A novel bacterium with electricity generation function and application thereof

Abstract

The application discloses a novel bacterium with electricity generation function and application thereof, and belongs to the technical field of microorganisms and bioenergy. A novel bacterium with stable electricity generation capacity is screened in papermaking wastewater at the end of discharging of a microbial fuel cell (MFC). Cupriavidus pauculus The 16s rRNA gene sequence of the bacterium is most homologous to known strains. Combining with the construction result of a phylogenetic tree, it is determined that the bacterium is a novel strain of the genus Cupriavidus, and is named PMWA1-3. Cupriavidus pauculus The bacterium can grow by taking phenolic organic pollutants, namely catechol, as the only carbon source, a MFC is constructed by using the bacterium, the highest electricity generation voltage is 291.7 mV, and the maximum power density reaches 202.2 mW / m2, which indicates that the bacterium has strong electricity generation capacity.

Description

Technical Field

[0001] This invention relates to the fields of microbial technology and bioenergy technology, specifically to a novel bacterium with electricity-generating function and its applications. Background Technology

[0002] The extraction of traditional fossil fuels is becoming increasingly difficult, reserves are gradually decreasing, and their prices are fluctuating and rising accordingly, placing heavy cost pressures on industrial production and people's lives. On the other hand, the massive consumption of energy has also triggered serious environmental problems, such as global warming caused by greenhouse gas emissions, further exacerbating the complexity of the energy crisis. Therefore, the search for clean and sustainable new energy sources has become a research hotspot.

[0003] Microbial fuel cells (MFCs), as a green energy device that directly converts the chemical energy of organic matter into electrical energy, are emerging in the field of new energy due to their unique advantages. Compared with traditional fuel cells, MFCs do not require harsh reaction conditions such as high temperature and high pressure, and can operate at normal temperature and pressure. During the energy conversion process, they produce almost no pollutants such as nitrogen oxides and sulfur oxides. Their fuel sources are wide-ranging, utilizing not only conventional organic matter such as glucose, but also degrading organic pollutants in domestic sewage, industrial wastewater, and agricultural waste, achieving the dual value of "turning waste into electricity".

[0004] Electrogenic microorganisms, as the core "catalysts" of MFCs, directly determine the energy conversion efficiency and operational stability of the battery. These microorganisms transfer electrons generated during metabolism from inside the cell to the anode surface through an electron transport chain on the cell membrane, forming an electric current. Currently discovered electrogenic microorganisms are mainly concentrated in the genus *Shewanella* (…). Shewanella ), genus Geobacter ( Geobacter ), Pseudomonas spp. Pseudomonas According to incomplete statistics, fewer than 100 species of microorganisms have been reported and verified to have the ability to generate electricity, representing a negligible proportion compared to the tens of thousands of microbial species in nature. This scarcity greatly limits the diversified development and application expansion of microbial fuel cells.

[0005] Due to the limited variety of electrogenic microorganisms available, it is often difficult to find suitable and highly efficient electrogenic strains when faced with different substrate types and environmental conditions, leading to significant limitations in the practical application of microbial fuel cells. Furthermore, these discovered electrogenic microorganisms also suffer from low electrogenic efficiency, limited environmental adaptability, and poor electrogenic performance under specific substrates. For example, while some strains of *Shewanella* can survive in various environments, their electrogenic efficiency is generally low; strains of *Geobacterium* show a significant decrease in electrogenic performance when using certain specific organic substances as substrates. These problems further restrict the practical application of microbial fuel cells. Therefore, screening for novel microorganisms with efficient and stable electrogenic performance is of significant practical importance. Summary of the Invention

[0006] In view of the above-mentioned prior art, the purpose of this invention is to provide a novel bacterium with power generation function and its application, so as to improve the power generation efficiency and stability of microbial fuel cells and solve the problems of limited types of existing power-generating microorganisms and their many performance deficiencies.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, this invention provides a strain of the genus *Copper-loving Bacteria*. A little copper-loving PMWA1-3, the strains of the genus *Copper-loving Bacteria* A little copper-loving PMWA1-3 was deposited on November 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 36474.

[0008] The copper-bearing bacteria strain A little copper-loving PMWA1-3 combines pollution control and power generation value. It is grown using catechol as the sole carbon source and has anodic power generation capability.

[0009] In a second aspect, the present invention provides an anodic electrogenic bacterial agent, wherein the anodic electrogenic bacterial agent contains the aforementioned copper-producing bacterium strain. A little copper-loving PMWA1-3.

[0010] The strain A little copper-loving PMWA1-3 exhibits activity in generating electricity at the anode.

[0011] The copper-bearing bacteria strain A little copper-loving PMWA1-3 is the core active ingredient in microbial fuel cells that enables anode power generation.

[0012] In a third aspect, the present invention provides the above-mentioned copper-loving bacteria strains. A little copper-loving Application of PMWA1-3 or the above-mentioned anodic electrogenic bacteria agent in the following (1) or (2): (1) Preparation of microbial fuel cells; (2) Degradation of phenolic pollutants.

[0013] The phenolic pollutant is catechol.

