| Autor: |
Dolya B; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Hryhorieva O; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Sorochynska K; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Lopatniuk M; Department of Pharmacy, Saarland University, 66123 Saarbrucken, Germany., Ostash I; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Tseduliak VM; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Sterndorff EB; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, 2800 Kongens Lyngby, Denmark., Jørgensen TS; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, 2800 Kongens Lyngby, Denmark., Gren T; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, 2800 Kongens Lyngby, Denmark., Dacyuk Y; Department of Mineral Geology and Geophysics, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Weber T; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, 2800 Kongens Lyngby, Denmark., Luzhetskyy A; Department of Pharmacy, Saarland University, 66123 Saarbrucken, Germany., Fedorenko V; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine., Ostash B; Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 79005 Lviv, Ukraine. |
| Abstrakt: |
Streptomyces albidoflavus J1074 is a popular platform to discover novel natural products via the expression of heterologous biosynthetic gene clusters (BGCs). There is keen interest in improving the ability of this platform to overexpress BGCs and, consequently, enable the purification of specialized metabolites. Mutations within gene rpoB for the β-subunit of RNA polymerase are known to increase rifampicin resistance and augment the metabolic capabilities of streptomycetes. Yet, the effects of rpoB mutations on J1074 remained unstudied, and we decided to address this issue. A target collection of strains that we studied carried spontaneous rpoB mutations introduced in the background of the other drug resistance mutations. The antibiotic resistance spectra, growth, and specialized metabolism of the resulting mutants were interrogated using a set of microbiological and analytical approaches. We isolated 14 different rpoB mutants showing various degrees of rifampicin resistance; one of them (S433W) was isolated for the first time in actinomycetes. The rpoB mutations had a major effect on antibiotic production by J1074, as evident from bioassays and LC-MS data. Our data support the idea that rpoB mutations are useful tools to enhance the ability of J1074 to produce specialized metabolites. |
