Limited proteolysis of pyranose 2-oxidase results in a stable and active complex

Abstract

Glucose oxidating enzymes have a tremendous potential for various energy, healthcare and environmental sensing applications. In this work, we studied the effect of reducing the size of pyranose 2-oxidase (POx) on stability and enzymatic activity of proteolyzed POx. Limited proteolysis of the POx was performed using trypsin to remove flexible structural regions without significant damage to the overall conformation and catalytic activity of the enzyme. Enzymatic activities of the modified and wild-type POx were measured by colorimetric coupled reaction assay and compared. The enzymatic activity of the modified POx showed 90% activity compared to the wild-type POx. This result indicates that reducing the size of the protein can be done without losing enzymatic activity and such enzymes potentially could provide a larger gain in electrochemical activity compared with wild-type enzymes.