Development of new instruments and methods for studying the molecular mechanism of enzymes

The US Department of Energy's Biosciences Program and the Pacific Northwest National Laboratory signed a three-year, $1.5 million contract to develop new instruments and methods for studying the molecular mechanisms of enzymes.

Intracellular Protein Nanomachines - Enzymes have many potential applications in energy, such as hydrogen manufacturing, fuel cell development, and environmental governance. But in order to get these applications, researchers must first fill in the basics of the process of enzymes. The new research program has set oxidoreductase as the research goal. Oxidoreductases are the basis of all life forms because oxidoreductases circulate intracellular reduction and oxidation reactions through intracellular electron transfer.

As a first step in the research, researchers at the Pacific Northwest National Laboratory plan to combine a chemical chemistry called "cyclic voltammetry" with single-molecule spectroscopy to develop a new electrochemistry single-molecule spectrometer. . The new device will enable researchers to conduct dynamic studies of basic enzyme redox reactions.

Because the enzyme has extracellular instability, it makes it difficult for researchers to study it. In a previous study, researchers at the Pacific Northwest National Laboratory invented a new method to induce enzymes into a matrix of nanostructures to increase enzyme stability and extend their lifespan.

The enzyme will be stable in the nanostructure matrix and then placed in a miniature electrochemical cell where it releases a controlled current. Since the tiny shaking of the current affects the enzyme's catalyzed reaction, the researchers will observe the individual enzyme molecules. Researchers will use the chemical information generated by the new electrochemistry single-molecule spectrometer to study the catalytic electron transfer process.

In order to obtain the necessary enzyme variants, the team will use a new cell-free approach instead of the traditional protein-making molecular approach. Unique mechanical instruments can produce up to 384 protein or protein variants a day.

Principal Investigator Eric Akerman said, "We expect that the results of this research will provide us with the basic knowledge necessary to understand the electron transfer in the catalyzed reaction. This research will include bioenergy and environmental governance. It has been applied in many areas." Other members of the research team also have lead researchers Lei Chenhong, Hu Dehong and Chuck Wentis.