A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
A BioBricks Metabolic Engineering Platform for the Biosynthesis of Anthracyclinones in Streptomyces coelicolor
Tekijät: Wang Rongbin, Nguyen Jennifer, Hecht Jacob, Schwartz Nora, Brown Katelyn V, Ponomareva Larissa V, Niemczura Magdalena, van Dissel Dino, van Wezel Gilles P, Thorson Jon S, Metsä-Ketelä Mikko, Shaaban Khaled A, Nybo S Eric
Kustantaja: American Chemical Society
Julkaisuvuosi: 2022
Journal: ACS Synthetic Biology
Tietokannassa oleva lehden nimi: ACS synthetic biology
Lehden akronyymi: ACS Synth Biol
Vuosikerta: 11
Numero: 12
Aloitussivu: 4193
Lopetussivu: 4209
ISSN: 2161-5063
eISSN: 2161-5063
DOI: https://doi.org/10.1021/acssynbio.2c00498
Verkko-osoite: https://pubs.acs.org/doi/10.1021/acssynbio.2c00498
Rinnakkaistallenteen osoite: https://research.utu.fi/converis/portal/detail/Publication/177538147
Actinomycetes produce a variety of clinically indispensable molecules, such as antineoplastic anthracyclines. However, the actinomycetes are hindered in their further development as genetically engineered hosts for the synthesis of new anthracycline analogues due to their slow growth kinetics associated with their mycelial life cycle and the lack of a comprehensive genetic toolbox for combinatorial biosynthesis. In this report, we tackled both issues via the development of the BIOPOLYMER (BIOBricks POLYketide Metabolic EngineeRing) toolbox: a comprehensive synthetic biology toolbox consisting of engineered strains, promoters, vectors, and biosynthetic genes for the synthesis of anthracyclinones. An improved derivative of the production host Streptomyces coelicolor M1152 was created by deleting the matAB gene cluster that specifies extracellular poly-β-1,6-N-acetylglucosamine (PNAG). This resulted in a loss of mycelial aggregation, with improved biomass accumulation and anthracyclinone production. We then leveraged BIOPOLYMER to engineer four distinct anthracyclinone pathways, identifying optimal combinations of promoters, genes, and vectors to produce aklavinone, 9-epi-aklavinone, auramycinone, and nogalamycinone at titers between 15-20 mg/L. Optimization of nogalamycinone production strains resulted in titers of 103 mg/L. We structurally characterized six anthracyclinone products from fermentations, including new compounds 9,10-seco-7-deoxy-nogalamycinone and 4-O-β-d-glucosyl-nogalamycinone. Lastly, we tested the antiproliferative activity of the anthracyclinones in a mammalian cancer cell viability assay, in which nogalamycinone, auramycinone, and aklavinone exhibited moderate cytotoxicity against several cancer cell lines. We envision that BIOPOLYMER will serve as a foundational platform technology for the synthesis of designer anthracycline analogues.
Ladattava julkaisu This is an electronic reprint of the original article. |  |
