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Team:Bielefeld-CeBiTec/Results/rMFC/ElectronTransfer
From 2014.igem.org
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Upon the expression of fumarate reductase in <i>E. coli</i> we analyzed the metabolic behavior under aerobic and anaerobic conditions. | Upon the expression of fumarate reductase in <i>E. coli</i> we analyzed the metabolic behavior under aerobic and anaerobic conditions. | ||
| - | Furthermore we characterize the impact on electrochemical behavior of E. coli. We show expression of fumarate reductase (frd) | + | Furthermore we characterize the impact on electrochemical behavior of E. coli. We show expression of fumarate reductase (frd) <a href="http://parts.igem.org/Part:BBa_K1465102">BBa_1465102</a> using SDS-PAGE in combination with MALDI-TOF/MS. |
| - | Activity could be shown with HPLC analysis. | + | Activity could be shown with HPLC analysis of fumarate consumption and succinate production. |
</p> | </p> | ||
<p> | <p> | ||
Revision as of 08:23, 12 October 2014
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Theory
Cytochroms
Generation of reduction equivalents
Short summary
Upon the expression of fumarate reductase in E. coli we analyzed the metabolic behavior under aerobic and anaerobic conditions. Furthermore we characterize the impact on electrochemical behavior of E. coli. We show expression of fumarate reductase (frd) BBa_1465102 using SDS-PAGE in combination with MALDI-TOF/MS. Activity could be shown with HPLC analysis of fumarate consumption and succinate production.
SDS-PAGE
The fumarate reductase could be detected in purified membrane and periplasmatic protein fraction. Proteins were fractioned by cold osmotic shock of E. coli KRX at different steps after induction of protein expression. SDS-PAGE shows the expression of fumarate reductase in E. coli KRX under control of T7 promotor (BBa_K1465102). Fumarate reductase consist of four subunits, two large catalytic and two smaller membrane associated subunits. The two catalytic subunits could be detected via SDS-PAGE.
Neutral Red
Bromphenol Blue
