Team:Bielefeld-Germany/Datapage

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Data Page


Overview

This page gives a basic overview about our our project Ecolectricity - currently available. A detailed description of our subprojects can be found in our project part of the Wiki.


Project description: The goal of our project is to generate electrical energy with a genetically modified Escherichia coli in a self-constructed fuel cell. Besides the design, construction and technical optimization of the fuel cell, we investigate different genetic approaches. Specific electron transfer proteins have been compiled from a variety of organisms, in order to gain an Escherichia coli Fuel Cell platform, which turns E. coli to an electro active organism. The main challenge is due to an efficient electron transfer from the bacteria to the electrode. Therefore we facilitate and improve electron donation by producing electron-shuttles, so called endogenous mediators, as well as permeabilizing the cell surface by integrating large membrane porins and providing a direct electron pathway by conductive transmembrane protein structures. All these electron transport elements increase electron transfer and bioelectricity generation.



Data for our favorite new parts

  1. <bbpart>BBa_K1172501</bbpart> - OprF – Outer membrane porin from Pseudomonas fluorescens: This outer membrane porin enhances membrane permeability and electron shuttle-mediated extracellular electron transfer for higher bioelectricity output.
  2. <bbpart>BBa_K1172901</bbpart> - Alanin-Racemase from E. coli: Catalizes the reversible isomerization from L-alanine into the enatiomer D-alanine. In an D-alanine auxotrphic strain it can be used as a antibiotic free selection marker.
  3. <bbpart>BBa_K1172303</bbpart> - Riboflavin synthesis gene cluster from Shewanella oneidensis: Short part description.
  4. <bbpart>BBa_K1172909</bbpart> - Biosafety-System araCtive: Biosafety-System taking advantage of the Alanin-Racemase and the tight repression of the arabinose promoter pBAD by the araC protein.


We have also characterized the following parts (Coding)

  1. <bbpart>BBa_K1172201</bbpart> - GldA - Glycerol dehydrogenase from Escherichia coli: Glycerol dehydrogenase can be used for endogenous mediator production of NADH. Higher amount of electron shuttles improve transfer of electrons from bacteria to the anode and makes the MFC more efficient.
  2. <bbpart>BBa_K1172301</bbpart> - NorM: Na+ Antiporter from Shewanella oneidensis: Short part description.
  3. <bbpart>BBa_K1172302</bbpart> - RibC: 6,7-dimethyl-8-ribityllumazine synthase alpha subunit RibC from Shewanella oneidensis: Short part description.
  4. <bbpart>BBa_K1172401</bbpart> - MtrCAB – Cytochromes from Shewanella oneidensis: Short part description.
  5. <bbpart>BBa_K1172904</bbpart> - Rnase Ba (Barnase) from Bacillus Amyloliquefaciens: Genetically modified variant that cleaves the intracellular RNA of the host organism.
  6. <bbpart>BBa_K1172916</bbpart> - Double plac-Promoter: Improved repression of the lac promoter by an addtional lacI-binding site, 1.5 whorl downstream of the natural binding site.


We have also characterized the following parts (Devices)


Endogenous mediator prodcution by glycerol dehydrogenase

  1. <bbpart>BBa_K1172203</bbpart> - gldA with T7 promoter and strong RBS: This glycerol dehydrogenase device is used to characterize the endogenous mediator production of NADH and seems to be the best part for efficient NADH production.
  2. <bbpart>BBa_K1172204</bbpart> - gldA with IPTG inducible lacI promoter and strong RBS: This glycerol dehydrogenase device is used to characterize the endogenous mediator production of NADH.
  3. <bbpart>BBa_K1172205</bbpart> - gldA with medium Anderson promoter and medium RBS: This glycerol dehydrogenase device is used to characterize the endogenous mediator production of NADH.

