Microbial synthesis reaction system based on electrochemical energy supply and application thereof

Abstract

The invention discloses a microbial synthesis reaction system based on electrochemical energy supply and application thereof, the system is provided with an anode chamber and a cathode chamber, a biomass derivative easy to oxidize and lose electrons is arranged in the anode chamber, the anode chamber and the cathode chamber are separated through a proton exchange membrane, H < + > transfer is allowed, and pH balance of the system is maintained; nAD < + > is converted into NADH through electrochemical reduction in the cathode chamber, the neutral environment is maintained through H < + >, the NADH in the cathode chamber is conveyed to microorganisms through a peristaltic pump, the microorganisms take in the NADH through transporter protein, and the NADH drives metabolism to synthesize target products such as rare sugar, rare sugar alcohol, biodegradable plastic monomers, high-activity natural products and microbial oil. Meanwhile, the NAD < + > is discharged out of the organisms through the transporter and flows back to the cathode chamber through the filter membrane, so that cyclic utilization is realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of advanced manufacturing technology of microorganisms, and in particular to a microorganism synthesis reaction system based on electrochemical energy supply and application thereof. BACKGROUND

[0002] Traditional microbial fermentation and other biological synthesis technologies rely on reducing carbon sources such as glucose as energy carriers to drive the synthesis of biomolecules through metabolic pathways. However, the existing technology has significant bottlenecks:

[0003] 1. Low carbon efficiency: In traditional synthetic biology, each molecule of glucose metabolism produces 6 molecules of CO2, resulting in about 30% carbon loss, exacerbating carbon emissions and resource waste.

[0004] 2. Strong energy dependence: A large amount of reducing carbon sources derived from sugars or fossil energy are consumed, which is high in cost and unsustainable.

[0005] 3. Reaction system limitations: The traditional electrochemical-biological coupling system has low efficiency and poor catalyst stability, making it difficult to adapt to complex biological synthesis pathways. In addition, although synthetic chemistry can precisely regulate molecular structure, it relies on harsh conditions such as high pressure / high temperature, which is high in energy consumption and pollution, and is difficult to meet the demand for green manufacturing.

[0006] For example, patent No. CN107244658A, a method for synthesizing hydrogen peroxide using a microbial electrochemical system, discloses a scheme that relies on reducing carbon sources such as glucose as energy sources to generate ATP and NADH through sugar metabolic pathways to drive the synthesis of target products. In this scheme, each molecule of glucose metabolism produces 6 molecules of CO2, resulting in about 30% carbon loss, exacerbating carbon emissions and resource waste, and the biological product is H2O2, which can be synthesized in large quantities and at low cost by industry. This scheme belongs to the typical carbon source energy supply type traditional biological synthesis technology.

[0007] For example, patent No. CN102340015A, a microbial electrochemical system capable of simultaneously producing electricity, hydrogen and treating wastewater, discloses a technical scheme that separates the pH gradient of the anode chamber and the cathode chamber through a diaphragm technology, increases the hydrogen evolution potential of the cathode, and maintains the activity of the anode electricity-producing microorganisms to oxidize organic matter in wastewater, which can simultaneously treat organic matter in wastewater at the anode and produce hydrogen at the cathode. Due to the increase in cathode potential, the reaction can occur spontaneously, and the output of electrical energy is low. The cost is high and unsustainable. This scheme belongs to the typical electrode direct energy supply type traditional electrochemical-biological coupling system.

[0008] For example, in the patent CN104328046B, a device and method for reducing carbon dioxide to produce acetic acid using a microbial electrochemical system are disclosed, which uses neutral red, methylene blue and other synthetic electron carriers or cobalt-based catalysts to mediate extracellular electron transfer. The efficiency of the electrochemical-biological coupling system in this scheme is low, the synthetic electron carrier has poor adaptability to the biological system, there is no uptake and transport channel, and the catalyst has poor stability and is easily oxidized by oxygen, which is difficult to adapt to complex biological synthesis pathways and difficult to meet the demand of green manufacturing. This scheme belongs to the traditional non-biological-biological interface type artificial electron carrier technology.

[0009] Based on the above problems, the inventors have conducted in-depth research on microbial synthesis reaction mechanism and electrochemical energy supply technology, and thus proposed an electrochemically energized microbial synthesis reaction system that can solve the above problems. SUMMARY

[0010] To overcome the above problems, the inventors have conducted intensive research and designed an electrochemically energized microbial synthesis reaction system and its application. The system is based on the deep integration of electrochemistry and synthetic biology, selects biocompatible electron carriers such as NAD + and NADH, uses monoMOF as a heterogeneous catalyst, and realizes efficient and high-selectivity conversion of oxidized NAD + to reduced NADH at the cathode through electrochemical reduction, i.e., "molecular charging", thereby replacing the traditional sugar metabolism to produce NADH and directly converting renewable electrical energy into biologically available reducing power, i.e., NADH. In addition, transport proteins are constructed on the cell membrane of model organisms to better match the biological system, avoid carbon loss during glucose metabolism, realize energy "storage-release" cycle, and adapt to the dynamic needs of microbial metabolism. In the anode chamber, biomass derivatives that are easily oxidized and lose electrons are configured, a proton exchange membrane is used to separate the anode chamber and the cathode chamber and allow H + transfer, and the pH balance of the system is maintained; in the cathode chamber, NAD + is converted to NADH through electrochemical reduction, H + is used to maintain a neutral environment, and a peristaltic pump is used to transport NADH from the cathode chamber to the microorganisms, which are then taken up by the microorganisms using transport proteins. NADH drives the synthesis of target products such as rare sugars, rare sugar alcohols, biodegradable plastic monomers, high-activity natural products, microbial oils, etc., while NAD + is removed from the organism by transport carriers and returned to the cathode chamber through a filter membrane for recycling; thus completing the invention.

