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An enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage

A biofuel cell and Escherichia coli technology, which is applied in biochemical fuel cells, battery electrodes, circuits, etc., achieves good application prospects, improves enzyme loading and activity, and has simple preparation methods

Active Publication Date: 2019-01-04
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects of the existing way of carrying enzymes on the anode, to provide an enzyme biofuel cell that uses trehalose produced by E. Surfactants are used to treat engineered E. coli cells to increase the permeability of cells, thereby avoiding the loss of enzymes caused by the method of osmotic shock extraction of enzymes, and improving the activity of enzymes and the amount of enzymes loaded

Method used

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  • An enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage
  • An enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage
  • An enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage

Examples

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Embodiment 1

[0026] This example illustrates the preparation method of the bioengineered Escherichia coli of the present invention.

[0027] Extract and amplify the trehalose hydrolase TreA gene from the genome of Escherichia coli str.K-12 substr.MG1655, carry out Nde I and XhoI double digestion of the TreA gene and the vector pet22b(+), and then carry out the digestion fragment The gel was recovered, T4 DNA ligase 16 NA was ligated overnight, and the recombinant plasmid pET22b(+)-TreA was obtained, and the plasmid was transformed into E.coli BL21 competent cells. A single colony was screened out on an ampicillin plate, and colony PCR and subsequent verification were performed. The successfully constructed engineered bacteria are preserved by glycerol cryopreservation, which is the bioengineered Escherichia coli.

Embodiment 2

[0029] This embodiment illustrates the whole cell permeabilization treatment method of bioengineered Escherichia coli of the present invention.

[0030] (1) Get the bacterial classification that embodiment 1 preserves, bacterial classification is inoculated in the LB liquid culture medium that adds ampicillin by volume ratio 1.5%, adds IPTG when measuring OD 600 is 0.6-0.8, keeps the IPTG final concentration of 0.5-1.5mmol / L, induce enzyme production for 7–9h, culture condition is 37°C, 200rpm, obtain fermentation broth; centrifuge to obtain wet bacteria, centrifuge condition is 4°C, 6000rpm, centrifugation time is 10min;

[0031] The formula of LB liquid medium with ampicillin is: peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / L, ampicillin antibiotic 0.1g / L.

[0032] (2) Add glycolipid, lipopeptide or fatty acid biosurfactant solution of 0.1-0.4g / L equal volume to the fermentation broth in step (1) to the wet thalline obtained in step (1), and 20-25°C, react at 100-2...

Embodiment 3

[0035] This example illustrates the preparation method of the enzyme cathode and enzyme anode in the biofuel cell of the present invention.

[0036] Cut two pieces of carbon felt into a geometric area of ​​8cm 2 rectangle. The carbon felt was ultrasonically cleaned with acetone, ethanol and ultrapure water for 10 min and dried to remove grease, dust or other debris on the surface.

[0037]Soak one of the carbon felts in the bioengineering Escherichia coli fermentation liquid obtained in Example 2 and let it stand at 35-40°C for 2-16 hours, take it out, rinse it with ultrapure water, dry it naturally, and set it aside;

[0038] Soak the cleaned anode carbon felt in engineering Escherichia coli fermentation broth and let it stand at 35-40°C for 2-16 hours, take it out and rinse it with ultra-pure water, and then put in an ultra-pure aqueous solution of 5-25g / L glucose oxidase Stand still at 35-40°C for 16 hours, take it out, wash it with ultrapure water, and store it at 4°C to...

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Abstract

The invention discloses an enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage. The invention successfully prepares a two-chamber enzyme biofuel cell capable of generating electricity by using trehalose from sewage by immobilizing engineering Escherichia coli containing trehalose hydrolase TreA and glucose oxidase GOx after whole-cell permeability treatment on an anode carbon felt and immobilizing bilirubin oxidase on a cathode carbon felt. The fuel cell assembled by the invention has a maximum output power of up to 16.2. M u.W / cm2, and the output current density of the fuel cell can still be maintained at 72% of the initial maximum output value after 60 hours of continuous power generation under the condition that the raw material is sufficient. The method for preparing the fuel cell of the invention is simple and low in cost, and the reaction is carried out at room temperature, and the prepare enzyme electrode has good catalytic performance and good application prospect.

Description

technical field [0001] The invention belongs to the technical field of preparation of enzyme fuel cells, in particular to a preparation method of an enzyme biofuel cell in which an anode is jointly modified by algalose hydrolase and glucose oxidase with high enzyme activity, good stability and high efficiency, and adopts the method The invention relates to a method for generating electricity by utilizing trehalose produced from sewage from different sources in an enzyme fuel cell with a dual-enzyme immobilized anode. Background technique [0002] Enzyme biofuel cell is a biofuel cell that uses oxidoreductase as a catalyst to catalyze the oxidation of fuel. Hydrogen ions and free electrons are generated at the anode, and hydrogen ions and free electrons are used at the cathode to reduce peroxide to water, thereby completing the flow of electrons. Chemical energy is converted into electrical energy. Compared with traditional chemical batteries, although due to the fragile nat...

Claims

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Application Information

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IPC IPC(8): H01M8/16H01M4/88
CPCH01M4/88H01M8/16Y02E60/50
Inventor 黄和徐阳雪江凌朱丽英
Owner NANJING UNIV OF TECH
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