Porins

  1. <bbpart>BBa_K1172502</bbpart> - oprF with T7 Promoter and strong RBS: This device is used to characterize the outer membrane porin expression and its effect on the cell surface permeability. This part is the most efficient OprF expression device leading to most improved bioelectricity output.
  2. <bbpart>BBa_K1172503</bbpart> - oprF with IPTG inducible lacI promoter and strong RBS: This device is used to characterize the outer membrane porin expression and its effect on the cell surface permeability.
  3. <bbpart>BBa_K1172504</bbpart> - oprF with medium Anderson promoter and weak RBS: This device is used to characterize the outer membrane porin expression and its effect on the cell surface permeability.
  4. <bbpart>BBa_K1172505</bbpart> - oprF with medium Anderson promoter and medium RBS: This device is used to characterize the outer membrane porin expression and its effect on the cell surface permeability.
  5. <bbpart>BBa_K1172507</bbpart> - oprF with strong Anderson promoter and strong RBS: This device is used to characterize the outer membrane porin expression and its effect on the cell surface permeability.

Endogenous mediator Riboflavin

  1. <bbpart>BBa_K1172304</bbpart> - Riboflavin synthesis gene cluster from Shewanella oneidensis with T7 promoter and strong RBS: riboflavine synthesis gene cluster, induced by rhamnose.
  2. <bbpart>BBa_K1172305</bbpart> - Riboflavin synthesis gene cluster from Shewanella oneidensis with medium Anderson promoter and medium RBS: riboflavine synthesis gene cluster.
  3. <bbpart>BBa_K1172306</bbpart> - Riboflavin synthesis gene cluster from Shewanella oneidensis with strong Anderson promoter and strong RBS: This device was used for extensive quantitative and qualitative proof of riboflavin overproduction.

Cytochromes

  1. <bbpart>BBa_K1172402</bbpart> - mtrCAB gene cluster with IPTG inducible lacI promoter and RBS: This device is coding for cytochromes mtaA, mtrB and mtrC. Leading to direct elctron transfer through the outer membran
  2. <bbpart>BBa_K1172403</bbpart> - mtrCAB with medium promoter and medium RBS: Short part description.
  3. <bbpart>BBa_K1172404</bbpart> - mtrCAB with strong promoter and strong RBS: Short part description.
  4. <bbpart>BBa_K1172405</bbpart> - mtrCAB with T7 promoter and strong RBS: Short part description.

Biosafety-System

  1. <bbpart>BBa_K1172902</bbpart> - Alanin-Racemase (alr) under the control of the ptac-promoter: Testing part for the complementation of the Alanine-Racemase (alr) in the Biosafety-Strain E. coli K-12 ∆alr ∆dadX to ensure that the Alanine-Racemase could be used as a antibiotic-free selection marker.
  2. <bbpart>BBa_K1172903</bbpart> - Alanine-Racemase (alr) with double terminator: Cloning intermediate of the construction of the Biosafety-Strains. As this part contains also a RBS this might be useful for other teams, who want the Alanine-Racemase (alr).
  3. <bbpart>BBa_K1172905</bbpart> - Part 1 of the Biosafety- System araCtive: Front part of the Biosafety-System araCtive containing: pRha - araC - alr - ter.
  4. <bbpart>BBa_K1172906</bbpart> - Part 2 of the Biosafety-System araCtive (pBAD Barnase): Second part of the Biosafety-System araCtive containing pBAD RNase Ba.
  5. <bbpart>BBa_K1172908</bbpart> - Part 1 of the Biosafety-System Lac of Growth: Front part of the Biosafety-System araCtive containing: pRha - lacI - alr - ter.
  6. <bbpart>BBa_K1172911</bbpart> - Biosafety-System Lac of Growth (lacI): Biosafety-System Lac of Growth by using the lacI gene as repressor and the lactose promoter.
  7. <bbpart>BBa_K1172912</bbpart> - Part 1 of the Biosafety-System TetOR alive: Front part of the Biosafety-System araCtive containing: pRha - TetR - alr - ter.
  8. <bbpart>BBa_K1172914</bbpart> - Part 2 of the Biosafety-System M with GFP (TetO GFP): Second part of the Biosafety-System araCtive containing TetO and GFP (<bbpart>BBa_E0040</bbpart>).
  9. <bbpart>BBa_K1172915</bbpart> - Biosafety-System TetOR alive (TetR): Biosafety-System TetOR alive by using the TetR as repressor and the Tet operator.
  10. <bbpart>BBa_K1172917</bbpart> - Double plac-Promoter with GFP: Improved repression of the lac promoter by an addtional lacI-binding site, 1.5 whorl downstream of the natural binding site.









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