Abstract
Genome sequencing has revealed that each Streptomyces contains a wide range of biosynthetic gene clusters (BGCs) and has the capability to produce more novel natural products than what is expected. However, most gene clusters for secondary metabolite biosynthesis are cryptic under normal growth conditions. In Streptomyces tsukubaensis, combining overexpression of the putative SARPs (Streptomyces antibiotic regulatory proteins) and bioactivity-guided screening, the silent gene cluster (tsu) was successfully activated and a novel bioactive anthracycline tsukubarubicin was further isolated and identified. Biological activity assays demonstrated that tsukubarubicin possessed much better antitumor bioactivities against various human cancer cell lines (especially the breast cancer cell lines) than clinically used doxorubicin. Moreover, the previously unreported gene cluster (tsu) for biosynthesis of tsukubarubicin was first characterized and detailed annotations of this gene cluster were also conducted. Our strategy presented in this work is broadly applicable in other Streptomyces and will assist in enriching the natural products for potential drug leads.
Key points
• Generally scalable strategy to activate silent gene clusters by manipulating SARPs.
• The novel anthracycline tsukubarubicin with potent antitumor bioactivities.
• Identification and annotation of the previously uncharacterized tsu gene cluster.
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Data availability
The datasets used during the current study are available from the corresponding author on reasonable request.
Code availability
Not applicable.
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We thank every people participating in this work.
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This research was sponsored by the NSFC key projects (no. 31520103901 and no. 31730002) and the National Key R&D Program of China (no. 2019YFA09005400).
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QBW carried out the experiments and wrote the manuscript. ZYL, XYZ, and YL assisted with the experiments (fermentation, construction of plasmids, and analysis of NMR data, respectively). QBW, YQL, and XAC conceived and designed the experiments. QBW and YQL revised the manuscript. All authors read and approved the final manuscript.
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Wu, QB., Chen, XA., Lv, ZY. et al. Activation and discovery of tsukubarubicin from Streptomyces tsukubaensis through overexpressing SARPs. Appl Microbiol Biotechnol 105, 4731–4741 (2021). https://doi.org/10.1007/s00253-021-11344-5
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DOI: https://doi.org/10.1007/s00253-021-11344-5