Bioelectrochemical method and apparatus for energy reclamation from nitrogen compounds

Abstract

Methods are described for treating aqueous solutions, including wastewater, to remove nitrogen-containing compounds using electrochemical processes. The method may be conducted electrolytically under an applied voltage or using endogenous current in a fuel cell arrangement. In some embodiments, energy is reclaimed in the form of hydrogen, methane, and other hydrocarbons or organic molecules. Microorganisms may be used as the catalyst for oxidation of the nitrogen-containing compound and/or reduction of hydrogen ions, carbon dioxide, or bicarbonate. Anaerobic or low-oxygen conditions may be used in the zone.

Claims

1. A method of treating an aqueous solution and producing methane using an electrolytic cell comprising an anode, a cathode, an aqueous electrolyte effecting ionic communication between the anode and the cathode, and operative microorganisms disposed relative to the anode and the cathode in the cell or on the anode wherein the anode is electrically coupled to the cathode, the method comprising: establishing a reactive process, including: supplying the aqueous solution to the electrolytic cell such that the aqueous solution becomes disposed in electrical communication with the anode, the aqueous solution comprising at least one nitrogen-containing compound; supplying an oxidant to the electrolytic cell such that the oxidant becomes disposed in electrical communication with the cathode; wherein the supplying of the aqueous solution and the supplying of an oxidant cooperate with effect that: (i) electrolytic oxidation of the at least one nitrogen-containing compound is facilitated at the anode by the operative microorganisms; and (ii) the oxidant is reduced at the cathode to produce methane; and collecting the produced methane; wherein the oxidant comprises carbon dioxide, and the electrolytic cell contains less than 100 μmol L.sup.−1 of free oxygen.

2. The method of claim 1, wherein the free oxygen is O.sub.2.

3. The method of claim 2, wherein the oxidation of the at least one nitrogen-containing compound is with effect that gaseous nitrogen is produced in a zone containing the anode.

4. The method of claim 1, wherein the microorganisms form a biofilm on the anode.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a Pourbaix diagram showing the relationship between pH and the redox potentials E.sub.h of ammonium oxidation (102), proton reduction (101), and proton reduction at +0.5 volts (103) versus a standard hydrogen electrode.

(2) FIG. 2 is a schematic electrolytic cell for carrying out an embodiment, in which ammonium is oxidized while producing hydrogen gas.

(3) FIG. 3A depicts nitrogen evolution in an electrolytic embodiment over two fed-batch cycles (304, 305) poised at +0.550 volts and three subsequent cycles (306, 307, 308) poised at +0.4 volts. All voltages are versus a standard hydrogen electrode. The results using microbial innocula derived from three separate locations on the ocean floor (301, 302, 303) are shown.

(4) FIG. 3B depicts hydrogen evolution in an electrolytic embodiment over two fed-batch cycles (304, 305) poised at +0.55 volts and the three subsequent cycles (306, 307, 308) poised at +0.4 volts. All voltages are versus a standard hydrogen electrode. The results using microbial innocula derived from three separate locations on the ocean floor (301, 302, 303) are shown.

(5) FIG. 4 is a Pourbaix diagram showing the relationship between pH and redox potentials E.sub.h of ammonium oxidation (101), proton reduction (102), CO.sub.2 reduction to methane (401), and proton reduction at +0.5 volts (103). All voltages are versus a standard hydrogen electrode.

(6) FIG. 5 is a schematic electrolysis reactor carrying out an embodiment, in which ammonium is oxidized while producing methane gas.

(7) FIG. 6A depicts the % Nitrogen removal in an embodiment using a carbon brush anode (601) or a graphite granule “drum” anode (603).

(8) FIG. 6B depicts the combined evolution of methane on the cathode and the anode in μmol/L/day using a carbon brush anode (601) or a graphite granule “drum” anode (603) in an embodiment.

(9) FIG. 6C depicts the evolution of nitrogen gas in μmol/L/day at the anode (203) or a cathode (604) of an embodiment.

(10) FIG. 7 depicts a fuel cell which includes an anode compartment and a cathode compartment.

DETAILED DESCRIPTION

(11) To gain a better understanding, the following illustrative embodiments are set forth. It will be understood that these embodiments are intended to describe examples only and are not intended to limit the scope of the present disclosure in any way.

