RESUMEN
The nature, the composition and the concentration of electrolytes is essential for electrocatalysis involving redox enzymes. Here, we discuss the effect of various electrolyte compositions with increasing ionic strengths on the stability and activity towards O2 reduction of the bilirubin oxidase from Myrothecium verrucaria (Mv BOD). Different salts, Na2SO4, (NH4)2SO4, NaCl, NaClO4, added to a phosphate buffer (PB) were evaluated with concentrations ranging from 100 mM up to 1.7 M. On functionalized carbon nanotube-modified electrodes, it was shown that the catalytic current progressively decreased with increasing salt concentrations. The process was reversible suggesting it was not related to enzyme leakage. The enzyme was then immobilized on gold electrodes modified by self-assembling of thiols. When the enzyme was simply adsorbed, the catalytic current decreased in a reversible way, thus behaving similarly as on carbon nanotubes. Enzyme mobility at the interface induced by a modification in the interactions between the protein and the electrode upon salt addition may account for this behavior. When the enzyme was covalently attached, the catalytic current increased. Enzyme compaction is proposed to be at the origin of such catalytic current increase because of shorter distances between the first copper site electron acceptor and the electrode.
Asunto(s)
Nanotubos de Carbono , Oxidación-Reducción , Catálisis , Oxígeno/metabolismo , Electrodos , Enzimas Inmovilizadas/metabolismoRESUMEN
How the redox proteins and enzymes involved in bioenergetic pathways are organized is a relevant fundamental question, but our understanding of this is still incomplete. This review provides a critical examination of the electrochemical tools developed in recent years to obtain knowledge of the intramolecular and intermolecular electron transfer processes involved in metabolic pathways. Furthermore, better understanding of the electron transfer processes associated with energy metabolism will provide the basis for the rational design of biotechnological devices such as electrochemical biosensors, enzymatic and microbial fuel cells, and hydrogen production factories. Starting from the redox complexes involved in two relevant bacterial chains, i.e., from the hyperthermophile Aquifex aeolicus and the acidophile Acidithiobacillus ferrooxidans, examination of protein-protein interactions using electrochemistry is first reviewed, with a focus on the orientation of a protein on an electrochemical interface mimic of a physiological interaction between two partners. Special attention is paid to current research in the electrochemistry of essential membrane proteins, which is one mandatory step toward the understanding of energy metabolic pathways. The complex and challenging architectures built to reconstitute a membrane-like environment at an electrode are especially considered. The role played by electrochemistry in the attempt to consider full bacterial metabolism is finally emphasized through the study of whole cells immobilized at electrodes as suspensions or biofilms. Before the performances of biotechnological devices can be further improved to make them really attractive, questions remain to be addressed in this particular field of research. We discuss the bottlenecks that need to be overcome in the future.