A BioBricks Metabolic Engineering Platform for the Biosynthesis of Anthracyclinones in Streptomyces coelicolor

• DOI: 10.1021/acssynbio.2c00498
• Publication Date: 2022-11-15T16:47:15Z
• Authors: Rongbin Wang; Jennifer Nguyen; Jacob Hecht; Nora Schwartz; Katelyn V. Brown; Larissa V. Ponoma...; Magdalena Niemczura; Dino van Dissel; Gilles P. van Wezel; Jon S. Thorson; Mikko Metsä-Ketelä; Khaled A. Shaaban; S. Eric Nybo
• Keywords: Metabolic Engineering; Streptomycetaceae; Synthetic Biology

Actinomycetes produce a variety of clinically indispensable molecules, such as antineoplastic anthracyclines. However, the actinomycetes are hindered in their further development genetically engineered hosts for synthesis new anthracycline analogues due to slow growth kinetics associated with mycelial life cycle and lack comprehensive genetic toolbox combinatorial biosynthesis. In this report, we tackled both issues via BIOPOLYMER (BIOBricks POLYketide Metabolic EngineeRing) toolbox: synthetic biology consisting strains, promoters, vectors, biosynthetic genes anthracyclinones. An improved derivative production host Streptomyces coelicolor M1152 was created by deleting matAB gene cluster that specifies extracellular poly-β-1,6-N-acetylglucosamine (PNAG). This resulted loss aggregation, biomass accumulation anthracyclinone production. We then leveraged engineer four distinct pathways, identifying optimal combinations genes, vectors aklavinone, 9-epi-aklavinone, auramycinone, nogalamycinone at titers between 15-20 mg/L. Optimization strains 103 structurally characterized six products from fermentations, including compounds 9,10-seco-7-deoxy-nogalamycinone 4-O-β-d-glucosyl-nogalamycinone. Lastly, tested antiproliferative activity anthracyclinones mammalian cancer cell viability assay, which nogalamycinone, aklavinone exhibited moderate cytotoxicity against several lines. envision will serve foundational platform technology designer analogues.