...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>The Journal of Antibiotics: An International Journal >A novel assay of bacterial peptidoglycan synthesis for natural product screening.
【24h】

A novel assay of bacterial peptidoglycan synthesis for natural product screening.

机译：一种用于天然产物筛选的细菌肽聚糖合成的新方法。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Although a large number of microbial metabolites have been discovered as inhibitors of bacterial peptidoglycan biosynthesis, only a few inhibitors were reported for the pathway of UDP-MurNAc-pentapeptide formation, partly because of the lack of assays appropriate for natural product screening. Among the pathway enzymes, D-Ala racemase (Alr), D-Ala:D-Ala ligase (Ddl) and UDP-MurNAc-tripeptide:D-Ala-D-Ala transferase (MurF) are particularly attractive as antibacterial targets, because these enzymes are essential for growth and utilize low-molecular-weight substrates. Using dansylated UDP-MurNAc-tripeptide and L-Ala as the substrates, we established a cell-free assay to measure the sequential reactions of Alr, Ddl and MurF coupled with translocase I. This assay is sensitive and robust enough to screen mixtures of microbial metabolites, and enables us to distinguish the inhibitors for D-Ala-D-Ala formation, MurF and translocase I. D-cycloserine, the D-Ala-D-Ala pathway inhibitor, was successfully detected by this assay (IC(50)=1.7 microg ml(-1)). In a large-scale screening of natural products, we have identified inhibitors for D-Ala-D-Ala synthesis pathway, MurF and translocase I.
机译：尽管已发现大量微生物代谢物可作为细菌肽聚糖生物合成的抑制剂，但仅报道了少数几种UDP-MurNAc-五肽形成途径的抑制剂，部分原因是缺乏适用于天然产物筛选的分析方法。在这些途径酶中，D-Ala消旋酶（Alr），D-Ala：D-Ala连接酶（Ddl）和UDP-MurNAc-三肽：D-Ala-D-Ala转移酶（MurF）作为抗菌目标特别吸引人，因为这些酶对于生长和利用低分子量底物至关重要。我们使用丹磺酰化的UDP-MurNAc-三肽和L-Ala作为底物，建立了一种无细胞测定法来测量Alr，Ddl和MurF以及转位酶I的顺序反应。该测定法灵敏且稳健，足以筛选微生物混合物代谢物，使我们能够区分D-Ala-D-Ala形成抑制剂，MurF和转位酶I.通过该测定法成功检测到D-Ala-D-Ala途径抑制剂D-环丝氨酸（IC（50） = 1.7微克ml（-1））。在对天然产物的大规模筛选中，我们确定了D-Ala-D-Ala合成途径，MurF和Translocase I的抑制剂。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号