[0011] Specifically, the present application aims to provide an electrochemically powered microbial synthesis reaction system, which comprises an anode chamber and a cathode chamber separated by a proton exchange membrane 1; an anode 2 is arranged in the anode chamber, and a cathode 3 is arranged in the cathode chamber;

[0012] An anode flow channel 4 is arranged in the anode chamber, and the anode flow channel 4 is filled with an anode substrate, and the anode substrate provides electrons;

[0013] Outside the anode chamber and the cathode chamber, the anode 2 and the cathode 3 are connected in series by a power source to form a loop;

[0014] A cathode flow channel 5 is arranged in the cathode chamber, and the cathode flow channel 5 is connected to a reactor 7 by a peristaltic pump 6; an electron carrier is arranged in the cathode flow channel 5 and the reactor 7, and the cathode 3 can continuously generate the electron carrier;

[0015] Microorganisms are also arranged in the reactor 7, which take up the electron carrier through transport proteins, and then drive microbial metabolism to synthesize target products through the electron carrier.

[0016] The proton exchange membrane 1 allows H + transfer, thereby maintaining the pH balance of the system and avoiding the inhibition of microbial activity caused by acid-base imbalance.

[0017] The electron carrier has biocompatibility;

[0018] Preferably, the electron carrier is selected from one or a mixture of more than one of NADH, NADH artificial analog, FADH2, and FADH2 artificial analog;

[0019] Preferably, when the electron carrier is NADH, NADH is oxidized to NAD + when the microorganisms metabolize to synthesize target products, and the cathode 3 continuously generates NADH based on NAD

[0020] When the electron carrier is FADH2, FADH2 is oxidized to FAD when the microorganisms metabolize to synthesize target products, and the cathode 3 continuously generates FADH2 based on FAD.

[0021] The anode 2 and the cathode 3 are both obtained by loading a monoMOF catalyst on a substrate; the thickness of the membrane on the anode 2 and the cathode 3 is 2-20 nm;

[0022] Preferably, the substrate is selected from any one or a mixture of more than one of a carbon / graphite substrate, a metal substrate, an ITO substrate, and a silicon substrate;

[0023] Preferably, the metal substrate is selected from any one of titanium, gold, silver, copper, nickel, platinum, cobalt, and stainless steel.

[0024] Wherein, by in-situ growth or chemical modification grafting method on carbon, metal and other substrates, directional assembly of monoMOF catalyst is realized.

[0025] Wherein, the anode substrate can lose electrons by oxidation;

[0026] Preferably, the anode substrate is a biomass derivative, more preferably, the anode substrate is 5-hydroxymethylfurfural (HMF), which is converted into 2,5-furan dicarboxylic acid (FDCA) by losing electrons and discharged from the outlet of the anode flow channel 4;

[0027] Further preferably, the industrial wastewater containing cellulose or starch is pretreated and input into the anode flow channel as the anode substrate.

[0028] Wherein, the reactor 7 is a batch fermentation tank or a microfluidic continuous flow reactor;

[0029] Preferably, a filter membrane 8 is arranged between the reactor 7 and the peristaltic pump 6, which allows the electron carrier to pass through and can intercept microorganisms.

[0030] Wherein, the power supply includes a solar panel or a wind turbine, and further includes an energy storage battery.

[0031] The application also provides an application of the above-mentioned electrochemically powered microbial synthesis reaction system in the production of high-value chemicals, including fragrances, amino acids, rare sugars, rare sugar alcohols, biodegradable plastic monomers, high-activity natural products, and microbial oils.

[0032] Wherein, the fragrances include vanillin, raspberry ketone, naringin, rose essential oil, artemisia oil, menthol, gamma-decalactone, 2-acetylpyrroline, benzaldehyde, ionone, citronellal, beta-caryophyllene, and algal beta-ionone;

[0033] The amino acids include L-glutamic acid sodium, L-lysine, L-threonine, L-tryptophan, L-arginine, L-proline, L-cysteine, L-valine, gamma-aminobutyric acid, L-theanine, L-selenium-methyl selenocysteine, and epsilon-polylysine;

[0034] The rare sugars include D-allulose, D-tagatose, L-ribose, D-allose, L-xylulose, D-turanose, L-fucose, D-isomaltulose, L-rhamnose, D-mannoheptulose, D-alloheptulose, and L-lyxose;