[0014] In a fourth aspect, the present invention provides a microbial fuel cell (MFC), the microbial fuel cell comprising an anode chamber, a cathode chamber, and a proton exchange membrane separating the two chambers, wherein the anode chamber contains an anolyte and the aforementioned *Copper-loving Bacteria* strain. A little copper-loving PMWA1-3.

[0015] strains of the genus Copper-loving bacteria A little copper-loving The PMWA1-3 MFC achieved a maximum power generation voltage of 291.7mV and a maximum power density of 202.2 mW / m². 2 This fully demonstrates that the strain possesses a strong anodic electrostatic capacity.

[0016] In a fifth aspect, the present invention provides a method using strains of the genus *Copper-loving Bacteria*. A little copper-loving The method for generating electricity from the anode using PMWA1-3 includes the following steps: Using Luria-Bertani medium as the anolyte, the isolated copper-loving bacteria strains were analyzed. Copper-loving a few PMWA1-3 is used as the anode microorganism, with nickel foam as the anode material and carbon paper as the cathode material to construct a microbial fuel cell, and an external resistor is connected to the battery.

[0017] The beneficial effects of this invention are: This invention discloses a novel bacterial strain with stable power generation capacity obtained from papermaking wastewater at the end of the discharge phase of a microbial fuel cell (MFC) through enrichment culture, plate separation, and pure culture screening. Morphological observation and molecular biological identification revealed that the strain exhibits round, smooth, and moist colonies; it is a Gram-negative short rod-shaped bacterium capable of growth using catechol, a phenolic organic pollutant, as its sole carbon source. Molecular biological analysis showed that the 16S rRNA gene sequence of this strain is similar to... Copper-loving a few The strain showed the highest homology. Based on the phylogenetic tree construction results, it was determined to be a new strain of the genus *Copper-loving Bacteria*, and named... A little copper-loving PMWA1-3. This strain was used to construct an MFC, achieving a maximum power generation voltage of 291.7 mV and a maximum power density of 202.2 mW / m², fully demonstrating its strong anodic power generation capability and providing a promising microbial resource for MFC power generation applications. Attached Figure Description

[0018] Figure 1 Copper-loving fungi A little copper-loving Gram-stained microscopic morphology of PMWA1-3.

[0019] Figure 2 : Phylogenetic tree.

[0020] Figure 3 Copper-loving fungi A little copper-loving PMWA1-3 growth curves with catechol as the sole carbon source (right 4-6 branches) and blank control (left 1-3 branches). Figure 4 Copper-loving fungi A little copper-loving PMWA1-3 constructs an evaluation system for the power generation performance of MFC; among which, Figure 4 A in the middle is a member of the genus *Copper-loving Bacteria*. A little copper-loving PMWA1-3 constructs the discharge curve of MFC. Figure 4 B belongs to the genus *Copper-loving Bacteria*. A little copper-loving PMWA1-3 constructs the polarization curves and power density curves of MFC. Detailed Implementation

[0021] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0022] The specific embodiments of the present invention will be described in further detail below with reference to examples. The following detailed descriptions are illustrative and intended to provide further explanation of this application, rather than limiting the scope of the invention.

[0023] Example 1: Strain screening and culture identification 1. Screening Method The strain was isolated from the anolyte sample at the end of discharge in a microbial fuel cell constructed using papermaking wastewater as the anolyte. Papermaking wastewater has a complex composition, containing a large amount of lignin, cellulose degradation products, and other recalcitrant organic matter. Microbial fuel cells operating in such an environment for a long time tend to accumulate microorganisms with special metabolic capabilities and power generation potential in their anolyte.

[0024] The screening medium formulation was: beef extract 3 g / L, peptone 10 g / L, NaCl 5 g / L, agar powder 15 g / L, pH 7.2–7.4, and cultured at 37°C for 24 h. Single colonies of varying morphology were picked from the plates and purified multiple times using the streak plate method until a pure culture was obtained. After culturing this strain on LB solid medium at 37°C for 24 hours, the colonies were round with neat edges, a smooth and moist surface, slightly raised, and a viscous texture. Gram staining confirmed that it was a Gram-negative bacterium, short rod-shaped (e.g., ...). Figure 1 (As shown).

[0025] PCR amplification was performed using universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') for the 16S rRNA gene. The amplified product was sequenced to obtain the nucleotide sequence shown in SEQ ID NO:1, which is 1429 bp in length, as follows: SEQ ID NO:1:

[0026] The sequence was submitted to the GenBank database, and BLAST alignment analysis revealed that it was similar to... Copper-loving a few The 16S rRNA gene sequence showed the highest homology. Combined with the phylogenetic tree construction results ( Figure 2 It was determined to be a new strain of the genus *Copper-loving Bacteria*, and named... A little copper-loving PMWA1-3. This strain was biopreserved, and the preservation information is as follows: Strain name: A little copper-loving ; Category Naming: A little copper-loving PMWA1-3; Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Cultures; The abbreviation for the depository institution is CGMCC. Address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Deposit date: November 3, 2025; Accession number: CGMCC No.36474.

