Development of a Burkholderia Bacterial Host for Natural Product Discovery and Production

Burkholderia bacteria are multifaceted organisms that are found in diverse ecological environments. They have emerged as a promising source of natural products with applications in medicine and agriculture. However, a challenge of working with Burkholderia strains is that many are pathogenic to animals or plants and some are hard-to-culture symbionts. Heterologous expression of biosynthetic gene clusters (BGCs) provide an opportunity to streamline and accelerate natural product discovery and production. My dissertation focuses on the development of a Burkholderia host, strain FERM BP-3421, as a platform to facilitate natural product discovery and production. The aims of my dissertation are to 1) test our host with model biosynthetic gene clusters, 2) apply our host to discover unknown natural products, and 3) gain a deeper understanding of our host through multi-omic approaches. As a proof-of-concept, we chose to express a lasso peptide BGC encoding the biosynthesis of capistruin. Our results demonstrate that our host, strain FERM BP-3421, can produce the model lasso peptide capistruin at up to 580 times more than the previously reported E. coli host. We next apply this host in genome mining efforts towards natural product discovery from Burkholderia sources. Lastly, we present the complete genome of FERM BP-3421, connect genomic and metabolomic information, and elucidate the methylome. We envision that FERM BP-3421 will serve as a good alternative to existing hosts towards discovery and production of natural products from Burkholderia bacteria.