[0035] The rare sugar alcohol includes allitol, erythritol, xylitol, allose, maltitol, sorbitol, lactitol, isomalt, mannitol, ribitol, tagatosyl, galactitol, erythrulose;

[0036] The biodegradable plastic monomer includes lactide, succinic acid, 5-hydroxymethyl furfural, 1,4-butanediol, FDCA, lactic acid, adipic acid, 3-hydroxypropionic acid, epsilon-caprolactone, glycolide, isosorbide, dimethyl terephthalate;

[0037] The high-activity natural product includes fucoidan sulfate, rare ginsenoside CK, paclitaxel, astaxanthin, lycopene, steviol glycoside, mogroside, artemisinic acid, ginkgolide B, ganoderic acid, huperzine A, breviscapine;

[0038] The microbial oil includes schizochytrium DHA algal oil, Yarrowia lipolytica ARA oil, Rhodotorula glutinis gamma-linolenic acid, Chlorella vulgaris EPA algal oil, Mortierella alpina arachidonic acid, Chlorella ellipsoidea astaxanthin oil, Starmerella bombicola microbial butter, Crypthecodinium cohnii DHA oil, Rhodotorula glutinis lycopene oil, Rhodotorula glutinis beta-carotene oil, Nannochloropsis gaduola squalene oil, marine bacteria wax ester.

[0039] The present application has the beneficial effects, including:

[0040] (1) The present application provides an electrochemically powered microbial synthesis reaction system and its application, which uses monoMOF as a heterogeneous electrocatalyst, efficiently and selectively converts oxidized NAD + into reduced NADH, directly converts renewable electrical energy into biologically available NADH with reducing power, and constructs a transport protein on the cell membrane of a model organism, which is more compatible with the biological system, avoids carbon loss in glucose metabolism, and significantly improves carbon efficiency; realizes energy "storage-release" cycle, adapts to the dynamic demand of microbial metabolism;

[0041] (2) The present application provides an electrochemically powered microbial synthesis reaction system and its application, which solves the compatibility problem of electrolyte and fermentation broth by separating the anode chamber and the cathode chamber through a proton exchange membrane;

[0042] (3) The present application provides an electrochemically powered microbial synthesis reaction system and its application, which regulates the NADH delivery rate through a peristaltic pump, synchronizes, and in addition, NADH is the core reducing power of microorganisms, supports multi-step product synthesis, so that the system adapts to complex reaction paths, thereby making the system universal;

[0043] (4) The application provides an electrochemically-powered microbial synthesis reaction system and application thereof, wherein the NADH in the system is reduced by an anode substrate to provide energy for the microbial synthesis reaction. + The circulation reduces carrier consumption, prolongs the service life of a platinum-based catalyst to more than 100 hours, improves system stability, reduces economic cost of the system, and makes the system have a higher market prospect.

[0044] (5) The application provides an electrochemically-powered microbial synthesis reaction system and application thereof, wherein the power supply can be a solar panel or a wind turbine, and the intermittent problem of renewable energy can be solved by combining with an energy storage battery, so that sustainable energy supply in all time periods is realized, and dependence on traditional carbon sources is reduced.

[0045] (6) The application provides an electrochemically-powered microbial synthesis reaction system and application thereof, wherein the anode substrate can be replaced by industrial wastewater, such as cellulose and starch in papermaking wastewater, so that the anode can be simultaneously used for wastewater treatment. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1 The application shows the overall structure of the electrochemically-powered microbial synthesis reaction system.

[0047] Figure 2 The application shows the nuclear magnetic resonance hydrogen spectrum of Me2DBB.

[0048] Figure 3 The application shows the nuclear magnetic resonance hydrogen spectrum of Me2DBB-RhCp*Cl.

[0049] Figure 4 The application shows the nuclear magnetic resonance hydrogen spectrum of H2DBB-RhCp*Cl.

[0050] Figure 5 The application shows the SEM image of Hf 12 DBB-RhCp*ClmonoMOF.

[0051] Figure 6 The application shows the SEM image of Rh monoMOF in-situ grown on a titanium felt, namely Rh monoMOF-Ti substrate.

[0052] Figure 7 The application shows the SEM image of Rh monoMOF in-situ grown on a carbon paper, namely Rh monoMOF-C substrate.

[0053] Figure 8 The application shows the ultraviolet standard concentration curve of NADH in the experimental example.

[0054] Figure 9 and Figure 10Electrochemical NADH oxidation of H2DBB-RhCp*Cl ligand + Reduction test results schematic diagram;

[0055] Figure 11 and Figure 12 Electrochemical NADH oxidation of Rh monoMOF-C + Reduction test results schematic diagram;

[0056] Figure 13 and Figure 14 Electrochemical NADH oxidation of Rh monoMOF-Ti + Reduction test results schematic diagram;

[0057] Figure 15 Schematic diagram showing E. coli membrane surface expressing NTT4;

[0058] Figure 16 Schematic diagram showing E. coli plasmid system for producing mannitol;

[0059] Figure 17 Schematic diagram showing E. coli NTT4 after purification, Coomassie brilliant staining of SDS-PAGE and WB results;

[0060] Figure 18 Schematic diagram showing DPE and RDH after purification, Coomassie brilliant staining of SDS-PAGE.

