Predictive Structure-Function Relationships for Enzymes Immobilized on Complex Surfaces
Abstract
Immobilization is frequently used to enhance the lifetime and stability of enzymes, yet a fundamental understanding of the interactions between enzymes, covalent linkers, and complex polymeric surfaces enabling accurate prediction of system performance is lacking. While prediction of enzyme behavior in simple monolayers is well described, these capabilities fall short on complex polymer surfaces used in electrochemical or optical biosensors. The stabilizing effects of enzyme immobilization results in enzymes with longer useful lifetimes; however, this can come at the expense of specific enzyme activity, due to mass transfer limitations caused by orientation effects, the complex interplay between enzyme-linker-substrate interactions, and immobilization related unfolding of enzyme structure. Current approaches for addressing enzyme immobilization focus mostly on solutions to specific problems, i.e., on a specific combination of enzyme, linker, and immobilization chemistry. A broadly applicable understanding of the molecular scale interactions between enzyme-linker-substrate is required to make accurate predictions of the best overall immobilization strategy. This project uses a combined experimental and computational approach to understand the fundamental interactions in enzyme-linker-substrate systems. By combining experimental and simulation data sets, we will identify how particular characteristics of the enzyme, the linker, and the polymeric surface influence structure and activity. This enzyme activity-linker-surface relationship can be used to predict the effect of immobilization strategy on the activity of the enzyme, with application to enzymes for specific detection of tributyl phosphate solvent used in nuclear fuel processing.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jun 29, 2016
- Source ID
- HDTRA11610023
Entities
People
- Mark Blenner
Organizations
- Clemson University
- Defense Threat Reduction Agency