(12) Definitions

(13) “microorganisms” includes bacteria archaea, and eukarya of the genera: Acidobacterium, Geothrix, Holophaga, Mycobacterium, Microbacterium, Marinobacter, Paludibacter, Petrimonas, Proteiniphilum, Sediminibacterium, Anaerolinea, Leptolinea, Caldilinea, Deinococcus, Thermus, Clostridium, Bacillus, Butyribacterium, Sporomusa, Acetobacterium, Acetogenium, Thermoanaerobacter, Anaerovorax, Desulfosporosinus, Proteiniborus, Faecalibacterium, Fastidiosipila, Hydrogenoanaerobacterium, Oscillibacter, Phascolarctobacterium, Turicibacter, Nitrospira, Nitrososphaera, Nitrosopumilus, Nitrobacter, Kuenenia, Brocardia, Nitrosomonas, Nitrosopumilus, Nitrosococcus, Nitrospina, Pirellula, Brevundimonas, Bradyrhizobium, Hyphomicrobium, Pedomicrobium, Xanthobacter, Methylosinus, Nordella, Rhodobium, Amaricoccus, Rhodobacter, Roseomonas, Botryococcus, Synechococcus, Synechocystis, Chloroflexus, Chlorobium, Sphingobium, Sphingomonas, Sphingopyxis, Chitinimonas, Ralstonia, Comamonas, Methylibium, Ottowia, Pelomonas, Herbaspirillum, Thiobacillus, Methylobacillus, Neisseria, Gallionella, Nitrosomonas, Azovibrio, Dechloromonas, Methyloversatilis, Propionivibrio, Kuenania, Thauera, Bdellovibrio, Desulfobacterium, Desulfobulbus, Desulfovibrio, Desulfuromonas, Geobacter, Anaeromyxobacter, Haliangium, Desulfobacca, Smithella, Syntrophus, Syntrophobacter, Syntrophorhabdus, Sulfurospirillum, Sulfuricurvum, Wolinella, Aeromonas, Haliea, Citrobacter, Methylobacter, Methylocaldum, Methylomonas, Methylosinus, Acinetobacter, Pseudomonas, Dokdonella, Thermomonas, Exilispira, Aminiphilus, Kosmotoga, Verrucomicrobia, Opitutus, Puniceicoccus, Tissierella, Sphingopyxis, Pseudoxanthomonas, Sterolibacterium, Brucella, Bosea, Brevundimonas, Singulisphaera, Azospira, Gemmatimonas, Sphingobacterium, Azonexus, Aquamicrobium, Petrimonas, Fontibacter, Arcobacter, Chryseobacterium, Megasphera, Truepera, Hydrogenophaga, Paracoccus, Stenotrophomonas, Methanobacterium, Methanobrevibacter, Methanothermobacter, Methanothermus, Methanomicrobium, Methanogenium, Methanoplanus, Methanoplanus, Methanolacinia, Methanocorpusculum, Methanofollis Methanolinea, Methanoculleus, Methanosphaerula, Methanolinea, Methanospirillum, Methanoregula, Methanofollis, Methanocalculus, Methanothrix, Methanosarcina, Methanosaeta, Methanosphaera, Halomethanococcus, Methanohalobium, Methanosalsum, Methanomethylovorans, Methanimicrococcus, Methanohalophilus, Methanolobus, Methanococcoides, Methanococcus, Methanoflorens, Methanohalophilus, Methanopyrus, Halobacterium, Thermococcus, Pyrococcus, Thermoproteus, and Saccharomyces.

(14) “nitrogen-containing compound” includes various organic and inorganic molecules containing nitrogen groups, such as amines, including without limitation ammonium, ammonium hydroxide, ethyl amine, hydroxyl amine, benzyl amine, and various organic nitrogen compounds including urea, trimethyl amine, ethyl amine, ethanolamine, or natural and artificial amino acids such as alanine, glutamine, arginine, cysteine, and serine. In some embodiments, the nitrogen-containing compound may also be drugs such as paracetamol, oxetacaine, chlorphenamine, chlorpromazine, amphetamine, clomipramine, or nortriptyline. Other nitrogen-containing compounds may be amides and alkylated amides such as acetamide, formamide, sulfonamide, phosphoramide, N-methylacetamide, or acrylamide. Additional nitrogen-containing compounds include glucosamines, for example glucosamine or N-acetylglucosamine or polymers of N-acetylglucosamine, such as chitin and peptidoglycan. Further nitrogen-containing compounds include nucleotides, deoxynucleotides, and purines or pyrimidines such as adenine, uridine, guanine, cytosine, and thymine. The nitrogen-containing compound may also be a polymer of the foregoing, such as polyamines, polyamides, or polynucleotides (e.g. RNA or DNA).

Example 1

Electrolytic Nitrogen Removal Coupled to Hydrogen Gas Production

(15) FIG. 1 is a Pourbaix diagram showing the relationship between pH on the X-axis and the redox potentials E.sub.h on the Y-axis of ammonium oxidation (101) and proton reduction (102). Also shown is the hypothetical offset potential (103) of proton reduction at +0.5 Volts versus a standard hydrogen electrode.

(16) Under the standard conditions (all concentrations, except protons, 1 M, gases 1 bar, temperature 25° C.) shown in FIG. 1, the oxidation of dissolved ammonium 101 (N.sub.2/NH.sub.4.sup.+) with protons 102 (H.sup.+/H.sub.2) as electron acceptor is not spontaneous. This is true for pH values (X-axis) between 0 and 14.

