Controlled assembly for well-defined 3D bioarchitecture using two active enzymes.

This paper reports that a bioarchitecture with two different active enzymes can be fabricated conveniently on a prepatterned surface by highly selective surface-templated layer-by-layer (LBL) assembly by coupling a bilayer of avidin/biotin-lactate oxidase (biotin-LOD) with a bilayer of avidin/biotin-horseradish peroxidase (biotin-HRP). The two different active enzymes can be utilized as excellent building blocks for the fabrication of well-defined 3D nanostructures with precise control of the position and height on the surface. In addition, the LBL assembled bienzyme structures are highly functional, and bioarchitectures based on LOD and HRP-mediated coupling reaction can be employed in a number of viable biosensing applications.