These tools will be tested with peptidic NP biosynthetic pathways involving cyclodipeptide synthases or non-ribosomal peptide synthetases. Moreover, this will allow the design of new pathways, and the production of new compounds, by the co-localization of enzymes originating from different pathways (combinatorial biosynthesis). This will position these enzymes close to each other, increasing the efficiency of natural biosynthetic pathways. We propose to develop novel systems to spatially organize biosynthetic enzymes fused with alphaRep. It is thus possible to generate, in a fast and efficient manner, a set of pairs of orthogonal proteins, each one interacting specifically with a single partner. It is possible to select alphaRep capable of specifically binding any target protein, natural or artificial, arbitrarily chosen. AlphaReps have a curved solenoid shape whose concave region forms a hypervariable surface of interaction. They can therefore be easily fused to other protein partners. AlphaReps are folded, extremely stable, and not prone to aggregation. We propose to use a new family of artificial proteins, alphaReps to develop new scaffolding tools. These scaffolding approaches require the development of powerful, modular and easy-to-use tools. This can be done by fusing catalytic domains with binding domains so that different enzymes bind to the same molecule (DNA, RNA, protein). Synthetic biology approaches aim at bringing together and spatially organizing in the cell the enzymes involved in the same pathway. However the enzymes of a metabolic pathway can be organized in space in order to improve the overall pathway’s efficiency. Each step can be performed independently of others. The biosynthesis of NPs involves an often complex succession of enzyme-catalyzed steps. However, synthetic biology of NPs is still in its infancy and novel and powerful tools are needed to facilitate its development. In the area of NPs, synthetic biology can contribute to the discovery of novel NPs by expressing NP biosynthetic gene clusters which are silent in laboratory conditions, to the improvement of NP biosynthetic yields, or to the biosynthesis of new NPs by the reprogramming of biosynthetic assembly lines. Synthetic biology, which aims at rationally modifying living organisms to give them new functions, can contribute to this search for bioactive molecules. ) new bioactive molecules are actively sought, notably among NPs. In many fields (anti-infectives, anti-tumor. The diversity of NPs has not been exhaustively explored nor exploited. Natural products (NPs) are small bioactive molecules produced by living organisms, often microorganisms.

Molecular scaffold protein