Engineering of an Extremely Thermostable Alpha/Beta Barrel Scaffold to Serve as a High Affinity Molecular Recognition Element for Use in Sensor Applications
Technical Report,01 Aug 2012,31 Jul 2015
Columbia University New York United States
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The overall goal of the project was to evolve a highly thermostable enzyme alcohol dehydrogenase D AdhD from Pyrococcus furiosus to bind an explosive molecule, RDX. The enzyme naturally catalyzes the nicotinamide cofactor-dependent oxidation or reduction of alcohols, aldehydes, ketones and carbohydrates. Directed evolution techniques were used to convert the enzyme into a simple binder of the small molecule RDX. Novel binders have been identified and characterized with sub-millimolar dissociation constants. These could be further developed for use in robust and low cost biosensors.