4.6 Article

Properties of Multidrug-Resistant Mutants Derived from Heterologous Expression Chassis Strain Streptomyces albidoflavus J1074

Journal

MICROORGANISMS
Volume 11, Issue 5, Pages -

Publisher

MDPI
DOI: 10.3390/microorganisms11051176

Keywords

Streptomyces albidoflavus J1074; rifampicin resistance; rpoB; specialized metabolites

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Mutations in the rpoB gene of Streptomyces albidoflavus J1074 have been found to increase its rifampicin resistance and enhance its ability to produce specialized metabolites. Through the isolation and analysis of 14 different rpoB mutants, the study showed that these mutations have a significant impact on the antibiotic production of J1074.
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 fi-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.

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