Enhanced productivity and functionality of Modified Ribosomally Produced Peptides (M-RIPPs)

Lead Research Organisation: University of Oxford
Department Name: Structural Biology

Abstract

Ribosomally Produced Peptides (RIPPs) are widely recognised as one of the most promising classes of compounds with the potential to treat many diseases including infection, cancer & inflammation. They are of great interest to the pharma industry, but are extremely costly to produce/modify - even in milligram amounts. Through the utilisation of cutting-edge techniques in combinatorial synthetic biology, this project sets out to achieve a world first; namely, to produce bespoke libraries of Modified RIPPs (M-RIPPs) in sufficient quantities to permit drug discovery screening. The project combines the fundamental knowledge of the natural processes involved in RIPP biosynthesis of the two premier UK academic groups active in the field with the applied expertise in industrial biosynthesis of a leading UK IB company. It will deliver a versatile yet robust technology platform for the production of M-RIPPs that will be transferred to a spinoff company to be formed around 18 months after project start.

Technical Summary

Ribosomally Produced Peptides (RIPPs) as a biosynthetic class contains many different families such as the cyanobactins, lanthipeptides, proteusins and lasso peptides amongst others. They have a range of biological activities and have a common biosynthesis in which a the core peptide, a small sequences within a larger precursor peptide is modified by tailoring enzymes. The modified core peptide is freed from the leader and additional signal sequences and often undergoes further modification (macrocycle formation, heterocycle oxidation, O/N prenylation) to produce the final modified peptide. The common RIPP biosynthetic pathway indicates that it should be possible to apply multiple types of chemical tailoring from different RIPP families to core peptides, thus generating hybrid molecules with features from multiple RIPPs. This project aims to generate such modified RIPPs (M-RIPPS) and overcome several barriers to their scaleable production. We will incorporate modifications common in cyanobactins (heterocycles, macrocycles, O/N prenylation), lanthipeptides (lanthionine and labionin bridges) and lass peptides. We will use this methodology to generate libraries of unique compounds with novel bioactivities. To assist with M-RIPPS that may not be producible using standard expression systems, we will also use one alternative expression system. We will scale up production to 1-3 L scale and improve downstream processing using a variety of methods, including the incorporation of a cyclic peptide exporter in the producing cells and cultivating them in a biphasic system to allow easy compound extraction and subsequent purification. The final step is technical marketing in consultation with big Pharma with a view to establishing a spinoff company based on this technology.

Planned Impact

As described in proposal submitted to IUK

Related Projects

Project Reference Relationship Related To Start End Award Value
BB/M028461/1 01/08/2015 31/08/2017 £330,521
BB/M028461/2 Transfer BB/M028461/1 01/09/2017 30/11/2018 £143,431
 
Description Our main break through was the discovery that the macrocyclase enzymes from plants and fungi could be used for biotechnology applications. These enzymes were known but how they handle their substrates was not. We were able to determine how they work and then modify the reaction so they could use other substrates. We have also discovered an approach that yields hybrid macrocycles. These molecules are potential drug leads.
Exploitation Route We are forming a company, Gyreox. Ingenza are exploiting the IP.
Sectors Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Our findings are been taken forward by Ingenza. Our major discovery was that it was possible to use the POP class of macrocyclases to replace the PatGmac class of macrocyclase. We were able from structural study evaluate the key determinants of recognition. This allows more practical and atom efficient substrates to be employed. They are being used to make biologically active macrocycles.
First Year Of Impact 2018
Sector Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Description Ingenza 
Organisation Ingenza Ltd
Country United Kingdom 
Sector Private 
PI Contribution Our task was to improve the macrocyclisation of peptides. Having identified the enzymes we modified their behaviour in order for them to work on more chemically relevant problems.
Collaborator Contribution Ingenza were able to construct polycistronic sequences with individual genes under the control of different promoters. This enabled the organisms
Impact We created new technology for macrocyclising peptides
Start Year 2017
 
Description Schools open day at Harwelll 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact The purpose is to showcase the work of the Harwell campus to regional school children. Each year we run this event changing the focus slightly. Pupils get hands on experiments, tours and talks. It is a very popular event for which we have to ration attendance.
Year(s) Of Engagement Activity 2017,2018
 
Description Visit to local School (Newbury) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact I spoke with prospective A level students about chemical biology and enzymes as part of their course choice. This was a chance for them to see chemistry in a more rounded way than taught at school. Around 12 to 20 pupils attended. I showed them how to use the web to look at enzymes and talked through mechanism. A show of hands indicated an increased interest in studying chemistry and an appreciation of its role in biology.
Year(s) Of Engagement Activity 2018