High-throughput characterization and engineering of nucleotide synthases for production of nucleoside analogues

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.

Eligibility

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.

Funding

At Manchester we offer a range of scholarships, studentships and awards at university, faculty and department level, to support both UK and overseas postgraduate researchers.

For more information, visit our funding page or search our funding database for specific scholarships, studentships and awards you may be eligible for.

Before you apply

We strongly recommend that you contact the supervisor(s) for this project before you apply.

How to apply

To be considered for this project you’ll need to complete a formal application through our online application portal.

When applying, you’ll need to specify the full name of this project, the name of your supervisor, how you’re planning on funding your research, supporting statement, details of your previous study, and names and contact details of two referees.

Your application will not be processed without all of the required documents submitted at the time of application, and we cannot accept responsibility for late or missed deadlines. Incomplete applications will not be considered.

If you have any questions about making an application, please contact our admissions team by emailing [email protected].

Equality, diversity and inclusion

Equality, diversity and inclusion is fundamental to the success of The University of Manchester, and is at the heart of all of our activities. We know that diversity strengthens our research community, leading to enhanced research creativity, productivity and quality, and societal and economic impact.

We actively encourage applicants from diverse career paths and backgrounds and from all sections of the community, regardless of age, disability, ethnicity, gender, gender expression, sexual orientation and transgender status.

We also support applications from those returning from a career break or other roles. We consider offering flexible study arrangements (including part-time: 50%, 60% or 80%, depending on the project/funder).

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