Recombinant plasmid, recombinant shewanella and electricity generation method of microbial fuel cell (MFC)

A technology for recombinant plasmids and Shewanella, applied in the direction of microorganism-based methods, biochemical equipment and methods, recombinant DNA technology, etc. Method limitations and other issues, to achieve the effect of increasing the thickness and anode bioload, increasing the electron transfer rate, and better transferring electrons

Active Publication Date: 2018-09-14
TIANJIN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficiency of these methods is still not high, and there are relatively few promoters available in Shewanella, and no systematic research results have been formed
At present, the transformation of Shewanella is not as mature as the technology of E. coli, and many transformation methods are still relatively limited. Therefore, the research or development of some biological methods or biological components that can work effectively in Shewanella is very important for the subsequent transformation and research of Shewanella. important

Method used

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  • Recombinant plasmid, recombinant shewanella and electricity generation method of microbial fuel cell (MFC)
  • Recombinant plasmid, recombinant shewanella and electricity generation method of microbial fuel cell (MFC)
  • Recombinant plasmid, recombinant shewanella and electricity generation method of microbial fuel cell (MFC)

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Experimental program
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Effect test

Embodiment 1

[0056] Embodiment 1: Construction of the recombinant plasmid of the present invention

[0057] Synthesize the promoter ptac or ptet gene sequence, synthesize and optimize the flavins gene sequence ribA, ribD, ribE, ribH, ribC (the gene is derived from Bacillus subtilis with RBS1);

[0058] Use the restriction site of PYYD (such as EcoRI, XbaI, SpeI, SdaI, two restriction sites of XbaI and SpeI are selected in this embodiment) and the bio-brick method to connect the ptac or ptet promoter and each flavins synthetic gene to the vector On PYYD, the recombinant plasmids (without the OprF gene) were obtained and named 3C5 (Ptac-ribADEHC-PYYD) and CT5 (Ptet-ribADEHC-PYYD) respectively. See the schematic diagram figure 2 (The arrow in the figure indicates the promoter).

[0059] Synthetic porin gene sequence OprF (gene derived from Pseudomonas aeruginosa, with RBS2), using the enzyme cutting site of PYYD (such as EcoRI, XbaI, SpeI, SdaI, the present embodiment selects two enzyme cut...

Embodiment 2

[0060] Embodiment 2: Adaptation comparison of different promoters and Shewanella

[0061] Synthesize the promoter gene sequence (pbad, ptac, ptet, LacUV5, T7, parcA), and use the four enzyme cutting sites (EcoRI, XbaI, SpeI, SdaI) of the biobrick to connect the promoter and the GFP gene (with RBS1) respectively On the vector PYYD, the plasmids Pbad-GFP-PYYD, Ptac-GFP-PYYD, Ptet-GFP-PYYD, LacUV5-GFP-PYYD, T7-GFP-PYYD, and ParcA-GFP-PYYD were obtained respectively. See the schematic diagram Figure 4 .

[0062] The Pbad sequence is shown in SEQ ID NO:12, the LacUV5 sequence is shown in SEQ ID NO:13, the T7 sequence is shown in SEQ ID NO:14, and the parcA sequence is shown in SEQ ID NO:13.

[0063] After ligation, each plasmid was introduced into Escherichia coli W3064 competent, and then combined with wild Shewanella MR-1 for transfer, and the constructed plasmid was transferred to Shewanella (transformation method can refer to Yang Y, Ding Y, Hu Y, etal.Enhancing bidirectiona...

Embodiment 3

[0065] Embodiment 3: the construction of the recombinant Shewanella of the present invention

[0066] 3C5 (Ptac-ribADEHC-PYYD), CT5 (Ptet-ribADEHC-PYYD), CT5-31O-PYYD and 3C5-31O-PYYD constructed in Example 1 were introduced into W3064 competent, and then mixed with Shewanella MR-1 Or combine the transfer of Shewanella with deposit number CGMCCNo.15432, transfer the constructed plasmid to Shewanella, and obtain the recombinant Shewanella of the present invention.

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Abstract

The invention relates to the technical field of biological energy, and discloses a recombinant plasmid, recombinant shewanella and an electricity generation method of a microbial fuel cell (MFC). A promoter suitable for the shewanella is selected to be connected with an exogenous electron carrier flavins gene, so that the shewanella is enabled to generate more flavis, and the electrochemical performance of the MFC is improved; furthermore, a mutant strain is combined with a porin oprF gene, so that the formation of a biofilm is further improved, the going in and out of the flavins in the shewanella can be promoted, the electron transfer rate is increased, and electrons are better transmitted. Furthermore, biocompatible graphene oxide (GO) is used and can be self-assembled into the three-dimensional multilayer biofilm together with the shewanella, so that the thickness of the biofilm and the anode biocapacity are further greatly improved; the operation is simple and fast, and the electrochemical performance of the MFC is effectively improved.

Description

technical field [0001] The invention relates to the technical field of bioenergy, and more specifically relates to a recombinant plasmid and a method for generating electricity by recombinant Shewanella and MFC. Background technique [0002] Energy shortage and environmental pollution are increasingly serious problems facing our country today. Therefore, energy development, environmental waste treatment and energy renewable utilization in the process have become a major challenge for the sustainable development of our modern society. Scientists are constantly looking for new technical solutions, among which Microbial Fuel Cell (MFC) is one of them, which is used to generate alternative energy and new devices for environmental waste management, and its importance is becoming increasingly apparent today. [0003] MFC is a device that uses electrogenic microorganisms as an anode catalyst to convert chemical energy in organic matter into electrical energy. The electricity-gener...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N1/21H01M8/16C12R1/01
CPCC12N15/74H01M8/16Y02E60/50Y02P70/50
Inventor 宋浩林童王磊曹英秀
Owner TIANJIN UNIV
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