[0061] BRIEF DESCRIPTION OF DRAWINGS

[0062] 1-proton exchange membrane, 2-anode, 3-cathode, 4-anode flow channel, 41-flow channel inlet, 42-flow channel outlet, 5-cathode flow channel, 6-peristaltic pump, 7-reactor, 8-filter membrane. DETAILED DESCRIPTION

[0063] The application will be further described below in detail by means of the accompanying drawings and examples. The features and advantages of the present application will become more apparent from these descriptions.

[0064] The term "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any implementation described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other implementations. Although various aspects of an implementation can be described herein as being a preferred or advantageous implementation, no inference should be drawn that other aspects necessarily are inferior or inferior to other aspects or implementations. Unless otherwise indicated, the drawings are not necessarily drawn to scale.

[0065] The present application provides a kind of electrochemically energized microbial synthesis reaction system based on, as shown in Figure 1 The system includes anode chamber and cathode chamber separated by proton exchange membrane 1, anode 2 is arranged in anode chamber, cathode 3 is arranged in cathode chamber;

[0066] An anode flow channel 4 is arranged in the anode chamber, and the anode flow channel 4 is filled with an anode substrate, through which electrons are provided;

[0067] Outside the anode chamber and the cathode chamber, the anode 2 and the cathode 3 are connected in series by a power supply to form a loop, thereby providing a path for the directional flow of electrons and continuously providing electrons for the cathode;

[0068] A cathode flow channel 5 is arranged in the cathode chamber, and the cathode flow channel 5 is connected to a reactor 7 by a peristaltic pump 6; an electron carrier is arranged in the cathode flow channel 5 and the reactor 7, and the cathode 3 can continuously generate the electron carrier; that is, the electron carrier loses electrons in the cathode chamber, regains electrons to become an electron carrier, and is pumped into the reactor 7 by the peristaltic pump 6, thereby realizing the recycling of the electron carrier;

[0069] Microorganisms are also arranged in the reactor 7, which take up the electron carrier through transport proteins, and then drive the synthesis of target products through the electron carrier.

[0070] In a preferred embodiment, the proton exchange membrane 1 allows H + transfer, thereby maintaining the pH balance of the system, avoiding the inhibition of microbial activity caused by acid-base imbalance, maintaining the pH value in the reactor 7 at 6.5-7.5 while avoiding the entry of electrochemical reagents into the reactor 7, and providing a suitable acid-base environment for the survival of microorganisms.

[0071] In a preferred embodiment, the electron carrier has biological compatibility and can meet the needs of specific microorganisms through electrochemical reduction;

[0072] Preferably, the electron carrier is selected from one or a mixture of more than one of NADH, an artificial analogue of NADH, FADH2, and an artificial analogue of FADH2; preferably NADH or FADH2;

[0073] When the electron carrier is NADH, NADH is oxidized to NAD + when microorganisms synthesize target products, and the cathode 3 continuously generates NADH based on NAD + ;

[0074] When the electron carrier is FADH2, FADH2 is oxidized to FAD when microorganisms synthesize target products, and the cathode 3 continuously generates FADH2 based on FAD.

[0075] The working process of the electron carrier is illustrated below using NADH as an example:

[0076] NAD +Using NADH as an electron carrier and monoMOF as a heterogeneous catalyst, oxidized NAD+ is efficiently and selectively reduced at the cathode via electrochemical reduction. + The process of converting NADH into reduced NADH, known as "molecular charging," replaces the traditional method of producing NADH through sugar metabolism. In this application, renewable electrical energy is directly converted into biologically usable reducing power (NADH), and transport proteins are constructed on the cell membranes of model organisms to better match the biological system and avoid carbon loss from glucose metabolism. This achieves an energy "storage-release" cycle that adapts to the dynamic metabolic needs of microorganisms.

[0077] NAD in this application + The full name is nicotinamide adenine dinucleotide (oxidized coenzyme I); the full name of NADH in this application is reduced nicotinamide adenine dinucleotide (reduced coenzyme I).

[0078] The construction of transport proteins on the cell membranes of model organisms described in this application can be based on existing technologies. For example, NTT4 is expressed on the surface of E. coli membranes to achieve NAD(H) transport. A schematic diagram of NTT4 expression on the surface of E. coli membranes is shown below. Figure 15 As shown, the plasmid system of Escherichia coli for the production of allicin is as follows: Figure 16 As shown; Coomassie smear staining and Western blot results of SDS-PAGE of purified E. coli NTT4 are as follows. Figure 17 As shown; Coomassie staining of SDS-PAGE purified from DPE and RDH as shown. Figure 18 As shown.

[0079] In a preferred embodiment, both the anode 2 and the cathode 3 are obtained by loading a monoMOF catalyst onto a substrate; the thickness of the films on the anode 2 and the cathode 3 is 2-20 nm.

[0080] Preferably, the substrate is selected from any one or a mixture of carbon / graphite substrate, metal substrate, ITO substrate, and silicon substrate;

[0081] Preferably, the metal substrate is selected from any one of titanium, gold, silver, copper, nickel, platinum, cobalt, and stainless steel.

