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

Overview

Journal Microorganisms

Specialty Microbiology

Date 2023 Jun 15

PMID 37317150

Authors

Borys Dolya

Olena Hryhorieva

Khrystyna Sorochynska

Maria Lopatniuk

Iryna Ostash

Vasylyna-Marta Tseduliak

Eva Baggesgaard Sterndorff

Tue Sparholt Jorgensen

Tetiana Gren

Yuriy Dacyuk

Tilmann Weber

Andriy Luzhetskyy

Victor Fedorenko

Bohdan Ostash

Affiliations

Soon will be listed here.

Abstract

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 for the β-subunit of RNA polymerase are known to increase rifampicin resistance and augment the metabolic capabilities of streptomycetes. Yet, the effects of mutations on J1074 remained unstudied, and we decided to address this issue. A target collection of strains that we studied carried spontaneous 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 mutants showing various degrees of rifampicin resistance; one of them (S433W) was isolated for the first time in actinomycetes. The mutations had a major effect on antibiotic production by J1074, as evident from bioassays and LC-MS data. Our data support the idea that mutations are useful tools to enhance the ability of J1074 to produce specialized metabolites.

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