to claim 1, wherein the microorganism seed sludge in the step 1 is derived from a fully mixed moderate temperature anaerobic fermentation system and is used for acclimatizing and enriching the electrogenic bacteria, the sludge is taken out and then placed in an anaerobic serum bottle for culture so as to exhaust organic matters in a sludge mixed solution, the daily methane production is lower than 5 mL/day, the sludge can be used, and the concentration of the sludge added into the anode chamber is 2.3g volatile solids/L.
8. The method for rapid domestication of enriched electrogenic bacteria as claimed in claim 1, wherein 50Mm/L of sodium bromoethane sulfonate (BES) is added to each anode chamber in step 1 as a methanogenic inhibitor to prevent methanogens in the anaerobic seed sludge from metabolizing sodium propionate or sodium acetate to produce methane.
9. The method for rapidly acclimatizing and enriching the electrogenic bacteria according to claim 1, wherein the anode culture solution for acclimatizing and enriching the electrogenic bacteria in the anode chamber in the step 1 comprises the following components: 500mg/L of ammonium chloride, 200mg/L of monopotassium phosphate, 40mg/L of sodium sulfate, 50mg/L of potassium chloride, 0.5mg/L of aluminum chloride, 10mg/L of calcium chloride, 70mg/L of magnesium chloride, 0.8mg/L of manganese chloride, 1.2mg/L of cobalt chloride, 0.5mg/L of nickel chloride, 3mg/L of EDTA-sodium, 3.2mg/L of ferrous sulfate, 1.1mg/L of copper chloride, 0.1mg/L of sodium manganate, 3.2mg/L of zinc sulfate and 0.2mg/L of boric acid.
10. The method for rapid domestication of enriched electrogenic bacteria as claimed in claim 1, wherein the concentration of potassium ferricyanide in the cathode chamber in step 1 is 50mM/L by using ferric ion as an electron acceptor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111535858.4A CN114314807A (en) | 2021-12-15 | 2021-12-15 | Method for rapidly domesticating and enriching electrogenic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111535858.4A CN114314807A (en) | 2021-12-15 | 2021-12-15 | Method for rapidly domesticating and enriching electrogenic bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114314807A true CN114314807A (en) | 2022-04-12 |
Family
ID=81053499
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111535858.4A Pending CN114314807A (en) | 2021-12-15 | 2021-12-15 | Method for rapidly domesticating and enriching electrogenic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114314807A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110055206A (en) * | 2018-01-18 | 2019-07-26 | 云南师范大学 | A kind of method of quick formation MEC hydrogen manufacturing anode microbial film |
| CN110127840A (en) * | 2019-04-24 | 2019-08-16 | 上海交通大学 | Apparatus for treating sewage based on aerobic particle mud bed reactor cathode microbial fuel cell |
| US20190319288A1 (en) * | 2017-04-11 | 2019-10-17 | Dalian University Of Technology | Preparation of a new type of composite anode and microbial fuel cell based on nitrogen doped biological carbon and porous volcanic rocks |
| CN110350226A (en) * | 2019-08-06 | 2019-10-18 | 农业农村部规划设计研究院 | A kind of microorganism electrolysis cell and its method for handling wood vinegar |
| CN111170599A (en) * | 2020-01-21 | 2020-05-19 | 河海大学 | Sludge MFC-anaerobic digestion coupling system and performance strengthening method thereof |
| CN111304124A (en) * | 2020-02-28 | 2020-06-19 | 五邑大学 | Compound microbial inoculum for strengthening propionic acid anaerobic degradation and construction method thereof |
-
2021
- 2021-12-15 CN CN202111535858.4A patent/CN114314807A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190319288A1 (en) * | 2017-04-11 | 2019-10-17 | Dalian University Of Technology | Preparation of a new type of composite anode and microbial fuel cell based on nitrogen doped biological carbon and porous volcanic rocks |
| CN110055206A (en) * | 2018-01-18 | 2019-07-26 | 云南师范大学 | A kind of method of quick formation MEC hydrogen manufacturing anode microbial film |
