著者

DECENZO MAUREEN KURANDA MIKE COHEN SETH BABIAK JOHN JIANG ZHI-DONG SUN DONGYU HICKEY MARK SANCHETI PRAVEEN BRADFORD PATRICIA A. YOUNGMAN PHIL PROJAN STEVE ROTHSTEIN DAVID M.

出版者

公益財団法人 日本感染症医薬品協会

雑誌

The Journal of Antibiotics

巻号頁・発行日

vol.55, no.3, pp.288-295, 2002

被引用文献数

25

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A screening system is described that can detect and confirm inhibitors of the late steps of cell wall biosynthesis. The primary high through-put screen monitors induction of &beta;-lactamase following exposure to samples, in an <i>Escherichia coli envA</i>- strain that carries the &beta;-lactamase gene from <i>Citrobacter freundii</i> on a plasmid. Positive samples were detected from compound libraries, from natural products libraries, and from fractions of natural products crude preparations. These samples were then subjected to <i>in vitro</i> assays that could detect the incorporation of soluble cell wall precursor into Lipid I, Lipid II, and polymerized cell wall, using a TLC system that was very accurate and unambiguous in detecting known cell wall inhibitors. One partially purified sample containing a novel antibacterial agent derived from natural products was found to inhibit the formation of Lipid I (50% inhibition at &le;62.5ng/ml), whereas another partially purified sample also derived from natural products inhibited transglycosylation into cell wall polymer (50% inhibition at &le;10&mu;g/ml). This screening system proved to be especially useful because it was sufficiently sensitive and robust to detect inhibitors among samples of crude preparations or varying states of purity.