Team:Bielefeld-CeBiTec/Results/rMFC/ElectronTransfer

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rMFC

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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_1465102 under controll of T7 promotor. Fumarate and succinate concentrations were detected with HPLC.

Neutral Red

Bromphenol Blue

Cytochroms



Reference

  • Iverson et al., 1999. Structure of the Escherichia coli Fumarate Reductase Respiratory Complex. Science, vol. 284, pp. 1961-1966

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