(17) TABLE-US-00001 TABLE 1 Gibbs Free Energy of the complete reactions for oxidation of various nitrogen-containing compounds using protons as the oxidant: ΔG°′ kJ Net reaction mol.sup.−1 Ammonium 2 NH.sub.4.sup.+ .fwdarw. N.sub.2 + 2 H.sup.+ + 3 H.sub.2 +40 3 NH.sub.4.sup.+ .fwdarw. N.sub.3.sup.− + 4 H.sup.+ + 4 H.sub.2 +407 2 NH.sub.4.sup.+ + H.sub.2O .fwdarw. N.sub.2O + 2 H.sup.+ + 4 H.sub.2 +420 2 NH.sub.4.sup.+ + 2 H.sub.2O .fwdarw. 2 NO + 2 H.sup.+ + 5 H.sub.2 +729 NH.sub.4.sup.+ + 2 H.sub.2O .fwdarw. NO.sub.2.sup.− + 2 H.sup.+ + 3 H.sub.2 +437 2 NH.sub.4.sup.+ + 3 H.sub.2O .fwdarw. N.sub.2O.sub.3 + 2 H.sup.+ + 6 H.sub.2 +933 2 NH.sub.4.sup.+ + 4 H.sub.2O .fwdarw. N.sub.2O.sub.4 + 2 H.sup.+ + 7 H.sub.2 +1,128 2 NH.sub.4.sup.+ + 4 H.sub.2O .fwdarw. 2 NO.sub.2 + 2 H.sup.+ + 7 H.sub.2 +1,130 NH.sub.4.sup.+ + 3 H.sub.2O .fwdarw. NO.sub.3.sup.− +2 H.sup.+ + 4 H.sub.2 +600 2 NH.sub.4.sup.+ + 5 H.sub.2O .fwdarw. N.sub.2O.sub.5 +2 H.sup.+ + 8 H.sub.2 +1,382 Ethyl Amine 2 H.sub.3C—CH.sub.2NH.sub.3.sup.+ + .fwdarw. N.sub.2 + 4 HCO.sub.3.sup.− + +187 12 H.sub.2O 6 H.sup.+ + 15 H.sub.2 2 H.sub.3C—CH.sub.2NH.sub.3.sup.+ + .fwdarw. 2 CN.sup.− + 2 HCO.sub.3.sup.− + +753 6 H.sub.2O 6 H.sup.+ + 10 H.sub.2 Alanine 2 H.sub.3C—HCNH.sub.3.sup.+— .fwdarw. N.sub.2 + 6 HCO.sub.3.sup.− + +15 COO.sup.− + 14 H.sub.2O 6 H.sup.+ + 15 H.sub.2 Glutamine CONH.sub.3.sup.+—H.sub.2C—H.sub.2C— .fwdarw. N.sub.2 + 5 HCO.sub.3.sup.− + HCNH.sub.3.sup.+—COO.sup.− + 12 H.sub.2O 6 H.sup.+ + 12 H.sub.2 Ammonium Hydroxide 2 NH.sub.4OH .fwdarw. N.sub.2 + 3 H.sub.2 + 2 H.sub.2O +34 Hydroxyl Amine 2 NH.sub.2OH .fwdarw. N.sub.2 + H.sub.2 + 2 H.sub.2O −232 Urea CO(NH.sub.3).sub.2.sup.2+ + 2 H.sub.2O .fwdarw. N.sub.2 + HCO.sub.3.sup.− + 3 −194 H.sup.+ + 3 H.sub.2

(18) An electron acceptor more positive than N.sub.2/NH.sub.4.sup.+ would ordinarily be needed to make the oxidation of ammonium chemically spontaneous. However, an applied electrode potential (103), for example set to +500 mV, can be used instead of a chemical electron acceptor to drive the reaction. At +500 mV, the oxidation of ammonium to N.sub.2 is spontaneous and hydrogen gas can be produced at the cathode.

(19) FIG. 2 is a schematic of the electrolytic cell used in Example 1 to generate hydrogen gas from an aqueous ammonium solution. In this Example, the cell is divided into a first compartment (206) containing an anode (203) and a second compartment (207) containing a cathode (204). The anode (203) and cathode (204) are partially submerged in an electrolyte (202). The electrolyte (202) also links the two compartments ionically, which were divided by an ion-exchange membrane (210) (Nafion™ 117, Chemours Company, New Castle, Del.). The anode (203) and cathode (204) were in electrical communication with one another via a DC power source (208), the electrolyte (202), and the ion-exchange membrane (210). In Example 1, the DC power source (208) was an AC/DC converter, which is a potentiostat. A reference electrode (209) is provided in order to assist the DC power supply (208) in maintaining a consistent voltage.

(20) Cells of this configuration are described in Siegert et al. “Comparison of Nonprecious Metal Cathode Materials for Methane Production by Electromethanogenesis”ACS Sustainable Chem. Eng., 2014, 2(4), pp 910-917, which is incorporated herein by reference in its entirety.

(21) Each of the reactor chambers (Adams & Chittenden, Berkely, Calif., USA) was sealed using a screw cap (GL45 Corning Screw Caps, Thomas Scientific, Swedesboro, N.J., USA) with a center hole having a septum. The septa were self-made 45 mm discs from 7 mm thick butyl rubber. The two reactor-halves were held together using 35/25 pinch clamps (Thomas Scientific, Swedesboro, N.J., USA).

