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Biochemistry
Electroquímico detección de deuterio cinético isótopo efecto en extracelular transporte de electr...
Electroquímico detección de deuterio cinético isótopo efecto en extracelular transporte de electr...
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1

Electroquímico detección de deuterio cinético isótopo efecto en extracelular transporte de electrones en la Shewanella oneidensis MR-1

Full Text
10,794 Views
09:00 min
April 16, 2018

DOI: 10.3791/57584-v

Yoshihide Tokunou1, Kazuhito Hashimoto2, Akihiro Okamoto2

1Department of Applied Chemistry,University of Tokyo, 2Global Research Center for Environment and Energy based on Nanomaterials Science,National Institute for Materials Science

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol outlines whole-cell electrochemical experiments to investigate proton transport's role in extracellular electron transport (EET) in Shewanella oneidensis MR-1. The study employs the deuterium kinetic isotope effect to assess the impact of proton transport on EET kinetics.

Key Study Components

Area of Science

  • Electrochemistry
  • Microbiology
  • Bioenergetics

Background

  • Shewanella oneidensis MR-1 is known for its ability to transfer electrons to electrodes.
  • Proton transport is crucial for the efficiency of extracellular electron transport.
  • The deuterium kinetic isotope effect is a valuable method for studying proton involvement in electron transport.
  • Using living bacteria provides insights into native proton management during EET.

Purpose of Study

  • To observe the contribution of proton transport to the rate of EET.
  • To utilize the deuterium kinetic isotope effect for examining proton transport's role.
  • To improve understanding of the mechanisms underlying bacterial electron transfer.

Methods Used

  • Whole-cell electrochemical experiments.
  • Detection of the deuterium kinetic isotope effect.
  • Use of living Shewanella oneidensis MR-1 cells.
  • Growth of bacterial cultures in LB medium.

Main Results

  • Proton transport significantly affects the kinetics of EET.
  • The deuterium kinetic isotope effect provides clear insights into proton involvement.
  • Noise from other cellular processes was effectively minimized.
  • The method reflects native proton management in EET.

Conclusions

  • Proton transport is a critical factor in the efficiency of EET in bacteria.
  • The protocol offers a reliable approach to study these processes in living cells.
  • Findings contribute to a deeper understanding of microbial electron transfer mechanisms.

Frequently Asked Questions

What is the main focus of this study?
The study focuses on the role of proton transport in extracellular electron transport in Shewanella oneidensis MR-1.
How does the deuterium kinetic isotope effect contribute to the research?
It allows for the examination of proton transport's contribution to electron transport kinetics.
Why use living bacteria instead of purified enzymes?
Using living bacteria reflects the native proton management during electron transport processes.
What challenges were faced during the experiments?
The main challenge was to minimize noise from other cellular processes that could interfere with results.
What are the implications of this research?
The findings enhance the understanding of microbial electron transfer mechanisms and their efficiency.

Aquí presentamos un protocolo de los experimentos electroquímicos celulares para estudiar la contribución del transporte de protones a la velocidad de transporte de electrones extracelular a través de los citocromos de la membrana externa de Shewanella oneidensis MR-1.

El objetivo general de este protocolo es observar la contribución del transporte de protones a la tasa de transporte de electrones extracelulares bacterianos mediante la detección del efecto del isótopo cinético de deuterio. Nuestro método revela el impacto del transporte de protones en la cinética del transporte de electrones extracelulares o EET de las bacterias a un electrodo a través de citocromos de tipo c de membrana externa. Nuestro ensayo refleja el manejo de protones nativos en EET porque utilizamos bacterias vivas en lugar de enzimas purificadas.

Sin embargo, nuestro reto consistía en eliminar los ruidos de otros procesos celulares. El efecto del isótopo cinético de deuterio, denominado KIE, es una de las mejores técnicas para examinar la contribución del transporte de protones en el proceso de transporte de electrones asociado. Para cultivar células Shewanella MR-1, transfiera una colonia de células cultivadas en una placa de agar a 15 mililitros de medio LB.

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