...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Highly-Active Recombinant Formate Dehydrogenase from Pathogenic Bacterium Staphylococcus aureus: Preparation and Crystallization
【24h】

Highly-Active Recombinant Formate Dehydrogenase from Pathogenic Bacterium Staphylococcus aureus: Preparation and Crystallization

机译：来自致病细菌金黄色葡萄球菌的高活性重组甲酸脱氢酶：制备和结晶

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

NAD(+)-dependent formate dehydrogenase from Staphylococcus aureus (SauFDH) is one of the key enzymes responsible for the survival of this pathogen in the form of biofilms. 3D structure of the enzyme might be helpful in the search for highly specific SauFDH inhibitors that can be used as antibacterial agents exactly against S. aureus biofilms. Here, we prepared a recombinant SauFDH in Escherichia coli cells with a yield of 1 g target protein per liter medium. The developed procedure for the enzyme purification allowed to obtain 400 mg of homogenous enzyme with 61% yield. The specific activity of the purified recombinant SauFDH was 20 U per mg protein, which was 2 times higher than the previously reported activities of formate dehydrogenases. We also found crystallization conditions in the course of two rounds of optimization and obtained 200- and 40-mu m crystals for the SauFDH apo- and holoenzymes, respectively. X-ray analysis using synchrotron X-ray sources produced diffraction data sufficient for solving the three-dimensional structures of the apo- and holoenzymes with the resolution of 2.2 and 2.7 angstrom, respectively. Crystals of the apo- and holoforms of SauFDH had different crystal space groups, which suggest coenzyme binding in the SauFDH holoenzyme.
机译：NAD（+） - 依赖于葡萄球菌（SAUFDH）的依赖性甲酸脱氢酶（SAUFDH）是负责该病原体以生物膜的形式存活的关键酶之一。酶的3D结构可能有助于寻找高度特异性的SAUFDH抑制剂，其可用作抗菌剂的抗菌剂与AUREUS生物膜完全相同。在此，我们在大肠杆菌细胞中制备了重组Saufdh，其产率为1g靶蛋白为每升培养基。酶纯化的开发方法使得获得400mg的均匀酶，产率61％。纯化的重组Saufdh的比活性为每Mg蛋白为20 u，比先前报告的甲酸脱氢酶的活性高2倍。我们还在两轮优化过程中发现了结晶条件，并分别获得了SAUFDH APO-和全酶的200-和40-mu m晶体。使用同步旋流X射线源的X射线分析产生了足以求解APO-和全酶的三维结构的衍射数据分别分别为2.2和2.7埃的分辨率。 Saufdh的APO-和全造型晶体的晶体具有不同的晶体空间组，这提出了Saufdh全酶中的辅酶结合。

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