(22) In other embodiments, the ion-exchange membrane (210) may be omitted. Likewise, the second compartment (207) may further include a collection means for collecting and storing hydrogen gas. Various other DC power sources (208) may also be substituted for the AC/DC converter, such as batteries, solar cells, and the like.

(23) In Example 1, the anode (203) was a cylindrical carbon fiber brush (4 cm×4 cm) made from carbonized polyacrylonitrile fibers sold as Panex 35™ (Zoltek in St. Louis, Mo., USA). Graphite blocks (2×2×0.32 cm) were used as cathode (204), which were sanded with 1,500 grit sandpaper and washed without further treatment. The current collector on the cathode (204) was a titanium wire inserted into the graphite block, through drill holes.

(24) Each electrode was connected to the DC power supply (208) by titanium wires. An Ag/AgCl reference electrode (209) (model RE-5B, BASi, West Lafayette, Ind., USA) was inserted through holes in rubber septa sealing the top of the first (206) compartment.

(25) The solution to be treated in Example 1 was 200 mL of artificial seawater containing 5 mM ammonium chloride and 30 mM sodium bicarbonate. This served as the electrolyte (202), which was added to the first (206) and second (207) compartment. The use of artificial seawater ensured that ammonium was the primary source of electrons and nitrogen in the solution being treated and sodium bicarbonate was the primary carbon source available. A head space of about 50 mL was left in each compartment (206, 207). In alternative embodiments, the electrolye (202) may be municipal wastewater or other nitrogen containing solutions in need of treatment and/or a second electrolyte, such as a suitable buffer, may also be used in the second (207) compartment.

(26) Trace nutrients, minerals, and other growth media were also added to the artificial seawater to support microbial growth. The resulting artificial seawater solution was as follows: 5 mM ammonium chloride 10 mL/L of the following trace element solution:

(27) TABLE-US-00002 Nitrilotracetic acid 1.5 g/L MgSO.sub.4 × 7H.sub.2O 3 g/L MnSO.sub.4 × 2H.sub.2O 0.5 g/L NaCl 1 g/L FeSO.sub.4 × 7H.sub.2O 1 g/L NiCl.sub.2 × 6H.sub.2O 0.2 g/L CoCl.sub.2 0.1 g/L CaCl.sub.2 × 2H.sub.2O 0.1 g/L ZnSO.sub.4 0.1 g/L CuSO.sub.4 × 5H.sub.2O 0.01 g/L AlK(SO.sub.4).sub.2 0.01 g/L H.sub.3BO.sub.3 0.01 g/L Na.sub.2MoO.sub.4 × 2H.sub.2O 0.01 g/L Na.sub.2SeO.sub.3 × 5 H.sub.2O 0.01 g/L Na.sub.2WO.sub.4 × 2 H.sub.2O 0.01 g/L 10 mL/L of the following vitamin solution:

(28) TABLE-US-00003 Pyridoxin × 2HCl 50 mg/L Thiamin × 2HCl 10 mg/L B.sub.12 (cyanocobalamine) 10 mg/L p-Aminobenzoic acid 10 mg/L Riboflavin 5 mg/L Nicotinic acid 5 mg/L Ca—D(+)-pantothenate 5 mg/L Lipoic (thioctic) acid 5 mg/L D(+)-biotin 2 mg/L Folic acid 2 mg/L Each litre of artificial seawater further contained:

(29) TABLE-US-00004 KH.sub.2PO.sub.4 0.20 g NaHCO.sub.3 0.25 g (30 mM) NH.sub.4Cl 0.25 g KCl 0.72 g KBr 0.09 g CaCl.sub.2 × 2H.sub.2O 1.40 g MgCl.sub.2 × 6H.sub.2O 5.7 g MgSO.sub.4 × 7H.sub.2O 6.8 g NaCl 26 g

(30) Further details are provided in Siegert M, Sitte J, Galushko A, Kruger M (2014a), “Starting up microbial enhanced oil recovery.” In: Schippers A, Glombitza F, Sand W (eds) Geobiotechnology II. Springer Berlin Heidelberg, pp 1-94, which is incorporated herein by reference in its entirety.

(31) The first compartment (206) of the electrolytic cell was also inoculated with ocean floor sediment from one of three different locations in the Atlantic Ocean, off the coast of Namibia. Three separate inocula (301, 302, 303) were collected from ocean sediments using gravity corers. The first inoculum (301) was collected at 25°45.060S and 13°04.200E at a water depth of 1,942 m and a sediment depth of 308 cm. The second inoculum (302) was collected at 26°22.178S and 11°53.492E in 3795.6 m water and 0-431 cm sediment depth. The third inoculum (303) was collected at 27°44.131S and 14°14.553E in 1249.3 water and 8-88 cm sediment depth. Live cultures were taken using 1 mL of gravity core sediment, diluted 1/5 with on-site seawater and stored over several years. One mL of the ⅕ (i.e. 20%) dilution was used to inoculate the first compartment (206). Without committing to a particular theory, it is believed that this mixed ocean floor inoculum contained a variety of microorganisms from various genera, including electrogenic species.

