- Dr Raine Thomson, The University of Queensland
- Prof. Elizabeth Gillam, The University of Queensland
Cytochrome P450 enzymes (P450s) can be used to perform a range of chemical transformations with exquisite selectivity that can be difficult or sometimes impossible to achieve with conventional synthetic chemistry. However, most enzymes are incompatible with the high temperatures encountered in industrial processes.
Ancestral protein reconstruction is a computational technique where you can try to predict an ancient protein sequence by building a phylogenetic tree based on evolutionarily-related sequences from the present day. Professor Elizabeth Gillam from The University of Queensland used this method to reconstruct the ancestors of several modern-day P450 enzymes. Her group discovered that these ancient enzymes can withstand much higher temperatures than their modern-day descendants, but can perform many of the same reactions. These thermostable P450s are really attractive as industrial biocatalysts and Prof. Gillam's group has developed methods for using them for high-temperature in vitro biocatalysis.
We want to take this a step further by introducing these thermostable enzymes into a thermostable bacterium, so that they can be used for high-temperature biocatalysis in live cells. There are several advantages to using these enzymes for biocatalysis in a thermostable bacterium including reduced costs and the opportunity to integrate the P450s into new biosynthetic pathways.
You will learn how to maintain, cultivate, and genetically manipulate a thermostable microorganism (with James Behrendorff at QUT).
You will also learn a specialised technique (carbon monoxide difference spectroscopy) for measuring the concentration of P450 enzyme in whole cells (with Dr Raine Thomson in Elizabeth Gillam's lab at UQ)
You will also have the opportunity to test your microbes for new catalytic functions enabled by their new P450 enzymes.
The aim of this project is to take the first steps towards developing thermophilic microbes capable of performing useful P450-mediated reactions for industrial biochemistry (e.g. drug development). Genetic transformation of the thermophilic bacterium and successful expression of the P450 enzymes are critical first steps.
There is potential to develop this VRES into an opportunity for further study focused on understanding and optimising P450 function in thermophiles.
Skills and experience
This project is suited to a student who enjoys practical work in the wet lab, and is interested in enzymes and developing futuristic biotechnology.
Students must have a strong theoretical and practical background in at least one of the following disciplines from their undergraduate studies:
- molecular biology
- synthetic biology.
Contact the supervisor for more information.