Presentation Information
[P01-082]Exploration of actinomycete secondary metabolites using SARP-type transcriptional activators
○Kotaro Kashihara1, Yuya Misaki1, Yang Liu1, Aiko Teshima1, Kenji Arakawa1 (1. Hiroshima University, Graduate School of Integrated Science for Life (Japan))
Keywords:
Genome mining,Secondary metabolites,Streptomyces,SARP-type activators
[Purpose] Secondary metabolite production in Streptomyces is generally controlled by transcriptional activators, so their overexpression in vivo is an effective method to activate silent secondary metabolism. In this study, we aimed to induce the production of “silent” secondary metabolites by overexpression of Streptomyces antibiotic production regulatory protein (SARP) genes coded on the chromosome of Streptomyces rochei 7434AN4.
[Method and results] Streptomyces rochei 7434AN4 has 40 BGCs and 15 SARP genes. First, we constructed the overexpressed strains, and they were cultivated in YM medium and overexpressed by thiostrepton. Later, the culture broth was extracted with ethyl acetate. As a result of metabolic analysis, we have found that the SRO_3163 overexpressed strain produced a novel cyclohexene-containing enamide(1). We have also found that the SRO_0732 overexpressed strain produced an azoxy-alkene compound, KA57A. This compound was isolated by gene disruption strain of Streptomyces rochei 7434AN4 in our previous study. Surprisingly, even though the BGC for KA57A is situated SRO_1819 to 1850, the production of KA57A was activated by overexpressing SRO_0732, whose genetic distance was 1.27 Mb.
[Conclusion] Here we focused on the function of SARP-type activator genes for activation of silent BGCs in Streptomyces rochei 7434AN4, and some compounds were detected in these overexpressed recombinants. Activation of SARP will be a strong strategy to occasional discovery of microbial natural products.
[Method and results] Streptomyces rochei 7434AN4 has 40 BGCs and 15 SARP genes. First, we constructed the overexpressed strains, and they were cultivated in YM medium and overexpressed by thiostrepton. Later, the culture broth was extracted with ethyl acetate. As a result of metabolic analysis, we have found that the SRO_3163 overexpressed strain produced a novel cyclohexene-containing enamide(1). We have also found that the SRO_0732 overexpressed strain produced an azoxy-alkene compound, KA57A. This compound was isolated by gene disruption strain of Streptomyces rochei 7434AN4 in our previous study. Surprisingly, even though the BGC for KA57A is situated SRO_1819 to 1850, the production of KA57A was activated by overexpressing SRO_0732, whose genetic distance was 1.27 Mb.
[Conclusion] Here we focused on the function of SARP-type activator genes for activation of silent BGCs in Streptomyces rochei 7434AN4, and some compounds were detected in these overexpressed recombinants. Activation of SARP will be a strong strategy to occasional discovery of microbial natural products.
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