(32) In operation, the power supply (208) was adjusted to maintain +400 mV or +550 mV at the anode (203), depending on the treatment. Potentials reported here are expressed versus a standard hydrogen electrode (SHE), which has an approximate offset potential to an Ag/AgCl electrode of about +0.2 volts.

(33) The cells were operated in fed-batch cycles in which the anode (203) was poised at +550 mV during the first two cycles (304, 305) and at +400 mV during the last three cycles (306, 307, 308) using a potentiostat as the power source (208). All voltages are versus a standard hydrogen electrode.

(34) While both compartments were batch-fed, only the first compartment (206) was inoculated with microorganisms. Re-inoculation of the first compartment (206) was carried out with a 10% volume of the solution from the previous batch cycle, resulting a 10-fold dilution series of the existing microorganisms in the first compartment (206). That is, at the end of one fed-batch cycle, 90 mL of solution from the first compartment (206) was discarded. The remaining 10 mL were mixed with 90 mL fresh solution to create a 1:9 mix, which was used to fill the first compartment (206) and the fed batch cycle was started anew. In contrast, the second compartment (207) was completely replenished with fresh electrolyte (202) on each feeding.

(35) Prior to the start of each fed-batch cycle, the headspace in each compartment (206, 207) was flushed using argon gas for at least 5 minutes. This created a substantially anaerobic environment in the electrolytic cell and allowed for more accurate measurements of nitrogen gas production. The anodic potentials used (i.e. +400 mV to +500 mV) were also too low to cause formation of secondary O.sub.2 within the cell through electrolysis of water, which can occur at higher potentials (i.e. +820 mV with Pt catalyst).

(36) Each batch cycle was operated at room temperature (i.e. between 20-30° C.) and standard atmospheric pressure. The total length of the experiment was 600 days.

(37) FIG. 3A depicts nitrogen evolution in the electrolytic cell over five fed-batch cycles (304, 305, 306, 307, 308) with ammonium as the only source of electrons and nitrogen. The results for three electrolytic cells are shown. Each cell was inoculated with one of the three ocean floor sediments (301, 302, 303).

(38) FIG. 3B depicts hydrogen evolution in the electrolytic cell over five fed-batch cycles (304, 305, 306, 307, 308) with ammonium as the only source of electrons and nitrogen. The results for three electrolytic cells are shown. Each cell (301, 302, 303) was inoculated with one of the three ocean floor sediments (301, 302, 303).

(39) The above experiment was repeated by the inventor, with similar results.

(40) The predicted half reactions are as follows:
2NH.sub.3+6e.sup.−.fwdarw.N.sub.2+6H.sup.+
6H.sup.++6e.sup.−.fwdarw.3H.sub.2

(41) Without necessarily committing to a particular theory, it is believed that the microorganisms formed a biofilm on the anode (203). In a first reaction zone, these microorganisms transferred electrons from the oxidation reaction (201) into the anode (203), which passed to the cathode (204) via the power supply (208) under the applied voltage. Simultaneously, hydrogen ions generated during the oxidation reaction (101) in the first reaction zone migrated across the membrane (210), and into the second compartment (207). In the second compartment (207), electrons arriving from the cathode (204) and hydrogen ions arriving from the first compartment (206) are consumed in a reduction reaction (205) at a second reaction zone around the cathode (204).

Example 2

Electrolytic Denitrification Coupled to Methane Gas Production

(42) FIG. 4 is a Pourbaix diagram showing the relationship between pH and redox potentials E.sub.h of ammonium oxidation (101), proton reduction (102), and CO.sub.2 reduction to methane (401). Also shown is the hypothetical offset potential of proton reduction (103) at +0.5 volts, for comparison.

