Ultra-Sensitive and Ultra-Fast Absorption Spectrometer for Micro-Droplet-based Enzyme Evolution Experiments

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Most biological processes are driven by the function of enzymes, these are proteins that bind to a chemical compound and through this interaction alter the chemical compound into a different chemical substance. Our cells are driven by a huge diversity of enzyme functions that dictate everything from how our cells move and grow to how cells convert energy into a usable form. Much of biological, medical and industrial science is focused on understanding how enzymes work and discovering new enzymes that can provide us with new functions for medical and industrial applications. However, our ability to study enzyme function is often limited to focused studies encompassing a handful of minor changes in the gene/protein and, furthermore, is focused on a relatively small pool of previously characterised enzymes. What we need now is new technologies that allow us to study and compare the function of enzymes and which allows us to make huge numbers of changes to that enzyme. Furthermore, such technologies should also allow us to screen huge numbers of protein in order to search a wide diversity of unknown genes for new types of enzyme. This can only be achieved by developing new technologies that allow us to simultaneously sample and investigate millions of genes and then conduct millions of enzyme function experiments in short time frames. The aim of this project is to develop new technologies that allow us to do this. Enzymes are formed of proteins which are encoded by genes on DNA, the nature of the genetic code recorded in DNA ultimately governs the nature of an enzyme and how it works. We will develop a new technology that will allow us to express thousands of fragments of DNA into protein all at once. We will then use cutting edge micro-plumbing to separate single cells each with different types of DNA/protein into a separate compartment as droplets. Using state of the art methods for studying chemical reactions we will build a new way of detecting chemical changes within each micro-compartment. This innovative and challenging combination of technology will allow us to screen huge numbers of variant enzymes and search DNA of unknown function for new gene functions. The power of the optofluidic technology proposed will allow us to take highly sensitive measurements surpassing current approaches. Specifically, the new optofluidic technology that will be utilised for detecting chemical changes will be around 1000 times more sensitive than current technologies used. By combining it with the droplet based compartmentation we will be able to screen 1000 independent experiments a second. Our proposed project will unlock the door to a range of new approaches for both investigating how enzyme evolution relates to enzyme function, discovery of new enzymes and down-stream the development of new antimicrobial drugs.