Enzymology of multi-enzyme systems on self-assembled surfaces Enzymology of multi-enzyme systems on self-assembled surfaces Surface-immobilized enzymes are widely used in a large number of diverse applications, from laundry detergents to biosensors. Enzyme systems associated with membranes and the cytoskeleton, in living systems, have shown us that enzymes associated with surfaces have additional abilities that soluble enzymes lack. Organized, surface-associated enzyme systems can catalyze whole reaction pathways much more efficiently that the same enzymes cannot achieve in solution. Currently, there are challenges that need to be overcome in order to incorporate surface-immobilized enzyme systems into useful applications. One is to improve the activities and stabilities of surface-bound enzymes by optimizing their orientations and conformations as well as their surface attachment distances. The other is to control the spatial relationships between the components of multi-enzyme cascades on surfaces aiming to maximize their catalytic efficiencies. Here, we propose to develop a novel approach for the design and assembly of enzyme systems on surfaces as well as for the optimization of their functions under a given set of conditions. We will use this system to perform a detailed mechanistic analysis of enzyme-catalyzed reaction pathways on solid or water-soluble nanostructured surfaces. This will be accomplished by creating enzyme-specific peptide ligands that bind to target enzymes, maintaining them in optimal orientations and conformations for enhanced function. Combining these peptides with surface attachment chemistries and DNA-based nanostructure will create an unprecedented opportunity to design and construct heterogeneous surfaces and nanoreactors that self-assemble with sets of enzymes to catalyze complex reaction pathways efficiently. Specifically, we aim to: 1) Explore peptide space to identify ligands that will bind to target enzymes and optimize their orientations on surfaces to achieve active functions. 2) Create and analyze single- and multi-enzyme systems on peptide-modified solid surfaces. 3) Create self-assembled DNAnanoscaffold surfaces, which will allow precise control over parameters including inter-enzyme orientation and spacing as well as the distances from enzymes to surfaces or scaffolds.
|Effective start/end date||12/1/10 → 11/30/13|
- National Science Foundation (NSF): $406,206.00
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