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Applied and Environmental Microbiology, October 2009, p. 6367-6372, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00019-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cell-Free Escherichia coli-Based System To Screen for Quorum-Sensing Molecules Interacting with Quorum Receptor Proteins of Streptomyces coelicolor{triangledown}

Yung-Hun Yang,1 Tae-Wan Kim,1 Sung-Hee Park,1 Kwangwon Lee,1 Hyung-Yeon Park,2 Eunjung Song,1 Hwang-Soo Joo,1 Yun-Gon Kim,1 Ji-Sook Hahn,1 and Byung-Gee Kim1*

School of Chemical and Biological Engineering, Seoul National University, Seoul, Republic of Korea,1 Department of Chemistry, In-ha University, In-Chon, Kyung-Gi Do, Republic of Korea2

Received 5 January 2009/ Accepted 4 August 2009

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 Escherichia coli-based expression system to identify QS molecules of Streptomyces. The binding of QS molecules to {gamma}-butyrolactone receptor ScbR was monitored by changes in the expression levels of the green fluorescent protein reporter in E. coli cell extract. Using this assay system, we could successfully confirm SCB1, a {gamma}-butyrolactone molecule in Streptomyces coelicolor, binding to its known receptor, ScbR. In addition, we have shown that N-hexanoyl-DL-homoserine lactone, one of the QS molecules in many gram-negative bacteria, can regulate ScbR and trigger precocious antibiotic production in S. coelicolor. Our new method can be applied to other strains for which a screening tool for QS molecules has not yet been developed.

* Corresponding author. Mailing address: School of Chemical and Biological Engineering, Seoul National University, Seoul, Republic of Korea. Phone: 82-2-880-6774. Fax: 82-2-873-6020. E-mail: byungkim{at}snu.ac.kr

{triangledown} Published ahead of print on 14 August 2009.

Applied and Environmental Microbiology, October 2009, p. 6367-6372, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00019-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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