PhD Studentship - High-throughput Characterization and Engineering of Nucleotide Synthases for Production of Nucleoside Analogues

This 3.5 year PhD is fully funded though CoEBio3; funding will cover fees and a tax free (depending on circumstances) stipend set at the UKRI rate (£19,237 for 2024/25).

Nucleotides are the building blocks of life, modulating diverse processes from protein production to cell signalling. It is therefore unsurprising that synthetic nucleoside analogues have been developed as therapeutics for the treatment of a wide range of diseases including cancers and viral infections. Nucleosides are challenging to chemically synthesize as they are structurally complex, contain multiple stereogenic centres, and often have poor solubility in organic solvents. In biology, nucleosides are synthesized efficiently and in a regio- and stereoselective manner by enzymes – thus, biocatalysis offers an attractive alternative to chemical approaches to produce modified nucleoside analogues.

In this project we will investigate different classes of nucleotide synthases. These will be evaluated using HT-MEK (High-Throughput Microfluidic Enzyme Kinetics), a state-of-the-art technology for expression, purification, and repeated quantitative measurement of 1,500 enzyme variants in a single experiment. This will allow us to quantitatively map the substrate specificity landscapes of large panels of nucleotide synthases and carry out systematic functional studies to reveal the molecular determinants of catalysis and specificity in these enzymes. This knowledge will inform subsequent engineering of the most promising synthases.

This is a multidisciplinary project involving state-of-the-art high-throughput technology and modern protein engineering methodologies. The student will gain diverse skills in molecular biology, in vitro expression, design and optimization of high-throughput biochemical assays, laboratory evolution, and analysis of large data sets (e.g. using computational notebooks, Python and Mathematica). The student will be based at the Manchester Institute of Biotechnology where they will work under the supervision of Dr Sarah Lovelock and Dr. Craig Markin. This project is supported by funding from CoEBio3, an industrially funded PhD training network.

Applicants should have, or expect to achieve, at least a 2.1 honours degree or a master’s in biochemistry, chemistry or a related discipline.

Before you apply please contact the supervsiors: Dr. Lovelock ([email protected] ) and Dr. Craig Markin ([email protected] ).