Transcriptome analysis of wild-type and afsS deletion mutant strains identifies synergistic transcriptional regulator of afsS for a high antibiotic-producing strain of Streptomyces coelicolor A3(2)
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- Transcriptome analysis of wild-type and afsS deletion mutant strains identifies synergistic transcriptional regulator of afsS for a high antibiotic-producing strain of Streptomyces coelicolor A3(2)
- Min Woo Kim; Bo-Rahm Lee; SungYong You; Eun-Jung Kim; Ji-Nu Kim; Eunjung Song; Yung-Hun Yang; Daehee Hwang; Byung-Gee Kim
- APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.102, no.7, pp.3242 - 3253
- Most secondary metabolism in Actinobacteria is controlled by multi-layered, gene-regulatory networks. These regulatory mechanisms
are not easily identified due to their complexity. As a result, when a strong transcriptional regulator (TR) governs
activation of biosynthetic pathways of target antibiotics such as actinorhodin (ACT), additional enhancement of the biosynthesis
is difficult in combination with other TRs. To find out any Bsynergistic transcriptional regulators (sTRs)^ that show an additive
effect on the major, often strong, transcriptional regulator (mTR), here, we performed a clustering analysis using the transcriptome
datasets of an mTR deletion mutant and wild-type strain. In the case of ACT biosynthesis in Streptomyces coelicolor,
PhoU (SCO4228) and RsfA (SCO4677) were selected through the clustering analysis, using AfsS (SCO4425) as a model mTR,
and experimentally validated their roles as sTRs. Furthermore, through analysis of synergistic effects, we were able to suggest a
novel regulation mechanism and formulate a strategy to maximize the synergistic effect. In the case of the double TR mutant
strain (ΔrsfA pIBR25::afsS), it was confirmed that the increase of cell mass was the major cause of the synergistic effect.
Therefore, the strategy to increase the cell mass of double mutant was further attempted by optimizing the expression of efflux
pump, which resulted in 2-fold increase in the cell mass and 24-fold increase in the production of ACT. This result is the highest
ACT yield from S. coelicolor ever reported.
(c) Springer-Verlag GmbH Germany, part of Springer Nature 2018
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