[0082] Preferably, the monoMOF catalyst is oriented assembled on a substrate such as carbon or metal by in-situ growth of monoMOF or by chemical modification grafting.

[0083] In a preferred embodiment, the monoMOF catalyst is selected as Hf. 12 -DBB-RhCp*ClmonoMOF, its synthesis process is as follows: where the Chinese name corresponding to DBB-RhCp*Cl is [chlorine-η] 5- [pentamethylcyclopentadienyl-(4,4'-bis(4-carboxyphenyl)-2,2'-bipyridine)] rhodium(III)]+;

[0084] Step 1, synthesis of Me2DBB; the synthetic route is as follows:

[0085]

[0086] The specific synthesis process is as follows:

[0087] Dissolve 5,5'-dibromo-2,2'-bipyridine (4.0 g, 12.75 mmol) 13 and 4-(methoxycarbonyl)phenylboronic acid (6.88 g, 38.25 mmol) in tetrahydrofuran (THF, 103 mL); the system is deoxygenated for 15 minutes;

[0088] Add an aqueous solution containing K2CO3 (8.8 g, 63.67 mmol) (21 mL) and Pd(PPh3)4 catalyst (1.5 g, 1.30 mmol);

[0089] Seal the reaction vessel and stir the reaction at 80°C for 3 days;

[0090] After cooling to room temperature, collect the solid by filtration; wash with THF and deionized water in turn; obtain the pure product Me2DBB. The nuclear magnetic resonance hydrogen spectrum of this Me2DBB is shown in Figure 2 .

[0091] Step 2, synthesis of Me2DBB-RhCp*Cl; the synthetic route is as follows:

[0092]

[0093] The specific synthesis process is as follows:

[0094] Add [RhCp*Cl]2Cl2 (1.0 mmol), Me2DBB (0.85 g, 2.0 mmol), methanol (25 mL) and chloroform (25 mL) into a 200 mL thick-walled reaction tube.

[0095] Degassing the reaction tube with N2, seal, and heat and stir at 120°C for 2 days.

[0096] After cooling to room temperature, remove the solvent under reduced pressure to obtain the pure Me2DBB-RhCp*Cl product as an orange solid (1.92 g, 2.0 mmol, 100%).

[0097] If the product is not pure, the solid can be treated with ethanol under ultrasonic, and then filtered to remove the remaining impurities. The nuclear magnetic resonance hydrogen spectrum of this Me2DBB-RhCp*Cl is shown in Figure 3 .

[0098] Step 3, synthesis of H2DBB-RhCp*Cl; the synthetic route is as follows:

[0099]

[0100] The specific synthesis process is as follows:

[0101] 100 mg of Me2DBB-Rh was added to 15 mL of a mixture of MeOH and THF in a molar ratio of 1:1, and the reaction solution was light orange; the ambient temperature in the above process was room temperature, and the reaction duration was 6 h;

[0102] 5 mL of a LiOH aqueous solution was further added to the reaction solution, the amount of LiOH in the aqueous solution was 10 mg, and after half an hour of reaction, the reaction solution became turbid;

[0103] After 4 h of reaction, cooling to room temperature, TLC monitoring, and substantially complete reaction, spin-drying, vacuum drying, H2DBB-RhCp*Cl was obtained;

[0104] 1 The H NMR sample was dissolved in deuterated DMSO, and the nuclear magnetic characterization is shown in Figure 4 .

[0105] Step 4, synthesis of Hf 12 -Rh;

[0106] The specific synthesis process is as follows:

[0107] A 5 mL glass bottle was added with: 0.5 mL of a HfCl4 solution (concentration 2.0 mg / mL, solvent N,N-dimethylformamide / DMF), 0.5 mL of a H2DBB-Rh solution (concentration 3.6 mg / mL, solvent DMF), 2 μL of trifluoroacetic acid (TFA), and 5 μL of deionized water; H2DBB-Rh in this application is a short form of H2DBB-RhCp*Cl

[0108] The mixture was placed in an 80°C oven and reacted for 24 h;

[0109] The generated orange precipitate Hf 12 -Rh was collected by centrifugation, and the precipitate was sequentially washed with DMF and ethanol. Hf 12 -Rh in this application is Hf 12 -DBB-RhCp*Cl MonoMOF;

[0110] Based on the recovery rate of hafnium elements, the yield determined by ICP-MS was 56%.

[0111] Figure 5 Hf 12- SEM image of H2DBB-RhCp*Cl monoMOF (Rh monoMOF for short).

[0112] In a preferred embodiment, the present application realizes the highest density packing of active sites on the unit area of the point electrode by assembling the molecular catalyst into a periodic structure monoMOF. How the monoMOF is combined with the electrode is a key issue. The commonly used Nafion TM The mixed drop-casting method is prone to cause the coating of active sites and easy to fall off from the electrode. Therefore, the present application grows the monoMOF in situ on the carbon substrate or metal substrate to realize the directional assembly of the catalyst.

