YAKHAK HOEJI (약학회지)

The Pharmaceutical Society of Korea (대한약학회)
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Screening of New Antibiotics Inhibiting Bacterial Enoyl-Acyl Carrier Protein Reductase (Fabl)

세균의 지방산 생합성 효소 (Enoyl-Acyl Carrier Protein Reductase, FabI)를 저해하는 새로운 항균물질의 스크리닝

  • 곽진환 (한동대학교 생물식품공학부)
  • Published : 2002.02.01
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Abstract

Enoyl-Acyl Carrier Protein Reductase (Fabl) of bacteria is hem as an important target for new antibacterial drugs and plays a determinant role in completing cycles of elongation in type-H fatty acid synthase system. In this study, a fabI gene from Staphylococcus aureus 6538p cloned in pET-l4b vector and FabI protein was over-produced in Escherichaia coli BL2l (DE3). $NH_2$-terminal His-tagged FabI protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metalaffinity chromatography Purified 6xHis-tagged FabI showed a catalytic activity on tram - 2 - octenoyl - N -acethlcysteamine by utilizing NADPH as a cofactor. For the discovery of new FabI inhibitors from chemical libraries, a target-oriented screening system using a 96-well plate was developed. About 10,000 chemical libraries from Korea Chemical Bank wore tested in this screening system, and 26 chemicals (0.25%) among them showed an inhibitory activity against FabI enzyme. This result showed that a new screening system can be used for the discovery of new FabI inhibitors.

Keywords

References

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