The U.S. Department of Energy’s Biosciences Program has partnered with the Pacific Northwest National Laboratory on a three-year, $1.5 million initiative aimed at advancing tools and techniques for investigating the molecular mechanisms of enzymes. This collaboration seeks to unlock new possibilities in energy production, environmental sustainability, and biotechnology by focusing on the fundamental processes that drive enzyme function. Enzymes play a critical role in various energy-related applications, such as hydrogen production, fuel cell development, and environmental remediation. However, to harness their full potential, scientists must first understand the intricate biochemical pathways they govern. The current research project centers on oxidoreductases—enzymes essential to all living organisms because they regulate intracellular redox reactions through electron transfer. As part of this effort, researchers at the Pacific Northwest National Laboratory plan to integrate a technique known as cyclic voltammetry with single-molecule spectroscopy to create an advanced electrochemistry-based single-molecule spectrometer. This innovative device will allow scientists to observe real-time redox reactions in enzymes, providing deeper insights into their dynamic behavior. One major challenge in studying enzymes is their instability outside the cellular environment. To address this, the team previously developed a novel method that embeds enzymes within nanostructured matrices, significantly enhancing their stability and longevity. With this improved stability, the enzymes can be placed in micro-scale electrochemical cells, where they generate controlled electrical currents. These minute fluctuations in current are closely linked to the enzyme’s catalytic activity, enabling researchers to monitor individual enzyme molecules in action. To gather the necessary enzyme variants, the team will employ a cutting-edge cell-free protein synthesis approach, which allows for the rapid production of up to 384 different proteins or variants per day. This high-throughput method accelerates the research process and supports more comprehensive studies. Principal Investigator Eric Akerman emphasized the importance of this work, stating, “We anticipate that the findings from this research will provide foundational knowledge about electron transfer in enzymatic reactions. This knowledge has broad implications for bioenergy and environmental technologies.” The research team also includes leading scientists Lei Chenhong, Hu Dehong, and Chuck Wentis, who bring expertise in chemistry, biology, and engineering to the project.

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