Team:Heidelberg/RFCs

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RFC 100

RFC 100: Standard for Synthesis of Customized Peptides by Non-Ribosomal Peptide Synthetases

With this RFC, we introduce a standardized framework for engineering customizable non-ribosomal peptide synthetases (NRPS) and their application for the in vivo and in vitro synthesis of short synthetic peptides of user-defined sequence. The framework consists of the NRPSDesigner, a software tool for the in silico design of user-defined NRPSs, a platform for standardized cloning and expression of NRPSs in different bacterial hosts and a quality control procedure for the easy validation of NRP production.

With this novel framework, we seek to overcome the two major challenges present in the synthetic peptides field today: making peptide production flexible and scalable at the same time. Chemical peptide synthesis is flexible and enables the incorporation of non-proteinogenic amino acids. However, it is relatively expensive and uneconomical when used for large-scale peptide production. In contrast, producing recombinant peptides may be cheaper, but their composition is mostly restricted to only 22 different building blocks.

Our framework combines the advantages of chemical and recombinant peptide synthesis: it enables the incorporation of hundreds of non-proteinogenic amino acids into custom peptides and is at the same time easily scalable. Our framework will highly simplify the production of custom peptides, including peptide antibiotics, detoxifying agents or specific peptide-based chelators applicable for recycling of metals from electronic waste. We thus believe, that RFC 100 represents a great foundational advance in the synthetic peptide production field.

Thanks to