Novel method for detection of butanolides in Streptomyces coelicolor culture broth, using a His-tagged receptor (ScbR) and mass spectrometry.
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.
Yang
Yung-Hun
YH
School of Chemical and Biological Engineering, Seoul National University, Shillim Dong, San 56-1, Kwan-ak Gu, Seoul, Korea.
Joo
Hwang-Soo
HS
Lee
Kwangwon
K
Liou
Kwang-Kyung
KK
Lee
Hei-Chan
HC
Sohng
Jae-Kyung
JK
Kim
Byung-Gee
BG
D015107:4-Butyrolactone / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006639:Histidine; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D048372:Streptomyces coelicolor / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*
D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; C446304:ScbR protein, Streptomyces coelicolor; D006639:Histidine; D015107:4-Butyrolactone