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A method for recombinant yeast strain and microbial mixed bacteria to produce electricity

The technology of yeast strain and Saccharomyces cerevisiae strain is applied in the field of bioenergy, which can solve the problems of narrow carbon source spectrum of Shewanella, complex sludge composition, unclear relationship between bacteria and carbon source, etc. Avoid competing carbon cloth electrodes, good electrochemical effect

Active Publication Date: 2020-12-18
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of sludge is very complex, and the relationship between many bacteria and carbon sources is not clear
In addition, the spectrum of carbon sources available to Shewanella is also narrow

Method used

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  • A method for recombinant yeast strain and microbial mixed bacteria to produce electricity
  • A method for recombinant yeast strain and microbial mixed bacteria to produce electricity
  • A method for recombinant yeast strain and microbial mixed bacteria to produce electricity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Construction of the recombinant Saccharomyces cerevisiae strain of the present invention

[0042] Using Saccharomyces cerevisiae BY4741 as the starting strain, refer to the patent 201510516170.X method to knock out the genes PDC1, PDC5, PDC6, ADHI, ADH4 to realize the knockout of the ethanol pathway of Saccharomyces cerevisiae, and at the same time insert the bovine LDH gene into the yeast genome to construct the Pathway for lactic acid production.

[0043] The xylose utilization pathway was constructed by introducing the XR, XDH and XKS genes derived from Pichia pastoris (shown in sequence as SEQ ID NO: 1-3), and inserting them into the delta site of the Saccharomyces cerevisiae genome by homologous recombination.

Embodiment 2

[0044] Example 2: Recombinant Saccharomyces cerevisiae-Shewanella (Shewanella Oneidensis, MR-1) combined with MFC to produce electricity

[0045] 1. Strain activation

[0046] The Shewanella Oneidensis (MR-1, purchased from ATCC) containing the empty plasmid PYYDT was taken out from the -80°C refrigerator, and cultured overnight in LB medium (containing kanamycin) at 30°C and 200 rpm. The overnight culture solution was transferred into a new LB medium containing kanamycin and IPTG inducer at a ratio of 1:100, 30°C, 200rpm, cultured for 10 hours, and measured OD 600 , calculate the volume (OD in MFC 600 =0.6) into the anolyte.

[0047] In Example 1, the recombinant yeast was stored in a -80°C refrigerator, transferred to YPD medium, 30°C, 200rpm, cultured for 48 hours, 1ml of culture medium was transferred to 100mlYPD, 30°C, 200rpm, cultivated for 36 hours Afterwards, calculate the volume, centrifuge at 4000rpm for 5 minutes, resuspend and pour in the anolyte (OD in the MFC)...

Embodiment 3

[0058] Example 3: Comparison of electricity production of recombinant Saccharomyces cerevisiae-Shewanella (Shewanella Oneidensis, MR-1) / Shewanella (Shewanella Oneidensis, 3C5) combined with MFC

[0059] With reference to the electricity generation method and electrochemical analysis method of embodiment 2, MFC electricity generation contrast is carried out, and the results are shown in Figure 6-7 .

[0060] Depend on Figure 6 It can be seen from the results that, compared with the effect of MFC power generation combined with Shewanella MR-1, the maximum output voltage increased to more than 600mV by using Shewanella 3C5 for MFC power generation, showing a very significant increase effect, and in the production There is no significant impact on the electrochemical effect of the electric cycle and the second cycle (change the liquid when the electricity in the first cycle drops to about 50mV, replace a quarter of the anolyte with new liquid, and replace all the catholyte).

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Abstract

The invention relates to the technical field of bio-energy and discloses a recombinant yeast strain and a microbial mixed bacteria electricity generation method. According to the recombinant yeast strain disclosed by the invention, genes PDC1, PDC5, PDC6, ADHI and ADH4 are knocked-out, and genes LDH, XR, XDH and XKS are inserted. Meanwhile, a yeast-Shewanella mixed bacteria system taking glucose and xylose as a carbon source is constructed, and by modifying brewer's yeasts, the glucose and xylose can be metabolized to produce lactic acid, and the carbon source and electron donors are providedfor shewanella in the MFC, so that available carbon source spectrums of the shewanella are increased, and the recombinant yeast strain has excellent performance on the electrochemical effect and laysa foundation for researching biomass by human beings and identifying new energy conversion manners.

Description

technical field [0001] The invention relates to the technical field of bioenergy, and more specifically relates to a method for producing electricity by recombining yeast strains and microbial mixtures. 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. The electricity-ge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12N1/20H01M8/16C12R1/865C12R1/01
CPCC12N1/20C12N9/0006C12N9/1205C12Y101/01027C12Y101/01028C12Y101/01307C12Y207/01017H01M8/16Y02E60/50
Inventor 宋浩林童白雪曹英秀李炳志元英进
Owner TIANJIN UNIV
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