University of Queensland/ Australia
The production of fuels and chemicals from renewable sources is a vital element for sustainable living in the foreseeable future. Here, we will optimise the properties of four enzymes in a previously designed non-natural and minimal reaction cascade required to convert glucose to isobutanol. Isobutanol is a chemical, solvent and, due to its low volatility and high energy density, it is an excellent biofuel. To achieve this goal, we will use structural and mechanistic insight in combination with bioinformatics tools (molecular design studies, ancient sequence reconstruction) and experimental approaches (high throughput techniques and robotics) to rationally and semi-rationally design enzymes that have increased catalytic activity and stability (Aim 1). Optimised enzymes will then be used to reconstitute the cascade and to design an optimal reaction setup to maximise its productivity and product (isobutanol) yield (Aim 2). The outcome will provide a blueprint for the cell-free enzymatic conversion of renewable raw materials into high-value chemicals.