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Applied and Environmental Microbiology, September 2005, p. 5050-5055, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5050-5055.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel Method for Detection of Butanolides in Streptomyces coelicolor Culture Broth, Using a His-Tagged Receptor (ScbR) and Mass Spectrometry

Yung-Hun Yang,1 Hwang-Soo Joo,1 Kwangwon Lee,2 Kwang-Kyung Liou,3 Hei-Chan Lee,3 Jae-Kyung Sohng,3 and Byung-Gee Kim1,2*

School of Chemical and Biological Engineering,1 Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University,2 Institute of Biomolecule Reconstruction Lab, Sunmoon University, Seoul, Korea3

Received 19 January 2005/ Accepted 5 April 2005

{gamma}-Butyrolactone derivative molecules in Streptomyces play a crucial role in cell density control, secondary metabolism, and cell differentiation. As their synthesis level in the cell is very low compared to those of similar N-acyl homoserine lactone molecules from gram-negative bacteria, it is very hard to analyze them even with several hundredfold concentration of the culture broth. We have developed a very quick and easy detection method using an affinity capture technique with His-tagged receptor proteins and electrospray tandem mass spectrometry. Using Streptomyces coelicolor as a model system, SCB1 was detected from only 100 ml of the culture broth after solvent extraction. This method can be further applied to detection and quantitative analysis of butanolides and inhibitor screening of the receptor molecules.

* Corresponding author. Mailing address: School of Chemical and Biological Engineering, Seoul National University, Shillim Dong, San 56-1, Kwan-ak Gu, Seoul, Korea. Phone: 82-2-880-6774. Fax: 82-2-873-6020. E-mail: byungkim{at}snu.ac.kr.

Applied and Environmental Microbiology, September 2005, p. 5050-5055, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5050-5055.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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