An electrogenic Clostridium beijerinckii and its construction method and application
A clostridium beijerinckii and fuel cell technology, applied in the field of environment and new energy, can solve the problems of unclear specific mechanism and weak microbial power generation ability, and achieve the effect of efficient power generation and improved power generation.
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Embodiment 1
[0028] Embodiment 1: Construction method of Clostridium beijerinckii Cbei_2996 gene inactivation mutant
[0029] (1) Design introns:
[0030] According to the Cbei_2996 gene sequence (as shown in SEQ ID No: 1) of Clostridium beijerinckii included in the NCBI database, that is, the base sequence (nuoA) encoding coenzyme Q oxidoreductase subunit A, a suitable insertion gene site is designed with the help of software point( http: / / www.clostron.com ), through the previous experimental screening and verification, it was selected to be inserted between the 62nd and 63rd bases of the Cbei_2996 gene sequence, and an intron sequence was generated. The synthesized intron was S-62, and its sequence was shown as SEQ ID NO: 2.
[0031] (2) Construction of Cbei_2996 insertion inactivation vector:
[0032] Use XhoI and BsrGI to double-enzyme-cut the vector pWJ (provided by Mr. Yang Sheng, Shanghai Academy of Biological Sciences, its sequence is shown in SEQ ID NO: 5, and the plasmid map ...
Embodiment 2
[0044] This example illustrates the electricity production experiment using 1 g / L glucose by Clostridium beijerinckii as an anode catalyst.
[0045] (1) Construction of microbial fuel electricity:
[0046] In this embodiment, a microbial fuel cell using Clostridium beijerinckii to generate electricity as an anode catalyst is established according to existing technologies and methods, as shown in the attached figure 2 As shown, it includes four parts: anode chamber, cathode chamber, proton exchange membrane and external circuit. The anode electrode and cathode electrode are PAN-based graphite soft felt (5×5cm), with titanium wire as the external circuit, the external resistance is 1000Ω, the proton exchange membrane is DuPont NafionN117, and the data collector is Keithley series.
[0047] (2) Culture medium formula:
[0048] YPS seed medium: yeast powder 3g / L, peptone 5g / L, glucose 1g / L, ammonium acetate 2g / L, sodium chloride 3g / L, magnesium sulfate heptahydrate 3g / L, potass...
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