[0113] Figure 6 SEM image of H2DBB-RhCp*Cl monoMOF (Rh monoMOF for short) grown in situ on a titanium felt substrate is shown,

[0114] Figure 7 SEM image of H2DBB-RhCp*Cl monoMOF grown in situ on a carbon paper substrate is shown

[0115] In a preferred embodiment, the anode substrate is capable of losing electrons;

[0116] Preferably, the anode substrate is a biomass derivative, more preferably, the anode substrate is 5-hydroxymethylfurfural (HMF), which is converted into 2,5-furan dicarboxylic acid (FDCA) by losing electrons and is discharged from the outlet of the anode flow channel 4;

[0117] Preferably, the anode flow channel 4 is provided with a flow channel inlet 41 and a flow channel outlet 42, and the anode substrate 5-hydroxymethylfurfural (HMF) is injected into the anode flow channel through the flow channel inlet 41, and the 2,5-furan dicarboxylic acid (FDCA) obtained by losing electrons is discharged from the flow channel outlet 42.

[0118] Further preferably, the industrial wastewater containing cellulose or starch is pretreated and then input into the anode flow channel as the anode substrate. The pretreatment can be in various ways, for example, biological fermentation method or hydrolysis method or acid treatment, etc., which converts the cellulose or starch into a biomass derivative.

[0119] In a preferred embodiment, the reactor 7 is a batch fermentation tank or a microfluidic continuous flow reactor; the corresponding reactor form can be selected according to the production efficiency of the target product.

[0120] Preferably, a filter membrane 8 is arranged between the reactor 7 and the peristaltic pump 6, which allows the electronic carrier to pass through and intercepts the microorganisms, thereby ensuring the recycling of the electronic carrier.

[0121] In the present application, the flow rate in the cathode flow channel 5 can be adjusted between 1-50 mL / min under the action of the peristaltic pump 6, so as to ensure that the supply rate of the electron carrier matches the production rate of the target product, avoid excess or insufficient reducing power, and make the reaction system adapt to the production needs of different types of target products.

[0122] In a preferred embodiment, the power supply includes a solar panel or a wind turbine, and further includes an energy storage battery. In the present application, the solar panel or the wind turbine can replace the fossil energy derived electric energy to achieve sustainable energy supply, reduce the dependence on traditional carbon sources, and through the configuration of an energy storage battery such as a lithium battery, the intermittent problem of renewable energy can be solved, such as power supply at night or no wind, to ensure the continuous biosynthesis in the reaction system.

[0123] In a preferred embodiment, the present application provides the application of the electrochemically powered microbial synthesis reaction system as described above in the production of high-value chemicals,

[0124] The high-value chemicals include fragrances, amino acids, rare sugars, rare sugar alcohols, biodegradable plastic monomers, high-activity natural products, and microbial oils.

[0125] Preferably, the fragrances include vanillin, raspberry ketone, naringin, rose essential oil, artemisia oil, menthol, γ-decalactone (milk aroma), 2-acetylpyrroline (popcorn aroma), benzaldehyde (bitter almond aroma), ionone (floral wood aroma), citronellal (citrus aroma), β-caryophyllene (spicy), algal β-ionone (oceanic fragrance);

[0126] The amino acids include L-glutamic acid sodium (monosodium glutamate), L-lysine, L-threonine, L-tryptophan, L-arginine, L-proline, L-cysteine, L-valine, γ-aminobutyric acid (GABA), L-theanine, L-selenium-methyl selenocysteine (anti-cancer activity), ε-polylysine (preservative);

[0127] The rare sugars include D- psicose, D-tagatose, L-ribose, D-allose, L-xylulose, D-turanose, L-fucose, D-isomaltulose, L-rhamnose, D-mannoheptulose, D-alloheptulose, L-lyxose;

[0128] The rare sugar alcohols include allitol, erythritol, xylitol, alititol, maltitol, sorbitol, lactitol, isomaltulose, mannitol, ribitol, tagatose, galactitol, erythrulose;

[0129] The biodegradable plastic monomers include lactide, succinic acid, 5-hydroxymethylfurfural (HMF), 1,4-butanediol (BDO), FDCA (2,5-furan dicarboxylic acid), lactic acid, adipic acid, 3-hydroxypropionic acid, epsilon-caprolactone, glycolide, dimethyl terephthalate, and the like;

[0130] The high-activity natural products include fucoidan sulfate, rare ginsenoside CK, paclitaxel, astaxanthin, lycopene, steviol glycoside, mogroside, artemisinic acid, ginkgolide B, ganoderic acid, huperzine A, scutellarin;

[0131] The microbial oils include schizochytrium DHA algal oil, Yarrowia lipolytica ARA oil, Rhodoturula glutinis gamma-linolenic acid, Chlorella vulgaris EPA algal oil, Mortierella alpina arachidonic acid, Chlorella ellipsoidea astaxanthin oil, Starmerella bombicola microbial butter, Crypthecodinium cohnii DHA oil, Rhodotorula lycopene oil, Rhodotorula glutinis beta-carotene oil, Nannochloropsis gaduola squalene oil, marine bacteria wax ester.

