Genetic approaches to improve clorobiocin production in Streptomyces roseochromogenes NRRL 3504

Streptomyces roseochromogenes NRRL 3504 is best known as a producer of clorobiocin, a DNA replication inhibitor from the aminocoumarin family of antibiotics. This natural product currently draws attention as a promising adjuvant for co-application with other antibiotics against Gram-negative multidr...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2022-02, Vol.106 (4), p.1543-1556
Main Authors: Melnyk, Sofia, Stepanyshyn, Anastasia, Yushchuk, Oleksandr, Mandler, Michael, Ostash, Iryna, Koshla, Oksana, Fedorenko, Victor, Kahne, Daniel, Ostash, Bohdan
Format: Article
Language:English
Subjects:
Anti-Bacterial Agents - pharmacology
Antibiotics
Applied Genetics and Molecular Biotechnology
Biomedical and Life Sciences
Biotechnology
Conjugation
DNA biosynthesis
E coli
Gene expression
Genetic aspects
Life Sciences
Microbial Genetics and Genomics
Microbiology
Multidrug resistance
Natural products
Novobiocin - analogs & derivatives
Plasmids
Streptomyces
Streptomyces - metabolism
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Summary:Streptomyces roseochromogenes NRRL 3504 is best known as a producer of clorobiocin, a DNA replication inhibitor from the aminocoumarin family of antibiotics. This natural product currently draws attention as a promising adjuvant for co-application with other antibiotics against Gram-negative multidrug-resistant pathogens. Herein, we expand the genetic toolkit for NRRL 3504 by showing that a set of integrative and replicative vectors, not tested previously for this strain, could be conjugally transferred at high frequency from Escherichia coli to NRRL 3504. Using this approach, we leverage a cumate-inducible expression of cluster-situated regulatory gene novG to increase clorobiocin titers by 30-fold (up to approximately 200 mg/L). To our best knowledge, this is the highest level of clorobiocin production reported so far. Our findings set a working ground for further improvement of clorobiocin production as well as for the application of genetic methods to illuminate the cryptic secondary metabolome of NRRL 3504. Key Points • Efficient system for conjugative transfer of plasmids into NRRL 3504 was developed. • Expression of regulatory genes in NRRL 3504 led to increase in clorobiocin titer. • Secondary metabolome of NRRL 3504 becomes an accessible target for genetic manipulations using the expanded vector set and improved intergeneric conjugation protocol.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-022-11814-4