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Recombinant shewanella algae capable of producing riboflavin and application of recombinant shewanella algae in electricity production

A technology of Shewanella and riboflavin, which is applied to the recombinant seaweed Shewanella and its application in electricity production, can solve the problems of high cost, low yield, unfavorable industrial application of riboflavin, etc., and achieve high yield , Improving power production capacity, and the effect of power production capacity improvement

Active Publication Date: 2020-08-18
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Shewanella algae can produce small amounts of flavins as electron transporters, but their own production is low
In addition, the ability to generate electricity can be greatly improved by adding riboflavin solution exogenously to the battery, but the cost of adding riboflavin exogenously is high, which is not conducive to its subsequent industrial application

Method used

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  • Recombinant shewanella algae capable of producing riboflavin and application of recombinant shewanella algae in electricity production
  • Recombinant shewanella algae capable of producing riboflavin and application of recombinant shewanella algae in electricity production
  • Recombinant shewanella algae capable of producing riboflavin and application of recombinant shewanella algae in electricity production

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Experimental program
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Effect test

Embodiment 1

[0038] Construction and expression of a riboflavin-producing recombinant algae Shewanella:

[0039] The five target genes ribA, ribC, ribD, ribE, and ribH in the flavin synthesis pathway were connected to the vector pyydt to obtain the recombinant plasmid R5, and its plasmid map is as follows: figure 1 shown.

[0040] The nucleotide sequence of the gene ribA is shown in SEQ ID NO.1;

[0041] The nucleotide sequence of the gene ribC is shown in SEQ ID NO.2;

[0042] The nucleotide sequence of the gene ribD is shown in SEQ ID NO.3;

[0043] The nucleotide sequence of the gene ribE is shown in SEQ ID NO.4;

[0044] The nucleotide sequence of the gene ribH is shown in SEQ ID NO.5;

[0045] The nucleotide sequence of the vector pyydt is shown in SEQ ID NO.6

[0046] The nucleotide sequence of the loaded recombinant plasmid R5 is shown in SEQ ID NO.7;

[0047] The resulting recombinant plasmid R5 was transformed into Escherichia coli WM3064 competent cells (commercial strains)...

Embodiment 2

[0051] Application of Recombinant Bacteria scs-R5 in Electricity Production

[0052] Streak activation of the constructed recombinant strain scs-R5 and the wild-type strain scs-1, and inoculate the recombinant strain scs-R5 in LB liquid medium containing kanamycin (concentration 25 μg / mL) to obtain first-grade seeds liquid.

[0053] Take 20 mL of the primary seed liquid and transfer it to 400 mL of anolyte (kanamycin concentration 25 μg / mL), add the inducer IPTG to a final concentration of 0.5 mM, and cultivate overnight to obtain the fermentation liquid 1.

[0054] The wild-type strain scs-1 was fermented by the same method, without adding antibiotics and inducers to the culture medium and anolyte, and fermented liquid 2 was obtained after overnight culture.

[0055] Anolyte configuration:

[0056] 1) 5XM9 solution: 30g / L anhydrous disodium hydrogen phosphate, 15g / L anhydrous potassium dihydrogen phosphate, 5g / L anhydrous ammonium chloride, 2.5g / L sodium chloride, accurately ...

Embodiment 3

[0065] Application of Recombinant Bacteria scs-R5 Combined with Anode Material in Electricity Production

[0066] Common anode materials in microbial fuel cells include carbon cloth, conductive cotton, stone-ground electrodes, etc. The electrode selected in the present invention is conductive cotton, which has a large specific surface area, which is very conducive to the attachment and reception of bacteria Electronics, in addition, in order to enhance the conductivity of conductive cotton, carbon nanotube conductive cotton composite electrode (CNT-CC electrode) is selected in this experiment. Percentage is 0.2%), add surfactant sodium dodecylbenzene sulfonate (SDBS), the mass percentage of SDBS is 1%, ultrasonic treatment 30min, obtain the dispersion liquid 2 that contains surfactant, soak conductive cotton in above-mentioned Put it in the dispersion liquid 2 for 30 minutes, take it out, put it in an oven to dry, and obtain a CNT-CC electrode, which can be used for assembling...

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Abstract

The invention discloses recombinant shewanella algae capable of producing riboflavin and an application of the recombinant shewanella algae in electricity production, wherein the recombinant shewanella algae is constructed by the following method: (1) connecting five target genes ribA, ribC, ribD, ribE and ribH in a flavin synthetic route to a vector pyydt to obtain a recombinant plasmid R5; and (2) transferring the recombinant plasmid R5 into a shewanella algae scs-1 CGMCC No.18696 strain, so as to obtain the recombinant shewanella algae scs-R5 for producing riboflavin. According to the recombinant bacterium disclosed by the invention, the yield of riboflavin serving as an electron transfer body is high, so that the electricity generation capacity is improved.

Description

technical field [0001] The invention belongs to the technical field of bioenergy, and in particular relates to a riboflavin-producing recombinant algal Shewanella strain and its application in electricity production. Background technique [0002] Microbial fuel cells are a clean and sustainable form of energy that uses microorganisms as biocatalysts to convert chemical energy in organic matter into electrical energy. The electrogenic bacteria oxidize and decompose organic matter at the anode of the battery to generate electrons and protons. The electrons pass through the anode and pass through the external circuit to the cathode to form a current. closed loop. Electron transfer at the anode is one of the main aspects that limit the ability of microorganisms to generate electricity. There are two electron transfer mechanisms confirmed by current studies: direct transfer and indirect transfer. Contact transfers electrons to the anode; indirect transfer refers to the process ...

Claims

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Application Information

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IPC IPC(8): C12N15/74C12N15/54C12N15/55C12N15/53C12N1/21H01M4/90H01M8/16C12R1/01
CPCC12N15/74C12N15/52C12N9/78C12N9/1205C12N9/1085C12N9/0006H01M4/90H01M4/9008H01M8/16C12Y305/04025C12Y207/01026C12Y205/01078C12Y305/04026C12Y101/01193Y02E60/50Y02P70/50
Inventor 宋浩陈媛媛李锋曹英秀
Owner TIANJIN UNIV
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