could give rise to a powerful biocatalytic technique for the preparation of synthetic alkaloids,
Objectives: In this project we will enable the synthesis of nicotine-like molecules through asymmetric arylation reactions catalysed by enzymes of the nicotine biosynthetic pathway.
Experimental Approach: (1): Reconstitution of the pathway for (S)-nicotine synthesis for scalable production: We have already established robust heterologous systems for the preparation of the four major constitutive enzymes of (S)-nicotine production: We will first aim to scale our current mg scale reactions using this system, and then also apply it to the synthesis of related natural alkaloids. (2): Mutant libraries of relevant enzymes for the transformation of non-natural nicotine-like alkaloids: We will create mutant libraries of both nicotine biosynthesis enzymes to develop mutants capable of the efficient transformation of substrate analogs with a view to constructing in vitro pathways for the production of pharmaceutically active compounds (3): Construction of an in vivo system in S. cerevisiae for the production of nicotine analogs from simple precursors using asymmetric biocatalytic cross coupling. Nicotine pathway genes, and associated transporters, will be introduced to an S. cerevisiae chassis. Substrate analogs will be introduced through de novo biosynthesis or media supplementation to enable in vivo formation of nicotine analogs.
Novelty: This project will demonstrate the first application of nicotine biosynthesis enzymes in ex vivo systems for the synthesis of new alkaloids.
Training: The student appointed to the project will be supervised in the Departments of Chemistry (Grogan, Fascione) and Biology (Lichman) at York. They will be trained in molecular biology, gene cloning, mutation and protein purification in the Grogan and Lichman labs. Organic synthesis and analysis will be performed in the Fascione lab.
About the programme
Join experts in industry and academia working to sustainably manufacture the complex and diverse molecules needed by modern society.
Industrial manufacturing is at a turning point. Many conventional production routes rely on non-renewable resources, harmful chemicals, and energy-intensive steps. Biocatalysis using engineered enzymes offers a proven solution.
Led by The University of Manchester in collaboration with AstraZeneca, The Universities of Bristol and York alongside other leading industrial partners, BioProcess aims to train the next generation of scientists in the skills needed to realise full the potential of biocatalysis, protein engineering and biomanufacturing for the UK bioeconomy.
