Principal Supervisor: Dr Binuraj Menon, School of Life Sciences
Co-supervisor: Professor Gregg Challis and Dr Christophe Corre
PhD project title: Development of novel halogenase enzymes for biopharmaceutical applications.
University of Registration: University of Warwick
Identification of new halogenated synthetic, natural and non-natural compounds; and further exploitation and synthesis of these compounds are of extreme importance in this modern era. This is due to the profound role of organo halides as pharmaceuticals, agrochemicals and valuable synthons in various organic reactions. As an organic synthetic intermediate, halogenated molecules are of particular importance in many metal-catalyzed cross-coupling reactions. Nature has evolved a number of biocatalysts to regioselectively halogenate a diverse range of biosynthetic precursors and secondary metabolites, and this unexplored repertoire is ever growing. Biosynthetic halogenation can occur over simple to extremely complex ring structures of natural compounds and in some cases it initiates the formation of complex structures and scaffolds. These reactions often range from simple aromatic substitutions to complex stereoselective C-H functionalization and activation of remote carbon centers. These reliable, facile and cleaner biosynthetic routes have potential utility and greater demand over traditional nonenzymatic halogenation chemistry that requires deleterious reagents and lacks regio-control. In the past few years we have identified a number of pharmaceutically important halogenase systems by genome mining in natural product pathways. In this project, we are planning to explore their enzyme structure, substrate scope along with their potential applications in organic synthesis. The ultimate aim is to incorporate these enzymes in to synthetic and biosynthetic pathways and into various natural product pathways for biotechnological and pharmaceutical applications.
- Menon, Binuraj R. K., Brandenburger, Eileen, Sharif, Humera H., Klemstein, Ulrike, Shepherd, Sarah A., Greaney, Michael F., Micklefield, Jason. 2017. RadH : a versatile halogenase for integration into synthetic pathways. Angewandte Chemie International Edition, 56 (39), pp. 11841-11845.
- Latham, Jonathan, Brandenburger, Eileen, Shepherd, Sarah A., Menon, Binuraj R. K., Micklefield, Jason. 2017. Development of halogenase enzymes for use in synthesis. Chemical Reviews.
- Latham, Jonathan, Henry, Jean-Marc, Sharif, Humera H., Menon, Binuraj R. K., Shepherd, Sarah A., Greaney, Michael F., Micklefield, Jason. 2016. Integrated catalysis opens new arylation pathways via regiodivergent enzymatic C-H activation. Nature Communications, 7.
- Menon, Binuraj R. K., Latham, Jonathan, Dunstan, Mark S., Brandenburger, Eileen, Klemstein, Ulrike, Leys, David, Karthikeyan, Chinnan, Greaney, Michael F., Shepherd, Sarah A., Micklefield, Jason. 2016. Structure and biocatalytic scope of thermophilic flavin-dependent halogenase and flavin reductase enzymes. Organic and Biomolecular Chemistry, 14 (39), pp. 9354-9361.
BBSRC Strategic Research Priority: Bioenergy and Industrial biotechnology
Techniques that will be undertaken during the project:
- Bioinformatics- analyzing gene cluster, protein structural prediction and metabolite prediction, protein-protein interactions
- Molecular biology – Manipulation with DNA (PCR, cloning, Gene knockouts etc), site-directed mutagenesis
- Protein expression and purification, optimization and analysis of protein quality
- Metabolic engineering:- pathway construction in different hosts and pathway optimization, separation and characterization of metabolites, analysis of natural products
- Enzymology: enzyme activity assays and assay development, enzyme kinetics (UV-Vis spectroscopy, Fluorescence spectroscopy. HPLC, GC, IR, Mass spec, NMR)
- Protein X-ray crystallography, crystallization and protein structure resolution
- Chemical synthesis:- of metabolites/intermediates, cross coupling chemistry and developing chemo-enzymatic reactions.