(43) TABLE-US-00005 TABLE 2 Gibbs Free Energy for complete reactions for the oxidation of various nitrogen-containing compounds using carbon dioxide as the oxidant: ΔG°′ kJ Net reaction mol.sup.−1 Ammonium 8 NH.sub.4.sup.+ + 3 HCO.sub.3.sup.− .fwdarw. 4 N.sub.2 + 3 CH.sub.4 + 5 H.sup.+ + 9 H.sub.2O −90 3 NH.sub.4.sup.+ + HCO.sub.3.sup.− .fwdarw. N.sub.3.sup.− + CH.sub.4 + 3 H.sup.+ + 3 H.sub.2O +271 2 NH.sub.4.sup.+ + HCO.sub.3.sup.− .fwdarw. N.sub.2O + CH.sub.4 + H.sup.+ + 2 H.sub.2O +284 8 NH.sub.4.sup.+ + 5 HCO.sub.3.sup.− .fwdarw. 8 NO + 5 CH + 3 H.sup.+ + 7 H.sub.2O +2,237 4 NH.sub.4.sup.+ + 3 HCO.sub.3.sup.− .fwdarw. 4 NO.sub.2.sup.− + 3 CH.sub.4 + 5 H.sup.+ + H.sub.2O +1,341 4 NH.sub.4.sup.+ + 3 HCO.sub.3.sup.− .fwdarw. 2 N.sub.2O3 + 3 CH.sub.4 + H.sup.+ + 3 H.sub.2O +1,459 8 NH.sub.4.sup.+ + 7 HCO.sub.3.sup.− .fwdarw. 4 N.sub.2O4 + 7 CH.sub.4 + H.sup.+ + 5 H.sub.2O +3,561 8 NH.sub.4.sup.+ + 7 HCO.sub.3.sup.− .fwdarw. 8 NO.sub.2 + 7 CH.sub.4 + H.sup.+ + 5 H.sub.2O +3,572 NH.sub.4.sup.+ + HCO.sub.3.sup.− .fwdarw. NO.sub.3.sup.− + CH.sub.4 + H.sup.+ +464 2 NH.sub.4.sup.+ + 2 HCO.sub.3.sup.− .fwdarw. N.sub.2O.sub.5 + 2 CH.sub.4 + H.sub.2O +1,111 Ethyl Amine 8 H.sub.3C—CH.sub.2NH.sub.3.sup.+ + .fwdarw. 4 N.sub.2 + 15 CH.sub.4 + −1,285 3 H.sub.2O HCO.sub.3.sup.− + 9 H.sup.+ 4 H.sub.3C—CH.sub.2NH.sub.3.sup.+ + .fwdarw. 4 CN.sup.− + 5 CH + 7 H.sup.+ + 3H.sub.2O +828 HCO.sub.3.sup.− Alanine 8 H.sub.3C—HCNH.sub.3.sup.+— .fwdarw. 4 N.sub.2 + 15 CH.sub.4 + −1,974 COO.sup.− + 11 H.sub.2O 9 HCO.sub.3.sup.− + 9 H.sup.+ Glutamine CONH.sub.3.sup.+—H.sub.2C-H.sub.2C— .fwdarw. N.sub.2 + 3 CH + 2 HCO.sub.3.sup.− + −507 HCN H.sub.3.sup.+—COO.sup.− + 3 H.sup.+ 3 H.sub.2O Ammonium Hydroxide 8 NH.sub.4OH + 3 HCO.sub.3.sup.− + .fwdarw. 4 N.sub.2 + 3 CH.sub.4 + 17 H.sub.2O −272 3 H.sup.+ Hydroxyl Amine 8 NH.sub.2OH + HCO.sub.3.sup.− + H.sup.+ .fwdarw. 4 N.sub.2 + CH.sub.4 + 11 H.sub.2O −1,063 Urea 4 CO(NH.sub.3).sub.2.sup.2+ .fwdarw. 4 N.sub.2 + 3 CH.sub.4 + HCO.sub.3.sup.− + −1,181 9 H.sup.+ + H.sub.2O

(44) As seen in FIG. 4 and Table 2, the oxidation of ammonium using carbon dioxide as the oxidant proceeds spontaneously (ΔG°′=−90 kJ mol.sup.−1), as follows:
8NH.sub.4.sup.++3HCO.sub.3.sup.−.fwdarw.3CH.sub.4+4N.sub.2+5H.sup.++9H.sub.2O

(45) Accordingly, one advantage of producing methane gas is that a power supply is not always required to drive the reaction. Nevertheless, the application of a potential can increase the kinetics of the reaction.

(46) FIG. 5 is a schematic of the electrolytic cell used in Example 2, which is substantially the same construction as in Example 1. However, in this example, the ion-exchange membrane (210) was omitted so as to allow free movement of microorganisms between the first (206) and second (207) compartments.

(47) In other embodiments, the ion-exchange membrane (210) may be included, as shown in FIG. 2. Likewise, the second compartment (207) may further include a collection means for collecting and storing methane.

(48) Two configurations were tested for the anode (203), with three replicate electrolytic cells in each configuration. In the first configuration, cylindrical carbon fiber brushes (4 cm×4 cm) made from carbonized polyacrylonitrile fibers sold as Panex 35™ (Zoltek in St. Louis, Mo., USA) were used as the anode (203). In the second configuration, a “drum” style anode (601) was used, which comprised a cylindrical titanium mesh basket (4 cm×4 cm, mesh size 40, Ti wire as current collector) with a closed top and bottom, in which the basket was filled with untreated graphite granules of 0.5-5 mm diameter and irregular shapes. Electrodes of this type are described by Siegert in PCT application No. PCT/IB2018/052671, filed April 2018, which is incorporated by reference herein. The cathodes (504) were untreated 4 cm×4 cm stainless steel brushes (type 304) purchased from Gordon Brush in Commerce, Calif., USA.

(49) In operation, the power supply (208) was adjusted to maintain +500 mV at the anode (203). Potentials reported here are expressed versus a standard hydrogen electrode (SHE), which has an approximate offset potential to an Ag/AgCl electrode of about +0.2 volts.

