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Team:Bielefeld-CeBiTec/Results/rMFC/ElectronTransfer
From 2014.igem.org
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| - | We cultivate E. coli KRX under anaerobic conditions to characterize activity of fumarate reductase frd (BBa_K1465102) under controll of T7 promotor. | + | We cultivate E. coli KRX under anaerobic conditions to characterize activity of fumarate reductase frd (BBa_K1465102) under controll of T7 promotor. Fumarate and succinate concentrations were detected with HPLC. |
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Revision as of 09:16, 12 October 2014
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Fumarate reductase
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_1465102). Fumarate reductase consist of four subunits, two large catalytic and two smaller membrane associated subunits (Iverson et al., 1999). The two catalytic subunits could be detected via SDS-PAGE.
Anaerobic cultivation
We cultivate E. coli KRX under anaerobic conditions to characterize activity of fumarate reductase frd (BBa_K1465102) under controll of T7 promotor. Fumarate and succinate concentrations were detected with HPLC.
Neutral Red
Bromphenol Blue
Cytochroms
Reference
