PhD Studentship: Protein design and evolution to create novel mono-oxygenases

The University of Manchester - Academic School: School of Chemistry

Supervisors

Principal Supervisor: Professor David Leys

Anticipated start date for project: Available to start in April 2017, July 2017 or September 2017

Closing date for applications: open until position is filled

Summary of Project

The chemical repertoire of enzymes is greatly enhanced by cofactors: small inorganic or organic molecules that are bound by the protein matrix to constitute the active holo-enzyme. Our group has recently discovered a new type of cofactor: a prenylated-flavin that has azomethine ylide properties (Refs 1 and 2). This cofactor is an integral part of the widespread ubiD/ubiX system. The latter is implicated in the non-oxidative reversible decarboxylation of aromatic substrates, and plays a pivotal role in bacterial ubiquinone biosynthesis or microbial biodegradation of aromatic compounds. Our data strongly suggests 1,3-dipolar cycloaddition chemistry supports reversible decarboxylation in these enzymes, the first example of an enzymatic 1,3-dipolar cycloaddition reaction. Our model for UbiD catalysis hints at new routes in alkene hydrocarbon production or aryl (de)carboxylation.

The current application builds ambitiously on these: we seek to explore and harness the unusual properties of the prenylated flavin, through targeted evolution of (monoxygenase) flavoenzymes to create artificial prFMN-dependent self-sufficient mono-oxygenases. Flavin-dependent mono-oxygenases are of particular interest, as they perform selective oxidation under ambient conditions. Despite the fact that 4a-(hydro)peroxyflavin intermediates are implicated in flavin monoxygenase catalysis, a peroxide driven “shunt” has not been shown to be viable in these enzymes. The production of prFMN offers a convenient route to combine existing flavin-dependent mono-oxygenase scaffolds with incorporation of a stable N5-alkylated flavin in vivo, aiming to create novel flavo-peroxygenases. Our approach seeks to harness these artificial prFMN-dependent enzymes in novel green routes to commodity chemicals.

A multidisciplinary approach is required to document enzyme mechanism to guide laboratory evolution experiments aimed at developing novel catalysts. This ambitious project seeks to use structural biology, advanced kinetics, analytical chemistry, spectroscopy and computational studies combined with state-of-the-art laboratory evolution studies to tackle the questions outlined above. The successful candidate will join the group of David Leys in the MIB (www.mib.ac.uk) working alongside other group members recruited on this ERC project.   

  1. Payne, K. A., et al.. (2015), Nature 522, 497-501
  2. White, M. D., et al. (2015), Nature 522, 502-506

Qualification

Applicants should have or expect a good II(i) honours degree (or an equivalent degree) in Chemistry or biochemistry.

Funding

This project is to be funded under the European Research Council (ERC) Programme. This Studentship is for 4 years covering tuition fees and a stipend of 14,296 for 2016/17. Eligibility restricted to UK/EU applicants.

Contact for further Information

For general admission information, please email: pg.chemistry@manchester.ac.uk.

Informal inquiries about the project should be sent to Prof. David Leys by email (David.Leys@manchester.ac.uk). Please note that to apply for this studentship you must submit the relevant information via the University’s online application form.

How to Apply

Complete an online application form using the following web link http://www.manchester.ac.uk/postgraduate/howtoapply/

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Type / Role:

PhD

Location(s):

Northern England