(50) The solution to be treated was domestic wastewater collected at Calgary's Fish Creek Wastewater Treatment Plant, usually during the morning hours. Pure wastewater was obtained from the primary clarifier. Total Kjeldah Nitrogen (“TKN”) concentrations varied from 43 to 120 mg/L (N) or 3-9 mM.

(51) In Example 2, the microorganisms were provided endogenously within the wastewater, which was enriched by a 1% inoculum of anaerobic digester sludge obtained from the same wastewater treatment plant. This wastewater solution was supplied to the first (206) and second (207) compartments of the electrolytic cell. In alternative embodiments, a different electrolyte, such as a suitable buffer, may instead be used in the second (207) compartment, in place of wastewater.

(52) Without committing to a particular theory, it is believed that the inoculum derived from the wastewater and anaerobic digester sludge contained a variety of microorganisms from various genera, including electrogenic and methanogenic species.

(53) Once the experiment was underway, no additional inoculum was used from the anaerobic digester. Instead, the electrolytic cells were batch fed using a 10% volume of solution from the previous batch cycle mixed with freshly collected wastewater, resulting a 10-fold dilution series. Each fed-batch cycle lasted until gas evolution ceased but no longer than ten days. Each batch cycle was operated at room temperature (i.e. between 20-30° C.) and standard atmospheric pressure.

(54) Each cell was operated as a closed system, such that oxidative decomposition of organic and other materials in the wastewater quickly created an anaerobic environment within the cells. The anodic potentials used (i.e. 0.5 volts) were also too low to cause formation of secondary O.sub.2 within the cell through electrolysis of water, which can occur at higher potentials (i.e. +0.82 volts with a Pt catalyst). The average O.sub.2 concentration within the wastewater was 42±34 μM with the error being the standard deviation, n=392 measurements. On average, there was a slight increase in O.sub.2 levels within the head space (1 μM/day) due to sampling and leakage. Nevertheless, O.sub.2 concentrations within the wastewater continued to decrease over the course of the experiment. The maximum observed O.sub.2 concentration within the wastewater was approximately 80 μM at one measurement. O.sub.2 concentrations of less than 100 μM, less than 50 μM, and less than 30 μM are also contemplated within the scope of the present disclosure.

(55) In Example 2, the oxidant was carbon dioxide, which was reduced to methane. The CO.sub.2 in the cathode reaction (205) may be in the form of bicarbonate ions within the wastewater to be treated (e.g. due to the previous oxidation of organic molecules in the wastewater) or can be supplied to the electrolytic cell by an outside source, such as CO.sub.2 sparging or gaseous supply to the head space in the second compartment (207).

(56) FIG. 6A depicts the % nitrogen removal using a carbon brush anode (603) or a graphite granule “drum” anode (601). Total nitrogen was determined using the TKN method.

(57) FIG. 6B depicts the combined evolution of methane from both compartments (206, 207) in μmol/L/day using a carbon brush anode (603) or a graphite granule “drum” anode (601) of the type described above. The cathodes were in all cases the steel brushes described above. Gases were measured using a gas chromatograph. During the first four fed-batch cycles, the second compartment (207) also produced hydrogen gas (not shown) which was consumed by methanogenic microorganisms and converted into methane gas. During the last fed-batch cycle, hydrogen gas accumulated only during the first day.

(58) FIG. 6C depicts the evolution of nitrogen gas in μmol/L/day at the anode (601) and cathode (604) of the electrolytic cells. A graphite granule “drum” style anode (601) was used in this experiment, which was performed in triplicate. As seen in FIG. 6C, there was more dinitrogen gas in the anode compartment than in the cathode compartment indicating an oxidative process, which in the case of wastewater can be the oxidation of organic nitrogen or ammonium.

(59) The predicted half reactions are as follows:
2NH.sub.3+6e.sup.−.fwdarw.N.sub.2+6H.sup.+
CO.sub.2+8e.sup.−+8H.sup.+.fwdarw.CH.sub.4+2H.sub.2O

(60) Without necessarily committing to a particular theory, it is believed that microorganisms populated both the anode (203) and the cathode (504) of the electrolytic cell. Electrons and hydrogen ions were transferred from the oxidation reaction (201) in a first reaction zone around the anode (203) to a second reaction zone around the cathode (504), in a similar manner to Example 1 above. In Example 2, microorganisms were also present in the second compartment (207), possibly as a biofilm on the cathode (504). It is believed that methanogens amongst the population of micoorganisms were responsible for the reduction reaction (401), in which incoming hydrogen ions from the electrolyte (202) were combined with incoming electrons from the cathode (504) in the presence of bicarbonate, at a second reaction zone in the second compartment (207), to produce methane and water. Intermediates in this process may include hydrogen gas, which is likely consumed by the methanogens when producing methane.