| CN110127840A (en) * | 2019-04-24 | 2019-08-16 | 上海交通大学 | Apparatus for treating sewage based on aerobic particle mud bed reactor cathode microbial fuel cell |
| CN110350226A (en) * | 2019-08-06 | 2019-10-18 | 农业农村部规划设计研究院 | A kind of microorganism electrolysis cell and its method for handling wood vinegar |
| CN111170599A (en) * | 2020-01-21 | 2020-05-19 | 河海大学 | Sludge MFC-anaerobic digestion coupling system and performance strengthening method thereof |
| CN111304124A (en) * | 2020-02-28 | 2020-06-19 | 五邑大学 | Compound microbial inoculum for strengthening propionic acid anaerobic degradation and construction method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 李凤祥等: "产电菌群及电子受体对微生物燃料电池性能的影响", 《应用生态学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yu et al. | A review on the applications of microbial electrolysis cells in anaerobic digestion | |
| Van Steendam et al. | Improving anaerobic digestion via direct interspecies electron transfer requires development of suitable characterization methods | |
| Zhao et al. | Evaluation on direct interspecies electron transfer in anaerobic sludge digestion of microbial electrolysis cell | |
| Lu et al. | Hydrogen production with effluent from an ethanol–H2-coproducing fermentation reactor using a single-chamber microbial electrolysis cell | |
| Yang et al. | Multiple anodic chambers sharing an algal raceway pond to establish a photosynthetic microbial fuel cell stack: voltage boosting accompany wastewater treatment | |
| Durruty et al. | Evaluation of potato-processing wastewater treatment in a microbial fuel cell | |
| Lee et al. | In situ integration of microbial electrochemical systems into anaerobic digestion to improve methane fermentation at different substrate concentrations | |
| KR101714431B1 (en) | Microbial electrolysis cell and method for producing hydrogen using the same | |
| Wang et al. | Microbial fingerprints of methanation in a hybrid electric-biological anaerobic digestion | |
| Ullery et al. | Anode acclimation methods and their impact on microbial electrolysis cells treating fermentation effluent | |
| CN105280940A (en) | Method for coking wastewater degradation and synchronous power generation by taking coking active bacterium as biocatalyst | |
| Dalvi et al. | Microbial fuel cell for production of bioelectricity from whey and biological waste treatment | |
| Uggetti et al. | Photosynthetic membrane-less microbial fuel cells to enhance microalgal biomass concentration | |
| CN103861463A (en) | Electrochemically assisted biological denitrification method of source separated urine | |
| Kim et al. | Electrode-attached cell-driven biogas upgrading of anaerobic digestion effluent CO2 to CH4 using a microbial electrosynthesis cell | |
| WO2023207134A1 (en) | Organic wastewater bod test device and method, and application | |
| Bai et al. | Acetate and electricity generation from methane in conductive fiber membrane-microbial fuel cells | |
| CN102786330A (en) | System for accelerating anaerobic composting of dewatered sludge by bioelectricity production | |
| CN107964552B (en) | Method for improving methane synthesis efficiency by coupling anaerobic digestion with MFC | |
| Duţeanu et al. | Microbial fuel cells–an option for wastewater treatment | |
| Addagada et al. | Tricks and tracks in resource recovery from wastewater using bio-electrochemical systems (BES): A systematic review on recent advancements and future directions | |
| Pang et al. | New insight into Na+-promoted microbial electrolysis cell towards hydrogen energy recovery: Feasibility and dual mechanism in salt-containing wastewater treatment | |
| Shankar et al. | Energy production through microbial fuel cells | |
| CN113387427A (en) | Diaphragm cathode and microbial electrolysis cell | |
| CN103864201A (en) | Method for microbial electrolytic preparation of hydrogen by use of source separated urine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220412 |
|
| RJ01 | Rejection of invention patent application after publication |