Project description
Harnessing nature's other path to peptide synthesis to generate new therapeutics
Normally, protein synthesis is accomplished with the help of ribosomes that stitch amino acids together to form the peptides that make a protein with the help of mRNA. Bacteria, fungi, and some marine microorganisms also have nonribosomal peptides, so called because their synthesis does not rely on ribosomes but rather on nonribosomal peptide synthetases (NRPSs). NRPSs are large, modular enzyme complexes whose manipulation and control could result in the production of novel new-to-nature peptides with important practical applications. SYNPEP has developed ways to do just that using low-cost bacterial cultures with excellent yields. They are now planning to expand their production capability and repertoire while exploiting screens for bioactivity with the goal of revolutionising peptide synthesis for new therapeutics.
Objective
Natural products (NPs) generated by microbial non-ribosomal peptide synthetases (NRPS) represent several very important and valuable clinical antibiotics, immune-suppressive and anti-cancer drugs. NPs have gone on to inspire several synthetic peptides that are used clinically, but contain amino acids (AAs) or other building blocks that are not found in nature. However, with >500 identified AAs and additional peptide modifications like glycosylation or cyclization, the chemical diversity in NRPS-derived peptides is far larger than proteins and has not yet been fully explored. The modular nature of NRPS suggests the possibility to manipulate them, subsequently leading to the production of non-natural NPs. With an eXchange Unit (XU) concept, developed in Photorhabdus, Xenorhabdus and Bacillus, we have recently identified efficient ways for NRPS manipulation enabling the de novo assembly of novel NRPS for the production of new-to-nature NPs in excellent production yields of >250 mg/L. Within SYNPEP we will expand this approach to other bacterial genera producing peptide NPs. We will identify unusual NRPS systems, analyse them bioinformatically, validate the function of novel NRPS units experimentally and combine high-throughput molecular biology, microfluidics for bioactivity screening, rapid NP identification and structure elucidation to produce potentially any peptide or a peptide library of 2-15 amino acids in <4 weeks, in a semi-automated manner. In contrast to chemical peptide synthesis this production pipeline is more economical, sustainable and scalable. The NPs are produced by bacterial cells in aqueous media using cheap energy sources and the bacterial cultures can be easily scaled up when larger NP amounts are needed. We will also develop NRPS units that accept synthetic building blocks currently not found in natural NRPS. These ‘synthetic’ NRPS units will enable the simplified chemical derivatization of the produced NPs for further compound diversification.
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Funding Scheme
ERC-ADG - Advanced GrantHost institution
60323 Frankfurt Am Main
Germany