The Journal of Antibiotics
Online ISSN : 1881-1469
Print ISSN : 0021-8820
ISSN-L : 0021-8820
Screening Systems for Detecting Inhibitors of Cell Wall Transglycosylation in Enterococcus
Cell Wall Transglycosylation Inhibitors in Enterococcus
NAGRAJ MANIPRAVEEN SANCHETZHI-DONG JIANGCOLLEEN MCNANEYMAUREEN DECENZOBEN KNIGHTIMARY STANKISMIKE KURANDADAVID M. ROTHSTEIN
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1998 Volume 51 Issue 5 Pages 471-479

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Abstract

We devised two screening systems to detect cell wall transglycosylation inhibitors. One screen utilizes a mutant of Enterococcus faecalis strain A256 that is dependent on vancomycin or moenomycin for growth. In the absence of transglycosylation inhibitors the strain fails to grow, while in the presence of inhibitors, cells are rescued. A second screening organism E. faecalis strain MDD212 utilizes a translational fusion of the lacZ gene to the vanH promoter in a derivative of E. faecalis that contains a vancomycin resistance determinant. Induction of β-galactosidase occurs when cells are exposed to inhibitors of transglycosylation. Our natural products drug source of fungal fermentations was tested with these screens. Several cultures that produced the same family of compounds, called the thielavins, were detected. Thielavin B inhibited the formation of peptidoglycan in an in vitro assay, suggesting that these screening systems can detect compounds that interfere with cell wall transglycosylation.

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© Japan Antibiotics Research Association
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