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)

被引:11
作者
Kim, Min Woo [1 ]
Lee, Bo-Rahm [2 ,3 ]
You, SungYong [4 ,5 ]
Kim, Eun-Jung [2 ,3 ]
Kim, Ji-Nu [2 ,3 ]
Song, Eunjung [2 ,3 ]
Yang, Yung-Hun [6 ]
Hwang, Daehee [7 ,8 ]
Kim, Byung-Gee [1 ,2 ,3 ]
机构
[1] Seoul Natl Univ, Interdisciplinary Program Biochem Engn, Seoul, South Korea
[2] Seoul Natl Univ, Inst Mol Biol & Genet, Sch Chem & Biol Engn, Seoul, South Korea
[3] Seoul Natl Univ, Bioengn Inst, Seoul, South Korea
[4] Cedars Sinai Med Ctr, Div Canc Biol & Therapeut, Dept Surg, Los Angeles, CA 90048 USA
[5] Cedars Sinai Med Ctr, Div Canc Biol & Therapeut, Dept Biomed Sci, Los Angeles, CA 90048 USA
[6] Konkuk Univ, Dept Microbial Engn, Coll Engn, Seoul, South Korea
[7] DGIST, Dept New Biol, Inst Basic Sci, Daegu, South Korea
[8] DGIST, Ctr Plant Aging Res, Inst Basic Sci, Daegu, South Korea
基金
新加坡国家研究基金会;
关键词
Combination of transcriptional regulators; Clustering analysis; Time-series transcriptome; Actinorhodin; Streptomyces coelicolor; SECONDARY METABOLITE BIOSYNTHESIS; GENE-EXPRESSION; PROTEIN; CLUSTER; DISCOVERY; LIVIDANS; ADPA; PHOP;
D O I
10.1007/s00253-018-8838-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 "synergistic 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 (Delta rsfA 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.
引用
收藏
页码:3243 / 3253
页数:11
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