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Yang, Yung-Hun,Song, Eunjung,Lee, Bo-Rahm,Kim, Eun-jung,Park, Sung-Hee,Kim, Yun-Gon,Lee, Chang-Soo,Kim, Byung-Gee American Society for Microbiology 2010 Applied and environmental microbiology Vol.76 No.11
<B>ABSTRACT</B><P>To elucidate the function of an unknown regulator in <I>Streptomyces</I>, differences in phenotype and antibiotic production between a deletion mutant and a wild-type strain (WT) were compared. These differences are easily hidden by complex media. To determine the specific nutrient conditions that reveal such differences, we used a multiwell method containing different nutrients along with bromothymol blue. We found several nutrients that provide key information on characterization conditions. By comparing the growth of wild-type and mutant strains on screened nutrients, we were able to measure growth, organic acid production, and antibiotic production for the elucidation of regulator function. As a result of this method, a member of the MarR-like regulator family, SCO5405 (AbsC), was newly characterized to control pyruvate dehydrogenase in <I>Streptomyces coelicolor</I>. Deletion of SCO5405 increased the pH of the culture broth due to decreased production of organic acids such as pyruvate and α-ketoglutarate and increased extracellular actinorhodin (ACT) production in minimal medium containing glucose and alanine (MMGA). This method could therefore be a high-throughput method for the characterization of unknown regulators.</P>
Yang, Yung-Hun,Kim, Tae-Wan,Park, Sung-Hee,Lee, Kwangwon,Park, Hyung-Yeon,Song, Eunjung,Joo, Hwang-Soo,Kim, Yun-Gon,Hahn, Ji-Sook,Kim, Byung-Gee American Society for Microbiology 2009 Applied and environmental microbiology Vol.75 No.19
<B>ABSTRACT</B><P>Quorum sensing (QS) is mediated by small molecules and involved in diverse cellular functions, such as virulence, biofilm formation, secondary metabolism, and cell differentiation. In this study, we developed a rapid and effective screening tool based on a cell-free <I>Escherichia coli</I>-based expression system to identify QS molecules of <I>Streptomyces</I>. The binding of QS molecules to γ-butyrolactone receptor ScbR was monitored by changes in the expression levels of the green fluorescent protein reporter in <I>E. coli</I> cell extract. Using this assay system, we could successfully confirm SCB1, a γ-butyrolactone molecule in <I>Streptomyces coelicolor</I>, binding to its known receptor, ScbR. In addition, we have shown that <I>N</I>-hexanoyl-dl-homoserine lactone, one of the QS molecules in many gram-negative bacteria, can regulate ScbR and trigger precocious antibiotic production in <I>S. coelicolor</I>. Our new method can be applied to other strains for which a screening tool for QS molecules has not yet been developed.</P>
Yang, Yung-Hun,Song, Eunjung,Kim, Ji-Nu,Lee, Bo-Rahm,Kim, Eun-Jung,Park, Sung-Hee,Kim, Woo-Seong,Park, Hyung-Yeon,Jeon, Jong-Min,Rajesh, Thangamani,Kim, Yun-Gon,Kim, Byung-Gee Springer-Verlag 2012 Applied microbiology and biotechnology Vol.96 No.1
<P>gamma-Butyrolactones in Streptomyces are well recognized as bacterial hormones, and they affect secondary metabolism of Streptomyces. gamma-Butyrolactone receptors are considered important regulatory proteins, and various gamma-butyrolactone synthases and receptors have been reported in Streptomyces. Here, we characterized a new regulator, SCO0608, that interacted with SCB1 (gamma-butyrolactone of Streptomyces coelicolor) and bound to the scbR/A and adpA promoters. The SCO0608 protein sequences are not similar to those of any known gamma-butyrolactone binding proteins in Streptomyces such as ScbR from S. coelicolor or ArpA from Streptomyces griseus. Interestingly, SCO0608 functions as a repressor of antibiotic biosynthesis and spore formation in R5 complex media. We showed the existence of another type of gamma-butyrolactone receptor in Streptomyces, and this SCO0608 was named ScbR-like gamma-butyrolactone binding regulator (SlbR) in S. coelicolor.</P>