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Electricity-generating Shewanella engineering strain, construction method and application

A technology of engineering strains and construction methods, applied in chemical instruments and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of inability to form thicker biofilms and weak adhesion of MR-1 cells, Achieve the effect of enhancing extracellular electron transfer and increasing power density

Inactive Publication Date: 2020-01-17
天津大学前沿技术研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the wild-type Shewanella strain MR-1 cells were weakly adherent and could not form thick and stable biofilms

Method used

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  • Electricity-generating Shewanella engineering strain, construction method and application
  • Electricity-generating Shewanella engineering strain, construction method and application
  • Electricity-generating Shewanella engineering strain, construction method and application

Examples

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Effect test

Embodiment 1

[0021] A method for constructing an engineering strain of electric-producing Shewanella, comprising the steps of: knocking out the target gene exeM of wild-type Shewanella MR-1 to obtain an engineering strain of electric-producing Shewanella ΔexeM, and the nucleoside of the gene exeM The acid sequence is represented by SEQ ID NO 15.

[0022] The specific construction method includes the following steps:

[0023] The wild-type Shewanella MR-1 target gene exeM was knocked out by att site-specific recombination and double crossover deletion.

[0024] (1) Primer design and gene amplification:

[0025] The genome sequence of Shewanella MR-1 was obtained from the NCBI database, and three pairs of primers were designed according to the target gene exeM and its upstream and downstream gene sequences, exeM-LF (SEQ ID NO.4) and exeM-LR (SEQ ID NO.5 ), exeM-5-O (SEQ ID NO.6) and exeM-5-I (SEQ ID NO.7), exeM-3-O (SEQ ID NO.8) and exeM-3-I (SEQ ID NO. 9) Amplify the upstream homologous ...

Embodiment 2

[0048] Example 2: Electricity production by engineering strain ΔexeM of Shewanella and wild-type Shewanella MR-1MFC

[0049] 1. Strain activation

[0050]The electrogenic Shewanella engineering strain ΔexeM and the wild-type Shewanella MR-1 were taken out from the -80°C refrigerator, and cultured overnight in LB medium at 30°C and 200 rpm respectively. The overnight culture solution was transferred into new LB medium at a ratio of 1% at 30°C, 200rpm, cultured for 10 hours, measured OD 600, calculated volume (OD 600 in MFC = 0.5), centrifuged at 5000rpm for 10 minutes, and used anolyte After resuspension, add to the MFC anode compartment.

[0051] 2. MFC power generation

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Abstract

The invention discloses an electricity-generating Shewanella engineering strain, a construction method and application. The construction method of the electricity-generating Shewanella engineering strain includes the following steps that a target gene exeM of a wild-type Shewanella oneidensis MR-1 is knocked out, an electricity-generating Shewanella engineering strain [delta]exeM is obtained, anda nucleotide sequence of the gene exeM is shown in SEQ ID NO15. The electricity-generating Shewanella engineering strain [delta]exeM constructed by the construction method is a gene-deficient strain,the strain can produce more extracellular DNA (eDNA), the eDNA promotes bacterial adhesion, aggregation and biofilm formation, furthermore, extracellular electron transfer is enhanced, and the power density of microbial fuel cells is increased.

Description

technical field [0001] The invention relates to the technical field of bioenergy, and more specifically relates to an engineering strain of electricity-producing Shewanella and its construction method and application. 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 cells (Microbial Fuel Cell, MFC) is one of them 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. [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21H01M8/16C12R1/01
CPCC07K14/195H01M8/16Y02E60/50
Inventor 宋浩李锋赵妍淑刘向
Owner 天津大学前沿技术研究院
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