[0027] Example 2: Carbon Source Verification MSM culture medium preparation: Dissolve 12g K₂HPO₄, 11g KH₂PO₄, 2g NaCl, 2g Na₂SO₄, and 0.2g MgSO₄•7H₂O in deionized water. Add 1 ml of a trace metal salt solution (0.02g MnCl・4H₂O, 0.3g FeCl₂•4H₂O, 0.0034g CuCl₂•2H₂O, 0.014g ZnCl₂, 0.0124g H₃BO₃, 0.038g CoCl₂•6H₂O, and 0.04g Na₂MoO₄•2H₂O dissolved in deionized water and bring the volume to 1L. Set aside at room temperature for later use. Adjust the pH to 7.0. Autoclave at 121℃ and dispense into 2g L containers for use. -1 Using ammonium chloride as a nitrogen source, and adding a carbon source (catechol) to verify its effect on... A little copper-loving Does it affect growth?

[0028] The test results indicate that, using MSM medium, it was preliminarily verified that it can utilize the phenolic organic pollutant catechol as the sole carbon source for growth (e.g., Figure 3 (As shown).

[0029] Example 3: Verification of Power Generation Function (1) Determination of discharge curve In order to investigate A little copper-lovingTo assess the power generation performance, a microbial fuel cell was constructed using Luria-Bertani medium as the anolyte and the strain isolated in Example 1 as the anolyte microorganism. A dual-chamber microbial fuel cell was used, with both the anode and cathode chambers having a volume of 300 mL, separated by a proton exchange membrane. The anode material was nickel foam (1 cm × 2.5 cm), and the cathode material was carbon paper (2 cm × 5 cm). An external 1000 Ω resistor was connected to the battery, and it was operated at 30°C. The output voltage of the battery was recorded using a data acquisition system, and a discharge curve was plotted with time on the x-axis and discharge voltage on the y-axis.

[0030] The MFC discharge voltage results collected by the data acquisition system during MFC operation show that... Copper-loving a few It exhibits distinct power generation characteristics. Figure 4 The discharge curve of the A-type circuit shows that its maximum discharge voltage can reach 291.7 mV during operation.

[0031] (2) Determination of polarization curve and power density curve When the MFC reaches the discharge stabilization period, the polarization curve is determined by artificially changing the external resistance. First, the circuit is opened for 1 hour to obtain the open-circuit voltage of the MFC. Then, a resistance box is connected, and the resistance is gradually reduced from 10000Ω to 100Ω. After each change in resistance value, the resistance is stabilized for a period of time, and the voltage data is recorded. Based on the obtained voltage data, the current density and power density are calculated, and the polarization curve and power density curve are plotted. Figure 4 The polarization and power density curves during the steady-state period of B show that the open-circuit voltage is 748 mV, and the power density exhibits a typical trend of first increasing and then decreasing, with the maximum power density reaching 202.2 mW / m².

[0032] The power generation performance evaluation results fully demonstrate that A little copper-loving The PMWA1-3 strain possesses strong electrogenic capabilities, providing a promising microbial resource for MFC electrogenic applications.

[0033] The above description is merely a preferred embodiment of this application and is not intended to limit the application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications made within the spirit and principles of this application are not permitted. Equivalent substitutions and improvements should all be included within the scope of protection of this application.

Claims

1. An occasional copper-loving fungus ( Cupriavidus pauculus PMWA1-3, characterized in that, The occasional copper-loving bacteria ( Cupriavidus pauculus PMWA1-3 was deposited on November 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36474.

2. The occasional copper-loving bacteria according to claim 1 ( Cupriavidus pauculus PMWA1-3, characterized in that, The occasional copper-loving bacteria ( Cupriavidus pauculus PMWA1-3 is grown using catechol as the sole carbon source and has anodic power generation capability.

3. An anode-generating bacterial agent, characterized in that, The anodic electrogenic bacteria agent contains the occasional copper-loving bacteria as described in claim 1. Cupriavidus pauculus )PMWA1-3.

4. The occasional copper-loving bacteria as described in claim 1 ( Cupriavidus pauculus The application of PMWA1-3 or the anodic electrogenic bacteria agent according to claim 3 in the following (1) or (2): (1) Preparation of microbial fuel cells; (2) Degradation of phenolic pollutants; The phenolic pollutant is catechol.

5. A microbial fuel cell, characterized in that, The microbial fuel cell includes an anode chamber, a cathode chamber, and a proton exchange membrane separating the two chambers. The anode chamber contains anolyte and the occasional copper-loving bacteria described in claim 1. Cupriavidus pauculus )PMWA1-3.

6. A method utilizing the occasional copper-loving bacteria described in claim 1 ( Cupriavidus pauculus The method for generating electricity at the anode using PMWA1-3 is characterized by, Includes the following steps: Using Luria-Bertani medium as the anolyte, the isolated occasional copper-loving bacteria ( Cupriavidus pauculus PMWA1-3 was used as the anode microorganism, with nickel foam as the anode material and carbon paper as the cathode material to construct a microbial fuel cell, and an external resistor was connected to the battery.

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