The ongoing increase in antimicrobial resistance combined with the low discovery of novel antibiotics is a serious threat to our health care. Genome mining has given new potential to the field of... Show moreThe ongoing increase in antimicrobial resistance combined with the low discovery of novel antibiotics is a serious threat to our health care. Genome mining has given new potential to the field of natural product discovery, as thousands of biosynthetic gene clusters (BGCs) are discovered for which the natural product is not known.Ribosomally synthesized and post-translationally modified peptides (RiPPs) represent a highly diverse class of natural products. The large number of different modifications that can be applied to a RiPP results in a large variety of chemical structures, but also stems from a large genetic variety in BGCs. As a result, no single method can effectively mine for all RiPP BGCs, making it an interesting source for new molecules.In this thesis, new methods are explored to mine genomes for the BGCs of novel RiPP variants, with a focus on discovering RiPPs that have new modifications. RRE-Finder is a new tool for the detection of RiPP Recognition Elements, domains that are often found in RiPP BGCs. DecRiPPter is another tool that employs machine learning models to discover new RiPP precursor genes encoded in the genomes. Both tools can be used to prioritize novel RiPP BGCs. Two candidate BGCs are characterized, one of which could be shown to specify a new RiPP, validating the approach. Show less
Streptomyces are multicellular bacteria that grow as branched filaments and are best known for producing the majority of our antibiotics, many immunosuppressant and anticancer compounds.... Show moreStreptomyces are multicellular bacteria that grow as branched filaments and are best known for producing the majority of our antibiotics, many immunosuppressant and anticancer compounds. Unfortunately their multicellular life style creates many problems for efficient industrial production. In a bioreactor, depending on the environment and the genetics, it can grow quickly as dispersed mycelia or aggregate in slow growing pellets. Either morphology has advantages and disadvantages, which can be product specific. For my thesis I studied the mechanism by which these filaments can aggregate into dense pellets. I found a small gene cluster that produces poly-1,6-N-acetylglucosamine, a bacterial glue which binds neighboring cells and required for pellet formation in S. coelicolor. Subsequently we can use these genes to control the morphology of streptomycetes in a liquid environment, tailoring it for production. My work has given us new insights in the mechanims through which streptomycetes aggregate, but also has the potential to make streptomycetes a more favorable host for industial production. Show less
