A synthetic biology approach to the production of terpenoids by pathway engineering.

Lead Research Organisation: University of Leeds
Department Name: Astbury Centre

Abstract

Synthetic biology integrates biochemistry and genetic engineering to artificially design and manipulate biological systems in organisms with the aim of remodelling their metabolic pathways to change their function and behaviours. This project will use synthetic biology to engineer natural product biosynthetic pathways. Natural products belong to an extensive family of diverse organic molecules with in excess of 200, 000 discovered and extracted from various sources and their importance is emphasised by their significance to the pharmaceutical industry for over 60 years, delivering novel antibiotics, hormones and anti-tumour agents to the therapeutic drug repertoire. Over the last 25 years, one quarter of pharmaceuticals are isolated natural products, or designed based on their structure.

Of the various classes of natural product, the isoprene family comprises over 40,000 distinct compounds and engineering of their biosynthetic pathways has already led to biotechnologically significant routes to isoprene-derived pharmaeuticals such as taxol and artemisinin. All isoprenes are derived from IPP (isopentenyl diphosphate) and DMPP (dimethylallyl diphosphate) via intermediates such as geranyl diphosphate (C10), farnesyl diphosphate (C15) and geranylgeranyl diphosphate (C20). Downstream enzymes then catalyse a range of chemical modifications to cyclise, oxidise, reduce, isomerise and hydroxylate the prenyldiphosphate intermediates to generate the respective terpenoid products. The tolerant nature of these terpene synthases presents engineering opportunities, modifying the enzyme structure to either synthesise different products or provide a preference for a desired product.

This project will use synthetic biology methods to optimise precursor concentrations either by native pathway optimisation or transportation of a heterologous pathway, in combination with heterologous expression of specific terpene synthase enzymes such the promiscuous y-humulene synthase, followed by structural studies and rational engineering of terpene synthases to bias the formation of desired terpenes from single substrate compounds.

Publications

10 25 50

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011151/1 30/09/2015 29/09/2023
1774730 Studentship BB/M011151/1 30/09/2016 30/11/2020 Emily Turri
 
Description So far the project has adapted an assay to be able to calculate kinetic parameters of terpene synthase. The remaining project will work to find residues that can be mutated to alter the product profile.
Exploitation Route The findings could be used to produce novel terpenes using the residues identified as potential hotspots.
Sectors Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description Astbury Conversation Public Engagement Event 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact As part of a larger public engagement event held at the the biannual Astbury conversation, my lab group presented our work alongside a a general more approachable overview of enzymes and how they relate to modern day society. The aim of the overall event was to make science more approachable to the general public and a selection of local sixth form students. This event was accompanied with a lecture from Professor Michael Levitt aimed at the general public. A specific aim was to encourage the sixth from students to pursue careers in science while encouraging the public to have interest or curiosity in science and its impact on our future. For this we provided experiments based around bio and non bio washing powder and the effect enzymes can have using an egg stain as the dirt. This stall was alongside stalls covering electron microscopy, GPCRs and sugar molecules.
Year(s) Of Engagement Activity 2018
URL https://astburyconversation.leeds.ac.uk/ehome/cte12345/previousevents/
 
Description Astbury Society 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact The astbury society creates social events alongside seminars to encourage collaboration between departments within the centre and invited speakers. The events include monthly seminars, annual lecture, conferences, cheese and wine and charity balls.
Year(s) Of Engagement Activity 2017,2018
URL http://www.astbury.leeds.ac.uk/about/society.php