[0132] Further specific elaboration of the technical effects that the technical solutions of the present application can produce in actual application are as follows:

[0133] (1) Breakthrough traditional bottlenecks, realize efficient and green manufacturing:

[0134] Carbon efficiency is improved: CO2 emission is reduced by more than 30%, and 0.3 tons of CO2 emission is reduced per ton of monosodium glutamate produced;

[0135] Product conversion rate is improved: the conversion rate of target products is increased by more than 30%, such as the conversion rate of glutamic acid, which is increased from 60% to more than 80%;

[0136] System stability is enhanced: stable operation time is more than 48 hours, which is 1 time higher than that of the traditional system (12-24 hours);

[0137] Complex path adaptation: support the production of complex products such as antibiotics and vaccines, for example, the production efficiency of penicillin can be increased by 40%.

[0138] (2) Reduce costs and improve industrialization feasibility:

[0139] Raw material cost is reduced: reduce dependence on sugar, reduce renewable energy cost, and raw material cost is reduced by 20%-30%; the raw material cost of monosodium glutamate is reduced from 1.2 yuan / kg to 0.8-0.9 yuan / kg;

[0140] Production efficiency is improved: continuous flow reactor and rate matching design, production efficiency is increased by more than 50%; the unit time yield of amino acid is increased from 1 ton / day to 1.5 tons / day;

[0141] Good industrialization compatibility: compatible with existing fermentation equipment, reducing modification cost by 40%-50%; modification cost of a certain monosodium glutamate plant reduced from 20 million yuan to 10-12 million yuan.

[0142] (3) Leading zero-carbon biological manufacturing and promoting green revolution:

[0143] Zero-carbon conversion: using renewable energy to realize "electricity to biomolecule" zero-carbon manufacturing, such as solar energy to monosodium glutamate and antibiotics;

[0144] Resource recycling: using industrial wastewater or biomass derivatives as anode, realizing waste resourceization, and producing 0.5 tons of target product from 1 ton of papermaking wastewater;

[0145] CO2 resourceization: combined with CO2 capture technology, converting industrial waste CO2 into biomolecules, capturing 0.7 tons of CO2 for producing 1 ton of acetic acid, reducing greenhouse gas emissions.

[0146] Experimental example

[0147] Platinum sheet as counter electrode, silver / silver chloride as reference electrode;

[0148] The electrode obtained by growing the experimental material in situ on carbon paper or titanium felt is directly used as the working electrode; some of the experimental materials used as comparison materials are directly drop-casted on carbon paper or titanium felt by mixing ink with Nafion.

[0149] Electrolyte: 100 mM Tris-HCl buffer (pH 7.2); substrate concentration: 1 mM NAD + The whole process was carried out under nitrogen protection. The product NADH was detected by ultraviolet quantification method. The UV standard concentration curve of NADH is shown in Figure 8 .

[0150] Experimental example 1

[0151] Performance test of H2DBB-RhCp*Cl ligand; that is, the experimental material is H2DBB-RhCp*Cl.

[0152] Due to the poor solubility of the ligand, H2DBB-RhCp*Cl was drop-casted on carbon paper as the working electrode by mixing ink with Nafion;

[0153] The test results are shown in Figure 9 and Figure 10 ,

[0154] Figure 9UV-Vis spectra of electrolyte before and after testing, electrolyte before testing has no NADH absorption peak, electrolyte after testing has obvious NADH absorption intensity, and the NADH concentration is obtained through a standard curve, and then the average reduction NAD of the ligand is converted + The faradic efficiency of the reduction of NADH is about 50% (as shown in Figure 10 ), which proves the effective catalytic ability of the ligand.

[0155] Experimental Example 2

[0156] ICP-MS quantification, H2DBB-RhCp*Cl (RhDBB-Nafion-C), Rh monoMOF (Rh monoMOF-Nafion-C), and Rh monoMOF (Rh monoMOF-C) grown in situ on carbon paper are used to test the NAD + reduction performance.

[0157] The test results are shown in Figure 11 and Figure 12 ;

[0158] Figure 11 UV-Vis spectra of electrolyte before and after testing, electrolyte before testing has no NADH absorption peak. The NADH concentration is obtained through a standard curve, and then the reduction NAD + of the Rh monoMOF grown in situ on carbon paper is about 12%, and the reduction NAD + of the RhDBB-Nafion-C is about 48% (as shown in Figure 12 ). The Rh monoMOF in the present application is a short name of DBB-RhCp*Cl MonoMOF.

[0159] Experimental Example 3

[0160] Rh monoMOF grown in situ on titanium felt and titanium felt without loaded catalyst are used to test the NAD + reduction performance.

[0161] The test results are shown in Figure 13 and Figure 14 ;

[0162] Figure 13 UV-Vis spectra of electrolyte before and after testing, electrolyte before testing has no NADH absorption peak. The NADH concentration is obtained through a standard curve, and then the reduction NAD +The faraday efficiency of the monoMOF catalyst is about 38%, and the titanium felt without catalyst has almost no catalytic effect.

[0163] The above experiments show that the in-situ growth of the monoMOF catalyst exhibits better performance than the drop-coating method, proving that the loading method can affect the subsequent catalytic performance. The materials loaded on different substrates exhibit different reaction performances, and the results show that the metal substrate is better.

