There is enormous potential for the use of ‘designer’ (engineered) enzymes in agriculture, industry, medicine and defense. However, to successfully engineer enzymes and harness their catalytic power, we must understand how their structure determines their function. Protein structure is classically defined in a static sense, but functioning enzymes are dynamic; their function is determined not by a single structure, but by an ensemble of different conformations of a structure. To be able to truly design and engineer enzymes to meet the future demands of society we must understand how the ‘conformational landscapes’ of enzymes modulate their activity and develop methodologies and approaches to be able to alter these landscapes as we wish.
This project aims to meet the challenges associated with studying and engineering conformational landscapes by using an innovative and multidisciplinary approach: cutting edge experimental and computational techniques will be used alongside both random and rational protein engineering approaches to determine the mechanisms by which mutations can affect activity by altering a conformational landscape.
(1) Characterizing conformational ensembles using X-ray crystallography and computer simulation.
(2) Engineering new conformational landscapes. Using random and directed mutagenesis to identify mutations that affect conformational change and enzyme activity.
(1) An enzyme from the Australian sheep blowfly (E3) that makes the insects resistant to pesticides.
(2) A recently discovered bacterial enzyme (TrzN) that breaks down a toxic herbicide called atrazine.
(3) A ‘de-novo’ designed enzyme that was created in the laboratory and catalyzes the Kemp elimination.