Example 3

Fuel Cell Nitrogen Removal

(61) Referring to FIG. 7, in another aspect, there is provided a fuel cell (700). The fuel cell (700) includes an anode compartment (702) and a cathode compartment (704). The anode compartment (702) is separated from the cathode compartment (704) by a separator (706) configured to permit selective permeation of ionic species therethrough. An anode (708) is disposed within the anode compartment (702) and a cathode (710) is disposed within the cathode compartment (704). The anode (708) and cathode (710) are electrically coupled to an external load (712) via an external circuit (714). A reference electrode (713) may be used to monitor the redox potential at either of the electrodes.

(62) Although control over the voltage may be more difficult, a fuel cell (700) is more efficient as it avoids two energy conversion steps typical of electrolytic arrangements, namely: (a) conversion of electricity into chemical energy; and (b) conversion of chemical energy back into electricity.

(63) Wastewater is supplied to the anode compartment (702) such that the wastewater becomes disposed within the anode compartment (702) in electrical communication with the anode (708). In some embodiments, for example, the wastewater includes nitrogen-containing or organic compounds that serve as the energy source for the fuel cell.

(64) Oxidant is supplied to the cathode compartment (704) such that the oxidant becomes disposed within the cathode compartment (704) in electrical communication with the cathode (710). Such an oxidant may be gaseous oxygen dissolved in solution but can be any other oxidizing compound. In some embodiments, for example, the oxidant includes O.sub.2, benzoyl peroxide, ferricyanide, manganese dioxide, or nitrate.

(65) Mediated by electrical communication, the oxidant reacts with a fuel source to create a cathodic reduction reaction with a redox potential more positive than the anodic oxidation reaction. Example compounds that may serve as the energy source for the fuel cell include nitrogen-containing compounds and organic compounds, which may be found in wastewater or may be supplied separately to the cathode compartment (704). Examples fuel sources include ammonium, benzylamine, ethanolamine, arginine, cysteine, serine, acetamide, N-methylacetamide, or other nitrogen-containing compounds.

(66) An example of such a reaction mediated by the fuel cell (700) is the oxidation of ammonium in the presence of molecular oxygen (2 NH.sub.4.sup.++3 O.sub.2.fwdarw.2 N.sub.2+4 H.sup.++6 H.sub.2O; ΔG°=−1,265 kJ mol.sup.−1). Various other nitrogen-containing compounds or organic molecules may be similarly oxidized to provide the required current.

(67) While: (i) the wastewater is disposed in electrical communication with the anode (708), and (ii) the oxidant is disposed in electrical communication with the cathode (710), electrical current is generated across the external circuit (714) of the fuel cell (700). Such current may be used to drive the anodic reaction and/or be used to power a useful external load (712) such as a resistor or battery.

(68) In some embodiments, a plurality of microorganisms may be disposed within the anode compartment (702) to assist the anodic reaction. In some embodiments, for example, at least a portion of such microorganisms may be coated on the anode (708) as a biofilm. These microorganisms may assist in nitrogen removal in the manner described in Examples 1 and 2 above.

(69) In yet another aspect, the electrochemical cell in Example 2 (see: FIG. 5) may itself be configured as a microbial fuel cell. As discussed, the oxidation of ammonium using carbon dioxide under anaerobic conditions is a spontaneous reaction (ΔG°′=−90 kJ mol.sup.−1). In this embodiment, the power supply (208) is replaced by a useful electrical circuit, and the voltage potential is generated by spontaneous oxidation of nitrogen-containing compounds by electrogenic microorganisms at the anode (203).

(70) In yet another aspect, the fuel cell (700) can be used for detecting target material within the wastewater, and the load (712) is an electrical current sensor. In this respect, in some embodiments, for example, the disposition of the wastewater within the fuel cell (700) is with effect that an electrical current is generated that is representative of the presence of a target material within the wastewater. The sensing of the current by the sensor can be used to indicate detection of the target material within the wastewater. In response to the sensing, a signal can be transmitted, representative of the detection of the target material. Examples of such targets include nitrogen-containing compounds.

(71) The detection of nitrogen-containing compounds may be used as a signal to begin treating the wastewater, suspending the treatment of the wastewater, increasing a stimulus applied to the wastewater for the treatment of wastewater, or decreasing a stimulus applied to the wastewater for the treatment of wastewater. The stimulus may include admixing the wastewater with a reagent, or disposing the wastewater within a reaction zone in the electrolytic cell, with effect that the aqueous solution is in electrical communication with an anode and the anode is disposed in electrical communication with a cathode and applying a voltage, and includes embodiments discussed herein.

(72) The embodiments of the present disclosure are intended to be examples only. Those of skill in the art may effect alterations, modifications and variations to the particular embodiments without departing from the intended scope of the present application.

(73) In particular, features from one or more of the above-described embodiments may be selected to create alternate embodiments comprised of a subcombination of features which may not be explicitly described above. In addition, features from one or more of the above-described embodiments may be selected and combined to create alternate embodiments comprised of a combination of features which may not be explicitly described above. Features suitable for such combinations and subcombinations would be readily apparent to persons skilled in the art upon review of the present application as a whole. The subject matter described herein and in the recited claims intends to cover and embrace all suitable changes in technology.