[0164] The above describes the present application in combination with preferred embodiments, but these embodiments are only exemplary and serve only to illustrate. On this basis, various substitutions and improvements can be made to the present application, and these all fall within the protection scope of the present application.

Claims

1. An electrochemically powered microbial synthesis reaction system, comprising: The system comprises an anode chamber and a cathode chamber separated by a proton exchange membrane (1); an anode (2) is arranged in the anode chamber, and a cathode (3) is arranged in the cathode chamber; An anode flow channel (4) is arranged in the anode chamber, and the anode flow channel (4) is filled with an anode substrate, and electrons are provided through the anode substrate; Outside the anode chamber and the cathode chamber, the anode (2) and the cathode (3) are connected in series by a power supply to form a loop; A cathode flow channel (5) is arranged in the cathode chamber, and the cathode flow channel (5) is connected with a reactor (7) through a peristaltic pump (6); an electron carrier is arranged in the cathode flow channel (5) and the reactor (7), and the cathode (3) can continuously generate the electron carrier; Microorganisms are also arranged in the reactor (7), and the microorganisms uptake the electron carrier through a transport protein, and then drive the metabolism of the microorganisms to synthesize a target product through the electron carrier.

2. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein the electron carrier is biocompatible. Said proton exchange membrane (1) allows H + Passing, thus maintaining the system pH balance, avoiding acid-base imbalance inhibiting microbial activity.

3. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein the electron carrier is selected from one or a mixture of more than one of NADH, an artificial analogue of NADH, FADH2, and an artificial analogue of FADH2.

4. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein the anode (2) and the cathode (3) are both obtained by loading a monoMOF catalyst on a substrate, and the thickness of the film on the anode (2) and the cathode (3) is 2-20 nm.

5. The electrochemically powered microbial synthesis reaction system according to claim 4, wherein the monoMOF catalyst is directionally assembled on a carbon or metal substrate by in-situ growth or chemical modification grafting. Preferably, when the electron carrier is NADH, the microorganism oxidizes NADH to NAD + when metabolizing the synthesis of the target product + The cathode (3) continuously generates NADH based on NAD 6. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein the anode substrate can be oxidized to lose electrons.

7. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein the reactor (7) is a batch fermentation tank or a microfluidic continuous flow reactor. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ Preferably, a filter membrane (8) is arranged between the reactor (7) and the peristaltic pump (6), which allows the passage of the electron carrier and is capable of intercepting microorganisms.

8. The electrochemically powered microbial synthesis reaction system according to claim 1, wherein, The power supply comprises a solar panel or a wind turbine, and further comprises an energy storage battery.

9. Use of the electrochemically powered microbial synthesis reaction system according to any one of claims 1 to 8 in the production of high-value chemicals, wherein, The high-value chemicals include fragrances, amino acids, rare sugars, rare sugar alcohols, biodegradable plastic monomers, high-activity natural products, and microbial oils.

10. Use of the electrochemically powered microbial synthesis reaction system according to claim 9 in the production of high-value chemicals, wherein, The fragrances include vanillin, raspberry ketone, naringin, rose essential oil, artemisia annua oil, menthol, gamma-decalactone, 2-acetyl pyrroline, benzaldehyde, ionone, citronellal, beta-caryophyllene, algal beta-ionone; The amino acids include L-glutamic acid sodium, L-lysine, L-threonine, L-tryptophan, L-arginine, L-proline, L-cysteine, L-valine, gamma-aminobutyric acid, L-theanine, L-selenium-methyl selenocysteine, epsilon-polylysine; The rare sugars include D-alloketose, D-tagatose, L-ribose, D-allose, L-xylulose, D-turanose, L-fucose, D-isomaltulose, L-rhamnose, D-mannoheptulose, D-alloheptulose, L-lyxose; The rare sugar alcohols include allitol, erythritol, xylitol, alititol, maltitol, sorbitol, lactitol, isomaltitol, mannitol, ribitol, tagatitol, galactitol, erythrulose; The biodegradable plastic monomers include lactide, succinic acid, 5-hydroxymethyl furfural, 1,4-butanediol, FDCA, lactic acid, adipic acid, 3-hydroxypropionic acid, epsilon-caprolactone, glycolide, isosorbide, dimethyl terephthalate; The high-activity natural products include fucoidan, rare ginsenoside CK, paclitaxel, astaxanthin, lycopene, steviol glycoside, mogroside, artemisinic acid, ginkolide B, ganoderic acid, huperzine A, breviscapine; The microbial oils include schizochytrium DHA algal oil, yarrowia lipolytica ARA oil, rhodosporidium paludigenum gamma-linolenic acid, chlorella EPA algal oil, mortierella alpina arachidonic acid, chlorella ovalis astaxanthin oil, starmerella bombicola microbial butter, crypthecodinium cohnii docosahexaenoic acid, rhodotorula lycopene oil, rhodotorula glutinis beta-carotene oil, botryococcus braunii squalene oil, marine bacteria wax ester.

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