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

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.

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.

As described in proposal submitted to IUK